NZ501110A - Random peptides that bind to gastro-intestinal tract (GIT) transport receptors and related drug delivery methods - Google Patents

Abstract

Proteins (e.g., random peptides and peptide motifs) capable of facilitating transport of an active agent through human or animal gastro-intestinal tissue, and derivatives (e.g., fragments and functionally active analogues) thereof, and nucleotide sequences coding for said proteins and derivatives are disclosed. The proteins facilitate transport of active agents from the lumenal side of the gastro-intestinal tract (GIT) into the systemic blood system and/or in targeting active agents to the GIT. Thus, by binding (covalently or noncovalently) a protein disclosed to an orally administered drug, the drug can be targeted to specific receptors, such as, HPT1, hEPT1, D2H and hSI, or transport pathways which are known to operate in the human GIT.

Description

RANDOM PEPTIDES THAT BIND TO GASTRO-INTESTINAL TRACT (GIT) TRANSPORT RECEPTORS AND RELATED METHODS This application claims priority to U.S. provisional application Serial No. 60/046,595 filed May 15, 1997, which is incorporated by reference herein in its entirety. 1. INTRODUCTION The present invention relates generally to random peptides capable of specific binding to gastro-intestinal tract (GIT) transport receptors. In particular, this 15 invention relates to peptide sequences and motifs, as well as derivatives thereof, which enhance drug delivery and transport through tissue, such as epithelial cells lining the lumenal side of the gastro-intestinal tract (GIT).

Production of peptides, derivatives and antibodies is also 20 provided. The invention further relates to pharmaceutical compositions, formulations and related methods. 2. BACKGROUND OF THE INVENTION 2.1. Peptide Libraries There have been two different approaches to the construction of random peptide libraries. According to one approach, peptides have been chemically synthesized in vitro in several formats. Examples of chemically synthesized libraries can be found in Fodor, S., et al., 1991, Science 30 251: 767-773; Houghten, R., et al., 1991, Nature 354: 84-86; and Lam, K., et al., 1991, Nature 354: 82-84.

A second approach to the construction of random peptide libraries has been to use the M13 phage, and, in particular, protein pill of M13. The viral capsid protein of 35 M13, protein III (pIII), is responsible for infection of bacteria. Several investigators have determined from mutational analysis that the 406 amino acid long pIII capsid Printed from Mimosa protein has two domains. The C-terminus anchors the protein to the viral coat, while portions of the N-terminus of pIII are essential for interaction with the E. coli pillm protein (Crissman, J.W. and Smith, G.P., 1984, Virology 132: 445-5 455). Although the N-terminus of the piII protein has shown to be necessary for viral infection, the extreme N-terminus of the mature protein does tolerate alterations. In 1985, George Smith published experiments reporting the use of the pIII protein of bacteriophage M13 as an experimental system 10 for expressing a heterologous protein on the viral coat surface (Smith, G.P., 1985, Science 228: 1315-1317). It was later recognized, independently by two groups, that the M13 phage piII gene display system could be a useful one for mapping antibody epitopes (De la Cruz, V., et al., 1988, 15 J. Biol. Chem. 263: 4318-4322; Parmley, S.F. and Smith, G.P., 1988, Gene 73: 305-318).

Parmley, S.F. and Smith, G.P., 1989, Adv. Exp. Med. Biol. 251: 215-218 suggested that short, synthetic DNA segments cloned into the pIII gene might represent a library 20 of epitopes. These authors reasoned that since linear epitopes were often ~6 amino acids in length, it should be possible to use a random recombinant DNA library to express all possible hexapeptides to isolate epitopes that bind to antibodies. Scott, J.K. and Smith, G.P., 1990, Science 249: 25 386-390 describe construction and expression of an "epitope library" of hexapeptides on the surface of M13. Cwirla, S.E., et al., 1990, Proc. Natl. Acad. Sci. USA 87: 6378-6382 also described a somewhat similar library of hexapeptides expressed as gene pIII fusions of M13 fd phage. PCT 30 Application WO 91/19818 published December 26, 1991 by Dower and Cwirla describes a similar library of pentameric to octameric random arruno acid sequences. Devlin et al. , 1990, Science, 249: 404-406, describes a peptide library of about 15 residues generated using an (NNS) coding scheme for 35 oligonucleotide synthesis in which S is G or C. Christian and colleagues have described a phage display library, Printed from Mimosa expressing decapeptides (Christian, R.B., et al., 1992, J. Mol. Biol. 227: 711-718).

Other investigators have used other viral capsid proteins for expression of non-viral DNA on the surface of 5 phage particles. For example, the major capsid protein pVIII was so used by Cesareni, G., 1992, FEBS Lett. 307: 66-70. Other bacteriophage than M13 have been used to construct peptide libraries. Four and six amino acid sequences corresponding to different segments of the Plasmodium 10 falciparum major surface antigen have been cloned and expressed in the filamentous bacteriophage fd (Greenwood, J., et al., 1991, J. Mol. Biol. 220: 821-827).

Kay et al., 1993, Gene 128: 59-65 (Kay) discloses a method of constructing peptide libraries that encode peptides 15 of totally random sequence that are longer than those of any prior conventional libraries. The libraries disclosed in Kay encode totally synthetic random peptides of greater than about 20 amino acids in length. Such libraries can be advantageously screened to identify peptides, polypeptides 20 and/or other proteins having binding specificity for a -variety of ligands. (See also U.S. Patent No. 5,498,538 dated March 12, 1996; and PCT Publication No. WO 94/18318 dated August 18, 1994.) A comprehensive review of various types of peptide 25 libraries can be found in Gallop et al., 1994, J. Med. Chem. 37:1233-1251.

Screening of peptide libraries has often been done using an antibody as ligand (Parmley and Smith, 1989, Adv. Exp. Med. Biol. 251:215-218; Scott and Smith, 1990, Science 249:386-390) . In many cases, the aiin of the screening is to identify peptides from the library that mimic the epitopes to which the antibodies are directed. Thus, given an available antibody, peptide libraries are excellent sources for identifying epitopes or epitope-like molecules of 35 that antibody (Yayon et al., 1993, Proc. Natl. Acad. Sci. USA 90:10643-10647) .

Printed from Mimosa McCafferty et al., 1990, Nature 348:552-554 used PCR to amplify immunoglobulin variable (V) region genes and cloned those genes into phage expression vectors. The authors suggested that phage libraries of V, diversity (D), 5 and joining (J) regions could be screened with antigen. The phage that bound to antigen could then be mutated m the antigen-binding loops of the antibody genes and rescreened. The process could be repeated several times, ultimately giving rise to phage which bind the antigen strongly. 10 Marks et al., 1991, J. Mol. Biol. 222:581-597 also used PCR to amplify immunoglobulin variable (V) region genes and cloned those genes into phage expression vectors.

Kang et al., 1991, Proc. Natl. Acad. Sci. USA 88:4363-4366 created a phagemid vector that could be used to 15 express the V and constant (C) regions of the heavy and light chains of an antibody specific for an antigen. The heavy and light chain V-C regions were engineered to combine in the periplasm to produce an antibody-like molecule with a functional antigen binding site. Infection of cells 20 harboring this phagemid with helper phage resulted in the incorporation of the antibody-like molecule on the surface of phage that carried the phagemid DNA. This allowed for identification and enrichment of these phage by screening with the antigen. It was suggested that the enriched phage 25 could be subject to mutation and further rounds of screening, leading to the isolation of antibody-like molecules that were capable of even stronger binding to the antigen.

Hoogenboom et al., 1991, Nucleic Acids Res. 19:4133-4137 suggested that naive antibody genes might be 30 cloned into phage display libraries. This would be followed by random mutation of the cloned antibody genes to generate high affinity variants.

Bass et al., 1990, Proteins: Struct. Func. Genet. 8:309-314 fused human growth hormone (hGH) to the carboxy 35 terminus of the gene III protein of phage fd. This fusion protein was built into a phagemid vector. When cells carrying the phagemid were infected with a helper phage, Printed from Mimosa about 10% of the phage particles produced displayed the fusion protein on their surfaces. These phage particles were enriched by screening with hGH receptor-coated beads. It was suggested that this system could be used to develop mutants 5 of hGH with altered receptor binding characteristics.

Lowman et al., 1991, Biochemistry 30:10832-10838 used an improved version of the system of Bass et al. described above to select for mutant hGH proteins with exceptionally high affinity for the hGH receptor. The 10 authors randomly mutagenized the hGH-pIII fusion proteins at sites near the vicinity of 12 amino acids of hGH that had previously been identified as being important in receptor binding.

Balass et al., 1993, Proc. Natl. Acad. Sci. USA 15 90:10638-10642 used a phage display library to isolate linear peptides that mimicked a conformationally dependent epitope of the nicotinic acetylcholine receptor. This was done by screening the library with a monoclonal antibody specific for the conformationally dependent epitope. The monoclonal 20 antibody used was thought to be specific to the acetylcholine receptor's binding site for its natural ligand, acetylcholine. 2.2. Drug Delivery Systems The common routes of therapeutic drug administration are oral ingestion or parenteral (intravenous, subcutaneous and intramuscular) routes of administration. Intravenous drug administration suffers from numerous limitations, including (i) the risk of adverse effects 30 resulting from rapid accumulation of high concentrations of drug, (ii) repeated injections which can cause patient discomfort; and (iii) the risk of infection at the site of repeated injections. Subcutaneous injection is not generally suitable for delivering large volumes or for irritating 35 substances. Whereas oral administration is generally more convenient, it is limited where the therapeutic agent is not efficiently absorbed by the gastrointestinal tract. To date, Printed from Mimosa the development of oral formulations for the effective delivery of peptides, proteins and macromolecules has been an elusive target. Poor membrane permeability, enzymatic instability, large molecular size, and hydrophilic properties 5 are four factors that have remained major hurdles for peptide and protein formulations (reviewed by Fix, J.A., 1996, J. Pharmac. Sci. 85:1282-1285). In order to develop an efficacious oral formulation, the peptide must be protected from the enzymatic environment of the gastrointestinal tract 10 (GIT), presented to the absorptive epithelial barrier in a sufficient concentration to effect transcellular flux (Fix, J.A., 1996, J. Pharmac. Sci. 85:1282-1285), and if possible "smuggled" across the epithelial barrier in an apical to basolateral direction.

Site specific drug delivery or drug targeting can be achieved at different levels, including (i) primary targeting to a specific organ, (ii) secondary targeting to a specific cell type within that organ and (iii) tertiary targeting where the drug is delivered to specific 20 intracellular structures {e.g., the nucleus for genes) (reviewed in Davis and Jllum, 1994, In: Targeting of Drugs 4, (Eds), Gregoriadis, McCormack and Poste, 183-194). At present there is a considerable amount of ongoing research work in the Drug Delivery Systems (DDS) area, and much of it 25 addresses (i) targeting delivery and (ii) the development of non-invasive ways of getting macromolecules, peptides, proteins, products of the biotechnology industry, etc. into the body (Evers, P., 1995, Developments in Drug Delivery: Technology and Markets, Financial Times Management Report). 30 It is generally accepted that targeted drug delivery is crucial to the improved treatment of certain diseases, especially cancer, and not surprisingly many of the approaches to targeted drug delivery are focused in the cancer area. Many anticancer drugs are toxic to the body as 35 well as to malignant cells. If a drug, or a delivery system, can be modified so that it "homes in" on the tumor, then by maximizing the drug concentration at the disease site, the Printed from Mimosa anti-cancer effect can be exploited to the full, while toxicity is greatly reduced. Tumors contain antigens which provoke the body to respond by producing antibodies designed to attach to the antigens and destroy them. Monoclonal 5 antibodies are being used as both delivery vehicles targeted to tumor cells (reviewed by Pietersz, G.A., 1990, Bioconjugate Chem. 1:89-95) and as imaging agents to carry molecules of drug or imaging agent to the tumor surface. 2.3. Transport Pathways The epithelial cells lining the lumenal side of the GIT are a major barrier to drug delivery following oral administration. However, there are four recognized transport pathways which can be exploited to facilitate drug delivery 15 and transport: the transcellular, paracellular, carrier-mediated, and transcytotic pathways. The ability of a conventional drug, peptide, protein, macromolecule or nano-or microparticulate system to "interact" with one of these transport pathways may result in increased delivery of that 20 drug or particle from the GIT to the underlying circulation.

In the case of the receptor-mediated, carrier-mediated or transcytotic transport pathways, some of the uptake signals have been identified. These signals include, inter alia, folic acid, which interacts with the folate 25 receptor, and cobalamin, which interacts with Intrinsic Factor. In addition, leucine- and tyrosine-based peptide sorting motifs or internalization sequences exist, such as YSKV, FPHL, YRGV, YQTI, TEQF, TEVM, TSAF, and YTRF (SEQ ID NOS:203, 204, 205, 206, 207, 208, 209, and 210, 30 respectively), which facilitate uptake or targeting of proteins using specific membrane receptors or binding sites to identify peptides that bind specifically to the receptor or binding site.

Non-receptor based assays to discover particular 35 ligands have also been used. For instance, a strategy for identifying peptides that alter cellular function by scanning whole cells with phage display libraries is disclosed in Fong Printed from Mimosa et al., Drug Development Research 33:64-70 (1994). However, because whole cells, rather than intact tissue or polarized cell cultures, are used for screening phage display-libraries, this procedure does not provide information 5 regarding sequences whose primary function includes affecting transport across polarized cell layers.

Additionally, Stevenson et al., Pharmaceutical Res. 12(9), S94 (1995) discloses the use of Caco-2 monolayers to screen a synthetic tripeptide combinatorial library for 10 information relating to the permeability of di- and tri-peptides.

A method of identifying a peptide which permits or facilitates the transport of an active agent through human or animal tissues has been developed (see U.S. patent 15 application Serial No. 08/746,411 filed November 8, 1996, which is incorporated by reference herein in its entirety). Phage from a random phage library is plated onto or brought into contact with a first side, preferably the apical side, of a tissue sample, either in vitro, in vivo or in situ, or 20 polarized tissue cell culture. The phage which is transported to a second side of the tissue opposite the first side, preferably the basolateral side, is harvested to select transported phages. The transported phages are amplified in a host and this cycle is repeated (using the transported 25 phage from the most recent cycle) to obtain a selected phage library containing phage which can be transported from the first side to the second side.

Discussion or citation of a reference hereinabove shall not be construed as meaning that such reference is 30 prior art to the present invention. 3. SUMMARY OF THE INVENTION The present invention relates generally to random peptides and peptide motifs capable of specific binding to 35 GIT transport receptors. Such proteins can be identified using any random peptide library, e.g., a chemically synthesized peptide library or a biologically expressed Printed from Mimosa peptide library. If a biological peptide expression library is used, the nucleic acid which encodes the peptide which binds to the ligand of choice can be recovered, and then sequenced to determine its nucleotide sequence and hence 5 deduce the amino acid sequence that mediates binding. Alternatively, the amino acid sequence of an appropriate binding domain can be determined by direct determination of the amino acid sequence of a peptide selected from a peptide library containing chemically synthesized peptides. In a 10 less preferred aspect, direct amino acid sequencing of a binding peptide selected from a biological peptide expression library can also be performed.

In particular, this invention relates to proteins (e.g., peptides) that are capable of facilitating transport 15 of an active agent through a human or animal gastrointestinal tissue, and derivatives (e.g., fragments) and analogs thereof, and nucleotide sequences coding for said proteins and derivatives.

Preferably, the tissue through which transport is 20 facilitated is of the duodenum, jejunum, ileum, ascending colon, transverse colon, descending colon, or pelvic colon. The tissue is most preferably epithelial cells lining the lumenal side of the GIT.

The proteins of the invention have use in 25 facilitating transport of active agents from the lumenal side of the GIT into the systemic blood system, and/or in targeting active agents to the GIT. Thus, for example, by binding (covalently or noncovalently) a protein of the invention to an orally administered drug, the drug can be 30 targeted to specific receptor sites or transport pathways which are known to operate in the human gastrointestinal tract, thus facilitating its absorption into the systemic system.

The invention also relates to derivatives and 35 analogs of the invention which are functionally active, i.e., they are capable of displaying one or more known functional activities associated with a full-length peptide. Such Printed from Mimosa functional activities include but are not limited to antigenicity (ability to bind or to compete with GIT transport receptor-binding peptides for binding to an anti-GIT transport receptor antibody) and ability to bind or 5 compete with full-length peptide for binding to a GIT transport receptor.

The invention further relates to fragments of (and derivatives and analogs thereof) GIT transport receptor-binding peptides which comprise one or more motifs of a GIT 10 transport receptor-binding peptide.

Antibodies to GIT transport receptor-binding peptides and GIT transport receptor-binding peptide derivatives and analogs are additionally provided.

Methods of production of the GIT transport 15 receptor-binding peptides, derivatives, fragments and analogs, e.g., by recombinant means, are also provided.

The present invention also relates to therapeutic methods, pharmaceutical compositions and formulations based on GIT transport receptor-binding peptides. Formulations of 20 the invention include but are not limited to GIT transport receptor-binding peptides or motifs and derivatives (including fragments) thereof; antibodies thereto; and nucleic acids encoding the GIT transport receptor-binding peptides or derivatives associated with an active agent. 25 Preferably, the active agent is a drug or drug-containing nano- or microparticle.

The GIT transport-receptor binding proteins of the invention can also be used to determine levels of the GIT transport receptors in a sample by binding thereto. 30 The GIT transport-receptor binding proteins can also be used to identify molecules that bind thereto, by contacting candidate test molecules under conditions conducive to binding, and detecting any binding that occurs. 4. DESCRIPTION OF THE FIGURES Figure 1. Figure 1 shows the human PEPT1 predicted amino acid sequence determined from the sequence of the cDNA clone Printed from Mimosa coding for human PEPT1 (SEQ ID NO:176) (Liang R. et al. J. Biol. Chem. 210 (12):6456-6463 (1995)), including the extracellular domain from amino acid 391 to 573 (Fei et al., Nature 368:563 (1994)).

Figures 2A-2C. Figures 2A-2C show the DNA sequence of the cDNA coding for the human intestinal peptide-associated transporter HPT1 and the corresponding putative amino acid sequence (bases 1 to 3345; Medline:94204643) (SEQ ID NOS: 177 and 178, respectively).

Figures 3A-3B. Figures 3A-3B show the putative Human Sucrase-isomaltase complex(hSI) amino acid sequence determined from the sequence of the cDNA clone coding for human sucrase-isomaltase complex (SEQ ID NO:179) (Chantret I., et al., Biochem. J. 285 (Pt 3):915-923 (1992).

Figures 4A-4B. Figures 4A-4B show the D2H nucleotide and deduced amino acid sequence for the human D2H transporter (SEQ ID NOS:180 and 181, respectively) (Wells, R.G. et al.,J. Clin. Invest. 90:1959-1963 (1993).

Figures 5A-5C. Figure 5A is a schematic summary of the 20 cloning of the DNA insert present in gene III of the phages selected from the phage display libraries into the expression vector pGex-4T-2. The gene insert in gene III of the phages was amplified by PCR using DNA primers which flank the gene insert and which contained recognition sequences for specific 25 restriction endonucleases at their extreme 5' sides. Alternatively, specific primers which amplify specific regions of the DNA inserts in gene III of the phages, and which contained recognition sequences for specific restriction endonucleases at their extreme 5' sides, were 30 used in PCR amplification experiments. Following amplification of the gene inserts, the amplified PCR fragments were digested with the restriction endonucleases Xhol and Notl. Similarly the plasmid pGex-4T-2, which codes for the reporter protein glutathione S-transferase (GST), was 35 digested with the restriction endonucleases Sail and Notl. The digested PCR fragments were ligated into the digested plasmid pGex-4T-2 using T4 DNA Ligase and the ligated Printed from Mimosa products were transformed into competent Escherichia coli, with selection of transformants on agar plates containing selection antibiotic. The selected clones were cultured, the plasmids were recovered and the in-frame sequence of the DNA 5 insert in the plasmids was confirmed by DNA sequencing. The correct clones were subsequently used for expression of the GST-fusion proteins (SEQ ID NO:182); Figure 5B shows the series of full-length P31 (designated P31) (SEQ ID NO:43) and truncated peptides derived from P31 (clones # 101, 102, 103 10 and 119), (SEQ ID NOS:183, 184, 185, and 186, respectively) full-length PAX2 (designated PAX2) (SEQ ID NO:55) and truncated peptides derived from PAX2 (clones # 104, 105, 106) (SEQ ID NOS:170, 187, and 188, respectively) and full-length DCX8 (DCX8) (SEQ ID NO:23) and series of truncated peptides 15 derived from DCX8 (clones # 107, 108, 109) (SEQ ID NOS:189, 190,,and 191, respectively) that were expressed as fusion proteins to GST. The construction of these GST-fusion proteins is shown in Figure 5A. Figure 5C shows the series of full-length P31 (designated P31) (SEQ ID NO:43) and 20 truncated peptides derived from P31 (clones # 103, 110, 119, 111, and 112) (SEQ ID NOS:185, 192, 193, 194, and 195, respectively), full-length PAX2 (designated PAX2) (SEQ ID NO:55) and truncated peptides derived from PAX2 (clones # 106, 113, 114, 115) (SEQ ID NOS:188, 196, 197, and 198, 25 respectively) and full-length SNilO (designated SNilO) (SEQ ID NO:4) and series of truncated peptides derived from SNilO (clones # 116, 117, 118) (SEQ ID NOS:199, 200, and 201, respectively) that were expressed as fusion proteins to GST. The construction of these GST-fusion proteins is shown in 30 Figure 5A. (Underlining and bold in Figs. 5A-5C are for orientation of the sequences.) Figures 6A-6B. Figures 6A-6B show the binding of GST and GST-fusion proteins to recombinant hSI and to fixed C2BBel fixed cells as detected by ELISA assays. Figure 6A shows the 35 binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from SNilO (designated GST-SNilO) and SNi34 (designated GST-SNi34) to Printed from Mimosa recombinant hSI. Figure 6B shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from SNilO (designated GST-SNilO) and SNi34 (designated GST-SNi34) to fixed C2BBel cells.

Figures 7A-7M. Figures 7A-7M show the binding of GST peptide and truncated fusion proteins to fixed Caco-2 cells, fixed C2BBel cells, and fixed A431 cells or to recombinant GIT transport receptors D2H, HPT1, hPEPTl or to BSA using increasing concentrations (expressed as fxg/ml on the X-axis) 10 of the control GST protein and the GST-fusion proteins, as detected by ELISA assays. Figure 7A shows the binding of the control protein GST, which does not contain a fusion peptide, and the series of GST-fusion proteins from P31 including the fusion to full-length P31 peptide (designated P31) (SEQ ID 15 NO:43) and clone # 101 (designated P31,101), clone # 102 (designated P31, 102) and clone # 103 (designated P31,103). Figure 7B shows the binding of the control protein GST, which does not contain a fusion peptide, and the series of GST-fusion proteins from PAX2 including the fusion to full-length 20 PAX2 peptide (designated PAX2) and clone # 104 (designated PAX2,104), clone # 105 (designated PAX2, 105) and clone # 106 (designated PAX2,106) (SEQ ID NOS:55, 170, 187, and 188, respectively). Figure 7C shows the binding of the control protein GST, which does not contain a fusion peptide, and the 25 series of GST-fusion proteins from DCX8 including the fusion to full-length DCX8 peptide (designated DCX8) and clone # 107 (designated DCX8,107), clone # 108 (designated DCX8, 108) and clone # 109 (designated DCX8,109) (SEQ ID NOS:23, 189, 190, and 191, respectively). Figure 7D shows the binding of the 30 control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from DCX8 (designated GST-DCX8) and DCX11 (designated GST-DCX11) to recombinant D2H. Figure 7E shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins 35 from DCX8 (designated GST-DCX8) and DCX11 (designated GST-DCX11) to fixed C2BBel cells. Figure 7F shows the binding of the control protein GST, which does not contain a fusion Printed from Mimosa peptide, and the GST-fusion proteins from P31 (designated GST-P31) and 5PAX5 (designated GST-5PAX5) to recombinant hPEPTl. Figure 7G shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-5 fusion proteins from P31 (designated GST-P31) and 5PAX5 (designated GST-5PAX5) to fixed C2BBel cells. Figure 7H shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from HAX42 (designated GST-HAX42) and PAX2 (designated GST-PAX2) 10 to recombinant HPT1. Figure 71 shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from HAX42 (designated GST-HAX42) and PAX2 (designated GST-PAX2) to fixed C2BBel cells. Figure 7J shows the binding of the control protein GST, which does 15 not contain a fusion peptide, and the GST-fusion proteins from P31 (designated GST-P31) and truncated derivatives clone # 101 (designated GST-P31-101), clone # 102 (designated GST-P31-102), clone # 103 (designated GST-P31-103) to either recombinant hPEPTl or to BSA.' Figure 7K shows the binding of 20 the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from P31 (designated GST-P31) and truncated derivatives clone # 101 (designated GST-P31-101), clone # 102 (designated GST-P31-102), clone # 103 (designated GST-P31-103) to either fixed C2BBel cells or 25 to fixed A431 cells. Figure 7L shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from PAX2 (designated GST-PAX2) and truncated derivatives clone # 104 (designated GST-PAX2-104), clone # 105 (designated GST-PAX2-105), clone # 106 30 (designated GST-PAX2-106) to either recombinant hPEPTl or to BSA. Figure 7M shows the binding of the control protein GST, which does not contain a fusion peptide, and the GST-fusion proteins from PAX2 (designated GST-PAX2) and truncated derivatives clone # 106 (designated GST-PAX2-106)to either 35 fixed Caco-2 cells or to fixed A431 cells.

Figures 8A-8D. Figure 8 shows the transport of GST or GST-peptide fusion derivatives across polarized Caco-2 cells in Printed from Mimosa an apical to basolateral direction as a function of time (1-4 hours) as detected by ELISA assays. Figure 8A shows the transport of either GST, the GST fusion to full-length P31 peptide (designated P31) (SEQ ID NO:43) and the GST clone 5 derivative clone # 103 (designated P31.103) across polarized Caco-2 cells in an apical to basolateral as a function of time (in hours) following initial administration of the proteins to the apical medium of polarized Caco-2 cells. The line designated No Protein corresponds to control assays in 10 which buffer control was applied to the apical medium of polarized Caco-2 cells followed by sampling of the basolateral medium as a function of time (hours) and assay for GST by the ELISA assay. Figure 8B shows the transport of either GST, the GST fusion to full-length PAX2 peptide 15 (designated PAX2) and the GST clone derivative clone # 106 (designated PAX2.106) across polarized Caco-2 cells in an apical to basolateral as a function of time (in hours) following initial administration of the proteins to the apical medium of polarized Caco-2 cells. The line designated 20 No Protein corresponds to control assays in which buffer control was applied to the apical medium of polarized Caco-2 cells followed by sampling of the basolateral medium as a function of time (hours) and assay for GST by the ELISA assay. Figure 8C shows the transport of either GST, the GST 25 fusion to full-length DCX8 peptide (designated DCX8), and the GST clone derivatives clone # 107 (designated DCX8.107) and clone # 109 (designated DCX8.109) across polarized Caco-2 cells in an apical to basolateral as a function of time (in hours) following initial administration of the proteins to 30 the apical medium of polarized Caco-2 cells. The line designated No Protein corresponds to control assays in which buffer control was applied to the apical medium of polarized Caco-2 cells followed by sampling of the basolateral medium as a function of time (hours) and assay for GST by the ELISA 35 assay. Figure 8D shows the amount of the GST and GST-fusion proteins (GST fusions to P31, P31-103, PAX2, PAX2.106, DCX8, DCX8-107, DCX8-109), used in the experiments shown in panels Printed from Mimosa A-C above, in the apical medium of the polarized Caco-2 cells as detected by ELISA assay.

Figures 9A-9B. Figures 9A-9B show the inhibition of GST-P31 binding to C2BBel fixed cells with varying concentration of 5 competitors while holding the concentration of GST-P31 constant at 0.015 /iM; the peptide competitors are ZElan024 which is the dansylated peptide version of P31 (SEQ ID NO:43) and ZElan044, ZElan049 and ZElan050 which are truncated, dansylated pieces of P31 (SEQ ID NO:43). Data is presented 10 as O.D. versus peptide concentration (Figure 9A) and as percent inhibition of GST-P31 binding versus peptide concentration (Figure 9B).

Figures 10A-10C. Figures 10A-10C present a compilation of the results of competition ELISA studies of GST-P31, GST-15 PAX2, GST-SNilO and GST-HAX42 versus listed dansylated peptides on fixed C2BBel cells ("Z" denotes e-amino dansyl lysine). The pi of the dansylated peptides is also included. Estimated ICS0 values are in /iM and where present, IC50 ranges .■.refer to results from multiple assays. If the IC50 value 20 could not be determined, a ">" or "<" symbol is used. *'The GST/C2BBel column shows GST protein binding to fixed C2BBel cells.

Figures 11A-11B. Figure 11A shows the transport of GST or GST-peptide fusion derivatives across polarized Caco-2 cells 25 in an apical to basolateral direction at 0, 0.5, 2 and 4 hours as detected by ELISA assays and described elsewhere in the text in full detail. The proteins used in the assay included GST, GST-P31 fusion, GST-5PAX5 fusion, GST-DCX8 fusion, GST-DCX11 fusion, GST-PAX2 fusion, GST-HAX42 fusion, 30 GST-SNi34 fusion and GST-SNilO fusion. The column designated No protein refers to control experiments in which buffer was applied to the apical medium of the cells and ELISA assay was performed on the corresponding basolateral medium of these cells at 0, 0.5, 2 and 4 hours post buffer addition. Figure 35 11B shows the internalization of GST or GST-peptide fusion derivatives within polarized Caco-2 cells following administration of the GST or GST-fusion protein derivatives Printed from Mimosa to the apical medium of polarized Caco-2 cells and subsequent recovery of the cells from the transwells and detection of the GST or GST fusions within the recovered cell lysates as detected by ELISA assays and as described elsewhere in the 5 text in full detail. The proteins used in the assay included GST, GST-P31 fusion, GST-5PAX5 fusion, GST-DCX8 fusion, GST-DCX11 fusion, GST-PAX2 fusion, GST-HAX42 fusion, GST-SNi34 fusion and GST-SNilO fusion. The column designated No protein refers to control experiments in which buffer was 10 applied to the apical medium of the cells and ELISA assay was performed on the corresponding cell lysates of these cells at the end of the experiment.

Figure 12. Figure 12 shows the binding of GST and GST-fusion proteins to fixed Caco-2 cells, and the corresponding 15 proteins following digestion with the protease Thrombin which cleaves at a recognition site between the GST portion and the fused peptide portion of the GST-fusion protein. The symbol refers to proteins which were not digested with thrombin and the symbol "+" refers to proteins which were digested 20 with thrombin prior to use in the binding assay. The binding of the proteins to the fixed Caco-2 cells was detected by ELISA assays.

Figures 13A-13B. Figures 13A-13B show binding of peptide-coated nanoparticles to fixed Caco-2 cells.

Figures 14A-14B. Figures 14A-14B show the binding of (A) dansylated peptide SNilO to the purified hSI receptor and BSA and (B) dansylated peptides and peptide-loaded insulin-containing PLGA particles to fixed C2BBel cells. Figure 14B depicts binding of dansylated peptides corresponding to P31 30 (SEQ ID NO:43), PAX2, HAX42, and SNilO to fixed C2BBel cells, as well as the insulin-containing PLGA particles adsorbed with each of these peptides. Data is presented with background subtracted.

Figures 15A-15B. Figure 15 shows the binding of peptide-35 coated particles to A) S100 and B) P100 fractions harvested from Caco-2 cells. The dilution series 1:2 - 1:64 represents particle concentrations in the range 0.0325-0.5 /ig/well.

Printed from Mimosa Data is presented with background subtracted. The particles are identified as follows: 939, no peptide; 1635, scrambled PAX2; 1726, P31 D-Arg 16-mer (ZElan053); 1756, HAX42; 1757, PAX2; 1758, HAX42/PAX2.

Figures 16A-16B. Figure 16 shows the binding of dansylated peptides to P100 fractions harvested from Caco-2 cells. Peptides were assayed in the range 0.0032-2.5 /xg/well. Data is presented with background subtracted. A) HAX42, P31 D-form (ZElan 053) and scrambled PAX2; B) PAX2, HAX42 and 10 scrambled PAX2.

Figures 17A-17B. Figures 17A and 17B show (A) the systemic blood glucose and (B)insulin levels following intestinal administration of control (PBS); insulin solution; insulin particles; all 8 peptides mix particles and study group 15 peptide-particles according to this invention (lOOiu insulin loading).

Figures 18A-18B. Figures 18A and 18B show the (A) systemic blood glucose and (B)insulin levels following intestinal administration of control (PBS); insulin solution; insulin 20 particles and study group peptide-particles according to this-invention (300iu insulin loading).

Figure 19. Figure 19 shows the enhanced plasma levels of leuprolide upon administration of P31 (SEQ ID NO:43) and PAX2 coated nanoparticles loaded with leuprolide relative to 25 subcutaneous injection. Group 1 was administered leuprolide acetate (12.5 /xg) subcutaneously. Group 2 was administered intraduodenally uncoated leuprolide acetate particles (600 fig, 1.5 ml). Group 3 was intraduodenally administered leuprolide acetate particles coated with PAX2 (600 ng; 1.5 30 ml). Group 4 was administered intraduodenally leuprolide acetate particles coated with P31 (SEQ ID NO:43) (600 pig, 1.5 ml) .

Figure 20. Figure 20 lists P31 (SEQ ID NO:43) known protein homologies.

Figures 21A-21C. Figures 21A-21C list DCX8 known protein homologies.

Figure 22. Figure 22 lists DAB10 known protein homologies.

Printed from Mimosa Figure 23. Figure 23 shows the DNA sequence (SEQ ID NO:211) and the corresponding amino acid sequence (SEQ ID NO:212) for glutathione S-transferase (Smith and Johnson, 1988, Gene 7:31-40).

. DETAILED DESCRIPTION OF THE INVENTION The present invention relates to proteins (e.g., peptides) that bind to GIT transport receptors and nucleic acids that encode such proteins. The invention further 10 relates to fragments and other derivatives of such proteins. Nucleic acids encoding such fragments or derivatives are also within the scope of the invention. The invention further relates to fragments (and derivatives and analogs thereof) of GIT transport receptor-binding peptides which comprise one or 15 more domains of the GIT transport receptor-binding peptides.

The invention also relates to derivatives of GIT transport receptor-binding proteins and analogs of the invention which are functionally active, i.e., they are capable of displaying one or more-known functional activities 20 associated with a full-length GIT transport receptor-binding peptide. Such functional activities include but are not limited to ability to bind to a GIT transport receptor, antigenicity [ability to bind (or compete with peptides for binding) to an anti-GIT transport receptor-binding peptide 25 antibody], immunogenicity (ability to generate antibody which binds to GIT transport receptor-binding peptide), etc.

Production of the foregoing proteins and derivatives, by, e.g., recombinant methods, is also provided.

Antibodies to GIT transport receptor-binding 30 proteins, derivatives and analogs, are additionally provided.

The present invention also relates to therapeutic and diagnostic methods and compositions based on GIT transport receptor-binding proteins and nucleic acids.

The invention is illustrated by way of examples infra.

Printed from Mimosa For clarity of disclosure, and not by way of limitation, the detailed description of the invention is divided into the subsections which follow. 5.1. GIT Transport Receptor-Binding Peptides, Derivatives and Analogs The invention relates to peptides that bind GIT transport receptors and derivatives (including but not limited to fragments) and analogs thereof. In specific embodiments, of the present invention, such peptides that bind to GIT transport receptor include but are not limited to those containing as primary amino acid sequences, all or part of ,,the amino acid sequences substantially as depicted in • Table 7 (SEQ ID NOS:1-55). Nucleic acids encoding such , c peptides, derivatives and peptide analogs are also provided. 15 In one embodiment, the GIT transport receptor-binding peptides are encoded by the nucleic acids having the nucleotide sequences set forth in Table 8 infra (SEQ ID NOS:56-109). Proteins whose amino acid sequence comprise, or 2Q alternatively, consist of SEQ ID NOS:1-55 or apportion thereof that mediates binding to a GIT transport receptor are provided.

The production and use of derivatives and analogs related to GIT transport receptor-binding peptides are within 25 the scope of the present invention. In a specific embodiment, the derivative or analog is functionally active, i.e., capable of exhibiting one or more functional activities associated with a full-length GIT transport receptor-binding peptide. For example, such derivatives or analogs which have the desired immunogenicity or antigenicity can be used, in immunoassays, for immunization, etc. A specific embodiment relates to a GIT transport receptor-binding peptide fragment that can be bound by an anti-GIT transport receptor-binding peptide antibody. In a preferred aspect, the derivatives or analogs have the ability to bind to a GIT transport receptor. Derivatives or analogs of GIT transport receptor-binding peptides can be tested for the desired activity by procedures Printed from Mimosa known in the art, including binding to a GIT transport receptor domain or to Caco-2 cells, in vitro, or to intestinal tissue, in vivo. (See the Examples infra.) In particular, derivatives can be made by altering 5 GIT transport receptor-binding peptide sequences by substitutions, additions or deletions that provide for functionally equivalent molecules. Due to the degeneracy of nucleotide coding sequences, other nucleotide sequences which encode substantially the same amino acid sequence may be used 10 in the practice of the present invention. These include but are not limited to nucleotide sequences which are altered by the substitution of different codons that encode a functionally equivalent amino acid residue within the sequence, thus producing a silent change. Likewise, the GIT 15 transport receptor-binding peptide derivatives of the invention include, but are not limited to, those containing, as a primary amino acid sequence, all or part of the amino acid sequence of a GIT transport receptor-binding peptide including altered sequences in which functionally equivalent 20 amino acid residues are substituted for residues within the sequence resulting in a silent change. For example, one or more amino acid residues within the sequence can be substituted by another amino acid of a similar polarity which acts as a functional equivalent, resulting in a silent 25 alteration. Substitutes for an amino acid within the sequence may be selected from other members of the class to which the amino acid belongs. For example, the nonpolar (hydrophobic) amino acids include alanine, leucine, isoleucine, valine, proline, phenylalanine, tryptophan and 30 methionine. The polar neutral amino acids include glycine, serine, threonine, cysteine, tyrosine, asparagine, and glutamine. The positively charged (basic) amino acids include arginine, lysine and histidine. The negatively charged (acidic) amino acids include aspartic acid and 35 glutamic acid.

In a specific embodiment of the invention, proteins consisting of or, alternatively, comprising all or a fragment Printed from Mimosa of a GIT transport receptor-binding peptide consisting of at least 5, 10, 15, 20, 25, 30 or 35 (contiguous) amino acids of the full-length GIT transport receptor-binding peptide are provided. In a specific embodiment, such proteins are not 5 more than 20, 30, 40, 50, or 75 amino acids in length. Derivatives or analogs of GIT transport receptor-binding peptides include but are not limited to those molecules comprising regions that are substantially homologous to GIT transport receptor-binding peptides or fragments thereof 10 (e.g., at least 50%, 60%, 70%, 80% or 90% identity) (e.g., over an identical size sequence or when compared to an aligned sequence in which the alignment is done by a computer homology program known in the art) or whose encoding nucleic acid is capable of hybridizing to a coding GIT transport 15 receptor-binding peptide sequence, under stringent, moderately stringent, or nonstringent conditions.

In a specific embodiment, the GIT transport receptor-binding derivatives of the invention are not known proteins with homology to the GIT transport receptor-binding 20 peptides of the invention or portions thereof.

The GIT transport receptor-binding peptide derivatives and analogs of the invention can be produced by various methods known in the art. The manipulations which result in their production can occur at the gene or protein 25 level. For example, the cloned GIT transport receptor-binding peptide gene sequence can be modified by any of numerous strategies known in the art (Maniatis, T., 1990, Molecular Cloning, A Laboratory Manual, 2d ed., Cold Spring Harbor Laboratory, Cold Spring Harbor, New York). The 30 sequence can be cleaved at appropriate sites with restriction endonuclease(s), followed by further enzymatic modification if desired, isolated, and ligated in vitro. In the production of the gene encoding a derivative or analog of GIT transport receptor-binding peptides, care should be taken to 35 ensure that the modified gene remains within the same translational reading frame uninterrupted by translational Printed from Mimosa stop signals, in the gene region where the desired GIT transport receptor-binding peptides activity is encoded.

Additionally, nucleic acid sequences encoding the GIT transport receptor-binding peptides can be mutated in 5 vitro or in vivo, to create and/or destroy translation, initiation, and/or termination sequences, or to create variations in coding regions and/or form new restriction endonuclease sites or destroy preexisting ones, to facilitate further in vitro modification. Any technique for mutagenesis 10 known in the art can be used, including but not limited to, chemical mutagenesis, in vitro site-directed mutagenesis (Hutchinson, C., et al., 1978, J. Biol. Chem 253:6551), use of TAB® linkers (Pharmacia), use of. PCR primers containing mutation(s) for use in amplification, etc. 15 Manipulations of GIT transport receptor-binding peptide sequences may also be made at the protein level. Included within the scope of the invention are GIT transport receptor-binding peptide fragments or other derivatives or analogs which are differentially modified during or after 20 translation or chemical synthesis, e.g., by glycosylation, acetylation, phosphorylation, amidation, derivatization by known protecting/blocking groups, proteolytic cleavage, linkage to an antibody molecule or other cellular ligand, etc. Any of numerous chemical modifications may be carried 25 out by known techniques, including but not limited to specific chemical cleavage by cyanogen bromide, trypsin, chymotrypsin, papain, V8 protease, NaBH4; acetylation, formylation, oxidation, reduction; metabolic synthesis in the presence of tunicamycin; etc. In a specific embodiment, the 30 amino- and/or carboxy-termini are modified.

In addition, GIT transport receptor-binding peptides and analogs and derivatives thereof can be chemically synthesized. For example, a peptide corresponding to all or a portion of a GIT transport receptor-binding 35 peptide which comprises the desired domain or which mediates the desired activity in vitro, can be synthesized by use of a peptide synthesizer. Furthermore, if desired, nonclassical Printed from Mimosa amino acids or chemical amino acid analogs can be introduced as a substitution or addition into the GIT transport receptor-binding peptide sequence. Non-classical amino acids include but are not limited to the D-isomers of the common 5 amino acids, a-amino isobutyric acid, 4-aminobutyric acid, Abu, 2-amino butyric acid, 7-Abu, e-Ahx, 6-amino hexanoic acid, Aib, 2-amino isobutyric acid, 3-amino propionic acid, ornithine, norleucine, norvaline, hydroxyproline, sarcosine, citrulline, cysteic acid, t-butylglycine, t-butylalanine, 10 phenylglycine, cyclohexylalanine, /3-alanine, fluoro-amino acids, designer amino acids such as /3-methyl amino acids, Ca-methyl amino acids, Nat-methyl amino acids, and amino acid analogs in general. Furthermore, the amino acid can be D (dextrorotary) or L (levorotary).

In a specific embodiment, the GIT transport receptor-binding peptide derivative is a chimeric, or fusion, peptide comprising a GIT transport receptor-binding peptide or fragment thereof (preferably consisting of at least a domain-or motif of the GIT transport receptor-binding 20 peptide, or at least 6, 10, 15, 20, 25, 30 or all amino acids of the GIT transport receptor-binding peptides or a binding portion thereof) joined at its amino- or carboxy-terminus via a peptide bond to an amino acid sequence of a different peptide. In one embodiment, such a chimeric peptide is 25 produced by recombinant expression of a nucleic acid encoding the protein (comprising a transport receptor-coding sequence joined in-frame to a coding sequence for a different protein). Such a chimeric product can be made by ligating the appropriate nucleic acid sequences encoding the desired 30 amino acid sequences to each other by methods known in the art, in the proper coding frame, and expressing the chimeric product by methods commonly known in the art. Alternatively, such a chimeric product may be made by protein synthetic techniques, e.g., by use of a peptide synthesizer. Chimeric 35 genes comprising portions of GIT transport receptor fused to any heterologous protein-encoding sequences may be constructed. A specific embodiment relates to a chimeric Printed from Mimosa protein comprising a fragment of GIT transport receptor-binding peptides of at least six amino acids.

In another specific embodiment, the GIT transport receptor-binding peptide derivative is a molecule comprising 5 a region of homology with a GIT transport receptor-binding peptide. By way of example, in various embodiments, a first protein region can be considered "homologous" to a second protein region when the amino acid sequence of the first region is at least 30%, 40%, 50%, 60%, 70%, 75%, 80%, 90%, or 10 95% identical, when compared to any sequence in the second region of an equal number of amino acids as the number contained in the first region or when compared to an aligned sequence of the second region that has been aligned by a computer homology program known in the art. For example, a 15 molecule can comprise one or more regions homologous to a GIT transport receptor-binding peptide domain (see infra) or a portion thereof.

The GIT transport receptor-binding proteins and derivatives thereof of the invention can be assayed for 20 binding activity by suitable in vivo or in vitro assays, e.g., as described in the examples infra and/or as will be known to the skilled artisan.

Other specific embodiments of derivatives and analogs are described in the subsection below and examples 25 sections infra. .2. Motifs/Derivatives of GIT Transport Receptor-Binding Peptides Containing One or More Domains of The Protein In a specific embodiment, the invention relates to 3 0 GIT transport receptor-binding peptide derivatives and analogs, in particular GIT transport receptor-binding peptide fragments and derivatives of such fragments, that comprise, or alternatively consist of, one or more domains of a GIT transport receptor-binding peptide. In particular, examples of such domains are identified in the examples infra.

Printed from Mimosa .3. Synthesis of Peptides The peptides and derivatives of the present invention may be chemically synthesized or synthesized using recombinant DNA techniques. .3.1. Procedure For Solid Phase Synthesis Peptides may be prepared chemically by methods that are known in the art. For example, in brief, solid phase peptide synthesis consists of coupling the carboxyl group of 10 the C-terminal amino acid to a resin and successively adding N-alpha protected amino acids. The protecting groups may be any known in the art. Before each new amino acid is added to the growing chain, the protecting group of the previous amino acid added to the chain is removed. The coupling of amino 15 acids to appropriate resins is described by Rivier et al., U.S. Patent No. 4,244,946. Such solid phase syntheses have been described, for example, by Merrifield, 1964, J. Am.

Chem. Soc. 85:2149; Vale et al., 1981, Science 213:13 94-1397; Marki et al., 1981, J. Am. Chem. Soc. 103:3178 and in U.S. 20 Patent Nos. 4,305,872 and 4,316,891. In a preferred aspect, an automated peptide synthesizer is employed.

By way of example but not limitation, peptides can be synthesized on an Applied Biosystems Inc. ("ABI") model 431A automated peptide synthesizer using the "Fastmoc" 25 synthesis protocol supplied by ABI, which uses 2-(lH-Benzotriazol-l-yl)-1,1,3,3,-tetramethyluronium hexafluorophosphate ("HBTU") (R. Knorr et al., 1989, Tet. Lett., 30:1927) as coupling agent. Syntheses can be carried out on 0.25 mmol of commercially available 30 4-(2',4'-dimethoxyphenyl-(9-fluorenyl- methoxycarbonyl)-aminomethyl)-phenoxy polystyrene resin ("Rink resin" from Advanced ChemTech) (H. Rink, 1987, Tet. Lett. 28:3787). Fmoc amino acids (1 mmol) are coupled according to the Fastmoc protocol. The following side chain 35 protected Fmoc amino acid derivatives are used: FmocArg(Pmc)OH; FmocAsn(Mbh)OH; FmocAsp(tBu) OH; FmocCys(Acm)OH; FmocGlu(cBu) OH; FmocGln(Mbh)OH; FmocHis(Tr)OH; Printed from Mimosa WO 98/51325 PCT/US98/10088 FmocLys (Boc) OH; FmocSer (cBu) OH; FmocThr ('Bu) OH; FmocTyr(tBu)OH. [Abbreviations: Acm, acetamidomethyl; Boc, tert-butoxycarbonyl; ^u, tert-butyl; Fmoc, 9-fluorenylmethoxycarbonyl; Mbh, 4,4'-dimethoxybenzhydryl; Pmc, 2,2,5,7,8-pentamethylchroman-6-sulfonyl; Tr, trityl] .

Synthesis is carried out using N-methylpyrrolidone (NMP) as solvent, with HBTU dissolved in N,N-dimethylformamide (DMF). Deprotection of the Fmoc group is effected using approximately 20% piperidine in NMP. At 10 the end of each synthesis the amount of peptide present is assayed by ultraviolet spectroscopy. A sample of dry peptide resin (about 3-10 mg) is weighed, then 20% piperidine in DMA (10 ml) is added. After 30 min sonication, the UV (ultraviolet) absorbance of the dibenzofulvene-piperidine 15 adduct (formed by cleavage of the N-terminal Fmoc group) is recorded at 301 nm. Peptide substitution (in mmol g"1) can be calculated according to the equation: A x v substitution = x 1000 7800 x w where A is the absorbance at 301 nm, v is the volume of 20% piperidine in DMA (in ml), 7800 is the extinction coefficient (in mol~1dm3cm"1) of the dibenzofulvene-piperidine adduct, and w is the weight of the peptide-resin sample (in mg). 25 Finally, the N-terminal Fmoc group is cleaved using % piperidine in DMA, then acetylated using acetic anhydride and pyridine in DMA. The peptide resin is thoroughly washed with DMA, CH2C12 and finally diethyl ether. 2Q 5.3.2. Cleavage And Deprotection By way of example but not limitation, cleavage and deprotection can be carried out as follows: The air-dried peptide resin is treated with ethylmethyl-sulfide (EtSMe), ethanedithiol (EDT), and thioanisole (PhSMe) for 35 approximately 20 min. prior to addition of 95% aqueous trifluoracetic acid (TFA). A total volume of approximately 50 ml of these reagents per gram of peptide-resin is used.

Printed from Mimosa The following ratio is used: TFA:EtSMe:EDT:PhSMe (10:0.5:0.5:0.5). The mixture is stirred for 3 h at room temperature under an atmosphere of N2. The mixture is filtered and the resin washed with TFA (2 x 3 ml). The 5 combined filtrate is evaporated in vacuo, and anhydrous diethyl ether added to the yellow/orange residue. The resulting white precipitate is isolated by filtration. See King et al., 1990, Int. J. Peptide Protein Res. 36:255-266 regarding various cleavage methods. .3.3. Purification of the Peptides Purification of the synthesized peptides can be carried out by standard methods including chromatography (e.g., ion exchange, affinity, and sizing column 15 chromatography, high performance liquid chromatography (HPLC)), centrifugation, differential solubility, or by any other standard technique. .3.4. Biological Peptide Libraries Biological peptide libraries can be used to express and identify peptides that bind to GIT transport receptors. According to this second approach, involving recombinant DNA techniques, peptides can, by way of example, be expressed in biological systems as either soluble fusion proteins or viral 25 capsid proteins. .3.4.1. Methods To Identify Binders: Construction Of Libraries In a specific embodiment, the peptides of the 2Q invention that specifically bind to GIT transport receptors are identified by screening a random peptide library by contacting the library with a ligand selected from among HPT1, hPEPTl, D2H, or hSI (or a molecule consisting essentially of an extracellular domain thereof or fragment of 35 the domain) to identify members of the library that specifically bind to the ligand.

Printed from Mimosa In a particular embodiment, a process to identify the peptides of the present method utilizes a library of recombinant vectors constructed by methods well known in the art and comprises screening a library of recombinant vectors 5 expressing inserted synthetic oligonucleotide sequences encoding extracellular GIT transport receptor domains, for example, attached to an accessible surface structural protein of a vector to isolate those members producing peptides that bind to HPT1, hPEPTl, D2H, or hSI. The nucleic acid sequence 10 of the inserted synthetic oligonucleotides of the isolated vector is determined and the amino acid sequence encoded can be deduced to identify a binding domain that binds the ligand of choice (e.g., HPT1, hPEPTl, D2H, or hSI).

The present invention encompasses a method for 15 identifying a peptide which binds to a ligand selected from among HPT1, hPEPTl, D2H, or hSI comprising: screening a library of random peptides with the ligand (or an extracellular domain or fragment thereof) under conditions conducive to ligand binding and isolating the peptide which 20 binds to the ligand. Additionally, the methods of the invention further comprise determining the nucleotide sequence encoding the binding domain of the peptide identified to deduce the amino acid sequence of the binding domain. .3.4.2. Preparation of Extracellular Domain Ligand In a specific embodiment, molecules consisting essentially of an extracellular domain of the desired GIT transport receptor or a fragment of an extracellular domain are used to screen a random peptide library for binding thereto. Preferably, a nucleic acid encoding the extracellular domain is cloned and recombinantly expressed, and the domain is then purified for use. The GIT transport 35 receptor is preferably selected from among HPT1, hPEPTl, D2H, or hSI.

Printed from Mimosa PCMJS98/10088 .3.4.3. Methods to Identify Binders: Screening Libraries Once a suitable random peptide library has been constructed (or otherwise obtained), the library is screened 5 to identify peptides having binding affinity for the GIT transport receptor, e.g., HPTl, hPEPTl, D2H, or hSI. In a preferred aspect, the library is a TSAR library (see U.S. Patent No. 5,498,538 dated March 12, 1996 and PCT Publication WO 94/18318 dated August 18, 1994, both of which are incorporated by reference herein in their entireties). Screening the libraries can be accomplished by any of a variety of methods known to those of skill in the art. See, e.g., the following references, which disclose screening of peptide libraries: Parmley and Smith, 1989, Adv. Exp. Med.

Biol. 251: 215-218; Scott and Smith, 1990, Science 249: 386-390; Fowlkes et al. , 1992; BioTechniques 13.: 422-427; Oldenburg et al., 1992, Proc. Natl. Acad. Sci. USA £9: 5393-5397; Yu et al., 1994, Cell 76: 933-945; Staudt et al., 1988, Science 241: 577-580; Bock et al., 1992, Nature 355: 564-566; 2Q Tuerk et al. , 1992, Proc. Natl. Acad. Sci. USA 89_: 6988.-6 992; Ellington et al., 1992, Nature 355: 850-852; U.S. Patent No. 5,096,815, U.S. Patent No. 5,223,409, and U.S. Patent No. 5,198,346, all to Ladner et al.; and Rebar and Pabo, 1993, Science 263: 671-673. See also PCT publication WO 94/18318, dated August 18, 1994.

One of ordinary skill in the art will recognize that, with suitable modifications, the screening methods described below would be suitable for a wide variety of biological expression libraries. 2Q Once a library has been constructed or otherwise obtained, the library is screened to identify binding molecules having specific binding affinity for a ligand for a GIT transport receptor preferably selected from among HPTl, hPEPTl, D2H, or hSI.

Screening the libraries can be accomplished by any of a variety of methods known to those of skill in the art. Exemplary screening methods are described in Fowlkes et al., Printed from Mimosa 1992, BioTechniques, 13:422-427 and include contacting the vectors with an immobilized target ligand and harvesting those vectors that bind to said ligand. Such useful screening methods, are designated "panning" methods. In 5 panning methods useful to screen the present libraries, the target ligand can be immobilized on plates, beads (such as magnetic beads), sepharose, beads used in columns, etc. If desired, the immobilized target ligand can be "tagged", e.g., using labels such as biotin, fluoroscein isothiocyanate, 10 rhodamine, etc. e.g. for FACS sorting. Panning is also disclosed in Parmley, S.F. and Smith, G.P., 1988, Gene 73: 305-318.

In a particular embodiment of the invention, the library can be screened with a recombinant receptor domain. 15 In another embodiment, the library can be screened successively with receptor domains and then on CaCO-2 cells.

For screening of the peptide libraries in vitro, the solvent requirements involved in screening are not limited to aqueous solvents; thus, nonphysiological binding 20 interactions and conditions different from those found in - i vivo can be exploited.

Screening a library can be achieved using a method comprising a first "enrichment" step and a second filter lift as follows. The following description is given by way of 25 example, not limitation.

Binders from an expressed library [e.g., m phage) capable of binding to a given ligand ("positives") are initially enriched by one or two cycles of panning or affinity chromatography. A microtiter well is passively 30 coated with the ligand (e.g., about 10 fig in 100 /xl) . The well is then blocked with a solution of BSA to prevent nonspecific adherence of the phage of the library to the plastic surface. For example, about 1011 phage particles expressing peptides are then added to the well and incubated for several 35 hours. Unbound phage are removed by repeated washing of the plate, and specifically bound phage are eluted using an acidic glycine-HCl solution or other elution buffer. The Printed from Mimosa eluted phage solution is neutralized with alkali, and amplified, e.g., by infection of E. coli and plating on large petri dishes containing Luria broth (LB) in agar. Amplified cultures expressing the binding peptides are then titered and 5 the process repeated. Alternatively, the ligand can be covalently coupled to agarose or acrylamide beads using commercially available activated bead reagents. The phage solution is then simply passed over a small column containing the coupled bead matrix which is then washed extensively and 10 eluted with acid or other eluant. In either case, the goal is to enrich the positives to a frequency of about > 1/105.

Following enrichment, a filter lift assay is conducted-. For example, when specific binders are expressed in phage, approximately 1-2 x 105 phage are added to 500 fil of 15 log phase E. coli and plated on a large Luria Broth-agarose plate with 0.7% agarose in broth. The agarose is allowed to solidify, and a nitrocellulose filter (e.g., 0.45 /x) is placed on the agarose surface. A series of registration marks is made with a sterile needle to allow re-alignment of 20 the filter and plate following development as described below. Phage plaques are allowed to develop by overnight incubation at 37 °C (the presence of the filter does not inhibit this process). The filter is then removed from the plate with phage from each individual plaque adhered in situ. 25 The filter is then exposed to a solution of BSA or other blocking agent for 1-2 hours to prevent non-specific binding of the ligand (or "probe").

The probe itself is labeled, for example, either by biotinylation (using commercial NHS-biotin) or direct enzyme 30 labeling, e.g., with horse radish peroxidase or alkaline phosphatase. Probes labeled in this manner are indefinitely stable and can be re-used several times. The blocked filter is exposed to a solution of probe for several hours to allow the probe to bind in situ to any phage on the filter 35 displaying a peptide with significant affinity to the probe. The filter is then washed to remove unbound probe, and then developed by exposure to enzyme substrate solution (in the Printed from Mimosa case of directly labeled probe) or further exposed to a solution of enzyme-labeled avidin (in the case of biotinylated probe). Positive phage plaques are identified by localized deposition of colored enzymatic cleavage product 5 on the filter which corresponds to plaques on the original plate. The developed filter is simply realigned with the plate using the registration marks, and the "positive" plaques are cored from the agarose to recover the phage. Because of the high density of plaques on the original plate, 10 it may be difficult to isolate a single plaque from the plate on the first pass. Accordingly, phage recovered from the initial core can be re-plated at low density and the process can be repeated to allow isolation of individual plaques.^nd hence single clones of phage.

Successful screening experiments are optimally conducted using 3 rounds of serial screening. The recovered cells are then plated at a low density to yield isolated colonies for individual analysis. The individual colonies are selected and used to inoculate LB culture medium 20 containing ampicillin. After overnight culture at 37°C, the cultures are then spun down by centrifugation. Individual cell aliquots are then retested for binding to the target ligand attached to the beads. Binding to other beads having attached thereto a non-relevant ligand, can be used as a 25 negative control.

One aspect of screening the libraries is that of elution. The following discussion is applicable to any system where the random peptide is expressed on a surface fusion molecule. It is conceivable that the conditions that 30 disrupt the peptide-target interactions during recovery of the phage are specific for every given peptide sequence from a plurality of proteins expressed on phage. For example, certain interactions may be disrupted by acid pH but not by basic pH, and vice versa. Thus, it may be desirable to test 35 a variety of elution conditions (including but not limited to pH 2-3, pH 12-13, excess target in competition, detergents, mild protein denaturants, urea, varying temperature, light, Printed from Mimosa presence or absence of metal ions, chelators, etc.) and compare the primary structures of the binding proteins expressed on the phage recovered for each set of conditions to determine the appropriate elution conditions for each 5 ligand/binding protein combination. Some of these elution conditions may be incompatible with phage infection because they are bactericidal and will need to be removed by dialysis (i.e., dialysis bag, Centricon/Amicon microconcentrators).

In a preferred embodiment, a phage display library 10 of random peptides is screened to select phage expressing peptides that bind to a GIT transport receptor. Preferably, a first step is to isolate a preselected phage library. The "preselected phage library" is a library consisting of a subpopulation of a phage display library. This subpopulation 15 can be formed by initially screening against either a target GIT transport receptor (or domain thereof) so as to permit the selection of a subpopulation of phages which specifically bind to the receptor. Alternatively, the subpopulation can be formed by screening against a target cell or cell type or 20 tissue type or tissue barrier of the gastro-intestinal tract, so as to permit the selection of a subpopulation of phages which either bind specifically to the target cell or target cell type or target tissue or target tissue barrier, or which binds to and/or is transported across (or between) the target 25 cell or target cell type or target tissue or target tissue barrier either in situ or in vivo. This preselected phage library or subpopulation of selected phages can also be rescreened against the target GIT transport receptor, permitting the further selection of a subpopulation of phages 30 which bind to the GIT transport receptor or target cell or target cell type or target tissue or target tissue barrier or which bind to and/or is transported across the target cell, target tissue or target tissue barrier either in situ or in vivo. Such rescreening can be repeated from zero to 30 times 35 with each successive "pre-selected phage library" generating additional pre-selected phage libraries.

Printed from Mimosa In a preferred embodiment, a preselected phage library binding a ligand that is a GIT transport receptor preferably selected from among HPTl, hPEPTl, D2H, or hSI is obtained by an in vitro screening step as described above, 5 and then the phage are optionally further characterized using in vitro assays consisting of binding phage directly to the receptor domain of interest or, alternatively, to Caco-2 cells or using in vivo assays. In another preferred embodiment, in vivo assays are used that measure uptake of 10 phage by intestinal tissue or, alternatively, through the GIT. In alternative embodiments, such further in vitro or in vivo assays can be used as the initial screening step.

■I In vivo assays that may be used are described in : the examples infra. .4. Generation of Antibodies to GIT Transport Receptor-Binding Peptides and Derivatives Thereof According to the invention, a GIT transport receptor-binding peptide, fragments or other derivatives, or 2q analogs thereof, may be used as an immunogen to generate antibodies which immunospecifically bind such an immunogen.

Such antibodies include but are not limited to polyclonal, monoclonal, chimeric, single chain, Fab fragments, and an Fab expression library.

Various procedures known in the art may be used for the production of polyclonal antibodies to a GIT transport receptor-binding peptide or derivative or analog. For the production of antibody, various host animals can be immunized by injection with the native GIT transport receptor-binding 2Q peptides, or a synthetic version, or derivative {e.g., fragment) thereof, including but not limited to rabbits, mice, rats, fowl, etc. Various adjuvants may be used to increase the immunological response, depending on the host species, including but not limited to Freund's (complete and incomplete), mineral gels such as aluminum hydroxide, surface active substances such as lysolecithin, pluronic polyols, polyanions, peptides, oil emulsions, keyhole limpet Printed from Mimosa heraocyanins, dinitrophenol, and potentially useful human adjuvants such as BCG (bacille Calmette-Guerin) and corynebacterium parvum.

For preparation of monoclonal antibodies directed 5 toward a GIT transport receptor-binding peptide or analog thereof, any technique which provides for the production of antibody molecules by continuous cell lines in culture may be used. For example, the hybridoma technique originally developed by Kohler and Milstein (1975, Nature 256:495-497), 10 as well as the trioma technique, the human B-cell hybridoma technique (Kozbor et al., 1983, Immunology Today 4:72), and the EBV-hybridoma technique to produce human monoclonal antibodies (Cole et al., 1985, in Monoclonal Antibodies and Cancer Therapy, Alan R. Liss, Inc., pp. 77-96). In an 15 additional embodiment of the invention, monoclonal antibodies can be produced in germ-free animals utilizing recent technology (PCT/US90/02545). According to the invention, human antibodies may be used and can be obtained by using human hybridomas (Cote et al., 1983, Proc. Natl. Acad. Sci. 20 U.S.A. 80:2026-2030) or by transforming human B cells with EBV virus in vitro (Cole et al., 1985, in Monoclonal Antibodies and Cancer Therapy. Alan R. Liss, pp. 77-96) . According to the invention, techniques developed for the production of "chimeric antibodies" (Morrison et al., 1984, 25 Proc. Natl. Acad. Sci. U.S.A. 81:6851-6855; Neuberger et al., 1984, Nature 312:604-608; Takeda et al., 1985, Nature 314:452-454) by splicing the genes from a mouse antibody molecule specific for GIT transport receptor-binding peptides together with genes from a human antibody molecule of 30 appropriate biological activity can be used.

According to the invention, techniques described for the production of single chain antibodies (U.S. Patent 4,946,778) can be adapted to produce GIT transport receptor-binding peptide-specific single chain antibodies. An 35 additional embodiment of the invention utilizes the techniques described for the construction of Fab expression libraries (Huse et al., 1989, Science 246:1275-1281) to allow Printed from Mimosa rapid and easy identification of monoclonal Fab fragments with the desired specificity for GIT transport receptor-binding peptides, derivatives, or analogs.

Antibody fragments which contain the idiotype of 5 the molecule can be generated by known techniques. For example, such fragments include but are not limited to: the F(ab')2 fragment which can be produced by pepsin digestion of the antibody molecule; the Fab' fragments which can be generated by reducing the disulfide bridges of the F(ab')2 10 fragment, the Fab fragments which can be generated by treating the antibody molecule with papain and a reducing agent, and Fv fragments.

In the production of antibodies, screening for the desired antibody can be accomplished by techniques known in 15 the art, e.g. ELISA (enzyme-linked immunosorbent assay). For example, to select antibodies which recognize a specific domain of a GIT transport receptor-binding peptide, one may assay generated hybridomas for a product which binds to a GIT transport receptor-binding peptide fragment containing such a 2 0 domain.

Antibodies specific to a domain of a GIT transport receptor-binding peptide are also provided.

The foregoing antibodies can be used in methods known in the art relating to the localization and activity of 25 the GIT transport receptor-binding peptide sequences of the invention, e.g., for imaging these peptides after in vivo administration (e.g., to monitor treatment efficacy), measuring levels thereof in appropriate physiological samples, in diagnostic methods, etc. For instance, 30 antibodies or antibody fragments specific to a domain of a GIT transport receptor-binding peptide or to a derivative of a peptide, such as a dansyl group or some other epitope introduced into the peptide, can be used to 1) identify the presence of the peptide on a nanoparticle or other substrate: 35 2) quantify the amount of peptide on the nanoparticle; 3)measure the level of the peptide in appropriate physiological samples; 4) perform immunohistology on tissue Printed from Mimosa samples; 5) image the peptide after in vivo administration; 6) purify the peptide from a mixture using an immunoaffinity column or 7) bind or fix the peptide to the surface of nanoparticle. This last use envisions attaching the antibody 5 (or fragment of the antibody) to the surface of drug-loaded nanoparticles or other substrate and then incubating this conjugate with the peptide. This procedure results in binding of the peptide in a certain fixed orientation, resulting in a particle that contains the peptide bound to 10 the antibody in such a way that the peptide is fully active.

Abtides (or Antigen binding peptides) specific to a domain of a GIT transport receptor-binding peptide or to a derivative of a peptide, such as a dansyl group or some other epitope introduced into the peptide, can be used for the same 15 seven purposes identified above for antibodies. .5. Assays of GIT Transport Receptor-Binding Peptides. Derivatives and Analogs The functional activity of GIT transport receptor-2Q binding peptides, derivatives and analogs can be assayed by various methods.

In a preferred embodiment, in which binding to a GIT transport receptor is being assayed, the binding can be assayed by in vivo or in vitro assays such as described in 25 the examples infra, or by other means that are known in the art.

In another embodiment, where one is assaying for the ability to bind or compete with full-length GIT transport receptor-binding peptide for binding to anti-GIT transport 2Q receptor-binding peptide antibody, various immunoassays known in the art can be used, including but not limited to competitive and non-competitive assay systems using techniques such as radioimmunoassays, ELISA (enzyme linked immunosorbent assay), "sandwich" immunoassays, 35 immunoradiometric assays, gel diffusion precipitin reactions, immunodiffusion assays, in situ immunoassays (using colloidal gold, enzyme or radioisotope labels, for example), western Printed from Mimosa blots, precipitation reactions, agglutination assays (e.g., gel agglutination assays, hemagglutination assays), complement fixation assays, immunofluorescence assays, protein A assays, and immunoelectrophoresis assays, etc. In 5 one embodiment, antibody binding is detected by detecting a label on the primary antibody. In another embodiment, the primary antibody is detected by detecting binding of a secondary antibody or reagent to the primary antibody. In a further embodiment, the secondary antibody is labelled. Many 10 means are known in the art for detecting binding in an immunoassay and are within the scope of the present invention.

Other methods will be known to the skilled artisan and are within the scope of the invention. .6. Uses The invention provides compositions comprising the GIT transport receptor-binding proteins of the invention bound to a material comprising an active agent. Such 20 compositions have use in targeting the active agent to the GIT and/or in facilitating transfer through the lumen of the GIT into the systemic circulation. Where the active agent is an imaging agent, such compositions can be administered in vivo to image the GIT (or particular transport receptors 25 thereof). Other active agents include but are not limited to: any drug or antigen or any drug- or antigen-loaded or drug- or antigen-encapsulated nanoparticle, microparticle, liposome, or micellar formulation capable of eliciting a biological response in a human or animal. Examples of drug-30 or antigen-loaded or drug- or antigen-encapsulated formulations include those in which the active agent is encapsulated or loaded into nano- or microparticles, such as biodegradable nano- or microparticles, and which have the GIT transport receptor-binding protein or derivative or analog 35 adsorbed, coated or covalently bound, such as directly linked or linked via a linking moiety, onto the surface of the nano-or microparticle. Additionally, the protein, derivative or Printed from Mimosa analog can form the nano- or microparticle itself or the protein, derivative or analog can be covalently attached to the polymer or polymers used in the production of the biodegradable nano- or microparticles or drug-loaded or drug-5 encapsulated nano- or microparticles or the peptide can be directly conjugated to the active agent. Such conjugations to active agents include fusion proteins in which a DNA sequence coding for the peptide is fused in-frame to the gene or cDNA coding for a therapeutic peptide or protein such that 10 the modified gene codes for a recombinant fusion protein.

In a preferred embodiment, the invention provides for treatment of various diseases and disorders by administration of a therapeutic compound (termed herein "Therapeutic"). Such "Therapeutics" include but are not 15 limited to: GIT transport receptor-binding proteins, and analogs and derivatives (including fragments) thereof (e.g., as described hereinabove) that bind to GIT transport receptors, bound to an active agent of value in the treatment or prevention of a disease or disorder (preferably a , 20 mammalian, most preferably human, disease or disorder). Therapeutics also include but are not limited to nucleic acids encoding the GIT transport receptor-binding proteins, analogs, or derivatives bound to such a therapeutic or prophylactic active agent. The active agent is preferably a 2 5 drug.

Any drug known in the art may be used, depending upon the disease or disorder to be treated or prevented, and the type of subject to which it is to be administered. As used herein, the term "drug" includes, without limitation, 30 any pharmaceutically active agent. Representative drugs include, but are not limited to, peptides or proteins, hormones, analgesics, anti-migraine agents, anti-coagulant agents, anti-emetic agents, cardiovascular agents, antihypertensive agents, narcotic antagonists, chelating agents, 35 anti-anginal agents, chemotherapy agents, sedatives, antineoplastics, prostaglandins, and antidiuretic agents.

Typical drugs include peptides, proteins or hormones such as Printed from Mimosa insulin, calcitonin, calcitonin gene regulating protein, atrial natriuretic protein, colony stimulating factor, betaseron, erythropoietin (EPO), interferons such as a, 0 or 7 interferon, somatropin, somatotropin, somatostatin, 5 insulin-like growth factor (somatomedins), luteinizing hormone releasing hormone (LHRH), tissue plasminogen activator (TPA), growth hormone releasing hormone (GHRH), oxytocin, estradiol, growth hormones, leuprolide acetate, factor VIII, interleukins such as interleukin-2, and analogs 10 thereof; analgesics such as fentanyl, sufentanil, butorphanol, buprenorphine, levorphanol, morphine, hydromorphone, hydocodone, oxymorphone, methadone, lidocaine, bupivacaine, diclofenac, naproxen, paverin, and analogs thereof; anti-migraine agents such as heparin, hirudin, and 15 analogs thereof; anti-coagulant agents such as scopolamine, ondansetron, domperidone, etoclopramide, and analogs thereof; cardiovascular agents, anti-hypertensive agents and vasodilators such as diltiazem, clonidine, nifedipine, verapamil, isosorbide-5-mononitrate, organic nitrates, agents 20 used in treatment of heart disorders and analogs thereof; sedatives such as benzodiazeines, phenothiozines and analogs thereof; narcotic antagonists such as naltrexone, naloxone and analogs thereof; chelating agents such as deferoxamine and analogs thereof; anti-diuretic agents such as 25 desmopressin, vasopressin and analogs thereof; anti-anginal agents such as nitroglycerine and analogs thereof; antineoplastics such as 5-fluorouracil, bleomycin and analogs thereof; prostaglandins and analogs thereof; and chemotherapy agents such as vincristine and analogs thereof. 30 Representative drugs also include but are not limited to antisense oligonucleotides, genes, gene correcting hybrid oligonucleotides, ribozymes, aptameric oligonucleotides, triple-helix forming oligonucleotides, inhibitors of signal transduction pathways, tyrosine kinase inhibitors and DNA 35 modifying agents. Drugs that can be used also include, without limitation, systems containing gene therapeutics, including viral systems for therapeutic gene delivery such as Printed from Mimosa adenovirus, adeno-associated virus, retroviruses, herpes simplex virus, sindbus virus, liposomes, cationic lipids, dendrimers, and enzymes. For instance, gene delivery viruses can be modified such that they express the targeting peptide 5 on the surface so as to permit targeted gene delivery.

In a preferred embodiment, a Therapeutic is therapeutically or prophylactically administered to a human patient.

Additional descriptions and sources of Therapeutics 10 that can be used according to the invention are found in various Sections herein. .7. Therapeutic/Prophylactic Administration, Compositions and Formulations The invention provides methods of treatment (and prophylaxis) by administration to a subject of an effective amount of a Therapeutic of the invention. In a preferred aspect, the Therapeutic is substantially purified. The subject is preferably an animal, including but not limited to 2Q animals such as cows, pigs, horses, chickens, cats, dogs, etc., and is preferably a mammal, and most preferably a human.

As will be clear, any disease or disorder of interest amenable to therapy or prophylaxis by providing a 2g drug in vivo systemically or by targeting a drug in vivo to the GIT (by linkage to a GIT transport-receptor binding protein, derivative or analog of the invention) can be treated or prevented by administration of a Therapeutic of the invention. Such diseases may include but are not limited 20 to hypertension, diabetes, osteoporosis, hemophilia, anemia, cancer, migraine, and angina pectoris, to name but a few.

Any route of administration known in the art may be used, including but not limited to oral, nasal, topical, intravenous, intraperitoneal, intradermal, mucosal, 25 intrathecal, intramuscular, etc. Preferably, administration is oral; in such an embodiment the GIT-transport binding protein, derivative or analog of the invention acts Printed from Mimosa advantageously to facilitate transport of the therapeutic active agent through the lumen of the GIT into the systemic circulation.

The present invention also provides therapeutic 5 compositions/formulations. In a specific embodiment of the invention, a GIT transport receptor-binding peptide or motif of interest is associated with a therapeutically or prophylactically active agent, preferably a drug or drug-containing nano- or microparticle. More preferably, the 10 active agent is a drug encapsulating or drug loaded nano- or microparticle, such as a biodegradable nano- or microparticle, in which the peptide is physically adsorbed or coated or covalently bonded, such as directly linked or linked via a linking moiety, onto the surface of the nano- or 15 microparticle. Alternatively, the peptide can form the nano-or microparticle itself or can be directly conjugated to the active agent. Such conjugations include fusion proteins in which a DNA sequence coding for the peptide is fused in-frame to the gene or cDNA coding for a therapeutic peptide or 20 protein, such that the modified gene codes for a recombinant fusion protein in which the "targeting" peptide is fused to the therapeutic peptide or protein and where the "targeting" peptide increases the absorption of the fusion protein from the GIT. Preferably the particles range in size from 200-600 25 nm.

Thus, in a specific embodiment, a GIT transport-binding protein is bound to a slow-release (controlled release) device containing a drug. In a specific embodiment, polymeric materials can be used (see Medical Applications of 30 Controlled Release, Langer and Wise (eds.), CRC Pres., Boca Raton, Florida (1974); Controlled Drug Bioavailability, Drug Product Design and Performance, Smolen and Ball (eds.), Wiley, New York (1984); Ranger and Peppas, J. Macromol. Sci. Rev. Macromol. Chem. 23:61 (1983); see also Levy et al., 35 Science 228:190 (1985); During et al., Ann. Neurol. 25:351 (1989); Howard et al., J. Neurosurg. 71:105 (1989)).

Printed from Mimosa The present invention also provides pharmaceutical compositions. Such compositions comprise a therapeutically effective amount of a Therapeutic, and a pharmaceutically acceptable carrier. In a specific embodiment, the term 5 "pharmaceutically acceptable" means approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term "carrier" refers to a diluent, adjuvant, excipient, or 10 vehicle with which the therapeutic is administered. Such pharmaceutical carriers can be sterile liquids, such as water and oils, including those of petroleum, animal, vegetable or synthetic origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred carrier 15 when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable pharmaceutical excipients include starch, glucose, lactose, 20 sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like The composition, if desired, can also contain minor amounts of wetting or emulsifying 25 agents, or pH buffering agents. These compositions can take the form of solutions, suspensions, emulsion, tablets, pills, capsules, powders, sustained-release formulations and the like. The composition can be formulated as a suppository, with traditional binders and carriers such as triglycerides. 30 Oral formulations can include standard carriers such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, sodium saccharine, cellulose, magnesium carbonate, etc. Examples of suitable pharmaceutical carriers are described in "Remington's Pharmaceutical Sciences" by E.W. 35 Martin. Such compositions will contain a therapeutically effective amount of the Therapeutic, preferably in purified Printed from Mimosa PCT/U S98/10088 form, together with a suitable amount of carrier so as to provide the form for proper administration to the patient.

The Therapeutics of the invention can be formulated as neutral or salt forms. Pharmaceutically acceptable salts 5 include those formed with free amino groups such as those derived from hydrochloric, phosphoric, acetic, oxalic, tartaric acids, etc., and those formed with free carboxyl groups such as those derived from sodium, potassium, ammonium, calcium, ferric hydroxides, isopropylamme, 10 triethylamine, 2-ethylamino ethanol, histidine, procaine, etc.

The amount of the Therapeutic of the invention which will be effective in the treatment of a particular disorder or condition will depend on the nature of the 15 disorder or condition, and can be determined by standard clinical techniques. In addition, in vitro assays may optionally be employed to help identify optimal dosage ranges. The precise dose to be employed in the formulation will also depend on the route of administration, and the 20 seriousness of the disease or disorder, and should be decided according to the judgment of the practitioner and each patient's circumstances. 6. EXAMPLES 25 6.1. Selection of GIT Receptor Targets The HPTl, hPEPTl, D2H, and hSI receptors were selected for cloning as GIT receptor targets based on several criteria, including: (1) expression on surface of epithelial cells in gastro-intestinal tract (GIT); (2) expression along 30 the length of small intestine (HPTl, hPEPTl, D2H); (3) expression locally at high concentration (hSI); (4) large putative extracellular domains facing into the lumen of the GIT; and (5) extracellular domains that permit easy access and bioadhesion by targeting particles.

The four recombinant receptor sites screened with the peptide libraries additionally have the following characteristics: Printed from Mimosa Receptor Characteristics D2H Transport of neutral/basic amino acids; a transport activating protein for a range of amino acid translocases hSI Metabolism of sucrose and other sugars; represents 9% of brush border membrane protein in Jejunum HPTl di/tri peptide transporter or facilitator of peptide transport hPEPTl di/tri peptide transporter Figures 1-4 (SEQ ID NOS:176, 178, 179, and 181, respectively) show the predicted amino acid sequences for hPEPTl, HPTl, hSI and D2H, respectively. 6.2. Cloning of Extracellular Domain of Selected Receptor Site The following receptor domains were cloned and expressed as His-tag fusion proteins by standard techniques: Domain (amino Receptor , acid residues) hPEPTl3 391-571 HPTlb 29-273 hSI0 272-667 D2Hd 387-685 a Liang et al. , 1995, J. Biol. Chem. 270:6456-6463 b Dantzig et al. , 1994, Association of Intestinal Peptide Transport with a Protein Related to the Cadherin Superfamily c Chantret et al. , Biochem. J. 285:915-923 d Bertran et al., J. Biol. Chem. 268:14842-14949 The receptor proteins were expressed as His-tag fusion proteins and affinity purified under denaturing conditions, using urea or guanidine HC1, utilizing the pET His-tag metal chelate affinity for Ni-NTA Agarose (Hochuli, E., Purification of recombinant proteins with metal chelate __ adsorbent, Genetic Engineering, Principals and Methods (J.K. 3 5 Setlow, ed.), Plenum Press, NY, Vol. 12 (1990), pp. 87-98).

Printed from Mimosa 6.3. Phage Libraries Three phage DC8, D3 8, and DC43 libraries expressing N-terminal pIII fusions in M13 were used to identify peptides that bind to the GIT receptors. The D38 and DC43 libraries 5 which are composed of 37 and 43 random amino acid domains, respectively, have been described previously (McConnell et al., 1995, Molecular Diversity, 1:165-176). The DC8 library is similar to the other two except that the random insert is 8 amino acids long flanked on each side by a cysteine residue 10 (i.e., CX8C) . 6.4. Biopanning Three rounds of biopanning on the GIT receptors were performed generally by standard methods (McConnell et 15 al. , 1995, Molecular Diversity, 1:165-176), using a mixture of the DC8 (1 x 1010 pfu) , D38 and DC43 (1 x 1011 pfu) phage libraries. After each round of panning the percentage of phage recovered was determined. Following the first two rounds of panning, the eluted phage were amplified overnight. 20 Phage from the third pan were plated out and 100 plaques were picked, amplified overnight and screened in an ELISA assay for binding to the relevant receptor and BSA. After data analysis, phage clones were identified which had high absorbance in the ELISA assay and/or a good ratio of binding 25 to target compared to binding to BSA. The Insulin Degrading Enzyme (IDE) and recombinant human tissue factor (hTF) were used as irrelevant controls. Several variations of the standard panning technique, discussed below, were used. Selection or panning methods followed one of two strategies. 30 The first strategy involved panning the mixed libraries on the specific GIT receptor adsorbed to a solid surface. The second strategy panned the libraries twice against the GIT receptor and then against Caco-2 cells (Peterson and Mooseker, 1992, J. Cell Science 102:581-600), Selection 35 methods are reflected in the clone nomenclature as described below: Printed from Mimosa S designates the clone was identified by binding to the hSI receptor domain.

D designates the clone was identified by binding to the D2H receptor domain.

P designates the clone was identified by binding to the PEPT1 receptor domain.

H designates the clone was identified by binding to the HPT-1 receptor domain.

Phage designated Ni are from a solid phase band GIT 10 receptor pan that used the standard procedure with the addition of Ni-NTA Agarose (Qiagen, Chatsworth, CA).

Receptor coated plates were blocked with 0.5% BSA/PBS containing 160/xl Ni-NTA agarose and libraries were panned in-the presence of 50/il Ni-NTA agarose. The receptor proteins 15 were expressed as His-tag fusions. The His-tag has a high affinity for Ni-NTA Agarose. Blocking the plate and panning in the presence of Ni-NTA agarose minimized phage binding to the His-tag portion of the recombinant receptor.

Phage with the designation AX were eluted with acid 20 and Factor Xa. Phage were first eluted by standard acid elution then Factor Xa (New England Biolabs, Beverly, MA: 1/ig protease in 300/xl of 20mM Tris-HCL, lOOmM NaCl, 2mM CaCl2) was added to the panning plate and incubated 2 hours. Phage from both elution methods were pooled together then plated. 25 Phage with the designation AB were eluted with acid and base. Phage were eluted first by standard acid elution then lOOmM triethylamine pH 12.1 was added to the panning plate for 10 minutes. Phage from both elution methods were pooled together then plated.

C designates panning on receptor followed by Caco-2 cells. First and second round pans were performed on the receptor and the third round pan was on snapwells of Caco-2 cells. DCX11, DCX8 and DCX33 were identified by two pans on D2H receptor, third pan on Caco-2 cells. The third round 35 Factor Xa eluate from the Caco-2 cells was screened by ELISA on D2H, BSA and fixed Caco-2 cells. For HCA3 the first two rounds of panning were performed on the HPT-1 receptor and Printed from Mimosa the third pan was on monolayers cultured on snapwells of Caco-2 cells.

Phage designated 5PAX were carried through five rounds of panning after which a number of phage were 5 sequenced prior to screening by ELISA. 6.5. Sequencing of Selected Phage The amino acid sequence of phage inserts demonstrating a good ratio of binding to receptor domains 10 and/or Caco-2 cells over background BSA binding were deduced from the nucleotide sequence obtained by sequencing (Sequenase®, U.S. Biochemical Corp., Cleveland, OH) both DNA strands of the appropriate region in the viral genome. The third round acid eluate was screened by ELISA on HPT-1, BSA 15 and Caco-2 fixed cells. Phage designated 5PAX were carried through five rounds of panning after which a number of phages were sequenced prior to screening by ELISA.

One well of a 24 well plate was coated with 10 - pig/ml of GIT receptor and the plate was incubated overnight 20-at 4°C. The plate was blocked with 0.5 BSA-PBS for one hour. A mixture of the DC8, D3 8 and DC43 phage libraries was added to the plate and the plate was incubated for 2 to 3 hours at room temperature on a rotator. After washing the well 10 times with 1% BSA plus 0.05% Tween 20 in PBS, the well was 25 eluted with 0.05m glycine, pH2. The phage was then eluted with 0.2M NaP04. The eluted phage was titered on agar plates; the remaining phage was amplified overnight. The next day the amplified phage was added to a second coated plate and the panning procedure was repeated as described above. The 30 eluted phage from the second pan as well as the amplified phage from the first pan was titered on agar plates.

Following amplification overnight of the phage from the second pan, the panning procedure was repeated as described above. The phage eluted from the third pan and the amplified 35 phage from the second pan were then titered overnight on agar plates. Isolated phage colonies were amplified overnight prior to use in an ELISA assay.

Printed from Mimosa 6.6. Receptor ELISA Procedure 96 well plates were coated overnight with GIT receptor, BSA and, optionally, IDE (insulin degrading enzyme, an irrelevant His-fusion protein)or hTF. The plates were 5 blocked for one hour with 0.5% BSA-PBS. After clarification, the amplified phage were diluted 1:100 in 1% BSA plus 0.05% Tween 20 in PBS and added to the plates. Following incubation of the plates on a rotator for 1 to 2 hours, the plates were washed 5 times with 1% BSA plus 0.05% Tween 20 in 10 PBS. Dilute anti-M13-HRP conjugate (anti-M13 antibody linked to horse radish peroxidase (HRP)) was added to all the wells and the plate was incubated for one hour on a rotator. After the plates were washed 5 times, as described above, TMB substrate was added to the wells. The plates were read at 15 650nm absorbance.

RECEPTOR ELISA RESULTS: Below are the results of ELISA assays which assessed the binding of phage panned on the hSI receptor to 20 microtiter plates coated with hSI and BSA. Table 1 shows the OD results as well as the ratio of hSI to BSA binding.

Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 1 PHAGE hSI BSA hSI/BSA S15 0 .478 0 . 053 9 S21 0 . 845 0 . 092 9 S22 0 .399 0 . 061 7 SNilO 0.57 0.051 11 SNi28 0 . 942 0 .113 8 SNi34 0 . 761 0 .115 7 SNi38 0 .466 0 . 076 6 SNi45 0 . 518 0 . 056 9 SNiAX2 0 . 383 0 . 065 6 SNiAX6 0 . 369 0.056 7 SNiAX8 0 . 342 0 . 068 BLANK 0 . 063 0 . 042 2 Below are the results of an ELISA which assessed the binding of phage panned on the D2H receptor to microtiter plates coated with D2H and BSA. Table 2 shows the OD results 20 as well as the ratio of D2H to BSA binding.

Table 2 Phage D2H BSA D2H/BSA DAB 3 0 .406 0 . 072 6 DAB 7 0.702 0 .09 8 DAB 10 0.644 0 .153 4 DAB18 0.467 0 . 085 DAB24 1.801 0 .441 4 DAB30 0.704 0 .121 6 DAX15 0.391 0 .101 4 DAX23 0.698 0 .153 DAX24 0.591 0 .118 DAX27 1.577 0 .424 4 BLANK 0 . 038 0 .037 1 Below are the results of an ELISA which assessed 35 the binding of phage panned for two rounds on the D2H receptor followed by a third round pan on Caco-2 snapwells. Binding to fixed Caco-2 cells, D2H and BSA was examined.

Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 3 shows the OD results as well as the ratio of D2H to BSA binding.

Table 3 PHAGE Caco-2 D2H BSA D2H/BSA DCX8 0 .498 0 .163 0 . 063 3 DCX11 0 .224 0.222 0.071 3 DCX26 0.114 0.956 0.213 4 DCX33 0.164 0.616 0.103 6 DCX36 0 .149 0.293 0 . 064 DCX3 9 0 .121 0.299 0 . 066 DCX42 0 .308 0.158 0.065 2 DCX45 0 .147 0.336 0 . 075 4 Blank 0.065 0.043 o o 1 Below are the results of an ELISA which assessed the binding of phage panned on the hPEPTl receptor to hPEPTl and BSA. Table 4 shows the OD results as well as the ratio of hPEPTl to BSA binding.

Table 4 PHAGE hPEPTl BSA PEPT1/BSA PAX 9 0 .312 0 . 079 4 PAX14 1.102 0.139 8 PAX15 0 .301 0.079 4 PAX16 0 . 648 0 .171 4 PAX17 0.514 0 . 095 PAX18 0.416 0.087 PAX35 0 .474 0 . 065 7 PAX38 0 .292 0.064 PAX40 0 .461 0.076 6 PAX43 0 .345 0.069 PAX4 5 0 .419 0.081 PAX46 0 .429 0.077 6 P31 0 . 807 0 . 075 11 P90 1.117 0.107 9 5PAX3 0 .173 0.04 4 5PAX5 0 .15 0.036 4 5PAX7 0.171 0.037 5PAX12 0 .227 o o 6 Blank 0 .102 0.039 3 Printed from Mimosa Table 5 shows the results of an ELISA which assessed the binding of phage panned on the HPT-1 receptor to HPT-1 and BSA. The table shows the OD results as well as the ratio of HPT-1 to BSA binding.

Table 5 PHAGE HPTl BSA HPT/BSA HAX9 0.382 0.075 HAX4 0 0 . 991 0.065 HAX42 0 .32 0.071 Table 6 shows the results of an ELISA which assessed the binding of phage panned for two rounds on the HPT-1 receptor followed by a third round pan on Caco-2 15 snapwells. Binding to fixed Caco-2 cells, HPT-1 and BSA was examined. The table shows the OD results as well as the ratio of HPT-1 to BSA binding.

Table 6 PHAGE Caco-2 HPTl BSA HPT1/BSA HCA3 0.406 0.048 0 . 038 1 CELL ELISA PROCEDURE Phage ELISA was used as described above with the following changes. Diluent and wash buffer was PBS containing 1%BSA and 0.05% Tween 20 and plates were washed five times at each wash step. Supernatant of infected bacterial cultures was diluted 1:100 and incubated with protein coated plates for 2-3 hours with mild agitation.

Anti-M13 Horseradish peroxidase (HRP) conjugate (Pharmacia, Piscataway, NJ) was diluted 1:8000.

Fixed Caco-2, C2BBel, and A431 cell plates were prepared by growing cells on tissue culture treated microtiter plates. When cells were confluent, plates were fixed with 10% formaldehyde, washed twice with PBS and stored with 0.5%BSA-PBS at -20°C. On the day of the assay, thawed Printed from Mimosa PCT /US98/10088 plates were treated with PBS containing 0.1% phenylhydrazine for one hour at 37°C followed by two PBS washes and blocking for One hour with 0.5%BSA-PBS. The standard ELISA procedure was followed at this point.

Phage which showed specificity to a GIT receptor was further characterized by ELISA on a variety of recombinant proteins. Phage which continued to exhibit GIT receptor specificity was sequenced.

TARGET Table 7 BINDING PHAGE INSERT SEQUENCES: hSI S15 521 522 SNilO SNi28 SN134 SN138 SNi45 SNiAX2 SNiAX4 SN1AX6 SNiAX8 SEQ. ID. NO. 1 2 3 4 6 7 8 9 11 12 RSGAYE S PDGRGGRSYVGGGGGCGNIGRKHNLWGLRTASPACWD S PRS FWPWSRHE S FGISNYLGCGYRTCISGTMTKS SPIYPRHS SSSSDWGGVPGKWRERFKGRGCGISITSVLTGKPNPCPEPKAA RVGQCTDSDVRRPWARSCAHQGCGAGTRNSHGCITRPLRQASAH SHSGGMNRAYGDVFRELRDRWNATSHHTRPTPQLPRGPN SPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSDY RGAADQRRGWSENLGLPRVGWDAIAHNSYTFTSRRPRPP SGGEVSSWGRVNDLCARVSWTGCGTARSARTDNKGFLPKHSSLR SDSDGDHYGLRGGVRCSLRDRGCGLALSTVHAGPPSFYPKLSSP RS LGNYGVTGTVDVTVL PM PGHANHLGV SSASSSDPPRR RTTTAKGCLLGSFGVLSGCSFTPTSPPPHLGYPPHSVN S PKLSSVGVMTKVTELPTEGPNAISIPISATLGPRNPLR D2H DAB 3 13 RWCGAELCNSVTKKFRPGWRDHANPSTHHRTPPPSQSSP DAB7 14 RWCGADDPCGASRWRGGNSLFGCGLRCSAAQS TP SGRIHSTSTS DAB 10 SKSGEGGDSSRGETGWARVRSHAMTAGRFRWYNQLPSDR DAB18 16 RSSANNCEWKSDWMRRACIARYANSSGPARAVDTKAAP DAB24 17 SKWSWSSRWGSPQDKVEKTRAGCGGSPSSTNCHPYTFAPPPQAG DAB30 18 SGFWEFSRGLWDGENRKSVRSGCGFRGSSAQGPCPVTPATIDKH DAX15 19 SESGRCRSVSRWMTTWQTQKGGCGSNVSRGSPLDPSHQTGHATT DAX23 REWRFAGPPLDLWAGPSLPSFNASSHPRALRTYWSQRPR DAX24 21 RMEDIKNSGWRDSCRWGDLRPGCGSRQWYPSNMRS SRDYPAGGH DAX27 22 SHPWYRHWNHGDFSGSGQSRHTPPESPHPGRPNATI Printed from Mimosa DCX8 23 RYKHDIGCDAGVDKKSSSVRGGCGAHSSPPRAGRGPRGTMVSRL DCX11 24 SQGSKQCMQYRTGRLTVGSEYGCGMNPARHATPAYPARLLPRYR DCX2 6 SGRTTSEISGLWGWGDDRSGYGWGNTLRPNYIPYRQATNRHRYT DCX33 26 RWNWTVLPATGGHYWTRSTDYHAINNHRPSIPHQHPTPI DCX36 27 SWSSWNWSSKTTRLGDRATREGCGPSQSDGCPYNGRLTTVKPRT DCX39 28 SGSLNAWQPRSWVGGAFRSHANNNLNPKPTMVTRHPT DCX42 29 RYSGLSPRDNGPACSQEATLEGCGAQRLMSTRRKGRNSRPGWTL DCX45 SVGNDKTSRPVSFYGRVSDLWNASLMPKRTPSSKRHDDG hPEPTl PAX9 31 RWPSVGYKGNGSDTIDVHSNDASTKRSLIYNHRRPLFP PAX14 32 RTFENDGLGVGRSIQKKSDRWYASHNIRSHFASMSPAGK PAX15 33 SYCRVKGGGEGGHTDSNLARSGCGKVARTSRLQHINPRATPPSR PAX16 34 SWTRWGKHTHGGFVNKSPPGKNATSPYTDAQLPSDQGPP PAX17 SQVDSFRNSFRWYEPSRALCHGCGKRDTSTTRIHNSPSDSYPTR PAX18 36 SFLRFQS PRFEDY SRTI SRLRNATNPSNVSDAHNNRALiA PAX35 37 RSITDGGINEVDLSSVSNVLENANSHRAYRKHRPTLKRP PAX3 8 38 S S KVS S PRDPTVPRKGGNVDYGCGHRS SARMPTSALS SITKCYT PAX40 39 RAS TQGGRGVAPE FGASVLGRGCGSATYYTNSTS CKDAMGHNYS PAX43 . 40 RWCEKHKFTAARCSAGAGFERDASRPPQPAHRDNTNRNA PAX45 41 SFQVYPDHGLERHALDGTGPLYAMPGRWIRARPQNRDRQ PAX46 42 SRCTDNEQCPDTGTRSRSVSNARYFSSRLLKTHAPHRP P31 43 SARDSGPAEDGSRAVRLNGVENANTRKSSRSNPRGRRHP P90 44 SSADAEKCAGSLLWWGRQNNSGCGSPTKKHLKHRNRSQTSSSSH 5PAX3 45 RPKNVADAYSSQDGAAAEETSHASNAARKSPKHKPLRRP 5PAX5 46 RGSTGTAGGERSGVLNLHTRDNASGSGFKPWYPSNRGHK 5PAX7 47 RWGWERSPSDYDSDMDLGARRYATRTHRAPPRVLKAPLP 5PAX12 48 RGWKCEGSQAAYGDKDIGRSRGCGSITKNNTNHAHPSHGAVAKI HPT-1 HAX9 49 SREEANWDGYKREMSHRSRFWDATHLSRPRRPANSGDPN HAX35 50 EWYSWKRSSKSTGLGDTATREGCGPSQSDGCPYNGRLTTVKPRK HAX40 51 REFAERRLWGCDDLSWRLDAEGCGPTPSNRAVKHRKPRPRSPAL HAX42 52 SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRRRPSAIPT HCA3 53 RHISEYSFANSHLMGGESKRKGCGINGSFSPTCPRSPTPAFRRT H40 54 S RES GMWGSWWRGHRLN STGGNANMNAS LPPDPPVSTP PAX2 55 STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 8 DNA Sequences for Clones used in in vivo Pan S15 (SEQ ID NO: 56) TCTCACTCCTCGAGATCCGGCGCTTATGAGAGTCCGGATGGTCGGGGGGGTCGGAGCTATG TGGGGGGCGGGGGTGGNTGTGGTAACATTGGTCGGAAGCATAACCTGTGGGGGCTGCGTAC CGCGTCGCCGGCCTGCTGGGACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 521 (SEQ ID NO: 57) TCTCACTCCTCGAGTCCTCGCTCTTTCTGGCCCGTTGTGTCCCGGCATGAGTCGTTTGGGA TCTCTAACTATTTGGGNTGTGGTTATCGTACATGTATCTCCGGCACGATGACTAAGTCTAG 1 CCCGATTTACCCTCGGCATTCGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 522 (SEQ ID NO: 58) TCTCACTCCTCGAGTAGTAGCTCCGATTGGGGTGGTGTGCCTGGGAAGGTGGTTAGGGAGC GCTTTAAGGGGCGCGGTTGTGGTATTTCCATCACCTCCGTGCTCACTGGGAAGCCCAATCC GTGTCCGGAGCCTAAGGCGGCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi 10 (SEQ ID NO: 59) TCTCACTCCTCGAGAGTTGGCCAGTGCACGGATTCTGATGTGCGGCGTCCTTGGGCCAGGT CTTGCGCTCATCAGGGTTGTGGTGCGGGCACTCGCAACTCGCACGGCTGCATCACCCGTCC TCTCCGCCAGGCTAGCGCTCATTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi 28 (SEQ ID NO: 60) TCTCACTCCTCGAGCCACTCCGGTGGTATGAATAGGGCCTACGGGGATGTGTTTAGGGAGC TTCGTGATCGGTGGAACGCCACTTCCCACCACACTCGCCCCACCCCTCAGCTCCCCCGTGG GCCTAATTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi 34 (SEQ ID NO: 61) TCTCACTCCTCGAGTCCGTGCGGGGGGTCGTGGGGGCGTTTTATGCAGGGTGGCCTTTTCG GCGGTAGGACTGATGGTTGTGGTGCCCATAGAAACCGCACTTCTGCGTCGTTAGAGCCCCC GAGCAGCGACTACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNI 38 (SEQ ID NO: 62) TCTCACTCCTCGAGGGGCGCCGCCGATCAGCGGCGGGGGTGGTCCGAGAACTTGGGGTTGC - CTAGGGTGGGGTGGGACGCCATCGCTCACAATAGCTATACGTTCACCTCGCGCCGCCCGCG CCCCCCCTCTAGA SNi 45 (SEQ ID NO: 63) TCTCACTCCTCGAGCGGTGGGGAGGTCAGCTCCTGGGGCCGCGTGAATGACCTCTGCGCTA GGGTGAGTTGGACTGGTTGTGGTACTGCTCGTTCCGCGCGTACCGACAACAAAGGCTTTCT TCCTAAGCACTCGTCACTCCGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi AX2 (SEQ ID NO: 64) Printed from Mimosa TCTCACTCCTCGAGTGATAGTGACGGGGATCATTATGGGCTTCGGGGGGGGGTGCGTTGTT CGCTTCGTGATAGGGGTTGTGGTCTGGCCCTGTCCACCGTCCATGCTGGTCCCCCCTCTTT TTACCCCAAGCTCTCCAGCCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi AX4 (SEQ ID NO: 65) TCTCACTCCTCGAGGAGCTTGGGTAATTATGGCGTCACCGGGACTGTGGACGTGACGGTTT TGCCCATGCCTGGCCACGCCAACCACCTTGGTGTCTCCTCCGCCTCTAGCTCTGATCCTCC GCGGCGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi AX6 (SEQ ID NO: 66) TCTCACTCCTCGAGAACTACGACGGCTAAGGGGTGTCTTCTCGGAAGCTTCGGCGTTCTTA GTGGGTGCTCATTTACGCCAACCTCTCCACCGCCCCACCTAGGATACCCCCCCCACTCCGT CAATTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA SNi AX8 (SEQ ID NO: 67) TCTCACTCCTCGAGCCCGAAGTTGTCCAGCGTGGGTGTTATGACTAAGGTCACGGAGCTGC CCACGGAGGGGCCTAACGCCATTAGTATTCCGATCTCCGCGACCCTCGGCCCGCGCAACCC GCTCCGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAB3 (SEQ ID NO: 68) TCTCACTCCTCGAGGTGGTGCGGCGCTGAGCTGTGCAACTCGGTGACTAAGAAGTTTCGCC CGGGCTGGCGGGATCACGCCAATCCCTCCACCCATCATCGTACTCCCCCGCCCAGCCAGTC CAGCCCTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 2Q DAB7 (SEQ ID NO: 69) TCTCACTCCTCGAGGTGGTGCGGCGCTGATGACCCGTGTGGTGCCAGTCGTTGGCGGGGGG GCAACAGCTTGTTTGGTTGTGGTCTTCGTTGTAGTGCGGCGCAGAGCACCCCGAGTGGCAG GATCCATTCCACTTCGACCAGCTCTAGAATCGAAGGTGCGCTAGACCTTCGAGA DAB10 (SEQ ID NO: 70) TCTCACTCCTCGAGTAAGTCCGGGGAGGGGGGTGACAGTAGCAGGGGCGAGACGGGCTGGG CGAGGGTTCGGTCTCACGCCATGACTGCTGGCCGCTTTCGGTGGTACAACCAGTTGCCCTC TGATCGGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAB18 (SEQ ID NO: 71) TCTCACTCCTCGAGGTCGAGCGCCAATAATTGCGAGTGGAAGTCTGATTGGATGCGCAGGG ,0 CCTGTATTGCTCGTTACGCCAACAGTTCGGGCCCCGCCCGCGCCGTCGACACTAAGGCCGC GCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAB24 (SEQ ID NO: 72) TCTCACTCCTCGAGTAAGTGGTCGTGGAGTTCGAGGTGGGGCTCCCCGCAGGATAAGGTTG AGAAGACCAGGGCGGGTTGTGGTGGTAGTCCCAGCAGCACCAATTGTCACCCCTACACCTT TGCCCCCCCCCCGCAAGCCGGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA Printed from Mimosa DAB30 (SEQ ID NO: 73) TCTCACTCCTCGAGTGGGTTCTGGGAGTTTAGCAGGGGGCTTTGGGATGGGGAGAACCGTA AGAGTGTCCGGTCGGGTTGTGGTTTTCGTGGCTCCTCTGCTCAGGGCCCGTGTCCGGTCAC GCCTGCCACCATTGACAAACACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAX15 (SEQ ID NO: 74) TCTCACTCCTCGAGTGAGAGCGGGCGGTGCCGTAGCGTGAGCCGGTGGATGACGACGTGGC AGACGCAGAAGGGCGGTTGTGGTTCCAATGTTTCCCGCGGTTCGCCCCTCGACCCCTCTCA CCAGACCGGGCATGCCACTACTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 1Q DAX23 (SEQ ID NO: 75) TCTCACTCCTCGAGGGAGTGGAGGTTTGCCGGGCCGCCGTTGGACCTGTGGGCGGGTCCGA GCTTGCCCTCTTTTAACGCCAGTTCCCACCCTCGCGCCCTGCGCACCTATTGGTCCCAGCG GCCCCGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAX24 (SEQ ID NO: 76) TCTCACTCCTCGAGGATGGAGGACATCAAGAACTCGGGGTGGAGGGACTCTTGTAGGTGGG GTGACCTGAGGCCTGGTTGTGGTAGCCGCCAGTGGTACCCCTCGAATATGCGTTCTAGCAG AGATTACCCCGCGGGGGGCCACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DAX27 (SEQ ID NO: 77) TCTCACTCCTCGAGTCATCCGTGGTACAGGCATTGGAACCATGGTGACTTCTCTGGTTCGG GCCAGTCACGCCACACCCCGCCGGAGAGCCCCCACCCCGGCCGCCCTAATGCCACCATTTC TAGAATCGAAGGTCGCGCTAGACCTTCGAG DCX8 (SEQ ID NO: 78) TCTCACTCCTCGAGATATAAGCACGATATCGGTTGCGATGCTGGGGTTGACAAGAAGTCGT CGTCTGTGCGTGGTGGTTGTGGTGCTCATTNGTCGCCACCCCGCGCCGGCCGTGGTCCTCG CGGCACGATGGTTAGCAGGCTTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DCX11 (SEQ ID NO: 79) TCTCACTCCTCGAGTCAGGGCTCCAAGCAGTGTATGCAGTACCGCACCGGTCGTTTGACGG TGGGGTCTGAGTATGGTTGTGGTATGAACCCCGCCCGCCATGCCACGCCCGCTTATCCGGC GCGCCTGCTGCCACGCTATCGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 3Q DCX26 (SEQ ID NO: 80) TCTCACTCCTCGAGTGGGCGGACTACTAGTGAGATTTCTGGGCTCTGGGGTTGGGGTGACG ACCGGAGCGGTTATGGTTGGGGTAACACGCTCCGCCCCAACTACATCCCTTATAGGCAGGC GACGAACAGGCATCGTTATACGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DCX33 (SEQ ID NO: 81) TCTCACTCCTCGAGGTGGAATTGGACTGTCTTGCCCGCCACTGGCGGCCATTACTGGACGC GTTCGACGGACTATCACGCCATTAACAATCACAGGCCGAGCATCCCCCACCAGCATCCGAC CCCTATCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 58 - Printed from Mimosa DCX36 (SEQ ID NO: 82) TCTCACTCCTCGAGTTGGTCGTCGTGGAATTGGAGCTCTAAGACTACTCGTCTGGGCGACA GGGCGACTCGGGAGGGTTGTGGTCCCAGCCAGTCTGATGGCTGTCCTTATAACGGCCGCCT TACGACCGTCAAGCCTCGCACGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA DCX39 (SEQ ID NO: 83) TCTCACTCCTCGAGTGGTAGTTTGAACGCATGGCAACCGCGGTCATGGGTGGGGGGCGCGT TCCGGTCACACGCCAACAATAACTTGAACCCCAAGCCCACCATGGTTACTNGTCACCCTAC CTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 1Q DCX42 (SEQ ID NO: 84) TCTCACTCCTCGAGGTATTCGGGTTTGTCCCCGCGGGACAACGGTCCCGCTTGTAGTCAGG AGGCTACCTTGGAGGGTTGTGGTGCGCAGAGGCTGATGTCCACCCGTCGCAAGGGCCGCAA CTCCCGCCCCGGGTGGACGCTCTCTAGAATCGAAGGTCGCGCTAGACCCTTCGAGA DCX45 (SEQ ID NO: 85) TCTCACTCCTCGAGCGTGGGGAATGATAAGACTAGCAGGCCGGTTTCCTTCTACGGGCGCG TTAGTGATCTGTGGAACGCCAGCTTGATGCCGAAGCGTACTCCCAGCTCGAAGCGCCACGA TGATGGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX2 (SEQ ID NO: 86) TCTCACTCCTCGAGTACTCCCCCCAGTAGGGAGGCGTATAGTAGGCCCTATAGTGTCGATA GCGATTCGGATACGAACGCCAAGCACAGCTCCCACAACCGCCGTNTGCGGACGCGCAGCCG CCCGAACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX9 (SEQ ID NO: 87) TCTCACTCCTCGAGATGGCCTAGTGTGGGTTACAAGGGTAATGGCAGTGACACTATTGATG TTCACAGCAATGACGCCAGTACTAAGAGGTCCCTCATCTATAACCACCGCCGCCCCNTCTT TCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX14 (SEQ ID NO: 88) TCTCACTCCTCGAGAACGTTTGAGAACGACGGGCTGGGCGTCGGCCGGTCTATTCAGAAGA AGTCGGATAGGTGGTACGCCAGCCACAACATTCGTAGCCATTTCGCGTCCATGTCTCCCGC TGGTAAGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 3Q PAX15 (SEQ ID NO: 89) TCTCACTCCTCGAGCTATTGTCGGGTTAAGGGTGGTGGGGAGGGGGGGCATACGGATTCCA ATCTGGCTAGGTCGGGTTGTGGTAAGGTGGCCAGGACCAGCAGGCTTCAGCATATCAACCC GCGCGCTACCCCCCCCTCCCGGTCTAGAATCGAAGGTC PAX16 (SEQ ID NO: 90) 3 5 TCTCACTCCTCGAGTTGGACTCGGTGGGGCAAGCACANTCATGGGGGGTTTGTGAACAAGT CTCCCCCTGGGAAGAACGCCACGAGCCCCTACACCGACGCCCAGCTGCCCAGTGATCAGGG TCCTCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA Printed from Mimosa PCT A3S98/10088 PAX17 (SEQ ID NO: 91) TCTCACTCCTCGAGTCAGGTTGATTCGTTTCGTAATAGCTTTCGGTGGTATGAGCCGAGCA GGGCTCTGTGCCATGGTTGTGGTAAGCGCGACACCTCCACCACTCGTATCCACAATAGCCC CAGCGACTCCTATCCTACACGCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX18 (SEQ ID NO: 92) TCTCACTCCTCGAGCTTTTTGCGGTTCCAGAGTCCGAGGTTCGAGGATTACAGTAGGACGA TCTNTCGGTTGCGCAACGCCACGAACCCGAGTAATGTCTCCGATGCGCACAATAACCGGGC CTTGGCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 1(J PAX35 (SEQ ID NO: 93) TCTCACTCCTCGAGGAGCATCACCGACGGGGGCATCAATGAGGTGGACCTGAGTAGTGTGT CGAACGTTCTTGAGAACGCCAACTCGCATAGGGCCTACAGGAAGCATCGCCCGACCTTGAA GCGTCCTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX38 (SEQ ID NO: 94) TCTCACTCCTCGAGTTCGAAGGTGAGCAGCCCGAGGGATCCGACGGTCCCGCGGAAGGGCG GCAATGTTGATTATGGTTGTGGTCACAGGTCTTCCGCCCGGATGCCTACCTCCGCTCTGTC GTCGATCACGAAGTGCTACACTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX40 (SEQ ID NO: 95) TCTCACTCCTCGAGAGCCAGTANGCAGGGCGGCCGGGGTGTTGCCCCTGAGTTTGGGGCGA GCGTTTTGGGTNGTGGTTGTGGTAGCGCCACTTATTACACGAACTCCACCAGCTGCAAGGA TGCTATGGGCCACAACTACTCGTCTAGAATCGAAGGTCGCGNTAGACCTTCGAGA PAX43 (SEQ ID NO: 96) TCTCACTCCTCGAGATGGTGCGAGAAGCACAAGTTTACGGCTGCGCGTTGCAGCGCGGGGG CGGGTTTTGAGAGGGANGCCAGCCGTCCGCCCCAGCCTGCCCACCGGGATAATACCAACCG TAATGCNTNTAGAATCGAAGGTCGCGCTAGACCTTCGAGA PAX45 (SEQ ID NO: 97) TCTCACTCCTCGAGTTTTCAGGTGTACCCGGACCATGGTCTGGAGAGGCATGCTTTGGACG GGACGGGTCCGCTTTACGCCATGCCCGGCCGCTGGATTAGGGCGCGTCCGCAGAACAGGGA CCGCCAGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 3() PAX46 (SEQ ID NO: 98) TCTCACTCCTCGAGCAGGTGTACGGACAACGAGCAGTGCCCCGATACCGGGANTAGGTCTC GTTCCGTTAGTAACGCCAGGTACTTTTCGAGCAGGTTGCTCAAGACTCACGCCCCCCATCG CCCTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA P31 (SEQ ID NO: 99) TCTCACTCCTCGAGTGCCAGGGATAGCGGGCCTGCGGAGGATGGGTCCCGCGCCGTCCGGT TGAACGGGGTTGAGAACGCCAACACTAGGAAGTCCTCCCGCAGTAACCCGCGGGGTAGGCG CCATCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA - 60 Printed from Mimosa P90 (SEQ ID NO: 100) TCTCACTCCTCGAGTTCCGCCGATGCGGAGAAGTGTGCGGGCAGTCTGTTGTGGTGGGGTA GGCAGAACAACTCCGGTTGTGGTTCGCCCACGAAGAAGCATCTGAAGCACCGCAATCGCAG TCAGACCTCCTCTTCGTCCCACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 5PAX3 (SEQ ID NO: 101) TCTCACTCCTCGAGACCGAAGAACGTGGCCGATGCTTATTCGTCTCAGGACGGGGCGGCGG CCGAGGAGACGTCTCACGCCAGTAATGCCGCGCGGAAGTCCCCTAAGCACAAGCCCTTGAG GCGGCCTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 5PAX5 (SEQ ID NO: 102) TCTCACTCCTCGAGAGGCAGTACGGGGACGGCCGGCGGCGAGCGTTCCGGGGTGCTCAACC TGCACACCAGGGATAACGCCAGCGGCAGCGGTTTCAAACCGTGGTACCCTTCGAATCGGGG TCACAAGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 5PAX7 (SEQ ID NO: 103) TCTCACTCCTCGAGGTGGGGGTGGGAGAGGAGTCCGTCCGACTACGATTCTGATATGGACT 15 TGGGGGCGAGGAGGTACGCCACCCGCACCCACCGCGCGCCCCCTCGCGTCTTGAAGGCTCC CCTGCCCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA 5PAX12 (SEQ ID NO: 104) TCTCACTCCTCGAGGCACTGGAAGTGCGAGGGCTCTCAGGCTGCCTACGGGGACAAGGATA TCGGGAGGTCCAGGGGTTGTGGTTCCATTACAAAGAATAACACTAATCACGCCCATCCTAG 2 CCACGGCGCCGTTGCTAAGATCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA HAX9 (SEQ ID NO: 105) TCTCACTCCTCGAGCCGCGAGGAGGCGAACTGGGACGGCTATAAGAGGGAGATGAGCCACC GGAGTCGCTTTTGGGACGCCACCCACCTGTCCCGCCCTCGCCGCCCCGCTAACTCTGGTGA CCCTAACTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA HAX40 (SEQ ID NO: 106) TCTCACTCNTCGAGAGAGTTCGCGGAGAGGAGGTTGTGGGGGTGTGATGACCTGAGTTGGC GTCTCGACGCGGAGGGTTGTGGTCCCACTCCGAGCAATCGGGCCGTCAAGCATCGCAAGCC CCGCCCACGCTCCCCCGCACTCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA HAX42 (SEQ ID NO: 107) 3 ° TCTCACTCNTNGAGTGATCACGCGTTGGGGACGAATCTGAGGTCTGACAATGCCAAGGAGC CGGGTGATTACAACTGTTGTGGTAACGGGAACTCTACCGGGCGAAAGGTTTTTAACCGTAG GCGCCCCTCCGCCATCCCCANTTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA HCA3 (SEQ ID NO: 108) TCTCACTCCTCGAGGCATATTTCTGAGTATAGCTTTGCGAATTCCCACTTGATGGGTGGCG 3 5 AGTCCAAGCGGAAGGGTTGTGGTATTAACGGCTCCTTTTCTCCCACTTGTCCCCGCTCCCC CACCCCAGCCTTCCGCCGCACCTCTAGAATCGAAGGTCGCGCTAGACCTTCGAGA Printed from Mimosa PCT/U S98/10088 H40 (SEQ ID NO: 109) TCTCACTCCTCGAGCCGGGAGAGCGGGATGTGGGGTAGTTGGTGGCGTGGTCACAGGTTGA ATTCCACGGGGGGTAACGCCAACATGAATGCTAGTCTGCCCCCCGACCCCCCTGTTTCCAC TCCGTCTAGAATCGAAGGTCGCGCTAGACCTTCGAG Peptide Motifs By comparison of the amino acid sequences of the clones binding GIT receptors, certain sequence similarities or "motifs" were recognized. These motifs can often represent the part of the sequence that is important for binding to the target. Table 9 identifies regions of sequence similarity or sequence motifs (in boldface) that were identified among GIT binding peptides (corresponding SEQ ID NOS. are shown in Table 7).

Table 9 SARDSGPAEDGSRAVRLNGVENANTRKSSRSNPRGRRHP RWPSVGYKGNGSDTIDVHSNDASTKRSLIYNHRRPLFP SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRK-VFNRRRPSAIPT STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN u RVGQCTDSDVRRPWARSCAHQGCGAGTRNSHGCITRPLRQASAH RGAADQRRGWSENLGLPRVGWDAIAHNSYTFTSRRPRPP RSGAYES PDGRGGRSYVGGGGGCGNIGRKHNLWGLRTASPACWD SPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSDY SKSGEGGDSSRGETGWARVRSHAMTAGRFRWYNQLPSDR SGFWEFSRGLWDGENRKSVRSGCGFRGSSAQGPCPVTPATIDKH RYKHDIGCDAGVDKKSSSVRGGCG-AHSS PPRAGRGPRGTMVSRL Phage Binding to Caco-2 Cells 3 0 Phage expressing presumed GIT binding peptide inserts were also assayed by ELISA on fixed Caco-2 or C2BBel cells as follows. Cells were plated at 1 x 105 cells/well on 100 nl culture media and incubated at 30°C in 5% C02 overnight. 100 ptl 25% formaldehyde was added to each well for 15 minutes. Contents of the wells were removed by inverting the plate. The plate was then washed 3 times with PEPT-1 HPTl P31 PAX 9 HAX42 PAX 2 hSI SNilO SNi38 S15 SNi 3 4 D2H DAB10 DAB30 DCX8 Printed from Mimosa WO 98/51325 PCT/US98/10088 DPBS. 0.1% phenylhydrazine DPBS solution was added to each well and incubated for 1 hr at 37°C. The plate was inverted and washed 3 times. The plate was blocked with 0.5% BSA-DPBS for 1 hr at room temperature. The plate was inverted and 5 washed 3 times with 1% BPT (PBS containing 1% BSA and 0.05% Tween20). Phage diluted with 1% BPT was added to wells containing fixed cells. Wells without phage added were used to determine background binding of the HRP conjugate. The plates were incubated 2-3 hours on a rotor at room 10 temperature. Plates were washed as before. Plates were incubated with dilute anti-M13-HRP antibody in 1% BPT for 1 hour at room temperature. Following washing, TMB substrate was added and absorbance of the plates were read at 650 nm. Table 10 shows the relative binding of phage encoding 15 peptides to fixed Caco-2 cells.

Table 10.

Relative binding of phage encoding peptides to fixed Caco-2 cells Fixed Caco-2 Phage cell binding SNilO ++ SNi34 + P31 ++ 5PAX5 ++ PAX2 + HAX42 + DCX8 +++ DCX11 + HI + M13mpll8 In vivo phage selection: Further selection of phage expressing peptides capable of binding to the GIT or transporting the GIT was done as follows. The purified library was resuspended in a Printed from Mimosa buffer, such as TBS or PBS, and introduced onto one side of a tissue barrier, e.g., injected into the duodenum, jejunum, ileum, colon or other in vivo animal site using, for instance, a closed loop model or open loop model. Following 5 injection, samples of bodily fluids located across the tissue barrier, e.g., samples of the portal circulation and/or systemic circulation, were withdrawn at predetermined time points, such as 0 to 90 minutes and/or 2 to 6 hours or more. An aliquot of the withdrawn sample (e.g., blood) was used to 10 directly infect a host, e.g., E. coli, in order to confirm the presence of phage. The remaining sample was incubated, e.g., overnight incubation with E. coli at 37°C with shaking. The amplified phage present in the culture can be sequenced-individually to determine the identity of peptides coded by 15 the phage or, if further enrichment is desired, can be precipitated using PEG, and resuspended in PBS. The phage can then be further precipitated using PEG or used directly for administration to another animal using a closed or open GIT loop model system. Portal or systemic blood samples are 20 collected and the phage transported into such circulation systems is subsequently amplified. In this manner, administration of the phage display library with, if desired, repeat administration of the amplified phage to the GIT of the animal, permitted the selection of phage which was 25 transported from the GIT to the portal and/or systemic circulation of the animal.

If desired, following administration of the phage display library to the tissue barrier (e.g., GIT) of the animal model, the corresponding region of the tissue barrier 30 can be recovered at the end of the procedures given above. This recovered tissue can be washed repeatedly in suitable buffers, e.g., PBS containing protease inhibitors and homogenized in, for example, PBS containing protease inhibitors. The homogenate can be used to infect a host, 35 such as E. coli, thus permitting amplification of phages which bind tightly to the tissue barrier (e.g., intestinal tissue). Alternatively, the recovered tissue can be Printed from Mimosa homogenized in suitable PBS buffers, washed repeatedly and the phage present in the final tissue homogenate can be amplified in E. coli. This approach permits amplification (and subsequent identification of the associated peptides) of 5 phages which either bind tightly to the tissue barrier (e.g., intestinal tissue) or which are internalized by the cells of the tissue barrier (e.g., epithelial cells of the intestinal tissue). This selection approach of phage which bind to tissues or which are internalized by tissues can be repeated.

Treatment of animal tissue barriers in vivo with phage display populations The purified phage display library (random or preselected) was diluted to 500 fil in PBS buffer and injected 15 into the closed (or open) intestinal loop model (e.g., rat, rabbit or other species). At time 0 and at successive time points after injection, a sample of either the portal circulation or systemic circulation was withdrawn. An aliquot of the withdrawn blood was incubated with E. coli, 20 followed by plating for phage plaques or for transduction units or for colonies where the phage codes for resistance to antibiotics such as tetracycline. The remainder of the withdrawn blood sample (up to 150 /x 1) was incubated with 250 )il of E. coli and 5 ml of LB medium or other suitable 25 growth medium. The E. coli cultures were incubated overnight by incubation at 37°C on a shaking platform. Blood samples taken at other time points (such as 15 min, 30 min, 45 min, 60 min, up to 6 hours) were processed in a similar manner, permitting amplification of phages present in the portal or 30 systemic circulation in E. coli at these times. Following amplification, the amplified phage was recovered by PEG precipitation and resuspended in PBS buffer or TBS buffer. The titer of the amplified phage, before and after PEG precipitation, was determined. The amplified, PEG 35 precipitated phage was diluted to a known phage titer (generally between 10s and 1010 phage or plaque forming units (p.f.u.) per ml) and was injected into the GIT of the animal Printed from Mimosa closed (or open) loop model. Blood samples were collected from portal and/or systemic circulation at various time points and the phage transported into the blood samples were amplified in E. coli as given above for the first cycle.

Subsequently, the phage was PEG-precipitated, resuspended, titered, diluted and injected into the GIT of the animal closed (or open) loop model. This procedure of phage injection followed by collection of portal and/or systemic blood samples and amplification of phage transported into 10 these blood samples can be repeated, for example, up to 10 times, to permit the selection of phages which are preferentially transported from the GIT into the portal and/or systemic circulation. 6.7. Transport of Phage From Rat Lumen Into the Portal and Systemic Circulation Phage from random phage display libraries as well as control phage were injected into the lumen of the rat gastro-intestinal tract {in situ rat closed loop model). 2q Blood was collected over time from either the systemic circulation or portal circulation and the number of phage which were transported to the circulation was determined by titering blood samples in E. coli.

The phage display libraries used in this study were 25 D3 8 and DC43 in which gene III codes for random 38-mer and 43-mer peptides, respectively. As a negative control, the identical phage M13mpl8, in which gene III does not code for a "random" peptide sequence, was used. Both the library phages D38 and DC43 were prepared from E. coli, mixed together, dialyzed against PBS, precipitated using PEG/NaCl and were resuspended in PBS buffer. The M13mpl8 control was processed in a similar manner. The titer of each phage sample was determined and the phage samples were diluted in PBS to approximately the same titers prior to injection into 35 the rat closed loop model.

For sampling from the systemic circulation, approximately 15 cm of the duodenum of Wistar rats was tied Printed from Mimosa off (closed loop model), approximately 0.5ml of phage solution was injected into the closed loop and blood (0.4ml) was sampled from the tail vein at various times. The time points used (in min) were: 0, 15, 30, 45, 60, 90, 120, 180, 5 240 and 300 minutes. For sampling from the portal circulation, the portal vein was catheterized, approximately 15 cm of the duodenum was tied off (closed loop model), 0.5ml of phage solution was injected into the closed loop and blood was sampled from the portal vein catheter at various times.

As the portal sampling is delicate, sampling times were restricted to 15, 30, 45 and 60 minutes, where possible. The volume of phage injected into each animal was as follows: The estimated number of transported phage has been adjusted 20 to account for differences in volume injected into each animal (using 0.5 ml as the standard volume). circulation, animals Rl, R2 and R3 received the control phage M13mpl8 and animals R4, R5, R6 and R7 received the test phage 25 D38/DC43 mix. To investigate transport into the portal circulation, animals R8, R9 and R10 received the control phage M13mpl8 and animals Rll, R12, R13 and R14 received the test phage D38/DC43 mix. Animal R15* received the combined phage samples from animals R4-R7 (see Table 11) which were 30 sampled from the systemic circulation on day one, followed by amplification in E. coli, PEG precipitation and resuspension in PBS. On subsequent analysis, the titer of this phage was found to be 100 times greater than the other phage samples used for animals R8-R14. Thus, the data presented for animal 35 R15* is adjusted down.

Animals (15) R1-R3 R4 R5-R15 Volume of Phage Injected 0.50 ml 0.43 ml 0.45 ml To investigate transport into the systemic 67 Printed from Mimosa Approximately 0.4 ml of the blood was collected at each time point in each model system. 30 /il of the collected blood (systemic) was mixed with 100 /il of the prepared E. coli strain K91Kan, incubated at 37°C for 30 min, and 5 plated out for plaque formation using Top Agarose on LB plates. Various negative controls were included in the titering experiments. The following day, the number of plaque forming units was determined. Similarly, 3 0 fil of the collected blood (portal) and serial dilutions (1:100, 1:1000) 10 thereof was mixed with 100 /il of the prepared E. coli strain K91Kan, incubated at 37°C for 30 min, and plated out for plaque formation using Top Agarose on LB plates. The following day, the number of plaque forming units was c determined.

In addition, approximately 300 ^1 of the collected blood from each time point (systemic and portal) was incubated with 5ml of prepared E. coli strain K91Kan in modified growth media containing 5mM MgCl2/MgS04 at 37°C overnight with shaking (to permit phage amplification). The samples were 20 centrifuged and the cell pellet was discarded. Samples of the phage supernatant were collected, serially diluted (10~2, 10"4, 10"6, 10"8) in TBS buffer, and plated for plaques in order to determine the number of plaque forming units present in the amplified phage samples.

Furthermore, an aliquot of phage was removed from the "amplified" supernatants obtained from test animals R4-R7 (samples from each time point were used), combined, and precipitated using PEG for two hours. The precipitated phage was resuspended in PBS buffer and was injected into closed 30 loop model of animal R15*, followed by portal sampling.

The number of phage transported from the closed loop model into the systemic circulation is presented in Table 11 hereafter. The number of phage transported from the closed loop model into the portal circulation is presented in 35 Table 12 hereafter. These numbers are corrected for phage input difference and for volume input differences. Clearly, more phage are present in the portal samples than in the Printed from Mimosa systemic samples, indicative of either hepatic or RES clearance and/or phage instability m the systemic circulation. In addition, the uptake of phage from the GIT into the portal circulation is quite rapid, with substantial 5 number of phages detected within 15 minutes. The results from the portal sampling experiments would also indicate that the kinetics of uptake of phage from the D38/DC43 libraries is quicker than that of the control phage. Thus, there may be preferential uptake of phage coding for random peptide 10 sequences from the GIT into the portal circulation. In the case of animals R13, R14 and R15*, the % of the phage transported into the titered blood sample within the limited time frame (30, 45 and 15 mins, respectively) was estimated as 0.13%, 1.1% and 0.013%, respectively.

TABLE 11 NUMBER OF PHAGE TRANSPORTED FROM THE CLOSED LOOP MODEL INTO THE SYSTEMIC CIRCULATION Time (min) Rl R2 R3 R4 R5 R6 R7 0 0 0 0 0 0 0 0 0 1 9 0 0 1 7 2 1 0 0 46 1 11 45 4 2 1 32 0 60 63 19 21 1 114 0 21 90 104 18 3 115 0 22 120 94 24 27 0 64 0 6 180 94 12 23 1 413 0 0 240 14 1 0 36 0 0 300 1 1 4 2 0 0 0 Total number of transported phage 382 83 124 8 820 2 87 Animals Rl, R2 and R3 received the control phage Animals R4, R5, R6 and R7 received the test phage mix.

M13mpl8. 35 D38/DC43 Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 12 NUMBER OF PHAGE TRANSPORTED FROM THE CLOSED LOOP MODEL INTO THE PORTAL CIRCULATION Time (mm) R8 R9 R10 Rll R12 R13 R14 R15* 6 3 1 19 231,000 1,000,000 ,000 1 26 - 0 60,000 272,000 1 555 1,240,000 - - - 420,000 - - - Animals R8, R9 and RIO received the control phage M13mpl8.

Animals Rll, R12, R13 and R14 received the test phage D38/DC4 3 mix.

Animal R15* received the combined phage samples 15 from animals R4-R7 (see Table 11) which were sampled from the systemic circulation on day one, followed by PEG precipitation and resuspension in PBS. On subsequent analysis, the titer of this phage was found to be 100 times greater than the other phage samples used for animals R8-R14. 20 Thus, the data measuring phage transport into the portal circulation for animal R15* is adjusted down.

These studies demonstrated that both the control phage and the D3 8/DC43 phages are transported over time from the lumen of the GIT into the portal and systemic 25 circulation, as demonstrated by titenng the phage transported to the blood in E. coli. More phage were transported from the test phage samples into the portal circulation than the corresponding control phage sample. In addition, the kinetics of transport of the test phage into 30 the portal circulation appeared to exceed that of the control phage. Phage from the D38/DC43 libraries which appeared in the systemic circulation of different animals (R4-R7) were pooled, amplified in E. coli, precipitated, and re-applied to the lumen of the GIT, followed by collection in the portal 35 circulation and titering in E. coli. These selected phage were also transported from the lumen of the GIT into the portal circulation. This in situ loop model may represent an Printed from Mimosa WO 98/51325 PCT/US98/10088 attractive screening model in which to identify peptide sequences which facilitate transport of phage and particles from the GIT into the circulation.

Using this screening model system, a number of 5 preselected phage libraries now exist, including a one pass systemic phage library from animals R4-R7, a one-pass portal library from animals R11-R14, and a two pass, rapid transport, systemic-portal phage library SP-2 from animal R15*. 6.8. Transport of Phage From Preselected Phage Libraries From the Rat Lumen Into the Portal and Systemic Circulation Four preselected phage libraries, GI-D2H, Gl-hSI, GI- HPT1 and GI-hPEPTl, were constructed by pooling phage previously selected by screening random phage display libraries D38 and DC43 using the HPTl, HPEPTl, D2H and hSI receptor or binding sites located in the GIT. The phage pools, preselected phage libraries are shown in Table 13.

Note that the sequences for PAX2, HAX1, HAX5, HAX6, HAX10, H10 and HAX44 are the same. Also, the sequence for HAX40 is the same as that for H44. The corresponding SEQ ID NOS. are shown in Table 7.

Table 13 PRESELECTED PHAGE LIBRARIES D2H HSI HPTl hPEPTl DAB 3 S15 HAX9 PAX2 (H10 DAB 7 S21 HAX35 PAX 9 DAB10 S22 HAX4 0 (H44) PAX14 DAB18 SNilO HAX42 PAX15 DAB24 SNi28 HCA3 PAX16 DAB3 0 SNi34 HAX1 PAX17 DAX15 SNi 3 8 HAX5 PAX18 DAX23 SNi45 HAX6 PAX35 DAX24 SNiAX2 HAX10 PAX38 DAX27 SNiAX6 H40 PAX4 0 DCX8 SNiAX8 M13mpl8 PAX43 DCX11 M13mpl8 PAX45 DCX26 PAX46 DCX33 P31 DCX3 6 P90 Printed from Mimosa DCX39 5PAX3 DCX42 5PAX5 DCX45 5PAX7 M13mpl8 5PAX12 H4 0 M13mpl8 Similar to methods described herein above, these preselected phage libraries together with the negative control phage M13mpl8 were injected into the rat closed loop model (6 animals per preselected phage library), blood was collected over time from the portal circulation via the portal vein and, at the termination of the experiment, a systemic blood sample was collected from the tail vein and the intestinal tissue region from the closed loop was collected.

In particular, phages selected in vitro to each receptor or binding site located in the GIT were amplified in E. coli, PEG-precipitated, resuspended in TBS and the titer of each phage sample was determined by plaquing in E. coli as described above. Subsequently, an equal number of each phage (8 x 108phage) for each receptor site was pooled into a preselected phage library together with the negative control phage M13mpl8 and each preselected phage library was administered to 6 Wistar rats per library (rats 1-6; GI-D2H, rats 7-12; Gl-hSI, rats 13-18; GI-hPEPTl, and rats 19-24; GI-HPT1). Using the in situ loop model described above, 0.5 ml of preselected phage library solution was injected into the tied-off portion of the duodenum/jejunum. Blood was collected into heparinized tubes from the portal vein at 0, 15, 30, 4 5 and 60 minutes. A blood sample was taken from the systemic circulation at the end of the experiment.

Similarly, the portion of the duodenum/jejunum used for phage injection was taken at the end of the experiment.

Thirty microliters of the collected portal blood (neat and 10~2, 10"4, 10~6 dilutions) was added to 30 fil E. coli K91Kan cells (overnight culture) and incubated at 37°C for 10 min. Subsequently, 3 ml of top agarose was added and the samples were plated for plaques. One hundred microliters of - 72 Printed from Mimosa the collected portal blood was added to 100/tl of E. coli K91Kan. Five milliliters of LB medium was then added and the samples were incubated at 37°C overnight in a rotating microbial incubator. The E. coli was removed by 5 centrifugation and the amplified phage supernatant samples were either titered directly or were PEG-precipitated, resuspended in TBS and titered. Following titration of the amplified phage, samples containing phage from each set of animals were combined, adjusting the titer of each sample to 10 the same titer, and were plated for plaques on LB agar plates (22cm2 square plates). Either 12,000 or 24,000 phage were plated for plaques.

Thirty microliters of the collected systemic blood (neat and 10~2, 10~4, 10"6 dilutions) was added to E. coli 15 K91Kan cells, incubated at 37°C for 10 mm. Three ml of top agarose was then added and the samples were plated for plaques. One hundred microliters of the collected systemic blood was added to lOOfil of E. coli K91Kan, incubated at 37°C for 10 min. Five milliliters of LB medium was then added and 20 the samples were incubated at 37°C overnight in a rotating microbial incubator. The E. coli was removed by centrifugation and the amplified phage supernatant samples were either titered directly or were PEG-precipitated, resuspended in TBS and titered. Following titration of the 25 amplified phage, samples containing phage from each set of animals were combined, adjusting the titer of each sample to the same titer, and were plated for plaques on LB agar plates (22cm2 square plates). Either 12,000 or 24,000 phage were plated for plaques.

The intestinal tissue portion used in each closed loop was excised. The tissue was cut into small segments, followed by 3 washings in sterile PBS containing protease inhibitors, and homogenized in an Ultra thorex homogeniser (Int-D samples). Alternatively, the tissue (in PBS 35 supplemented with protease inhibitors) was homogenized in an Ultra Thorex homogenizer, washed 3 times in PBS containing protease inhibitors and resuspended in PBS containing Printed from Mimosa protease inhibitors (Int-G samples). In each case, serial dilutions (neat and 10~2, 10"4, 10"s dilutions) of the tissue homogenate was titered in E. coli. In addition, an aliquot (100/^1) of the tissue homogenate was added to lOOpil of 5 E. coli K91Kan, incubated at 37°C for 10 min, followed by addition of 5ml of LB medium and incubation overnight at 37°C in a rotating microbial incubator.

The phage amplified from the portal blood, systemic blood and intestinal tissue was plated for plaques. The 10 plaques were transferred to Hybond-N Nylon filters, followed by denaturation (1.5M NaCl, 0.5M NaOH), neutralization (0.5M TRIS-HC1, pH7.4, 1.5M NaCl), and washing in 2X SSC buffer. The filters were air-dried, and the DNA was cross-linked to the filter (UV crosslinking: 2min, high setting). The 15 filters were incubated in pre-hybridization buffer (6X SSC, 5X Denhardt's solution, 0.1% SDS, 20/ig/ml yeast tRNA) at 40°C-45°C for at least 60 min.

Synthetic oligonucleotides, (22-mers), complimentary to regions coding for the receptor or binding 20 sites used to create the preselected phage library, were synthesized (see Table 14 below).

Table 14 OLIGONUCLEOTIDES USED IN IN VIVO SCREEN CLONE NAME OLIGO SEQ.

ID. NO.

SI 5 5'TCCGGACTCTCATAAGCGCCGG3' 111 521 5' ACAACGGGCCAGAAAGAGCGAG3' 112 522 B'ACACCACCCCAATCGGAGCTAC3' 113 SNilO 5' TCAGAATCCGTGCACTGGCCAA3' 114 SNi28 5'GCCCTATTCATACCACCGGAGT3' 115 SNi 3 4 5'CATCAGTCCTACCGCCGAAAAG3' 116 SNi 3 8 5' CGTATAGCTATTGTGAGCGATG3' 117 SNi45 5'ACGCGCGGAACGAGCAGTACCA3' 118 SNiAX2 5'CCATAATGATCCCCGTCACTAT3' 119 SNiAX6 5' AGACACCCCTTAGCCGTCGTAG3' 120 SNi AX 8 S'AGCTCCGTGACCTTAGTCATAA3' 121 Printed from Mimosa PCT /US98/10088 CLONE NAME 0LI60 SEQ.

ID. NO.

DAB3 5'TGCACAGCTCAGCGCCGCACCA 3' 122 DAB7 5'ACGGGTCATCAGCGCCGCACCA 3' 123 DAB10 5'TGTCACCCCCCTCCCCGGACTT 3' 124 DAB18 5'ACTCGCAATTATTGGCGCTCGA 3' 125 DAB 2 4 5'GTCTTCTCAACCTTATCCTGCG 3' 126 DAB3 0 5'AAAGCCCCCTGCTAAACTCCCA 3' 127 DAX15 5'CTGCGTCTGCCACGTCGTCATC 3' 128 DAX23 5'GTTAAAAGAGGGCAAGCTCGGA 3' 129 DAX24 5'CCGAGTTCTTGATGTCCTCCAT 3' 13 0 DAX27 5'TCCAATGCCTGTACCACGGATG 3' 131 DCX8 5' TCGCAACCGATATCGTGCTTAT3' 132 DCX11 5'TGCATACACTGCTTGGAGCCCT3' 133 DCX26 5' GAAATCTCACTAGTAGTCCGCC3' 134 DCX33 5' GCGGGCAAGACAGTCCAATTCC3' 135 DCX36 5'GAGCTCCAATTCCACGACGACC3' 136 DCX3 9 5'GGTTGCCATGCGTTCAAACTAC3' 137 DCX42 5'TCCCGCGGGGACAAACCCGAAT3' 138 DCX45 5' CTGCTAGTCTTATCATTCCCCA3' 139 PAX2 5' CTATCGACACTATAGGGCCTAC3' 140 PAX 9 5'TACCCTTGTAACCCACACTAGG3' 141 PAX14 S'TTCTTCTGAATAGACCGGCCGA3' 142 PAX15 5'CCACCACCCTTAACCCGACAAT3' 143 PAX16 5'AGGGGGAGACTTGTTCACAAAC3' 144 PAX17 5'CGGCTCATACCACCGAAAGCTA3' 145 PAX18 5'ATCGTCCTACTGTAATCCTCGA3' 14 6 PAX35 5' GACACACTACTCAGGTCCACCT3' 147 PAX38 5' CCATAATCAACATTGCCGCCCT3' 148 PAX4 0 5'CAAAACGCTCGCCCCAAACTCA3' 14 9 PAX43 5'GTAAACTTGTGCTTCTCGCACC3' 150 PAX45 5'CCATGGTCCGGGTACACCTGAA3' 151 PAX46 S'GTTACTAACGGAACGAGACCTA3' 152 P31 5'TGTTGGCGTTCTCAACCCCGTT3' 153 P90 5'ACAACCGGAGTTGTTCTGCCTA3' 154 5PAX3 5'TAAGCATCGGCCACGTTCTTCG3' 155 5PAX5 5' TTATCCCTGGTGTGCAGGTTGA3' 156 Printed from Mimosa CLONE NAME OLIGO SEQ.

ID. NO. 5PAX7 ' TATCAGAATCGTAGTCGGACGG3' 157 5PAX12 ' CTTTGTAATGGAACCACAACCC3' 158 HAX9 ' CGGTGGCTCATCTCCCTCTTAT3' 159 HAX35 ' ATCAGACTGGCTGGGACCACAA3' 160 HAX4 0 ' CACAACCTCCTCTCCGCGAACT3' 161 HAX42 ' AGATTCGTCCCCAACGCGTGAT3' 162 HCA3 ' GGGAATTCGCAAAGCTATACTC3' 163 H40 ' CCCCGTGGAATTCAACCTGTGA3' 164 M13 (positive) ' GTCGTCTTTCCAGACGT3' 165 M13 (negative) ' CTTGCATGCCTGCAGGTCGAC3' 166 The oligonucleotides (5pmol) were 5'end labelled with 32P-ATP and T4 polynucleotide kinase and approximately 2.5pmol of 15 labelled oligonucleotide was used in hybridization studies. Hybridizations were performed at 40-45°C overnight in buffer containing 6X SSC, 5X Denhardt's solution, 0.1% SDS, 20/zg/ml yeast tRNA and the radiolabeled synthetic oligonucleotide, followed by washings (20-30 min at 40-45°C) in the following 20 buffers: (i) 2X SSC / 0.1% SDS, (ii) IX SSC / 0.1% SDS, (iii) 0.IX SSC / 0.1% SDS. The filters were air-dried and exposed for autoradiography for 15 hours, 24 hours or 72 hours.

Hybridization data indicated that all the oligonucleotide probes bound specifically to their phage 2 5 target except for the HAX9 probe which apparently was not labeled. A negative control probe that hybridized only to M13mpl8 DNA showed a weak to negative signal in all samples tested (data not shown).

Hybridization data for pools from each receptor 30 group of rats was compiled. Tables 15, 16, 17 and 18 show a representative compilation of autoradiograph signals of the HSI, D2H, HPTl and hPEPTl receptor groups. These Tables show the phage absorption and uptake from the closed loop GIT model to portal and systemic circulation and phage 3 5 absorption/internalization to intestinal tissue. In these Tables, Int-G refers to intestinal tissue homogenized prior Printed from Mimosa to washing and recovery while Int-D refers to intestinal tissue washed prior to homogenization and phage recovery. In all cases, leading phage candidates were present in more than one animal.

Table 15 SUMMARY OF AUTORADIOGRAPH SIGNALS OF HSI ANIMAL STUDY Phage Portal Int.-G Int.-D S15 + + + /- + /- S21 - - - S22 - -/ + - SNi-10 ++ + /+ + + + + SNi-28 - - - SNi-34 ++ - - SNi-38 ++ - - SNi-45 - - - SNiAX-2 - - - SNiAX-6 - - - SNiAX-8 - - - M13 ++++++ ++++++ ++++++ M13 nd* + - *not detected Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 16 SUMMARY OF AUTORADIOGRAPH SIGNALS OF D2H ANIMAL STUDY Phage Portal Int.-G Int.-D DAB 3 +++ +/- -/+ DAB 7 ++ + + -/+ DAB10 ++++++ +/- -/+ DAB18 - - - DAB 2 4 - - - DAB30 ++++ ++ +++ DAX15 - - - DAX23 -/+ + -/+ DAX24 - - - DAX27 - + - DCX8 +++++ +/- - DCX11 ++++++ + + -/+ DCX2 6 - " - DCX33 +++ + + ++ DCX3 6 - - - DCX3 9 - -/+ - DCX42 - - -/+ DCX4 5 - ++ - M13 (+) +++++ +++++ +++++ M13 (-) +/- -/+ ~ Table 17 SUMMARY OF AUTORADIOGRAPH SIGNALS OF HPTl ANIMAL STUDY Phage Int.-G Portal Systemic H4 0 _ ++++ HAX9 ND ND ND HAX35 - + - HAX4 0 - - - HAX42 - ++ ++ HCA3 - - - PAX2 - ++ + ++++ M13(+) ++++++ ++++++ ++++++ M13 (-) - --/ + - Printed from Mimosa Table 18 SUMMARY OF AUTORADIOGRAPH SIGNALS OF hPEPTl ANIMAL STUDY Phage Int.-G Portal Systemic PAX2 . ++ _ PAX 9 + + +++ - PAX14 - ++ - PAX15 -/ + - - PAX16 - - - PAX17 + ++/ + - PAX18 - - - PAX35 - - - PAX3 8 -/ + - - PAX4 0 + +++ - PAX43 + - - PAX45 - - - PAX46 - +++ - P31 ++ ++++ ++ 5PAX3 + + / + ++ - 5PAX5 - - ++ 5PAX7 + + + - - 5PAX12 +++ + ++ - H40 ++ ++ - M13(+) ++++++ ++++++ ++++++ M13 (-) - - - Apart from the synthetic oligonucleotide to HAX9, all oligonucleotides were initially confirmed to be radiolabeled, as determined by hybridization to the corresponding phage target (eg., phage S15 hybridized to the oligonucleotide 25 S15). In addition, under the experimental conditions used, the oligonucleotides essentially did not hybridize to the negative control phage template M13mpl8. Two oligonucleotides were synthesized to the phage M13mpl8: (1) a positive oligonucleotide which hybridizes to a conserved 30 sequence in both M13mpl8 and each of the GIT receptor or GIT binding site selected phages [designated M13 (positive)]; and (2) a negative oligonucleotide which only hybridizes to a sequence unique to the multiple cloning site of phage M13mpl8 and which does not hybridize to any of the GIT receptor or 35 gix binding site selected phages.

Printed from Mimosa PCT/U S98/10088 In the case of the hSI pool of phages, only four phages were transported from the closed loop model into the portal circulation: phages S15, SNi-10, SNi-34 and SNi-38. The other phages, S21, S22, SNi-28, SNi-45, SNiAX-2, SNiAX-6 and 5 SNiAX-8, were not transported from the GIT into the portal circulation. In addition, phages SNi-10 and to a lesser extent phages S15 and S22 were found in the intestine samples or fractions, whereas the other phages were not. There was a very low presence (<0.1%) of the phage M13mpl8 in the Int-G 10 samples. These results show that phages can be further selected from pre-selected libraries, permitting the identification of phages which are transported from the GIT closed loop into the portal circulation or phages which bind to or are internalized by intestinal tissue.

In the case of the D2H pool of phages, there was a rank order by which phages were transported from the GIT closed loop model into the portal circulation, with phages DCX11 and DAB10 preferably transported, followed by phages DCX8, DAB30, DAB3 and DAB7. A number of phages from this pool were not 20 transported into the portal circulation, including phages DAB18, DAB24, DAX15, DAX24, DAX27, DCX26, DCX3 6, DCX3 9, DCX42, DCX45. There is a very low level of transport of phage DAX23 from the GIT into the portal circulation. Similarly, only some of the phages were found in the intestinal samples 25 fractions, including phages DAB3 0, DCX3 3, DAB7, DCX11, DCX45 and to a much lesser extent phages DAB3, DAB10, DCX8, DCX3 9, DCX42. Some phages were not found in the intestinal samples, including phages DAB18, DAB24, DAX15, DAX24, DCX26, and DCX36. There was a very low presence (<0.1%) of the phage 30 Ml3mpl8 in the Int-G samples. These results showed that phages can be further selected from pre-selected libraries, permitting the identification of phages which are transported from the GIT closed loop into the portal circulation or phages which bind to or are internalized by intestinal 35 tissue.

In the case of the HPTl pool of phages, there was a rank order by which phages were transported from the GIT closed Printed from Mimosa loop model into the portal or systemic circulation. Phage PAX2 (which was used at a 4X concentration relative to the other phages in this pool) followed by phage HAX42 was found in the portal and systemic circulation; phage H40 was found 5 in the systemic circulation only. None of the phages in this pool were found in the intestine samples or fractions. Phage M13mpl8 was not found in the intestine fractions or systemic circulation, with very low incidence (<0.001%) in the portal circulation. These results show that phages can be further 10 selected from pre-selected libraries, permitting the identification of phages which are transported from the GIT closed loop into the portal and/or systemic circulation or phages which bind to or are internalized by intestinal tissue.

In the case of the hPEPTl pool of phages, the phages PAX2 and H40 were also included in this pool. A number of phages from this pool were found in the portal circulation, including phages P31 (SEQ ID NO:43), PAX4 6, PAX9, H40, PAX17, PAX4 0, PAX2, PAX14, 5PAX3 and 5PAX12. A number of phages 20 were not found m the portal blood including the negative control phage M13mpl8, PAX15, PAX16, PAX18, PAX35, PAX38, PAX43, PAX45, P90, 5PAX5 and 5PAX7. The only phage found in the systemic circulation were phages 5PAX5 and P31 (SEQ ID NO:43). In addition, there was preferential binding of some 25 phages to the intestine, including phages 5PAX12, 5PAX7, 5PAX3, H40, P31 (SEQ ID NO:43), PAX9, and to a lesser extent phages PAX38 and PAX15. Some phages were not found in the intestine samples, including the negative control phage M13mpl8 and the phages PAX2, PAX14, PAX16, PAX18, PAX35, 30 PAX45, PAX46, P90 and 5PAX5. These results show that phages can be further selected from pre-selected libraries, permitting the identification of phages which are transported from the GIT closed loop into the portal and/or systemic circulation or phages which bind to or are internalized by 35 intestinal tissue.

Printed from Mimosa Further Characterization of Select Sequences Following initial screening of the four recombinant receptor sites (hPEPTl, HPTl, D2H, hSI) of the gastrointestinal tissue, with the phage display libraries, a 5 series of phage were isolated which showed preferential binding to the respective target receptor sites in comparison to negative control protein BSA protein and the recombinant protein recombinant human tissue factor (hTF) (which, like the recombinant receptors of the gastrointestinal tissue, 10 contained a poly-histidine tag at its NH2-terminal end) . In subsequent experiments same titers of the selected phage which bound to each target receptor site were combined into a single pool ii.e., one pool of HPTl binding phage, one pool of hPEPTl binding phage, one pool of D2H binding phage, and 15 one pool of hSI binding phage). Each pool was supplemented with an equivalent titer of the negative control phage M13mpl8. These phage pools were injected into a closed duodenal loop region of rat intestinal tissue and subsequently phage was harvested and recovered which was 20 bound to and retained by the intestinal tissue and/or was absorbed from the intestinal loop into the portal and/or systemic circulation. In addition, a selection of the initial phages which bound to the target recombinant receptor site were analyzed for binding to either fixed Caco-2 cells 25 and/or to fixed C2BBel cells. The selection of the final lead peptide sequences was based on the ability of the phage, coding for that peptide sequence (1) to bind to the target recombinant receptor site in vitro in preference to its binding to the negative control proteins BSA and/or hTFs, (2) 30 to bind to rat intestinal tissue following injection into a closed duodenal loop of rat intestinal tissue in preference to the negative control phage M13mpl8, (3) to be absorbed from rat intestinal tissue into either the portal and/or systemic circulation following injection into a closed 35 duodenal loop of rat intestinal tissue in preference to the negative control phage M13mpl8, and (4) to bind to either fixed Caco-2 cells or fixed C2BBel cells in phage binding Printed from Mimosa studies in preference to the negative control phage M13mpl8. Peptides were also selected with consideration to the ease of chemical synthesis. 6.9. GST Fusion Proteins of GIT Targeting Peptides Construction of GST Fusion Proteins of GI Targeting Peptides Glutathione S-transferase (GST) vectors encoding fusion proteins of GI targeting peptides were constructed in the vector pGEX4T-2 (source, Pharmacia Biotech, Piscataway, NJ). Briefly, single-strand DNA from the clones of interest were amplified by the polymerase chain reaction. The amplified DNA was then cleaved with the restriction enzymes Xhol and Notl and then ligated into Sall/NotI cleaved pGEX4T-2. Following transformation, the DNA sequence for each construct was verified by sequencing.

For construction of the truncated versions of the GST fusion proteins, where the inserted sequence was less than 45 base pairs, overlapping oligonucleotides containing cohesive Sail and Notl termini, and encoding the sequence of o n interest, were annealed and then ligated directly into Sall/NotI cleaved pGEX4T-2. Following transformation, the DNA sequence for each construct was verified.

A diagrammatic representation of the various GST fusion protein constructs that have been synthesized is indicated in Figures 5A-5C.

Expression and Purification of GST Fusion Proteins Escherichia coli BL21 cells containing GST fusion protein constructs were grown overnight in 2X YT media 3 0 containing 100 /xg/ml ampicillin (2X YT/amp) . Overnight cultures were diluted 1:100 in 2X YT broth (100 ml), and cells were grown to an A600 of 0.5 at 30°C, induced with ImM isopropyl-l-thio-B-D-galactopyranoside, and grown for an additional 3 h. Cells were harvested by centrifugation and 3 5 resuspended in 5 ml of PBS containing a mixture of the proteinase inhibitors (Boehringer/Mannheim). Cells were Printed from Mimosa sonicated on ice, and the cell lysates were centnfuged at 12,000 x g for 10 minutes at 4°C. Supernatant fractions were reacted for 30 minutes at room temperature with 2 ml of a 50% slurry of glutathione-Sepharose® 4B, washed 3 times with 1.5 5 ml of PBS (at room temperature), and the bound GST fusion proteins were eluted by reaction for 10 minutes at room temperature with 3 X 1ml of 10 mM reduced glutathionein 50 mM Tris HCl pH 8.0. Protein was quantified by the Bio-Rad protein assay followed by characterization by SDS-10 polyacrylamide gel electrophoresis.

ELISA of GST fusion peptides The standard ELISA procedure was modified as follows. GST proteins were diluted to an appropriate 15 concentration in PBS containing 1%BSA and 0.05% Tween20 (1%BPT), titered and incubated one hour at room temperature. Following five washes an anti-GST monoclonal antibody was added (Sigma, St. Louis Clone GST-2 diluted 1:10,0 00 in 1%BPT) and incubated one hour. After five more washes goat 20 anti-mouse IgG2b-HRP was added (Southern Biotechnology Associates Inc., Birmingham, AL, diluted 1:4000 in 1%BPT) and incubated one hour. After five washes plates were developed with TMB peroxidase substrate (Kirkegard and Perry, Gaithersburg, MD). All data is presented with background 25 binding subtracted.

Figure 6 shows the binding of GST-SNilO, GST-SNi34 and GST alone to the hSI receptor and to fixed C2BBel cells.

GST Fusion Proteins of Selected GIT Targeting Peptides Results show that GST-DXB8, GST-PAX2, GST-P31, GST- SNilO and GST-SNi34 bound fixed Caco-2 or C2BBel cells (Figures 7 and 8) relative to GST control binding.

GST-HAX42, GST-5PAX5, all showed weak to moderate binding relative to GST control.

Interestingly, P31 truncation 103-GST fusion protein bound almost as well as full-length P31 (SEQ ID NO:43) to fixed Caco-2 cells (A). This suggests the portion Printed from Mimosa PCMJS98/10088 of the P31 sequence (SEQ ID NO:43) responsible for binding resides in this portion. PAX2.107 bound similarly to full-length PAX2; therefore, this portion most likely contains the amino acid sequence responsible for binding (B). In 5 preliminary assays, none of the DCX8 truncations bound similarly to full-length DCX8 to Caco-2 cells suggesting the binding region spans more than one of these pieces.

Inhibition of Binding by Synthetic Peptides 10 Binding of GST-P31 to fixed C2BBel Cells The standard ELISA procedure was modified as follows. GST fusion proteins and peptides were diluted to an appropriate concentration in PBS containing 1% BSA and 0.05%' Tween 20. Peptides were titered, a constant concentration of 15 diluted GST protein was added to titered peptides and the mixture was incubated one hour at room temperature.

Following five washes, an anti-GST monoclonal antibody was added (Sigma, St. Louis Clone GST-2 diluted 1:10,000 in 1% BPT) and incubated one hour. After five more washes goat 20 anti-mouse IgG2b-HRP was added (Southern Biotechnology Associates Inc., Birmingham, AL, diluted 1:4000 in 1% BPT) and incubated one hour. After five washes plates were developed with TMB peroxidase substrate (Kirkegard and Perry, Gaithersburg, MD). All data is presented with background 25 binding subtracted.

Figures 9A and 9B show the inhibition of GST-P31 binding to C2BBel fixed cells. The peptide competitors are ZElan024 which is the dansylated peptide version of P31 (SEQ ID NO:43) and ZElan044, ZElan049 and ZElan050 which are 30 truncated, dansylated pieces of P31 (SEQ ID NO:43). Data is presented as O.D. vs. peptide concentration and as percent inhibition of GST-P31 binding vs. peptide concentration. Uncompeted GST-P31 binding was considered as 100% binding. IC50 values are estimates using the 50% line on the percent 35 inhibition graph.

GST-P31 and GST-PAX2 exhibited no crossreactive binding to ZElan024 (P31) (SEQ ID NO:43) and ZElan018 (PAX2) Printed from Mimosa at the 0.5 /ig/ml concentration used in competition assays GST-HAX42 exhibited crossreactivity to ZElan018 (PAX2) and ZElan021 (HAX42) at the 5 £ig/ml concentration used in competition assays.

Figures 10A-10C present a compilation of data generated by competition ELISA of GST-P31, GST-PAX2, GST-SNilO and GST-HAX42 versus various dansylated peptides on fixed C2BBel cells. IC50 values are in /iM and include ranges determined from multiple assays. The GST/C2BBel column is a 10 summary of GST protein binding to fixed C2BBel cells.

Binding to fixed Caco-2 Cells Caco-2 cells were fixed, treated with phenylhydrazine and blocked as described above. Synthetic 15 peptides (100/xg/ml) were applied in duplicate to Caco-2 cells and serially diluted down the 96-well plate. The corresponding GST-peptide fusion protein (10/ig) was added to each well and the plates were incubated for 2h at room temperature with agitation. Binding of the GST-peptide 20 fusion proteins to the cells was assayed using the ELISA technique described above. GST-P31 binding was inhibited by ZElan024, ZElan028 and ZElan031 as well as the two D forms ZElan053 and ZElan054. GST-PAX2 binding was inhibited by ZElan032, ZElan033, and ZElan035. GST-HAX42 binding was not 25 inhibited by ZElan021 (full length HAX42) but it was inhibited by ZElan018 (PAX2) and ZElan026 and ZElan038 (scrambled PAX2 peptides).

Transport and Uptake of GST-Peptide 20 Fusions into Live Caco-2 Cells Transport and uptake of GST-peptide fusions and deletion derivatives across cultured polarized Caco-2 monolayers over 4 hours in HBSS buffer was examined using an anti-GST ELISA assay. In another experiment, transport and uptake of GST-peptide fusions and deletion derivatives across Printed from Mimosa cultured polarized Caco-2 monolayers over 24 hours in serum-free medium (SFM) was examined using an anti-GST ELISA assay.

Materials Buffered Hank's balanced salt solution (bHBSS) = lx HBSS (Gibco CN.14065-031) supplemented with 0.011M glucose (lg/1), 25 mM Hepes (15 mM acid (3.575g/l; Sigma CN.H3375); lOmM base (2.603g/l; Sigma CN.H1016)].

Chloroguine. Made up as lOmM solution in water 10 [Sigma CN C6628] Lysate buffer: 30 mM Tris-HCl pH8.0; ImM EDTA Serum-free medium (SFM) is normal medium without serum.

Method a) 4h HBSS study: Transepithelial electrical flux (TER) across the Caco-2 monolayers grown on snapwells (passage 33; 23 days old) was measured to confirm monolayer integrity before beginning the experiment. The medium was removed and the cells were washed once with bHBSS. bHBSS containing 100/xM chloroquine was added and the cells were incubated for 2h at 37°C. The bHBSS+chloroquine was replaced with 0.5ml bHBSS containing GST-peptide fusions (100/xg/ml) and the cells were incubated as before. Basolateral samples 25 were removed at the following times: 0, 0.5h, 2h, and 4h. At 4h, TER was measured, the apical medium was sampled and the apical reservoir was washed 6 times with HBSS. The cells were allowed to lyse for lh on ice in lysate buffer, after which, lysate sample was collected. All samples were stored 30 at -70°C until assay by anti-GST ELISA. Before analysis, samples were normalized for protein content relative to each other using a BioRad protein assay. b) 24h SFM study: Transepithelial electrical flux (TER) across the Caco-2 monolayers grown on snapwells (passage 33; 23 days old) was measured to confirm monolayer integrity before beginning the experiment. The medium was removed and the cells were washed once with SFM. SFM Printed from Mimosa PCT/U S98/10088 containing GST-peptide fusions (100/xg/ml) was added to the cells which were incubated at 37°C for 24h at 5% C02. After 24 hours, TER readings were taken, and samples from the basolateral and apical reservoirs were removed. The apical 5 reservoir was washed 6 times with PBS. The cells were allowed to lyse for lh on ice in lysate buffer, after which lysate sample was collected. All samples were stored at -70° until assay by anti-GST ELISA. Before analysis, samples were normalized for protein content relative to each other using a 10 BioRad protein assay.

Results All of the GST-peptide fusions and controls examined were transported across live Caco-2 monolayers. 15 Full-length GST-P31 and GST-DCX8, but not truncations of these molecules had a higher flux than GST alone.

Internalization of GST-peptide fusions into polarized Caco-2 cells was investigated in two experiments. In experiment 1, 15/xg of GST-peptide fusion was applied in 20 bHBSS and internalized GST-peptide was recovered by lysing the cells after 4h. In experiment 2, lOfig of GST-peptide was applied in either a) bHBSS (lysate recovered after 4h), or b) serum-free medium (lysate recovered after 24h).

Figure 11A describes complete transport of GST-25 peptide across a polarized Caco-2 monolayer and does not necessarily refer to internalization, i.e., the GST-peptide was recovered from the basolateral reservoir of a snapwell but the proteins could have crossed the barrier by the paracellular route.' Effect of Thrombin Cleavage on Binding of GST-Peutide Fusions to Fixed Caco-2 Cells Binding of intact and thrombin-cleaved GST-peptide fusions to fixed Caco-2 cells was compared. Reduced binding 35 of the thrombin-cleaved GST-peptide fusions relative to intact fusions indicates that the peptide component of the fusion, and not the GST domain, mediates binding.

Printed from Mimosa Method Confluent Caco-2 monolayers grown in 96-well plates (p38) were fixed and treated with 0.1% phenylhydrazine before blocking with 0.1% BSA in PBS. Thirty micrograms of each 5 GST-peptide was treated with bovine thrombin (1/z/ml; 0.4 NIH units; Sigma CN.T9681) for 18h at room temperature in 20mM Tris-HCl pH8.0, 150mM NaCl, 2.5mM CaCl2. Controls were similarly treated without addition of thrombin. Ten micrograms of each GST-peptide fusion was removed for PAGE 10 analysis, and 10/ig of fusions were added in duplicate to the fixed Caco-2 cells before 5-fold serial dilutions (1% BPT diluent). The fusions were allowed to bind for lh at room temperature. Following 6 washes with 1% BPT, binding was assayed by ELISA.

Results Results are shown in Figure 12.

Conclusions: PAGE analysis confirmed that the GST-peptide fusions were effectively cleaved with thrombin. Cleavage with thrombin significantly reduced detection of binding of GST-P31.103, GST-PAX2.106, GST-DCX8, GST-SNilO to fixed Caco-2 cells, indicating that the peptide component, and not the 25 GST domain, mediates binding. 6.10. Synthesis of Peptides 6.10.1. Procedure For Solid Phase Synthesis Peptides may be prepared by methods that are known 30 in the art. For example, in brief, solid phase peptide synthesis consists of coupling the carboxyl group of the C-terminal amino acid to a resin and successively adding N-alpha protected amino acids. The protecting groups may be any known in the art. Before each new amino acid is added to 35 the growing chain, the protecting group of the previous amino acid added to the chain is removed. The coupling of amino acids to appropriate resins is described by Rivier et al., Printed from Mimosa U.S. Patent No. 4,244,94 6. Such solid phase syntheses have been described, for example, by Merrifield, 1964, J. Am.

Chem. Soc. 85:2149; Vale et al., 1981, Science 213:1394-1397; Marki et al., 1981, J. Am. Chem. Soc. 103:3178 and in U.S.

Patent Nos. 4,305,872 and 4,316,891. In a preferred aspect, an automated peptide synthesizer is employed.

By way of example but not limitation, peptides can be synthesized on an Applied Biosystems Inc. ("ABI") model 431A automated peptide synthesizer using the "Fastmoc" 10 synthesis protocol supplied by ABI, which uses 2-(lH-Benzotriazol-l-yl)-1,1,3,3,-tetramethyluronium hexafluorophosphate ("HBTU") (R. Knorr et al., 1989, Tet. Lett., 30:1927) as coupling agent. Syntheses can be carried out on 0.25 mmol of commercially available 15 4-(2',4'-dimethoxyphenyl-(9-fluorenyl- methoxycarbonyl)-aminomethyl)-phenoxy polystyrene resin ("Rink resin" from Advanced ChemTech) (H. Rink, 1987, Tet. Lett. 28:3787). Fmoc amino acids (1 mmol) are coupled according to the Fastmoc protocol. The following side chain 20 protected Fmoc amino acid derivatives are used: FmocArg(Pmc)OH; FmocAsn(Mbh)OH; FmocAsp ^Bu) OH; FmocCys(Acm)OH; FmocGlu C^Bu) OH; FmocGln(Mbh)OH; FmocHis(Tr)OH; FmocLys (Boc) OH; FmocSer TBu) OH; FmocThr CBu) OH; FmocTyr (tBu) OH. [Abbreviations: Acm, acetamidomethyl; Boc, 25 tert-butoxycarbonyl,- ^u, tert-butyl; Fmoc, 9-fluorenylmethoxycarbonyl; Mbh, 4,4'-dimethoxybenzhydryl; Pmc, 2,2,5,7,8-pentamethylchroman-6-sulfonyl; Tr, trityl].

Synthesis is carried out using N-methylpyrrolidone (NMP) as solvent, with HBTU dissolved in 30 N,N-dimethylformamide (DMF). Deprotection of the Fmoc group is effected using approximately 20% piperidine in NMP. At the end of each synthesis the amount of peptide present is assayed by ultraviolet spectroscopy. A sample of dry peptide resin (about 3-10 mg) is weighed, then 20% piperidine in DMA 35 (10 ml) is added. After 30 min sonication, the UV (ultraviolet) absorbance of the dibenzofulvene-piperidine adduct (formed by cleavage of the N-terminal Fmoc group) is Printed from Mimosa recorded at 301 nm. Peptide substitution (in mmol g"1) can be calculated according to the equation: A x v substitution = x 1000 7800 x w where A is the absorbance at 301 nm, v is the volume of 20% piperidine in DMA (in ml), 7800 is the extinction coefficient (in mol "Mm3 cm"1) of the dibenzofulvene-piperidine adduct, and w is the weight of the peptide-resin sample (in mg) .

Finally, the N-terminal Fmoc group is cleaved using 20% piperidine in DMA, then acetylated using acetic anhydride and pyridine in DMA. The peptide resin is thoroughly washed with DMA, CH2Cl2 and finally diethyl ether. 6.10.2. Cleavage and Deprotection By way of example but not limitation, cleavage and deprotection can be carried out as follows: The air-dried peptide resin is treated with ethylmethyl-sulfide (EtSMe), ethanedithiol (EDT) ,( and thioanisole (PhSMe) for 20 approximately 20 min. prior to addition of 95% aqueous trifluoracetic acid (TFA). A total volume of approximately 50 ml of these reagents are used per gram of peptide-resin. The following ratio is used: TFA:EtSMe:EDT:PhSme (10:0.5:0.5:0.5). The mixture is stirred for 3 h at room 2g temperature under an atmosphere of N2. The mixture is filtered and the resin washed with TFA (2 x 3 ml). The combined filtrate is evaporated in vacuo, and anhydrous diethyl ether added to the yellow/orange residue. The resulting white precipitate is isolated by filtration. See 2Q King et al., 1990, Int. J. Peptide Protein Res. 36:255-266 regarding various cleavage methods. 6.10.3. Purification of the Peptides Purification of the synthesized peptides can be carried out by standard methods including chromatography (e.g., ion exchange, affinity, and sizing column chromatography, high performance liquid chromatography Printed from Mimosa (HPLC)), centrifugation, differential solubility, or by any other standard technique. 6.10.4. Conjugation of Peptides to Other Molecules The peptides of the present invention may be linked to other molecules (e.g., a detectable label, a molecule facilitating adsorption to a solid substratum, or a toxin, according to various embodiments of the invention) by methods that are well known in the art. Such methods include the use of homobifunctional and heterobifunctional cross-linking molecules.

The homobifunctional molecules have at least two reactive functional groups, which are the same. The reactive functional groups on a homobifunctional molecule include, for example, aldehyde groups and active ester groups. Homobifunctional molecules having aldehyde groups include, for example, glutaraldehyde and subaraldehyde. The use of glutaraldehyde as a cross-linking agent was disclosed by 2Q Poznansky et al., 1984, Science 223:1304-1306.

Homobifunctional molecules having at least two active ester units include esters of dicarboxylic acids and N-hydroxysuccinimide. Some examples of such N-succinimidyl esters include disuccinimidyl suberate and dithio-bis-(succinimidyl propionate), and their soluble bis-sulfonic Z D acid and bis-sulfonate salts such as their sodium and potassium salts. These homobifunctional reagents are available from Pierce, Rockford, Illinois.

The heterobifunctional molecules have at least two different reactive groups. Some examples of heterobifunctional reagents containing reactive disulfide bonds include N-succinimidyl 3-(2-pyridyl-dithio)propionate (Carlsson et al., 1978, Biochem J. 173:723-737), sodium S-4-succinimidyloxycarbonyl-alpha-methylbenzylthiosulfate, and 35 4-succinimidyloxycarbonyl-alpha-methyl-(2-pyridyldithio)toluene. N-succinimidyl 3-(2-pyridyldithio)propionate is preferred. Some examples of Printed from Mimosa heterobifunctional reagents comprising reactive groups having a double bond that reacts with a thiol group include succinimidyl 4-(N-maleimidomethyl)cyclohexahe-l-carboxylate and succinimidyl m-maleimidobenzoate.

Other heterobifunctional molecules include succinimidyl 3-(maleimido)propionate, sulfosuccinimidyl 4-(p-maleimido-phenyl)butyrate, sulfosuccinimidyl 4-(lima leimi dome thy 1-eye lohexane) -1-carboxylate, maleimidobenzoyl-N-hydroxy-succinimide ester. The sodium sulfonate salt of 10 succinimidyl m-maleimidobenzoate is preferred. Many of the above-mentioned heterobifunctional reagents and their sulfonate salts are available from Pierce.

Additional information regarding how to make and use these as well as other polyfunctional reagents may be 15 obtained from the following publications or others available in the art: Carlsson et al., 1978, Biochem. J. 173:723-737; Cumber et al., 1985, Methods in Enzymology 112:207-224; Jue et al., 1978, Biochem 17:5399-5405; Sun et al., 1974, Biochem. 13:2334-2340; Blattler et al., 1985, Biochem. 20 24:1517-152; Liu et al., 1979, Biochem. 18:690-697; Youle and Neville, 1980, Proc. Natl. Acad. Sci. USA 77:5483-5486; Lerner et al., 1981, Proc. Natl. Acad. Sci. USA 78:3403-3407; Jung and Moroi, 1983, Biochem. Biophys. Acta 761:162; Caulfield et al., 1984, Biochem. 81:7772-7776; Staros, 1982, 25 Biochem. 21:3950-3955; Yoshitake et al., 1979, Eur. J. Biochem. 101:395-399; Yoshitake et al., 1982, J. Biochem. 92:1413-1424; Pilch and Czech, 1979, J. Biol. Chem. 254:3375-3381; Novick et al., 1987, J. Biol. Chem. 262:8483-8487; Lomant and Fairbanks, 1976, J. Mol. Biol. 104:243-261, Hamada 30 and Tsuruo, 1987, Anal. Biochem. 160:483-488; Hashida et al., 1984, J. Applied Biochem. 6-56-63.

Additionally, methods of cross-linking are reviewed by Means and Feeney, 1990, Bioconjugate Chem. 1:2-12. 6.10.4.1. Biotinvlation of Peptides Methods of biotinylating peptides are well known in the art. Any convenient method may be employed in the Printed from Mimosa practice of the invention. For example, the following procedure was used. Ten micrograms of peptide was dissolved in 100 nl of 0.1 % acetic acid. PBS (900/xl) and 3.3 mg of biotin-LC-NHS (Pierce, Rockford, IL) was added. Following 5 incubation for 30 minutes at room temperature the biotinylated peptides were purified over a Superose 12 column (Pharmacia, Piscataway, NJ). 6.10.5. Synthetic Peptides Tables 19, 20 and 21 provide the primary structure for various synthetic peptides manufactured in the practice of the present invention.

Table 19 Seq Peptide Sequence ID name No ELAN005 H2N-C-K(dns)- FITKALGISYGRKKRRQRRRPPQGSQTHQVSLSKQ-CONH2 ELAN006 Ac-CLNGGVKMYVESVDRYVC-CONH2 FITC- Ac-CLNGGVK(FITC)MYVESVDRYVC-CONH2 ELAN006 ELAN006ii H2N-C-K(dns)-RLNGGVSMYVESVDRYVCR-C0NH2 167 ELAN00 7 H2N-RIAGL.PWYRCRTVAFETGMQNTQLCSTIVQLSFTPEE- COOH 193 ELAN007ii H2N-KKRIAGLPWYRCRTVAFETGMQNTQLCSTIVQLSFTPEE- CONH2 bZElan008 biotin-K(dns)SARDSGPAEDGSRAVRLNGVENANTRKSSR (P31) SNPRGRRHP-COOH bZElan009 biotin-K(dns)SSADAEKCAGSLLWWGRQNNSGCGSPTKKH LKHRNRSQTSSS SHG-COOH 168 ELAN010 H2N-REFAERRLWGCDDLSWRLDAEGCGPTPSNRAVKHRKPRPR SPAL-COOH bZElanOlO biotin-K (dns)REFAERRLWGCDDLSWRLDAEGCGPTPSNR AVKHRKPRPRS PAL-COOH 169 ELAN012 H2N- SGSHSGGMNRAYGDVFRELRDRWYATSHHTRPTPQLPRGPN- COOH bELAN012 biotin- SGSHSGGMNRAYGDVFRELRDRWYATSHHTRPTPQLPRGPN- COOH ZElan012 h2n- K(dns)SGSHSGGMNRAYGDVFRELRDRWYATSHHTRPTPQLP RGPN-COOH Printed from Mimosa 249 250 251 252 ELAN013 ELAN014 bZElan014 ZElan014 ZElan015 (DCX11) ZElan016 (SNilO) bZElan017 ZElan017 ZElan018 (PAX2) ZElan019 (5PAX5) ZElan020 (CY0 9) bZElan020 (CY09) ZElan021 (HAX42) ZElan022 (SNi34) ZElan023 (DCX8) ZElan024 (P31) ZElan025 (DAB10) ZElan026 (PAX2/con trol) bELAN027 (PAX2) 18C21 Fmoc-Z16N23 16C23 h2n- SGSPPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSD y-conh, h2n- SHSGGMNRAYGDVFRELRDRWNATSHHTRPTPQLPRGPNS- CONH2 biotin- K(dns)SHSGGMNRAYGDVFRELRDRWNATSHHTRPTPQLPRG PNS-CONH, h2n- K(dns)SHSGGMNRAYGDVFRELRDRWNATSHHTRPTPQLPRG PNS-CONH2 H2N- K(dns)SQGSKQCMQYRTGRLTVGSEYGCGMNPARHATPAYPA RLLPRYR-CONH2 h2n- K(dns)RVGQCTDSDVRRPWARS CAHQGCGAGTRNSHGCITRP LRQASAH-CONH2 biotin-K(dns)SGSGRVGQCTDSDVRRPWARSCA-CONH2 H2N-K(dns)RVGQCTDSDVRRPWARSCA-CONH2 h2n- K(dns)STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSR png-conh2 h2n- K(dns)RGSTGTAGGERSGVLNLHTRDNASGSGFKPWYPSNRG hk-conh2 H2N-K(dns)SGSGLYANPGMYSRLHSPA-CONH2 biotin-K(dns)SGSGLYANPGMYSRLHSPA-CONH2 H2N- K(dns)SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRR rpsaipt-conh2 h2n- K(dns)SPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPP ssdy-conh2 h2n- K(dns)RYKHDIGCDAGVDKKSSSVRGGCGAHSSPPRAGRGPR GTMVSRL-CONH2 h2n- K(dns)SARDSGPAEDGSRAVRLNGVENANTRKSSRSNPRGRR hpgg-conh2 h2n- K(dns)SKSGEGGDSSRGETGWARVRSHAMTAGRFRWYNQLPS DR-CONH, h2n- K(dns)SEANLDGRKSRYSSPRRNSSTRPRTSPNSVHARYPST DHD-CONH2 biotin- SGSGSTPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN G-CONH2 H2N-DTNAKHSSHNRRLRTRSRPNG-CONH2 Fmoc-K(dns)RVGQCTDSDVRRPWARSCAHQG-COOH h2n- CGAGTRNSHGCITRPLRQASAHG-CONH2 Printed from Mimosa Z16C23 ZElan028 (P31 fragment) ZElan029 (P31 fragment) ZElan030 (P31 fragment) ZElan031 (P31 fragment) ZElan032 (PAX2 fragment) ZElan033 (PAX2 fragment) ZElan034 (PAX2 fragment) ZElan035 (PAX2 fragment) ZElan036 (SNilO fragment) ZElan037 (SNilO fragment) ZElan03 8 (PAX2/con trol) ZElan039 (P31 fragment) ZElan040 (P31 fragment) ZElan041 (P31 fragment) ZElan042 (P31 fragment) ZElan043 (P31 fragment) ZElan044 (P31 fragment) ZElan045 (P31 fragment) H2N-K(dns)CGAGTRNSHGCITRPLRQASAHG-CONH2 H2N-K(dns)ENANTRKSSRSNPRGRRHPG-CONH2 H2N-K(dns)TRKSSRSNPRG-CONH, H,N-K(dns)ENANTRKSSRSNPRG-CONH, H,N-K(dns)TRKSSRSNPRGRRHPG-CONH, H,N-K(dns)TNAKHS SHNRRLRTRSRPN-CONH, H,N-K(dns)TNAKHSSHNRRLRTR-CONH, H2N-K(dns)SSHNRRLRTRSRPN-CONH, H,N-K(dns)SSHNRRLRTR-CONH, H2N-K(dns)VRRPWARSCAHQGCGAGTRNS-CONH2 H2N-K(dns)CTDSDVRRPWARSC-CONH, H2N- K(dns)SRANTDGRKSRYSSPRRNSSTEPRLSPNSVHARYPST DHD-CONH2 H2N-K(dns)ENANTRKSSR-CONH2 H,N-K(dns)SNPRGRRHPG-CONH, H2N-K(dns)ENANT-CONH2 H2N-K(dns)ANTRKS-CONH, H2N-K(dns)TRKSS-CONH, H,N-K(dns)RKSSR-CONH, H2N-K(dns)KSSRSN-CONH, - 96 Printed from Mimosa ZElan046 (P31 fragment) ZElan047 (P31 fragment) ZElan048 P31 fragment) ZElan049 (P31 fragment) ZElan050 (P31 fragment) ZElan051 (HepC) ZElan052 (HepC) ZElan053 (P31 analog) ZElan054 (P31 analog) ZElan055 (PAX2 fragment) ZElan056 (PAX2 fragment) ZElan057 (PAX2 fragment) ZElan058 (PAX2 fragment) ZElan059 (PAX2 analog) ZElan060 (HAX42 fragment) ZElan061 (HAX42 fragment) ZElan062 (HAX42 fragment) ZElan063 (HAX42 fragment) ZElan064 (HAX42 fragment) H2N-K(dns)SSRSNPG-CONH2 H2N-K(dns)RSNPRG-CONH2 H2N-K(dns)SNPRG-CONH2 H2N-K(dns)PRGRRH-CONH2 H2N-K(dns)RRHPG-CONH2 H2N-K(dns)KSSRGN-C0NH2 H2N-K(dns)KTSERSQPRGRRQPG-CONH2 H2N-K(dns)TrKSSrSNPrGrrHPG-CONH2 H2N-K(dns)TRKSSrSNPRGrRHPG-CONH2 H2N-K(dns)TNAKHSSHN-CONH2 H2N-K(dns)RRLRTRSRPN-CONH2 H2N-K(dns)RRLRTRSR-CONH2 H2N-K(dns)RRLRTR-CONH2 H2N-K(dns)rrLrTrSrPN-CONH2 H2N-K(dns)SDHALGTNLRSDNAKEPGDYNCCGNG-CONH2 H2N-K(dns)GDYNCCGNGNSTGRKVFNRRRPSAIPT-CONH2 H2N-K(dns)SDHALGTNLRSDNAKEPG-CONH2 H2N-K(dns)GDYNCCGNGNSTG-CONH2 H2N-K(dns)RKVFNRRRPSAIPT-CONH2 Printed from Mimosa 55 52 ZElan065 (HAX42 fragment) ZElan066 (HAX42 fragment) ZElan067 (HAX42 fragment) Elan018 (PAX2 no dns) Elan021 (HAX42 no dns) ZElan070 (HAX42 fragment) ZElan071 (HAX42 fragment) ZElan072 (HAX42 fragment) ZElan073 (PAX2 fragment) ZElan074 (PAX2 fragment) ZElan075 (PAX2 fragment) ZElan076 (PAX2 fragment) ZElan077 (PAX2 fragment) ZElan078 (PAX2 fragment) ZElan079 (PAX2 fragment) ZElanoso (PAX2 fragment) ZElan081 (PAX2 fragment) ZElan082 (PAX2 fragment) Elan035 H,N-K(dns)RKVFNRRRPS-CONH, h,n-k(dns)nrrrpsaipt-conh, H,N- K (dns)NRRRPS-CONH, h2n- stppsreaysrpysvdsdsdtnakhsshnrrlrtrsrpng-conh2 h2n-sdhalgtnlrsdnakepgdynccgngnstgrkvfnrrrps aipt-conh2 h2n-k(dns)sdhalgtnlrsdnakepgdynccgngnst-conh2 h2n-k (dns) nlrsdnakepgdynccgngnstgrkvfnr-conh2 h2n-k(dns)pgdynccgngnstgrkvfnrrpsaipt-conh, h2n-k(dns)ashnrrlrtr-conh, h,n-k(dns)sahnrrlrtr-conh, H2N-K(dns)SSANRRLRTR-CONH, H,N-K(dns)S SHARRLRTR-CONH, h,n-k(dns)sshnarlrtr-conh, h,n-k(dns)sshnralrtr-conh2 h,n-K(dns)sshnrrartr-conh, H2N-K(dns)SSHNRRLATR-CONH, h,n-k(dns)sshnrrlrar-conh, h2n-k(dns)sshnrrlrta-conh, h2n-sshnrrlrtr-conh2 Printed from Mimosa ZElan083 H2N- (PAX2/con K(dns)GRNHDWSSNTHKSYRSPRSASYPRLSNDRTDRTEPA trol) pss-conh2 ZElan084 H2N-K(dns)RNTRNKTSRLSANPHRSHR-CONH2 (PAX2/con trol) Elan032Z H2N-TNAKHSSHNRRLRTRSRPN K(dns)-CONH2 (PAX2 fragment) Elan057Z H2N-RRLRTRSRK(dns)-CONH2 (PAX2 fragment) TABLE 20 Name Description Sequence ZElan087 HAX42-1 (20 mer) H2N-K(dns)SDHALGTNLRSDNAKEPGDY ZElan088 HAX42-2 (20 mer) H2N-K(dns)SDNAKEPGDYNCCGNGNSTG ZElan089 HAX42-3 (15 mer) H2N-K(dns)SDHALGTNLRSDNAK ZElan090 HAX42-4 (15 mer) H2N-K (dns)EPGDYNCCGNGNSTG ZElan091 HAX42-5 (14 mer) H2N-K(dns)PGDYNCCGNGNSTG ZElan092 HAX42-6 (10 mer) H2N-K(dns)PGDYNCCGNG ZElan093 HAX42-7 (10 mer) H2N-K(dns)NCCGNGNSTG ZElanlOO P31 16 mer cyclic H2N-K(dns)Lys-TRKSSRSNPRGRRHPG I 1 ZElanlOl P31 16 mer cyclic D form H2N-K(dns)Lys-TrKS SrSNPrGrrHPG 1 1 ZElanl03 PAX2 15 mer cyclic H2N-K(dns)Lys-TNAKHSSHNRRLRTR 1 1 ZElanl03A PAX2 15 mer cyclic (internal) H2N-K(dns)TNAKHSSCNRRCRTR 1 1 ZElanl04 PAX2 15 mer cyclic (internal) H2N-K(dns)TNAKHSSCNRRLRCR 1 1 ZElanl05 PAX2 Ala Scan 1 H2N-K(dns)ANAKHSSHNRRLRTR ZElanl06 PAX2 Ala Scan 2 H2N-K(dns)TAAKNSSHNRRLRTR ZElanl07 PAX2 Ala Scan 3 H2N-K(dns)TNGKNSSHNRRLRTR ZElanl08 PAX2 Ala Scan 4 H2N-K(dns)TNAAHSSHNRRLRTR ZElanl09 PAX2 Ala Scan 5 H2N-K(dns)TNAKASSHNRRLRTR ZElanllO PAX2 Ala Scan 6 H2N-K(dns)TNAKHASHNRRLRTR ZElanlll PAX2 Ala Scan 7 H2N-K(dns)TNAKHSAHNRRLRTR ZElanll2 PAX2 Ala Scan 8 H2N-K(dns)TNAKHSSANRRLRTR Printed from Mimosa t WO 98/51325 PCT/US98/10088 ZElanll3 PAX2 Ala Scan 9 H2N-K(dns)TNAKHSSHARRLRTR ZElanll4 PAX2 Ala Scan 10 H2N-K(dns)TNAKHSSHNARLRTR ZElanll5 PAX2 Ala Scan 11 H2N-K(dns)TNAKHSSHNRALRTR ZElanll6 PAX2 Ala Scan 12 H2N-K(dns)TNAKHSSHNRRARTR ZElanll7 PAX2 Ala Scan 13 H2N-K(dns)TNAKHSSHNRRLATR ZElanll8 PAX2 Ala Scan 14 H2N-K(dns)TNAKHSSHNRRLRAR ZElanll9 PAX2 Ala Scan 15 H2N-K(dns)TNAKHSSHNRRLRTA ZElanl23 PAX2 15 mer cyclic D form H2N-K(dns)Lys-TNAKHSSHNrrLrTr ZElanl24 PAX2 15 mer D form H2N-K(dns)TNAKHSSHNrrLrTr ZElanl25 PAX2 10 mer cyclic H2N-K(dns)Lys-SSHNRRLRTR 1 1 ZElanl26 PAX2 10 mer cyclic D form H2N-K(dns)Lys-SSHNrrLrTr 1 1 ZElanl27 PAX2 10 mer cyclic H2N-K(dns)Lys-TNAKHSSHNR 1 1 ZElanl28 PAX2 10 mer cyclic D form H2N-K(dns)Lys-TNAKHSSHNr 1 1 ZElanl29 PAX2 15 mer H2N-K(dns)TNAKHSSHNRRLRTR ZElanl30 HAX42 14 mer Ala Scan 1 H2N-K(dns)AGDYNCCGNGNSTG ZElanl31 HAX42 14 mer Ala Scan 2 H2N-K(dns)PADYNCCGNGNSTG ZElanl32 HAX42 14 mer Ala Scan 3 H2N-K(dns)PGAYNCCGNGNSTG ZElanl33 HAX42 14 mer Ala Scan 4 H2N-K(dns)PGDANCCGNGNSTG ZElanl34 HAX42 14 mer Ala Scan 5 H2N-K(dns)PGDYACCGNGNSTG ZElanl35 HAX42 14 mer Ala Scan 6 H2N-K(dns)PGDYNACGNGNSTG ZElanl36 HAX42 14 mer Ala Scan 7 H2N-K(dns)PGDYNCAGNGNSTG ZElanl37 HAX42 14 mer Ala Scan 8 H2N-K(dns)PGDYNCCANGNSTG ZElanl38 HAX4 2 14 mer Ala Scan 9 H2N-K(dns)PGDYNCCGAGNSTG ZElanl39 HAX42 14 mer Ala Scan 10 H2N-K(dns)PGDYNCCGNANSTG ZElanl40 HAX42 14 mer Ala Scan 11 H2N-K(dns)PGDYNCCGNGASTG ZElanl41 HAX42 14 mer Ala Scan 12 H2N-K(dns)PGDYNCCGNGNATG ZElanl42 HAX42 14 mer Ala Scan 13 H2N-K(dns)PGDYNCCGNGNSAG ZElanl43 HAX42 14 mer Ala Scan 14 H2N-K(dns)PGDYNCCGNGNSTA Printed from Mimosa WO 98/51325 PCT/US98/10088 GST fusion proteins of GIT peptides are shown in Table 21.

Printed from Mimosa Table 21 Source Clone # GST Fusion Sequence SEQ ID NO.

DCX11 98 gst-SQGSKQCMQYRTGRLTVGSEYGCGMNPARHATPAYPARLLPRYR 213 HAX42 99 gst-SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRRRPSAIPT 214 SNi 3 4 100 gst-SPCGGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSDY 215 5PAX5 97 gst-RGSTGTAGGERSGVLNLHTRDNASGSGFKPWYPSNRGHK 216 SNi 2 8 84 gst-SHSGGMNRAYGDVFRELRDRWNATSHHTRPTPQLPRGPN 217 SNi 2 8 85 gst-SHSGGMNRAY 218 SNi28 86 gst-GDVFRELRDR 219 SNi 2 8 87 gst-WNATSHHTRP 220 SNi 2 8 88 gst-TPQLPRGPN 221 SNi 2 8 89 gst-GDVFRELRDRWNATSHHTRP 222 SNi 2 8 90 gst-WNATSHHTRPTPQLPRGPN 223 SNi 2 8 91 gst-GDVFRELRDRWNATSHHTRPTPQLPRGPN 224 SNi 2 8 92 gst-SHSGGMNRAYGDVFRELRDRWNATSAATRPTPQLPRGPN 225 P31 93 gst-SARDSGPAEDGSRAVRLNGVENANTRKSSRSNPRGRRHP 226 P31 101 gst-SARDSGPAEDGSRAVRLNG 227 P31 102 gst-DGSRAVRLNGVENANTRKSSR 228 P31 103 gst-ENANTRKSSRSNPRGRRHP 229 P31 110 gst-ENANTRKSSR 230 P31 111 gst-RKSSRSNPRG P31 112 gst-SNPRGRRHP 232 P31 119 gst-TRKSSRSNPRG 233 PAX2 94 gst-STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN 234 PAX2 104 gst-STPPSREAYSRPYSVDSDSD 235 PAX2 105 gst-SRPYSVDSDSDTNAKHSSHNR 236 PAX2 106 gst-TNAKHSSHNRRLRTRSRPN 237 PAX2 113 gst-TNAKHSSHN 238 PAX 2 114 gst-SSHNRRLRTR 239 PAX2 115 gst-RRLRTRSRPN 240 SNilO 96 gst-RVGQCTDSDVRRPWARSCAHQGCGAGTRNSHGCITRPLRQASAH 241 SNilO 116 gSt-RVGQCTDSDVRRPWARSCA 242 SNilO 117 gst-VRRPWARSCAHQGCGAGTRNS 243 SNilO 118 gst-GTRNSHGCITRPLRQASAH 244 DCX8 95 gst-RYKHDIGCDAGVDKKSSSVRGGCGAHSSPPRAGRGPRGTMVSRL 245 DCX8 107 gst-RYKHDIGCDAGVDKKSSSVRGGCG 246 DCX8 108 gst-GCDAGVDKKSSSVRGGCGAHSSPPRA 247 DCX8 109 gst-GAHSSPPRAGRGPRGTMVSRL 248 u ts> t/l *0 g c in 00 o o 00 00 6.10.6. Peptide Stability The relative stability for ZElan031, ZElan053 and ZElan054 was determined in simulated intestinal fluid (SIF) SIF was made by dissolving lOOmg of pancreatm (Sigma cat#P-5 1625, lot# 122H0812)in 8.4ml of phosphate stock solution, adjusting the pH to 7.5 with 0.2N NaOH and adjusting the volume to 10ml with water.

Peptide (3.25mg) was dissolved in 3.25 ml of 10,000 fold diluted SIF solution at 37°C. Aliquots (0.7ml) of the 10 digestion solution were then withdrawn at clmin, lh, 3h, and 21h or 24h. The samples were quickly passed through a syringe filter (Millipore Millex-GV 0.22/xm, part# SLGV025LS, lot# H2BM95250) and 300/xL of the filtered solution was immediately injected onto a Hewlett-Packard HPLC system equipped with a 15 C-8 column (Applied Biosystems column and guard column: column- p/n 0711-0023 Spheri-5 ODS 5^m, 220x4.6mm). The products were eluted at 1.5ml/min using an acetonitrile-water gradient. The major fluorescent peaks were collected, lyopholized and identified by MS analysis.

The HPLC gradient used was: Time (min) Solvent Mixture 0 95% H20-5% acetonitrile (0.1%TFA) 95% H20-5%acetonitrile (0.1%TFA) 85% H20-15% acetonitrile (0.1%TFA) linear solvent change 40 0% H20-100% acetonitrile (0.1%TFA) " 45 95% H20-5% acetonitrile (0.1%TFA) 52 95% H20-5%acetonitrile (0.1%TFA) w As shown in Table 22, the relative stability (to SIF) for the three peptides was found to be ZElan053>ZElan054>ZElan031. Enzymatic cleavage of the peptide was found to occur at arginine and/or lysine as expected. The replacement of 1-amino acids with their D-amino acid analogs significantly reduced the rate of proteolysis at these residues.

Printed from Mimosa TABLE 22 Peptide Percent Remaining at; Rel.

Stab. 1 m 1 h 3 h 24 h ZElan031 100 38.7 0 0 3 ZElan054 97.4 58.2 11.6 2.7 2 ZElan053 100 98 .3 98.1 94 . 0 1 7. CHARACTERIZATION OF PEPTIDE-COATED PARTICLES Binding of Peptide-Coated PLGA Nanoparticles to Fixed Caco-2 Cells Binding of nanoparticles coated with targeting peptides to fixed Caco-2 cells was investigated using an 15 ELISA assay based on reaction of antibody with the dansyl moiety present on the peptides. Isoelectric points of selected synthetic peptides are shown in Table 23 (corresponding SEQ ID NOS. are shown in Table 7). Corresponding dansylated synthetic GIT binding peptides are given m Table 24.

TABLE 23 Peptide Sequence Pi P31 SARDSGPAEDGSRAVRLNGVENANTRKSSRSNPRGRRHP 12 .26 5PAX5 RGSTGTAGGERSGVLNLHTRDNASGSGFKPWYPSNRGHK 11 .49 SNilO RVGQCTDSDVRRPWARSCAHQGCGAGTRNSHGCITRPLRQASAH .45 SNi 3 4 SPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSDY 8.

DCX11 SQGSKQCMQYRTGRLTVGSEYGCGMNPARHATPAYPARLLPRYR .44 DCX8 RYKHDIGCDAGVDKKSSSVRGGCGAHSSPPRAGRGPRGTMVSRL 11 .03 HAX42 SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRRRPSAIPT 9. 62 PAX2 STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN 11 .26 Printed from Mimosa WO 98/51325 PCT/US98/10088 TABLE 24 Peptide Sequence p31 h2N-k(dns)sardsgpaedgsravrlngvenantrkssrsnprgrrhpgg-conh2 PAX5 H2N-K(dns)RGSTGTAGGERSGVLNLHTRDNASGSGFKPWYPSNRGHK-CONHj SNilO H2N-K(dns)RVGQCTDSDVRRPWARSCAHQGCGAGTRNSHGCITRPLRQASAH-CONHj SNi34 H2N-K(dns)SPCGGSWGRFMQGGLFGGRTDGCGAHRNRTSASLEPPSSDY-CONHj DCX11 H2N-K(dns)SQGSKQCMQYRTGRLTVGSEYGCGMNPARHATPAYPARLLPRYR-CONHj DCX8 H2N-K(dns)RYKHDIGCDAGVDKKSSSVRGGCGAHSSPPRAGRGPRGTMVSRL-CONH2 HAX42 H2N-K (dns) SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRRRPSAIPT-CONH2 PAX2 H2N-K(dns)STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPNG-CONH2 DAB10 H2N-K(dns)SKSGEGGDSSRGETGWARVRSHAMTAGRFRWYNQLPSDR-CONHj Method: Confluent Caco-2 monolayers grown in 96-well plates (p38) were fixed and treated with 0.1% phenylhydrazine before blocking with 0.1% BSA in PBS. Control and dansyl peptide-15 coated nanoparticles were resuspended in sterile water at lOmg/ml and stirred with a magnet for lh at room temperature. Samples consisted of: (1) blank nanoparticle control, (2) scrambled PAX2-coated nanoparticles, (3) PAX2-coated nanoparticles, (4) HAX42-coated nanoparticles, 20 (5) PAX2/HAX42-coated nanoparticles, and (6) 8 peptide-coated nanoparticles.

Nanoparticles were added to the cells at lOmg/ml in 100/il 1%BSA-PBS (no Tween80 is used in this assay) and 2-fold serially-diluted. The 96-well plates were incubated for lh 2 5 at room temperature. The plates were washed 5 times with 1%BSA-PBS and 100/xl of anti-dansyl antibody (Cytogen DB3-226.3; 0.5 pig/ml; batch May 1997) was added per well and the plates incubated lh at room temperature. The wells were washed 5 times with 1%BSA-PBS; 100^1 of goat anti-mouse A:HRP o 0 antibody (Southern Biotechnology CN. 1060-05; 1:10,000) was added per well, and the plates incubated lh at room temperature. After washing 5 times with 1%BSA-PBS, 100^1 of TMB peroxidase substrate (KPL CN. 50-76-00) was added to the wells and the optical density at 650nm was measured after 15 ^ minutes.

Printed from Mimosa As shown in Figures 13A-B, a decreasing anti-dansyl ELISA response was observed for nanoparticles coated with PAX2, HAX2, PAX2+HAX2, and a mixture of 8 targeting peptides, when decreasing amounts of the nanoparticles were applied to 5 fixed Caco-2 cells. No concentration effect was observed for blank nanoparticles or nanoparticles coated with a scrambled version of PAX2 peptide. Nanoparticles coated with PAX2, HAX2, PAX2+HAX2, and the 8 peptide mix, showed increased response relative to blank nanoparticles or nanoparticles 10 coated with a scrambled version of PAX2 peptide. The OD values were low relative to those normally observed for GST-peptide fusion binding to fixed Caco-2 cells.

Table 25 below shows the insulin potency and level 15 of peptides coated onto the particles (measured by fluorescense) for formulation 1 particles (formulation by the coacervation method given below).

Table 25 Peptide Blend Insulin Peptide mg/g fi 1/mg PAX2 60.7 3. .51 HAX42 55.9 2. , 93 PAX2 SCRAMBLED 57 . 7 1. .26 P31 67 . 0 1, .22 5PAX5 52.7 2 , . 83 SNilO 59.5 1. .75 SNi34 61.5 4 , . 03 DCX8 59.1 1. .87 DAB10 55 . 9 1 .99 ELISA of dansylated peptides and insulin coated PLGA particles The standard ELISA procedure was modified as 35 follows. Peptides and particles were diluted to an appropriate concentration in PBS containing 1%BSA (particles were sonicated to achieve a homogeneous solution), titered Printed from Mimosa and incubated one hour at room temperature. Following five washes with PBS containing 1%BSA, an in-house IgGlX anti-dansyl monoclonal antibody was added (diluted to l^g/ml in 1%BSA-PBS) and the plates were incubated for one hour. After 5 five more washes goat anti-mouse X-HRP was added (Southern Biotechnology Associates Inc., Birmingham, AL, diluted 1:10,000 in 1%BSA-PBS) and the plates were incubated one hour. After five washes, plates were developed with TMB peroxidase substrate (Kirkegard and Perry, Gaithersburg, MD).

All data is presented with background binding subtracted. Tween 2 0 was not added to the diluent or the washes when insulin coated PLGA particles were included in the assay.

Figures 14A-14B show the binding of the dansylated peptide SNilO to hSI and BSA. 8. BINDING OF SYNTHETIC PEPTIDES AND PEPTIDE-COATED PARTICLES TO S100 AND P100 FRACTIONS DERIVED FROM CACO-2 CELLS 8.1. Detection of Binding to Membrane (P100) and CvtQBollc (S100) fractions Caco-2 cell membrane (P100) and cytosolic (S100) fractions were prepared using a modification of the method -c. described in Kinsella, B. T., O'Mahony, D. J. and G. A. 2 5 FitzGerald, 1994, J. Biol. Chem. 261(47): 29914-29919. Confluent Caco-2 cell monolayers (grown in 75 cm2 flasks for up to 1 week at 37°C and 5% C02) were washed twice in Dulbecco's PBS (DPBS) and the cells were harvested by centrifugation at 1000 rpm after treatment with 10 mM EDTA-DPBS. The cells were washed 3 times in DPBS and the final cell pellet was resuspended in 3 volumes of ice cold HED buffer (20 mM HEPES (pH 7.67), 1 mM EGTA, 0.5 mM dithiothreitol, 1 mM phenylmethylsulphonyl fluoride (PMSF)). The cells were allowed to swell for 5 min on ice prior to homogenization for 30 sec. The homogenates were centrifuged at 40,000 rpm for 45 min at 4°C. The supernatant (S100) was Printed from Mimosa removed and the pellet (P100) was resuspended in HEDG buffer (20 mM HEPES (pH 7.67), 1 mM EGTA, 0.5 mM dithiothreitol, 100 mM NaCl, 10% glycerol, 1 mM PMSF). Protein concentrations were determined using the Bradford assay (Bradford, M. M., 5 1976, Anal. Biochem. 72: 248-254).

Binding of peptide and/or peptide-coated PLGA particles to membrane (P100) and cytosolic (S100) fractions was assessed by detection of the dansyl moiety incorporated in the peptide. Costar ninety six well ELISA plates were 10 coated with S100 and P100 fractions (100 ixg/ml in 0.05 M NaHC03) overnight at 4°C. The plates were blocked with 0.5% bovine serum albumin in DPBS for 1 h at room temperature and washed 3 times in 1% BSA-DPBS. Peptide-coated particles or peptides were dispersed in the same buffer and added to the 15 plates at concentrations in the range 0.0325 - 0.5 mg/well. After 1 h at room temperature the plates were washed 5 times m 1% BSA-DPBS and 100 /xl of anti-dansyl antibody (Cytogen DB3-226.3; 0.5 /ig/ml) was added per well. The plates were incubated for 1 h at room temperature. The wells were washed 20 3 times in 1% BSA-DPBS and 100 fil of goat anti-mouse IgGA:HRP antibody (Southern Biotechnology 1060-05; 1:10,000) was added per well. The plates were incubated for 1 h at room temperature. After washing 3 times in 1% BSA-DPBS 100 fil of TMB substrate (3,3',5',5-tetramethylbenzidine; Microwell 25 Peroxidase Substrate System (Kirkegaard and Perry Laboratories 50-76-00)) was added and the optical density was measured at 650 nm at various time intervals. 8.2. Binding of Peptide-Coated PLGA particles A novel assay system is provided by the instant invention for detection of binding of peptide-coated PLGA particles to membrane (P100) and cytosolic (S100) fractions derived from live Caco-2 cells. The absorbance readings obtained using this assay system were substantially higher 35 than those obtained using similar peptide-coated PLGA particle concentrations on fixed Caco-2 cells. This greater sensitivity together with the derivation of the S100 and P100 Printed from Mimosa fractions from live Caco-2 cells suggests that this assay may be the assay system of choice for detection of peptide-coated PLGA particle binding. The assay was concentration dependent and peptide/particle correlation permitted differentiation 5 between specific and non-specific binding interactions.

Binding of peptide-coated PLGA particles was assessed using S100 and P100 fractions derived from live Caco-2 cells as described above. The fractions were coated onto 96-well plates at 10^g/well in 0.05 M NaHC03 and peptide-coated PLGA 10 particles were assayed by ELISA at concentrations in the range 0.0325 - 0.5 mg/well.

Figures 15A and 15B illustrate the data obtained on S100 and P100 fractions respectively for particles coated with no peptide, scrambled PAX2 (control), P31 D-Arg 16-mer 15 (ZElan053), HAX42, PAX2 and HAX42/PAX2. Using particle concentrations of 0.0325 - 0.5 mg/well all test peptide-coated PLGA particles exhibited greater binding to both the S100 and P100 fractions than the scrambled PAX2 coated control particles. All particles except P31 D-Arg 16-mer 20 (ZElan053) exhibited greater binding to the P100 fraction than the S100 fraction. Greater binding of the P31 D-Arg 16-mer (ZElan053) coated particles to the S100 fraction may be indicative of non-specific binding due to the D-Arg modification of the P31 peptide (SEQ ID NO:43). 25 Binding of PLGA particles coated with varying concentrations of PAX2 peptide ranging from 0.05 - 5.0 mg/g was assessed using a) fixed Caco-2 cells (P35) and b) S100 and P100 fractions (Caco-2 P33). The particles were assayed at concentrations in the range 0.03125 - 0.0625 mg/well. ' 30 Using a particle concentration of 0.0625 mg/well, all PAX2 coated particles except those coated at 0.05 mg/g exhibited greater binding to fixed Caco-2 cells than the scrambled PAX2 coated control particles. There appeared to be a concentration effect with increasing PAX2 peptide 35 concentration resulting in improved Caco-2 cell binding (in the range 0.05 - 1.0 mg/g). However all absorbance readings Printed from Mimosa were low and binding of the PAX2 (5 mg/g) was not consistent with this pattern.

Using particle concentrations of 0.03125 - 0.0625 mg/well all test peptide coated particles except PAX2 (0.05 5 mg/g) exhibited comparable or greater binding to both the S100 and P100 fractions than the scrambled PAX2 coated control particles. All particles exhibited greater binding to the P100 fraction than the S100 fraction. Binding to both the S100 and P100 fractions was directly proportional to the 10 concentration of the PAX2 peptide on the particle. The absorbance readings obtained using this assay system were substantially higher than those obtained on the fixed Caco-2 cells.

The effect of blocking solution on binding of peptide-15 coated PLGA particles to P100 fractions (Caco-2 P35) was assessed using 1% bovine serum albumin (BSA) and 1% milk powder blocking solutions to assess background binding. The following particles were assayed at concentrations in the range 0.03125 - 0.0625 mg/well: no peptide; scrambled PAX2; 20 and a range of PAX2 coated particles having peptide concentrations from 5-0.05 mg/g. As previously observed using 1% BSA, all test peptide coated particles except PAX2 coated at 0.05 mg/g exhibited comparable or greater binding to the P100 fractions than the scrambled PAX2 coated control 25 particles. Binding to P100 fractions was directly proportional to the concentration of the PAX2 peptide on the particle (although in this instance PAX2 (5 mg/g) exhibited slightly lower binding than PAX2 (1 mg/g)). A similar trend was observed using 1% milk powder and a particle 30 concentration of 0.0625 mg/well. However all absorbance readings were low when 1% milk powder was used and the binding pattern was not detectable using particles at a concentration of 0.0625 mg/well.

Non-specific binding of peptide-coated PLGA particles to 35 plastic was also assessed using 1% BSA and 1% milk powder blocking solutions. The binding pattern observed above could be detected when BSA was used; however, absorbance readings - Ill - Printed from Mimosa WO 98/51325 PCT/US98/10088 were substantially lower and binding of particles PAX2 (0.1 and 0.05 mg/g respectively) was not detectable. When 1% milk powder was used, all absorbance readings were low and no binding pattern was detectable. BSA was chosen for blocking 5 in subsequent assays. 8.3. Comparison of Peptide-Coated Particle and Synthetic Peptide Binding to P100 fractions Binding of dansylated peptides to P100 fractions was assessed to determine if peptide binding was predictive of peptide-coated particle binding. Figure 16 illustrates the data obtained for the dansylated peptides A) HAX42, P31 D-form and scrambled PAX2 and B) PAX2, HAX42 and scrambled PAX 2.

Two consecutive assays produced substantial variations in absorbance readings. Initially, the HAX42 peptide exhibited strong binding when compared to the scrambled PAX2 control. The P31 D-form peptide (ZElan053) exhibited binding at the highest dilution only. ,In the repeat assay, HAX42 2Q also exhibited significant binding compared to the scrambled PAX2 control. However, the scrambled PAX2 control and HAX42 produced relatively high absorbance values compared to those obtained in the previous assay. The PAX2 peptide was indistinguishable from the scrambled PAX2 control.

Peptide/particle binding correlation is summarized as follows in Table 26: TABLE 26 Peptide/particle assay correlation Peptide Assay correlation HAX42 + PAX2 +/- P31 D-form - Scrambled +/- PAX2 negative Peptide/particle binding correlated well for the HAX42 peptide. In contrast, no correlation could be detected - 112 Printed from Mimosa WO 98/51325 PCT/US98/10088 for the P31 D-form (ZElan053) peptide. Since the P31 D-form peptide-coated particles exhibited greater binding to the S100 fraction than the P100 fraction (unlike the other test peptides) it appears that the particle binding interaction 5 was non-specific or that some other molecule was competing for binding to the P100 fraction but not to the S100 fraction. Thus the peptide/particle assay correlation may be useful for distinguishing between specific and non-specific binding interactions. The scrambled PAX2 control produced 10 variable results so that it was difficult to assess the PAX2 binding correlation. 8.4. Determination of HAX42 and* PAX2 Binding Motif Sequences Peptides and GST fusion proteins of HAX42, PAX2 and various derivatives were assayed using peptide ELISA to PI00 membrane fractions derived from Caco-2 cells. The GST-PAX2 protein and PAX2 peptide data indicate that a core binding motif lies in the amino acid sequence TNAKHSSHNRRLRTR (SEQ ID 20 NO: ) otherwise named GST-106 and ZElan033. Similarly, the HAX42 peptide data suggest that a core binding motif for HAX42 lies in the amino acid sequence PGDYNCCGNCNSTG (SEQ ID NO: ), otherwise named ZElan091.

The peptides and proteins were analyzed by a 25 dansylated peptide ELISA method in which 96 well plates were coated overnight at 4°C with 100/il/well coating protein (normally 100^g/ml P100 membrane fraction) in 0.05M carbonate buffer pH9.6. Nonspecific binding was blocked using 200/il/well, 2% Marvel/PBS for 2 hours at 37°C prior to 30 incubation with dansylated peptides. The plates were washed three times with PBS/0.05% Tween 20 and after each subsequent incubation step. The peptides were diluted in blocking solution at a starting concentration of 100/xg/ml and diluted 1:2 downwards, 100/zl/well, followed by incubation at room 35 temperature for 1 hour, exactly. A buffer blank control was included to ensure that background binding to plastic was not due to the antibodies used in the assay system. To detect the Printed from Mimosa dansylated peptides, a mouse anti-dansyl antibody (DB3, Cytogen Corp.) at 1:1340 dilution in blocking buffer and lOOjul/well was added followed by incubation at room temperature for 1 hour. The plates were then incubated with 5 an anti-mouse X-HRP conjugated antibody (Southern Biotech 1060-05) at a 1:10,000 dilution m blocking solution, 100/xl/well for 1 hour at room temperature. Plates were developed using 75^1/well Bionostics TMB substrate and incubated for approximately 10 minutes. The developing 10 reaction was stopped using Bionostics Red Stop solution (25/zl/well) , and the optical density of the plates was read at 650nm.

GST-PAX2 Peptides - Relative Binding to PlOO Fractions 15 After subtraction of the GST-peptide binding to plastic from PlOO binding values, the binding of GST-PAX2 peptides were represented as a ratio of GST-HAX42 binding to PlOO, which was given the arbitrary value of 1.00. The following ratios were determined from binding to PlOO of GST-peptides 20 at a peptide concentration of 20/xg/ml. Bold denotes positive binding to the PlOO membrane fraction.

Table 27 GST-peptide Value __ GST-HAX42 1.00 GST-PAX2 1.79 GST-104 0.01 GST-105 -0.08 GST-106 2.71 GST-113 0.26 GST-114 0.17 GST-115 0.36 GST 0.48 Printed from Mimosa WO 98/51325 PCT/US98/10088 Table 2 8 GST-peptide Amino Acid Sequence GST-PAX2 STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPN GST-104 STPPSREAYSRPYSVDSDSD GST-105 STPPSREAYSRPYSVDSDSDTNAKHSSHN GST-106 TNAKHSSHNRRLRTRSRPN GST-113 TNAKHSSHN GST-114 SSHNRRLRTRSRPN GST-115 RRLRTRSRPN PAX2 Peptides - Relative Binding to PlOO Fractions ZElan021, full length HAX42, was given the arbitrary-value of 1.00 for binding to PlOO at a given peptide concentration determined from the signal-to-noise ratio data. PAX2 and its derivatives are given as a ratio of HAX42 value .c to reflect their binding abilities to PlOO membrane fractions .L3 derived from a Caco-2 cell line as shown in Table 29. Table 30 provides a line-up of the PAX2 peptides showing the positive binding peptides in boldface. The GST-PAX2 peptide and PAX2 peptide data agree, demonstrating that a binding 2Q motif is in the amino acid sequence TNAKHSSHNRRLRTR (GST-106 and ZElan033).

Printed from Mimosa PCT /US98/10088 TABLE 29 Binding value Binding Binding Binding at value Binding value value 50/tg/ml PAX2 at value at at (Jackson peptide /xg/ml at 20^g/ml 50 fig/ml 50/jg/ml Ab) ZElan018 -0.33 1.07 0.95 1. 01 ZElan032 1.43 2.87 0.95 1. 06 ZElan033 0.35 1.57 0.80 0. 66 ZElan035 0.12 0.43 0.81 0. 77 ZElan055 0.99 0.73 1.10 0. 59 ZElan056 0 . 00 0.16 0.21 0. 21 ZElan057 0.08 0.56 0 ZElan058 0.05 0.47 0. 16 ZElan073 0.07 -0.11 0 49 0.66 ZElan074 0.06 0.82 0. 52 0.71 ZElan075 0.13 0.52 0. 38 0.47 ZElan076 0.08 1.00 0. 41 0.60 ZElan077 0.20 0.76 0. 54 0.73 ZElan078 0.11 0.87 0. 69 0.68 ZElan079 0.31 0.97 0. 68 0.83 ZElanOSO 0.23 0.84 0. 45 0.67 ZElan081 0.01 0.89 0 47 ZElan082 0.00 0.92 0. 40 ZElan083 0.43 0.63 1.03 0. 88 ZElan084 1.06 0.93 1.16 0. 77 Binding value at 50^g/ml (Southern Ab) 0.49 0.48 0.32 42 52 47 53 38 Printed from Mimosa Table 30 PAX2 Peptide Amino acid sequence ZElan018 H2N-K(dns)STPPSREAYSRPYSVDSDSDTNAKHSSHNRRLRTRSRPNG -CONH, ZElan032 K,N-K(dns)TNAKHS SHNRRLRTRSRPN-CONH2 ZElan033 H2N-K(dns)TNAKHS SHNRRLRTR-CONH, ZElan034 HjN-K(dns)SSHNRRLRTRSRPN-CONH2 ZElan035 H2N-K (dns) SSHNRRLRTR-CONH2 ZElan055 HjN-K(dns)TNAKHSSHN-CONHj ZElan056 H2N-K(dns)RRLRTRSRPN-CONH2 ZElan057 H2N-K(dns)RRLRTRSR-CONH, ZElan058 H2N-K(dns)RRLRTR-CONH2 ZElan059 H2N-K(dns)rrLrTrSrPN-CONH2 ZElan073 H2N-K(dns)ASHNRRLRTR-CONH2 ZElan074 H2N-K(dns)SAHNRRLRTR-CONHj ZElan075 H2N-K(dns)S SANRRLRTR-CONH2 ZElan076 H2N-K(dns)S SHARRLRTR-CONH2 ZE1an 0 7 7 H2N-K(dns)S SHNARLRTR-CONH2 ZElan07B H2N-K(dns)S SHNRALRTR-CONH2 ZElan079 H2N-K(dns)S SHNRRARTR-CONH2 ZElan080 H2N-K(dns)S SHNRRLATR-CONH2 ZElan081 H2N-K(dns)SSHNRRLRAR-CONH2 ZElan082 H2N-K(dns)SSHNRRLRTA-CONH2 SCRAMBLED PAX2 PEPTIDES: ZElan083 H2N-K(dns)GRNHDWSSNTHKSYRSPRSASYPRLSNDRTDRTEPAPSS-CONHj ZElanO 8 4 H2N-K(dns)RNTRNKTSRLSANPHRSHR-CONH, SEQ ID NO: HAX42 Peptides - Relative Binding to PlOO Fractions ZElan021, full length HAX42, was given the arbitrary-value of 1.00 for binding to PlOO at a given peptide concentration determined from the signal-to-noise ratio data. HAX42 and its derivatives are given as a ratio of HAX42 value to reflect their binding abilities to PlOO membrane fractions derived from a Caco-2 cell line as shown in Table 31. Table 32 provides a line-up of the HAX42 peptides showing the positive binding peptides in boldface. A core binding motif appears to lie in the amino acid sequence PGDYNCCGNCNSTG (ZElan091).

Printed from Mimosa TABLE 31 HAX42 Binding Binding Binding Binding Binding peptide value at value at value at value at value at /*g/ml 50fig/ml 50 fig/ml /ig/ml 25pg/ml ZElan021 1.00 1.00 1. ,00 1.00 1.00 ZElanOeO 0.44 0.56 0, ,43 ZElanOSl 0.20 0.60 0 38 ZElan062 0.11 0.42 0, .34 ZElan065 0.00 0.54 0, .30 ZElan067 0.08 0.52 0 40 ZElan070 0.59 0.97 0 39 ZElan071 1.22 0.89 0 .75 ZElan072 0.83 0.61 0 .88 ZElan087 0.46 0.44 ZElan088 2.21 1.41 ZElan089 0.55 0.44 ZElan090 2.06 1.54 ZElan091 2.02 1.37 ZElan092 1.41 1.90 ZElan093 1.88 1.37 Binding value at 25/tg/ml 1.00 1.63 0.49 2.16 1.20 0.91 1.33 HAX42 Peptide ZElan021 ZElan060 ZElan06l ZE1anO 6 2 ZElan065 ZElan067 ZElan07 0 ZElan071 ZElan072 ZElan087 ZElan0B8 ZElan089 ZElan090 ZElan091 ZElan092 ZElan093 Table 32 Amino acid sequence HjN-K(dns)SDHALGTNLRSDNAKEPGDYNCCGNGNSTGRKVFNRRRPSAIPT-CONH2 HjN-K(dns)SDHALGTNLRSDNAKEPGDYNCCGNG-CONH, HjN-K(dns) GNGNSTGRKVFNRRRPSAIPT-CONH2 h2n-k(dns)sdhalgtnlrsdnakepg-conh2 H2N-K(dns)RKVFNRRRPS-CONH, H,N-K(dns)NRRRPS-CONH, HjN-K(dns)SDHALGTNLRSDNAKEPGDYNCCGNGNST-CONH2 H2N-K(dns)NLRSDNAKEPGDYNCCGNGNSTGRKVFNR-CONH, H,N-K(dns)PGDYNCCQNGNSTGRKVFNRRRPSAIPT-CONH, H,N- K (dns)SDHALGTNLRSDNAKEPGDY-CONH, HjN-K(dns)SDNAKEPGDYNCCGNGNSTG-CONH, HjN-K (dns) SDHALGTNLRSDNAK-CONH2 - CONH2 HjN-K (dns) EPGDYNCCGNGNSTG H2N-K (dns) PGDYHCCGNGNSTG-CONH, H,N-K(dns)PGDYNCCGNG-CONH, H,N-K(dns)NCCGNGNSTG-CONH, 9. FORMULATIONS General Method for Preparation of Coacervated Particles.

Solid particles containing a Therapeutic as defined herein are prepared using a coacervation method. The are particles are formed from a polymer and have a particle size of between about lOnm and 500 /*m, most preferably 50 to 800 nm. In addition the particles contain targeting ligands which are incorporated into the particles using a number of methods.

The organic phase (B) polymer of the general method given above may be soluble, permeable, impermeable, Printed from Mimosa biodegradable or gastroretentive. The polymer may consist of a mixture of polymer or copolymers and may be a natural or synthetic polymer. Representative biodegradable polymers include without limitation polyglycolides; polylactides; poly(lactide-co-glycolides), including DL, L and D forms; copolyoxalates; polycaprolactone; polyesteramides; polyorthoesters; polyanhydrides; polyalkylcyanoacrylates; polyhydroxybutyrates; polyurethanes; albumin; casein; citosan derivatives; gelatin; acacia; celluloses; polysaccharides; 10 alginic acid; polypeptides; and the like, copolymers thereof, mixtures thereof and stereoisomers thereof. Representative synthetic polymers include alkyl celluloses; hydroxalkyl celluloses; cellulose ethers; cellulose esters; nitrocelluloses; polymers of acrylic and methacrylic acids 15 and esters thereof; dextrans; polyamides; polycarbonates; polyalkylenes; polyalkylene glycols; polyalkylene oxides; polyalkylene terephthalates; polyvinyl alcohols; polyvinyl ethers; polyvinyl esters; polyvinyl halides; poyvinylpyrrolidone; polysiloxanes and polyurethanes and co-20 polymers thereof.

Typically, particles are formed using the following general method: An aqueous solution (A) of a polymer, surface active agent, surface stabilising or modifying agent or salt, 25 or surfactant preferably a polyvinyl alcohol (PVA) or derivative with a % hydrolysis 50 - 100% and a molecular weight range 500 - 500,000, most preferably 80-100% hydrolysis and 10,000-150,000 molecular weight, is introduced into a vessel. The mixture (A) is stirred under low shear 30 conditions at 10- 2000 rpm, preferably 100-600 rpm. The pH and/or ionic strength of this solution may be modified using salts, buffers or other modifying agents. The viscosity of this solution may be modified using polymers, salts, or other viscosity enhancing or modifying agents. 35 A polymer, preferably poly(lacide-co-glycolide), polylactide, polyglycolide or a combination thereof or in any enantiomeric form or a covalent conjugate of the these - -119 - Printed from Mimosa polymers with a targeting ligand is dissolved in water miscible organic solvents to form organic phase (B). Most preferably, a combination of acetone and ethanol is used in a range of ratios from 0:100 acetone: ethanol to 100: 0 5 acetone: ethanol depending upon the polymer used.

Additional polymer(s), peptide(s) sugars, salts, natural/biological polymers or other agents may also be added to the organic phase (B) to modify the physical and chemical properties of the resultant particle product. 10 A drug or bioactive substance may be introduced into either the aqueous phase (A) or the organic phase (B). A targeting ligand may also be introduced into either the aqueous phase (A) or the organic phase (B) at this point.

The organic phase (B) is added into the stirred 15 aqueous phase (A) at a continuous rate. The solvent is evaporated, preferably by a rise in temperature over ambient and/or the use of a vacuum pump. The particles are now present as a suspension (C). A targeting ligand may be introduced into the stirred suspension at this point. 20 A secondary layer of polymer(s), peptide(s) sugars, salts, natural/biological polymers or other agents may be deposited on to the pre-formed particulate core by any suitable method at this stage.

The particles (D) are then separated from the 25 suspension (C) using standard colloidal separation techniques, preferably by centrifugation at high 1 g' force, filtration, gel permeation chromatography, affinity chromatography or charge separation techniques. The supernatant is discarded and the particles (D) re-suspended 30 in a washing solution (E) preferably water, salt solution, buffer or organic solvent(s). The particles (D) are separated from the washing liquid in a similar manner as previously described and re-washed, commonly twice. A targeting ligand may be dissolved in washing solution (E) at the final washing 35 stage and may be used to wash the particles (D).

Printed from Mimosa The particles may then be dried. Particles may then be further processed for example, tabletted, encapsulated or spray dried.

The release profile of the particles formed above 5 may be varied from immediate to controlled or delayed release dependent upon the formulation used and/or desired.

Drug loading may be in the range 0-90% w/w. Targeting ligand loading may be in the range 0-90% w/w.

Specific examples include the following examples: EXAMPLE 1: Peptide added at the final washing stage Products Bovine Insulin loaded nanoparticles Aim: To prepare a 2g batch of insulin loaded nanoparticles at a theoretical loading of 50mg/g and with the 15 peptide ZElan018 added.

Formulation Details RG504H (Lot no. 250583) 2.Og Acetone 45ml Ethanol: 5ml PVA (aq. 5%w/v) 400ml Bovine Insulin (Lot no. 86H0674) lOOmg Peptide: PAX2 (ZElan018) 10mg/50ml dH20 Experimental details: The 5% w/v PVA solution was prepared by heating water until near boiling point, adding PVA and stirring until cool. The organic phase was prepared by adding acetone, 4 5ml, and ethanol, 5ml, together. The polymer solution was prepared by adding RG504H, 2g, to the organic phase and 30 stirring until dissolved. The IKA™ reactor vessel was set up, all seals greased and the temperature was set at 25°C. The PVA solution, 400ml, was added into the reactor vessel and stirred at 400 rpm.

Bovine insulin, lOOmg, was added into the stirring PVA 35 solution. Using clean tubing and a green needle, the polymer solution was slowly dripped in the stirring PVA solution with the peristaltic pump set at 40. The solvent was allowed to Printed from Mimosa evaporate by opening the ports and allowing the dispersion to stir overnight at 400 rpm.

The suspension was centrifuged in a Beckman Ultracentrifuge™ with swing-out rotor at 12,500 rpm, 4°C. The 5 supernatant was decanted and discarded. The "cake" of particles was broken up and dH20 (200mls) was added to wash the particles. The centrifugation and washing steps were repeated twice.

The peptide solution, (ZElan018, lOmg in 50ml dH20) was prepared and added to the particles for a final washing stage. The suspended particles were centrifuged as before. The supernatant liquid was decanted, the 'cake' broken up, and the particles were dried in the vacuum oven.

The particles were ground, placed in a securitainer and sent for analysis. The weight of particles recovered was 1.45g. A SEM showed discrete, reasonably spherical particles in the 300-500nm size range. The potency was 49.2mg/g (98.0% of label claim). Peptide loading was 2.42 fig/mg (48.4% of label claim).

EXAMPLE 2: Peptide added at the beginning of manufacture Product: Bovine Insulin loaded nanoparticles Aim: To prepare a 2g batch of insulin loaded nanoparticles at a theoretical loading of 50mg/g and with the V peptide ZElan018 added at the beginning of manufacture.

Formulation Details RG504H (Lot no. 250583) 2. Og Acetone 45ml Ethanol: 5ml PVA(aq. 5%w/v) 400ml Bovine Insulin (Lot no. 65H0640) lOOmg Peptide: PAX2 (ZElan018ii) lOmg Experimental details: The 5% w/v PVA solution was prepared by heating water until near boiling point, adding PVA and stirring until cool. The organic phase was prepared by adding acetone, Printed from Mimosa 45ml, and ethanol, 5ml, together. The polymer solution was prepared by adding RG504H (polyactide-co-glycolide, Boehringer Ingelheim), 2g, to the organic phase prepared in step above and stirring until dissolved. The IKA™ reactor 5 vessel was set up, all seals greased and the temperature was set at 25°C. The PVA solution, 400ml, was added into the reactor vessel and stirred at 400 rpm.

Bovine insulin, lOOmg, was added into the stirring PVA solution. PAX2 (ZElan018ii, lOmg) was added to the 10 stirring PVA solution. Using clean tubing and a green needle, the polymer solution was slowly dripped into the stirring PVA solution with the peristaltic pump set at 40. The solvent was allowed to evaporate by opening the ports and allowing the dispersion to stir overnight at 400 rpm. The 15 suspension was centrifuged in a Beckman Ultracentrifuge™ with swing-out rotor at 12,500 rpm, 4°C. The supernatant was decanted and discarded.

The "cake" of particles was broken up and dH20 (200ml) was added to wash the particles. The centrifugation 20 and washing steps were repeated twice. The 'cake' was broken up and the particles were dried m the vacuum oven.

The particles were ground, placed in a securitainer and sent for analysis. The weight of the particles recovered was 1.6g. The potency was 47.3mg/g (94.6% of label claim). 25 Peptide loading was 1.689/xg/mg (33.8% of label claim).

EXAMPLE 3 Peptide added 1 hour before centrifugation Product: Bovine Insulin loaded nanoparticles Aim: To prepare a lg batch of insulin loaded nanoparticles at a theoretical loading of 50mg/g and with the peptide ZElan018 added 1 hour before centrifugation.

Formulation Details RG504H (Lot no. 250583) 1.Og Acetone 22.5ml Ethanol: 2.5ml PVA(aq. 5%w/v) 200ml Bovine Insulin (Lot no. 65H0640) 50mg Printed from Mimosa Peptide: PAX2 (ZElan018) 5mg Experimental details: The 5% w/v PVA solution was prepared by heating 5 water until near boiling point, adding PVA and stirring until cool. The organic phase was prepared by adding acetone, 22.5ml, and ethanol, 2.5ml, together. The polymer solution was prepared by adding RG504H, lg, to the organic phase prepared above and stirring until dissolved. The IKA™ 10 reactor vessel was set up, all seals greased and the temperature was set at 25°C. The PVA solution, 200ml, was added into the reactor vessel and stirred at 400 rpm.

Bovine insulin, 50mg, was added into the stirring PVA solution. Using clean tubing and a green needle, the 15 polymer solution was slowly dripped in the stirring PVA solution with the peristaltic pump set at 40. The solvent was allowed to evaporate by opening the ports and allowing the dispersion to stir overnight at 400 rpm.

PAX2 (ZElan018 5mg) was added to the stirring 20 particle suspension. After 1 hr, the suspension was centrifuged in a Beckman Ultracentrifuge™ with swing-out rotor at 12,500 rpm, 4°C. The supernatant was decanted and discarded. The "cake" of particles was broken up and dH20 (200ml) was added to wash the particles. The centrifugation 25 and washing steps were repeated twice.

The 'cake' was broken up and the particles were dried in the vacuum oven. The particles were ground, placed in a securitainer and sent for analysis. Potency was 20.75mg/g (41.5% of label claim). Peptide loading was 30 1.256^g/mg (25.12 % of label claim).

EXAMPLE 4: Leuprolide acetate loaded nanoparticles Aim: To prepare a 3g batch of leuprolide-acetate loaded nanoparticles at a theoretical loading of 20mg/g and with the 35 peptide ZElan024 added.

Formulation Details RG504H (Lot no. 271077) 3.Og Printed from Mimosa Acetone Ethanol: PVA(aq. 5%w/v) Leuprolide acetate (Lot no. V14094) 5 Peptide: P31 (ZElan024) 67.5ml 7.5ml 600ml 60mg 15mg/5 0ml dH20 Experimental details: The PVA solution was prepared and the organic phase was prepared by adding acetone, 67.5ml, and ethanol, 7.5ml, 10 together. The polymer solution was prepared by adding RG504H, 3g, to the organic phase prepared above and stirring until dissolved. The IKA™ reactor vessel was set up, all seals greased and the temperature was set at 25°C. The PVA solution, 600ml, was added into the reactor vessel and 15 stirred at 400 rpm.

Leuprolide acetate, 60mg, was added into the stirring PVA solution. Using clean tubing and a green needle, the polymer solution, was slowly dripped in the stirring PVA solution with the peristaltic pump set at 40. 20 The solvent was allowed to evaporate by opening the ports and allowing the dispersion to stir overnight at 400 rpm. The suspension was centrifuged in a Beckman Ultracentrifuge™ with swing-out rotor at 15,000 rpm, 4°C. The supernatant was decanted and retained for analysis.

The "cake" of particles was broken up and dH20 200ml) was added to wash the particles. The centrifugation and washing steps were repeated twice.

The peptide solution (P31 (SEQ ID NO:43), 15mg in 50ml dH20) was prepared and added to the particles for a final 30 washing stage. The suspended particles were centrifuged as before. The supernatant liquid was decanted, and the particles were dried in the vacuum oven.

The particles were ground, placed in a securitainer and sent for analysis. The weight of particles recovered was 35 1.87g. SEM showed discrete, reasonably spherical particles in the 300-500nm size range. The potency was 4.7mg/g (23.4% of label claim) . Peptide loading was 1.76/xg/mg.

Printed from Mimosa WO 98/51325 PCT/US98/10088 EXAMPLE 5: Peptide added bv 'spiking' polymer phase with nolvmer-peptide conjugate Product: Bovine Insulin loaded nanoparticles Aim: To prepare a 3g batch of insulin loaded nanoparticles at a theoretical loading of 50mg/g and with the polymer-peptide conjugate PLGA-ZElan019 added.

Formulation Details RG504H (Lot no. 271077) 2.85g RG504H-ZElan019 conjugate 0.15g (5PAX5-conjugate) Acetone 67.5ml Ethanol: 7.5ml PVA(aq. 5%w/v) 600ml >~ Bovine Insulin(Lot no. 86H0674) 150mg Experimental details: The 5% w/v PVA solution was prepared by heating water until near boiling point, adding PVA and stirring until cool. The organic phase was prepared by adding acetone, 20 67.5ml, and ethanol, 7.5ml, together. The polymer solution was prepared by adding RG504H and the polymer-peptide conjugate to the organic phase and stirring until dissolved.

The IKA™ reactor vessel was set up, all seals greased and the temperature was set at 25°C. The PVA 25 solution, 400ml, was added into the reactor vessel and stirred at 400 rpm.

Bovine insulin, lOOmg, was added into the stirring PVA solution. Using clean tubing and a green needle, the polymer solution, was slowly dripped in the stirring PVA 30 solution with the peristaltic pump set at 40. The solvent was allowed to evaporate by opening the ports and allowing the dispersion to stir overnight at 400 rpm.

The suspension was centrifuged in a Beckman Ultracentrifuge™ with swing-out rotor at 12,500 rpm, 4°C. 35 The supernatant was decanted and discarded. The "cake" of particles was broken up and dH20 (200ml) was added to wash the Printed from Mimosa WO 98/51325 PCT/US98/10088 particles. The centrifugation washing step was repeated twice.

The 'cake' was broken up and the particles were dried in the vacuum oven. The particles were ground, placed 5 in a securitainer and sent for analysis. The weight of particles recovered was 2.8g. The potency was 53.1mg/g 106.2% of label claim). Peptide loading was 4.02 /zg/mg (80.4% of label claim). 10. ANIMAL STUDIES Study 1 An open-loop study in which the test solution was injected directly into the ileum was done. Wistar rats (300-3 50g) were fasted for 4 hours and anaesthetized by intramuscular administration 15 to 20 minutes prior to administration of the test solution with a solution of ketamine [0.525 ml of ketamine (100 mg/ml) and 0.875 ml of acepromazine maleate-BP ACP (2mg/ml)]. The rats were then injected with a test solution (injection volume: 1.5ml PBS) intra-duodenally at 2-3 cm below the pyloris. The test solution contained either PLGA particles manufactured according to the coacervation procedure given above with or without targeting peptides or by the "spiked" method given above. Insulin (fast-acting bovine; 28.1 iu/mg) was incorporated in the particles at 5% drug loading for a total of lOOiu insulin (70 mg particles) or 300iu insulin (210 mg particles). Blood glucose values for the rats were measured using a Glucometer™ (Bayer; 0.1 to 33.3 m/mol/L); plasma insulin values were measured using a Phadeseph RIA Kit™ (Upjohn Pharmacia; 3 to 240 /xU/ml-assayed in duplicate) . Systemic and portal blood was sampled.

Study groups included animals receiving test solutions containing particles coated with the following peptides shown in Table 33.

Printed from Mimosa Table 33 Study Group Receptor Peptide I hSI SNilO SNi34 II hPEPTl P31 5PAX5 III HPTl PAX2 HAX42 IV D2H DCX8 DCX11 V ("spiked") hPEPTl P31-PLGA conjugate 5PAX5-PLGA conjugate Control groups included: 1) PBS control (1.5ml) Open-Loop; 2) Insulin solution (liu/0.2ml) subcutaneous; 3) Insulin particles - no peptide (liu/0.2ml) subcutaneous; 4) Insulin particles/all 8 peptides mix (liu/0.2ml) subcutaneous; 5) Insulin loaded particles/peptide control (scrambled 5PAX5) (100iu/l.5ml) Open-Loop; 6) Insulin loaded particles/peptide control (scrambled 5PAX5) (300iu/l.5ml) Open-Loop; 7) Control particles (insulin-free)/all 8 peptide mix (equivalent 100iu/1.5ml) Open-Loop; and 8) Control particles (insulin-free) /all 8 peptide mix (equivalent 300iu/1.5ml) Open-Loop.

The following describes the pharmacokinetics for 300iu-loading: Target Receptor HPTl Spiked hPEPTl PAX2 scrambled Mix-8 hPEPTl D2H hSI * based on area under the curve (AUC) (l-4h), base-line adjusted, relative to subcutaneous insulin solution liu ** Fold increase in AUC compared to insulin particles: 300iu Figures 17A and 17B show the systemic blood glucose and insulin levels following intestinal administration of control (PBS); insulin solution; insulin particles; all 8 F%* Fold-increase** Stat. .37 17.0 <0.001 4 . 94 7.5 0.005 3 .50 3.6 NS 2.00 2 . 0 NS 1.60 1. 5 NS 1.57 1.4 NS 0.54 0.9 NS Printed from Mimosa peptides mix particles and study group peptide-particles (lOOiu). Figures 18A and 18B show the systemic blood glucose and insulin levels following intestinal administration of control (PBS); insulin solution; insulin particles and study 5 group peptide-particles (300iu).

HPTl targeted peptide coated particles provided the most potent enhancement of the delivery of insulin over subcutaneous injection of insulin followed by hPEPTl spiked > PAX2 scrambled > mix-8 > hPEPTl > D2H > uncoated particles > hSI > solution. In a repeat study, the uncoated particles containing insulin gave similar profiles but the HPTl-peptide targeted particles gave a reduced profile (3-fold). The insulin-free PLGA particles and the all-8 mix particles did not show an effect on the basal insulin or glucose levels.

The HPTl targeting particles, the PEPT1 spiked, targeting particles, and the PEPT1 targeting particles also reduced blood glucose levels indicative that the insulin delivered was bioactive. The other targeting particles were also shown to reduce blood glucose levels although not to the same extent as the HPTl and PEPT1 spiked particles. No histological differences were observed in the small intestine for any of the formulations evaluated.

Study 2 A second open-loop study, similar to study 1 above, was undertaken with the following treatment groups as shown in Table 34.

Table 34 Group Dose Description Number Insulin (iu) 1 PBS control 2a 1 subcutaneous, bovine insulin 2b 2 subcutaneous, bovine insulin 2c 3 subcutaneous, bovine insulin 2d 4 subcutaneous, bovine insulin 2e subcutaneous, bovine insulin Printed from Mimosa 2f subcutaneous, bovine insulin 2g 4 subcutaneous, human insulin 3 300 uncoated insulin particles 4 100 HAX42/PAX2 with 300 iu particle loading 300 HAX42/PAX2 (40mer) particles 6 300 HAX42 (40mer) particles 7 300 HAX42 particles + 10-fold excess free HAX42 (40mer) 8 300 PAX2 (40mer) particles 9 300 PAX2 freeze-dried (40mer) particles 300 PAX2 scrambled particles III (4 0mer) 11 300 PAX2 scrambled particles IV (19mer) 12 300 5PAX5/P31 (40mer) particles 13 300 P31 (40mer) particles 14 300 5PAX5 (40mer) particles 300 HAX42 (27mer) particles 16 300 PAX2 (20mer) particles 17 300 P31 (20mer) particles 18 300 PAX2 (15mer) particles 19 300 P31 (15mer) particles 300 P31 D-form 1(5 D-arginine)(16mer) particles 21 300 P31 D-form 11(2 D-arginine)(16mer) particles 22 300 HAX42 (lOmer) Availability of insulin following administration was assessed relative to a 1 and 2 0iu subcutaneous dose because the response to increasing subcutaneous doses of bovine insulin does not increase linearly over the range of 1 to 20iu. Data up to three hours post-dosing was available for most animals. Therefore, availability was first assessed using individual AUC(0-3h) data estimated from baseline-subtracted data for which data up to 3 hours was available. This approach may lead to an underestimation of the availability as some animals that gave a high response often did not survive for 3 hours and, therefore, were excluded from the analyses. In an attempt to capture as much of these high responses observed at the earlier timepoints as possible, the mean baseline-subtracted plasma concentration Printed from Mimosa data was used to estimate an AUC for each group. Table 35 shows the results based on this second approach (AUC(0-3h) calculated from the mean plasma concentration data).

Table 35 Group Dose iu Mean AUC(0.3h) F vs. 1 iu F vs. 1 0 2.14 2a 1 875.27 100.00 28 86 2b 2 2439.36 139.35 40 22 2c 3 3671.44 139.82 40 36 2d 4 6912.18 197 .43 56 98 2e 27224.41 311.04 89 77 2f 60651.28 346.47 100 .00 2g 4 14255.49 407.17 117 .52 3 300 10677.78 4.07 1. 17 3 -Rat43 300 4645.06 1.77 0 . 51 4 100 3527.18 4 .03 1. 16 300 27112.26 .33 2 . 98 6 300 33091.68 12.60 3 . 64 7 300 9303.09 3 .54 1. 02 8 300 34241.83 13 . 04 3 . 76 9 300 10968.83 4 .18 1. 21 300 27692.78 .55 3 . 04 11 . 300 3004.29 1.14 0 . 33 12 300 18852.61 7.18 2 . 07 13 300 20278.43 7.72 2 . 23 14 300 17400.38 6.63 1. 91 300 16775.69 6.39 1. 84 16 300 14217.47 .41 1. 56 17 300 8197.97 3 .12 0 . 90 18 300 25050.59 9.54 2 . 75 19 300 7927.96 3 . 02 0 . 87 300 21519.57 8.20 2 . 37 21 300 6322.41 2 .41 0 . 69 22 300 12553.01 4 .78 1. 38 The data for group 3 (uncoated insulin particles) are 30 expressed with and without Rat 43. This animal had an atypically high response to these uncoated particles and, therefore, may have biased the data for this group.

This data shows that a combination of peptide-coated particles (HAX42/PAX2 or 5PAX5/P31) shows no greater 35 availability than particles coated with the individual peptides. Further, peptide-coated particles have a greater availability than uncoated peptides. Scrambling the 40mer Printed from Mimosa PAX2 peptide did not result in a loss of bioavailability. Scrambling the PAX2 peptide and reducing the size to 19mer resulted in a loss of bioavailability although this loss may be attributed in part to the reduction in peptide size.

Reducing peptide size resulted in loss of bioavailability. The D-form of P31 (ZElan053) had increased bioavailability possibly due to greater resistance to peptide breakdown. A competitive excess of peptide resulted in a loss of bioavailability, and freeze drying caused a loss in 10 bioavailability. By way of example, measurement of blood glucose levels showed that the HPTl and hPEPTl targeting particles incorporating HAX42, PAX2, P31 (SEQ ID NO:43), and P31 D-form (ZElan053) reduced blood glucose levels indicating that the insulin delivered was bioactive. 15 In further studies, insulin was recovered from the targeting particles following particle formation by dissolution and analyzed by electrophoresis in non-denaturing sodum dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). The analysis of the insulin by non-20 denaturing SDS-PAGE and also by western blot transferred to membranes and subsequent screening with an antibody to insulin, indicated that the insulin was intact, with no evidence of degradation, dimerization, or aggregation during the process of particle formation.

Study 3 An intraduodenal open loop model study was carried out on Wistar rats (300-350g). Group 1 was administered leuprolide acetate (12.5 fig) subcutaneously. Group 2 was 30 administered intraduodenally uncoated leuprolide acetate particles (600 /xg, 1.5 ml). Group 3 was intraduodenally administered leuprolide acetate particles coated with PAX2 (600 ng; 1.5 ml). Group 4 was administered intraduodenally leuprolide acetate particles coated with P31 (SEQ ID NO:43) 35 (600 fig, 1.5 ml). Figure 19 shows the leuprolide plasma concentration following administration to these four groups. Both the P31 (SEQ ID NO:43) and the PAX2 coated leuprolide Printed from Mimosa particles administered intraduodenally provided enhanced plasma levels of leuprolide relative to subcutaneous injection.

Homologies of GIT transport-binding peptides to known proteins are shown m Figures 20, 21A-F, and 22 A-D.

The present invention is not to be limited in scope by the specific embodiments described herein. Indeed, 10 various modifications of the invention m addition to those described herein will become apparent to those skilled in the art from the foregoing description and accompanying figures. Such modifications are intended to fall within the scope of the appended claims.

Various publications are cited herein, the disclosures of which are incorporated by reference in their entireties.

Printed from Mimosa PCT/U S98/10088 SEQUENCE LISTING (1) GENERAL INFORMATION (l) APPLICANTS: CYTOGEN CORPORATION and ELAN CORPORATION, pic (ll) TITLE OF THE INVENTION: RANDOM PEPTIDES THAT BIND TO GASTROINTESTINAL TRACT (GIT) TRANSPORT RECEPTORS AND RELATED METHODS (lil) NUMBER OF SEQUENCES: 265 (lv) CORRESPONDENCE ADDRESS- (A) ADDRESSEE. Pennie & Edmonds LLP (B) STREET. 1155 Avenue of the Americas (C) CITY New York (D) STATE: New York (E) COUNTRY: USA (F) ZIP: 10036 (v) COMPUTER READABLE FORM: (A) MEDIUM TYPE: Diskette (B) COMPUTER. IBM Compatible 15 (C) OPERATING SYSTEM: DOS (D) SOFTWARE. FastSEQ Version 2.0 (vi) CURRENT APPLICATION DATA- (A) APPLICATION NUMBER- (B) FILING DATE: (C) CLASSIFICATION: (viil) ATTORNEY/AGENT INFORMATION: (A) NAME: Misrock, S Leslie (B) REGISTRATION NUMBER: 10,872 (C) REFERENCE/DOCKET NUMBER: 1101-209-228 (ix) TELECOMMUNICATION INFORMATION (A) TELEPHONE: 212-790-9090 (B) TELEFAX. 212-86 9-9741 (C) TELEX: 66141 PENNIE (2) INFORMATION FOR SEQ ID NO.l: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:l: Arg Ser Gly Ala Tyr Glu Ser Pro Asp Gly Arg Gly Gly Arg Ser Tyr 10 15 Val Gly Gly Gly Gly Gly Cys Gly Asn lie Gly Arg Lys His Asn Leu 25 30 Trp Gly Leu Arg Thr Ala Ser Pro Ala Cys Trp Asp 35 35 40 (2) INFORMATION FOR SEQ ID NO:2- (i) SEQUENCE CHARACTERISTICS: Printed from Mimosa WO 98/51325 PCT/US98/10088 (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:2: Ser Pro Arg Ser Phe Trp Pro Val Val Ser Arg His Glu Ser Phe Gly 10 15 lie Ser Asn Tyr Leu Gly Cys Gly Tyr Arg Thr Cys lie Ser Gly Thr 25 30 Met Thr Lys Ser Ser Pro lie Tyr Pro Arg His Ser 35 40 (2) INFORMATION FOR SEQ ID NO:3. (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 44 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:3: Ser Ser Ser Ser Asp Trp Gly Gly Val Pro Gly Lys Val Val Arg Glu 10 15 Arg Phe Lys Gly Arg Gly Cys Gly lie Ser lie Thr Ser Val Leu Thr 25 30 Gly Lys Pro Asn Pro Cys Pro Glu Pro Lys Ala Ala 35 40 (2) INFORMATION FOR SEQ ID NO:4: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE, amino acid (C) STRANDEDNESS (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:4.

Arg Val Gly Gin Cys Thr Asp Ser Asp Val Arg Arg Pro Trp Ala Arg 10 15 Ser Cys Ala His Gin Gly Cys Gly Ala Gly Thr Arg Asn Ser His Gly 25 30 Cys lie Thr Arg Pro Leu Arg Gin Ala Ser Ala His 30 35 40 (2) INFORMATION FOR SEQ ID NO:5: (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 39 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.5.

Printed from Mimosa Ser His Ser Gly Gly Met Asn Arg Ala Tyr Gly Asp Val Phe Arg 1 Leu Arg Asp Arg 20 Trp Asn Ala Thr Ser 25 His His Thr Arg Pro 30 Thr Gin Leu Pro 35 Arg Gly Pro Asn (2) INFORMATION FOR SEQ ID NO:6. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 41 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:6 Ser Pro Cys Gly Gly Ser Trp Gly Arg Phe Met Gin Gly Gly Leu Phe 10 15 Gly Gly Arg Thr Asp Gly Cys Gly Ala His Arg Asn Arg Thr Ser Ala 25 30 Ser Leu Glu Pro Pro Ser Ser Asp Tyr 35 40 (2) INFORMATION FOR SEQ ID NO:7 (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS (D) TOPOLOGY: unknown 2 0 (") MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO 7: Arg Gly Ala Ala Asp Gin Arg Arg Gly Trp Ser Glu Asn Leu Gly Leu 10 15 Pro Arg Val Gly Trp Asp Ala lie Ala His Asn Ser Tyr Thr Phe Thr 25 30 Ser Arg Arg Pro Arg Pro Pro 25 35 (2) INFORMATION FOR SEQ ID NO:8. (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 8- Ser Gly Gly Glu Val Ser Ser Trp Gly Arg Val Asn Asp Leu Cys Ala 10 15 Arg Val Ser Trp Thr Gly Cys Gly Thr Ala Arg Ser Ala Arg Thr Asp 20 25 30 Asn Lys Gly Phe Leu Pro Lys His Ser Ser Leu Arg 35 40 (2) INFORMATION FOR SEQ ID NO.9: Printed from Mimosa (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO-9.

Ser Asp Ser Asp Gly Asp His Tyr Gly Leu Arg Gly Gly Val Arg Cys 10 15 Ser Leu Arg Asp Arg Gly Cys Gly Leu Ala Leu Ser Thr Val His Ala 25 30 Gly Pro Pro Ser Phe Tyr Pro Lys Leu Ser Ser Pro 35 40 (2) INFORMATION FOR SEQ ID NO:10: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO 10: Arg Ser Leu Gly Asn Tyr Gly Val Thr Gly Thr Val Asp Val Thr Val 10 15 Leu Pro Met Pro Gly His Ala Asn His Leu Gly Val Ser Ser Ala Ser 25 30 Ser Ser Asp Pro Pro Arg Arg 20 35 (2) INFORMATION FOR SEQ ID NO-11: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH 3 8 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:ll: Arg Thr Thr Thr Ala Lys Gly Cys Leu Leu Gly Ser Phe Gly Val Leu 10 15 Ser Gly Cys Ser Phe Thr Pro Thr Ser Pro Pro Pro His Leu Gly Tyr 20 25 30 Pro Pro His Ser Val Asn 35 (2) INFORMATION FOR SEQ ID NO.12: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO.12: Printed from Mimosa WO 98/51325 PCT/US98/10088 Ser Pro Lys Leu Ser Ser Val Gly Val Met Thr Lys Val Thr Glu Leu 1 Pro Thr Glu Gly 20 Pro Asn Ala lie Ser 25 He Pro lie Ser Ala 30 Thr Leu Gly Pro Arg 35 Asn Pro Leu Arg (2) INFORMATION FOR SEQ ID NO.13. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:13: Arg Trp Cys Gly Ala Glu Leu Cys Asn Ser Val Thr Lys Lys Phe Arg 10 15 Pro Gly Trp Arg Asp His Ala Asn Pro Ser Thr His His Arg Thr Pro 25 30 Pro Pro Ser Gin Ser Ser Pro 35 (2) INFORMATION FOR SEQ ID NO:14: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 44 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (n) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION:,SEQ ID NO.14: Arg Trp Cys Gly Ala Asp Asp Pro Cys Gly Ala Ser Arg Trp Arg Gly 10 15 Gly Asn Ser Leu Phe Gly Cys Gly Leu Arg Cys Ser Ala Ala Gin Ser 25 30 Thr Pro Ser Gly Arg lie His Ser Thr Ser Thr Ser 35 40 (2) INFORMATION FOR SEQ ID NO 15: (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 3 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:15 Ser Lys Ser Gly Glu Gly Gly Asp Ser Ser Arg Gly Glu Thr Gly Trp 10 15 Ala Arg Val Arg Ser His Ala Met Thr Ala Gly Arg Phe Arg Trp Tyr 20 25 30 Asn Gin Leu Pro Ser Asp Arg 35 (2) INFORMATION FOR SEQ ID NO: 16.

Printed from Mimosa (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 38 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:16 Arg Ser Ser Ala Asn Asn Cys Glu Trp Lys Ser Asp Trp Met Arg Arg 10 15 Ala Cys lie Ala Arg Tyr Ala Asn Ser Ser Gly Pro Ala Arg Ala Val 25 30 Asp Thr Lys Ala Ala Pro 35 (2) INFORMATION FOR SEQ ID NO:17: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH 44 ammo acids (B) TYPE, ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ll) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:17- Ser Lys Trp Ser Trp Ser Ser Arg Trp Gly Ser Pro Gin Asp Lys Val 10 15 Glu Lys Thr Arg Ala Gly Cys Gly Gly Ser Pro Ser Ser Thr Asn Cys 25 30 His Pro Tyr Thr Phe Ala Pro Pro Pro Gin Ala Gly 20 35 40 (2) INFORMATION FOR SEQ ID NO:18- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:18: Ser Gly Phe Trp Glu Phe Ser Arg Gly Leu Trp Asp Gly Glu Asn Arg 10 15 Lys Ser Val Arg Ser Gly Cys Gly Phe Arg Gly Ser Ser Ala Gin Gly 20 25 30 0 Pro Cys Pro Val Thr Pro Ala Thr lie Asp Lys His 35 40 (2) INFORMATION FOR SEQ ID NO:19: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:19: Printed from Mimosa 0 Ser Glu Ser Gly Arg Cys Arg Ser Val Ser Arg Trp Met Thr Thr Trp 10 15 Gin Thr Gin Lys Gly Gly Cys Gly Ser Asn Val Ser Arg Gly Ser Pro 25 30 Leu Asp Pro Ser His Gin Thr Gly His Ala Thr Thr 40 (2) INFORMATION FOR SEQ ID NO:20: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 39 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS (D) TOPOLOGY, unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION- SEQ ID N0:20 Arg Glu Trp Arg Phe Ala Gly Pro Pro Leu Asp Leu Trp Ala Gly Pro 10 15 Ser Leu Pro Ser Phe Asn Ala Ser Ser His Pro Arg Ala Leu Arg Thr 25 30 Tyr Trp Ser Gin Arg Pro Arg 35 (2) INFORMATION FOR SEQ ID NO:21: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE, ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown 0 (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:21.

Arg Met Glu Asp lie Lys Asn Ser Gly Trp Arg Asp Ser Cys Arg Trp 10 15 Gly Asp Leu Arg Pro Gly Cys Gly Ser Arg Gin Trp Tyr Pro Ser Asn 25 30 Met Arg Ser Ser Arg Asp Tyr Pro Ala Gly Gly His 5 35 40 (2) INFORMATION FOR SEQ ID NO:22: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 36 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS- (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:22: 0 Ser His Pro Trp Tyr Arg His Trp Asn His Gly Asp Phe Ser Gly Ser 10 15 Gly Gin Ser Arg His Thr Pro Pro Glu Ser Pro His Pro Gly Arg Pro 20 25 30 Asn Ala Thr lie 35 (2) INFORMATION FOR SEQ ID NO: 23: 140 - Printed from Mimosa (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23: Arg Tyr Lys His Asp lie Gly Cys Asp Ala Gly Val Asp Lys Lys Ser 10 15 Ser Ser Val Arg Gly Gly Cys Gly Ala His Ser Ser Pro Pro Arg Ala 25 30 Gly Arg Gly Pro Arg Gly Thr Met Val Ser Arg Leu 35 40 (2) INFORMATION FOR SEQ ID NO:24: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 44 amino acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ll) MOLECULE TYPE- peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:24: Ser Gin Gly Ser Lys Gin Cys Met Gin Tyr Arg Thr Gly Arg Leu Thr 10 15 Val Gly Ser Glu Tyr Gly Cys Gly Met Asn Pro Ala Arg His Ala Thr 25 30 Pro Ala Tyr Pro Ala Arg Leu Leu Pro Arg Tyr Arg 35 40 (2) INFORMATION FOR SEQ ID NO:25. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:25.

Ser Gly Arg Thr Thr Ser Glu lie Ser Gly Leu Trp Gly Trp Gly Asp 10 15 Asp Arg Ser Gly Tyr Gly Trp Gly Asn Thr Leu Arg Pro Asn Tyr lie 20 25 30 Pro Tyr Arg Gin Ala Thr Asn Arg His Arg Tyr Thr 35 40 (2) INFORMATION FOR SEQ ID NO-26: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO:26: Printed from Mimosa WO 98/51325 PCT/US98/10088 Arg Trp Asn Trp Thr Val Leu Pro Ala Thr Gly Gly His Tyr Trp Thr 1 Arg Ser Thr Asp 20 Tyr His Ala lie Asn 25 Asn His Arg Pro Ser 30 lie Pro His Gin His 35 Pro Thr Pro lie (2) INFORMATION FOR SEQ ID NO:27: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 44 ammo acxds (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (n) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO 27 Ser Trp Ser Ser Trp Asn Trp Ser Ser Lys Thr Thr Arg Leu Gly Asp 10 15 Arg Ala Thr Arg Glu Gly Cys Gly Pro Ser Gin Ser Asp Gly Cys Pro 25 30 Tyr Asn Gly Arg Leu Thr Thr Val Lys Pro Arg Thr 40 (2) INFORMATION FOR SEQ ID NO:28: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 37 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:28 Ser Gly Ser Leu Asn Ala Trp Gin Pro Arg Ser Trp Val Gly Gly Ala 10 15 Phe Arg Ser His Ala Asn Asn Asn Leu Asn Pro Lys Pro Thr Met Val 25 30 Thr Arg His Pro Thr 25 35 (2) INFORMATION FOR SEQ ID NO.29. (l) SEQUENCE CHARACTERISTICS (A) LENGTH: 44 ammo acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:29- Arg Tyr Ser Gly Leu Ser Pro Arg Asp Asn Gly Pro Ala Cys Ser Gin 10 15 Glu Ala Thr Leu Glu Gly Cys Gly Ala Gin Arg Leu Met Ser Thr Arg 20 25 30 Arg Lys Gly Arg Asn Ser Arg Pro Gly Trp Thr Leu 35 40 (2) INFORMATION FOR SEQ ID NO:30: Printed from Mimosa (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNES S: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO:30: Ser Val Gly Asn Asp Lys Thr Ser Arg Pro Val Ser Phe Tyr Gly Arg 10 15 Val Ser Asp Leu Trp Asn Ala Ser Leu Met Pro Lys Arg Thr Pro Ser 25 30 Ser Lys Arg His Asp Asp Gly 35 (2) INFORMATION FOR SEQ ID NO:31. (l) SEQUENCE CHARACTERISTICS- (A) LENGTH: 38 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:31: Arg Trp Pro Ser Val Gly Tyr Lys Gly Asn Gly Ser Asp Thr lie Asp 10 15 Val His Ser Asn Asp Ala Ser Thr Lys Arg Ser Leu lie Tyr Asn His 25 30 Arg Arg Pro Leu Phe Pro 20 35 (2) INFORMATION FOR SEQ ID NO:32: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:32: Arg Thr Phe Glu Asn Asp Gly Leu Gly Val Gly Arg Ser lie Gin Lys 10 15 Lys Ser Asp Arg Trp Tyr Ala Ser His Asn lie Arg Ser His Phe Ala 20 25 30 Ser Met Ser Pro Ala Gly Lys 35 (2) INFORMATION FOR SEQ ID NO:33: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (II) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:33: Printed from Mimosa PCT /US98/10088 ) Ser Tyr Cys Arg Val Lys Gly Gly Gly Glu Gly Gly His Thr Asp Ser 1 Asn Leu Ala Arg 20 Ser Gly Cys Gly Lys 25 Val Ala Arg Thr Ser 30 Arg Leu Gin His lie 35 Asn Pro Arg Ala Thr 40 Pro Pro Ser Arg (2) INFORMATION FOR SEQ ID NO:34: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH' 39 amino acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:34 Ser Trp Thr Arg Trp Gly Lys His Thr His Gly Gly Phe Val Asn Lys 10 15 Ser Pro Pro Gly Lys Asn Ala Thr Ser Pro Tyr Thr Asp Ala Gin Leu 25 30 Pro Ser Asp Gin Gly Pro Pro 35 (2) INFORMATION FOR SEQ ID NO:35: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 44 amino acids (B) TYPE- ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown 3 (n) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.35: Ser Gin Val Asp Ser Phe Arg Asn Ser Phe Arg Trp Tyr Glu Pro Ser 10 15 Arg Ala Leu Cys His Gly Cys Gly Lys Arg Asp Thr Ser Thr Thr Arg 25 30 lie His Asn Ser Pro Ser Asp Ser Tyr Pro Thr Arg 5 35 40 (2) INFORMATION FOR SEQ ID NO:36: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown 0 (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36: Ser Phe Leu Arg Phe Gin Ser Pro Arg Phe Glu Asp Tyr Ser Arg Thr 10 15 lie Ser Arg Leu Arg Asn Ala Thr Asn Pro Ser Asn Val Ser Asp Ala 20 25 30 His Asn Asn Arg Ala Leu Ala 35 (2) INFORMATION FOR SEQ ID NO:37 - 144 Printed from Mimosa (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 3 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION- SEQ ID NO:37.

Arg Ser lie Thr Asp Gly Gly lie Asn Glu Val Asp Leu Ser Ser Val 10 15 Ser Asn Val Leu Glu Asn Ala Asn Ser His Arg Ala Tyr Arg Lys His 25 30 Arg Pro Thr Leu Lys Arg Pro 35 (2) INFORMATION FOR SEQ ID NO:38: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 44 amino acids (B) TYPE amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:38: Ser Ser Lys Val Ser Ser Pro Arg Asp Pro Thr Val Pro Arg Lys Gly 10 15 Gly Asn Val Asp Tyr Gly Cys Gly His Arg Ser Ser Ala Arg Met Pro 25 30 Thr Ser Ala Leu Ser Ser lie Thr Lys Cys Tyr Thr 35 40 (2) INFORMATION FOR SEQ ID NO:39: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE amino acid (C) STRANDEDNESS.

(D) TOPOLOGY- unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:39: Arg Ala Ser Thr Gin Gly Gly Arg Gly Val Ala Pro Glu Phe Gly Ala 10 15 Ser Val Leu Gly Arg Gly Cys Gly Ser Ala Thr Tyr Tyr Thr Asn Ser 20 25 30 Thr Ser Cys Lys Asp Ala Met Gly His Asn Tyr Ser 35 40 (2) INFORMATION FOR SEQ ID NO-.40: (l) SEQUENCE CHARACTERISTICS (A) LENGTH: 3 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:40: Printed from Mimosa Arg Trp Cys Glu Lys His Lys Phe Thr Ala Ala Arg Cys Ser Ala Gly 10 15 Ala Gly Phe Glu Arg Asp Ala Ser Arg Pro Pro Gin Pro Ala His Arg 25 30 Asp Asn Thr Asn Arg Asn Ala 35 (2) INFORMATION FOR SEQ ID NO:41: (l) SEQUENCE CHARACTERISTICS- (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO-41: Ser Phe Gin Val Tyr Pro Asp His Gly Leu Glu Arg His Ala Leu Asp 10 15 Gly Thr Gly Pro Leu Tyr Ala Met Pro Gly Arg Trp lie Arg Ala Arg 25 30 Pro Gin Asn Arg Asp Arg Gin 35 (2) INFORMATION FOR SEQ ID NO:42 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 38 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (il) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO:42: Ser Arg Cys Thr Asp Asn Glu Gin Cys Pro Asp Thr Gly Thr Arg Ser 10 15 Arg Ser Val Ser Asn Ala Arg Tyr Phe Ser Ser Arg Leu Leu Lys Thr 25 30 His Ala Pro His Arg Pro 25 35 (2) INFORMATION FOR SEQ ID NO:43: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 3 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:43: Ser Ala Arg Asp Ser Gly Pro Ala Glu Asp Gly Ser Arg Ala Val Arg 10 15 Leu Asn Gly Val Glu Asn Ala Asn Thr Arg Lys Ser Ser Arg Ser Asn 20 25 30 Pro Arg Gly Arg Arg His Pro 35 (2) INFORMATION FOR SEQ ID NO:44: Printed from Mimosa (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO 44 Ser Ser Ala Asp Ala Glu Lys Cys Ala Gly Ser Leu Leu Trp Trp Gly 10 15 Arg Gin Asn Asn Ser Gly Cys Gly Ser Pro Thr Lys Lys His Leu Lys 25 30 His Arg Asn Arg Ser Gin Thr Ser Ser Ser Ser His 35 40 (2) INFORMATION FOR SEQ ID NO.45: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE. amino acid (C) STRANDEDNESS (D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:45: Arg Pro Lys Asn Val Ala Asp Ala Tyr Ser Ser Gin Asp Gly Ala Ala 10 15 Ala Glu Glu Thr Ser His Ala Ser Asn Ala Ala Arg Lys Ser Pro Lys 25 30 His Lys Pro Leu Arg Arg Pro 35 (2) INFORMATION FOR SEQ ID NO:46: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:46: Arg Gly Ser Thr Gly Thr Ala Gly Gly Glu Arg Ser Gly Val Leu Asn 10 15 Leu His Thr Arg Asp Asn Ala Ser Gly Ser Gly Phe Lys Pro Trp Tyr 20 25 30 Pro Ser Asn Arg Gly His Lys 35 (2) INFORMATION FOR SEQ ID NO:47: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 3 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:47: Printed from Mimosa PCT /US98/10088 3 Arg Trp Gly Trp Glu Arg Ser Pro Ser Asp Tyr Asp Ser Asp Met 1 Leu Gly Ala Arg 20 Arg Tyr Ala Thr Arg 25 Thr His Arg Ala Pro 30 Pro Val Leu Lys 35 Ala Pro Leu Pro (2) INFORMATION FOR SEQ ID NO:48: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:48: Arg Gly Trp Lys Cys Glu Gly Ser Gin Ala Ala Tyr Gly Asp Lys Asp 10 15 lie Gly Arg Ser Arg Gly Cys Gly Ser lie Thr Lys Asn Asn Thr Asn 25 30 His Ala His Pro Ser His Gly Ala Val Ala Lys lie 35 40 (2) INFORMATION FOR SEQ ID NO:49: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS (D) TOPOLOGY unknown g (n) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:49: Ser Arg Glu Glu Ala Asn Trp Asp Gly Tyr Lys Arg Glu Met Ser His 10 15 Arg Ser Arg Phe Trp Asp Ala Thr His Leu Ser Arg Pro Arg Arg Pro 25 30 Ala Asn Ser Gly Asp Pro Asn 5 35 (2) INFORMATION FOR SEQ ID NO:50: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown 0 (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:50.

Glu Trp Tyr Ser Trp Lys Arg Ser Ser Lys Ser Thr Gly Leu Gly Asp 10 15 Thr Ala Thr Arg Glu Gly Cys Gly Pro Ser Gin Ser Asp Gly Cys Pro 20 25 30 Tyr Asn Gly Arg Leu Thr Thr Val Lys Pro Arg Lys 35 40 (2) INFORMATION FOR SEQ ID NO:51: - 14 8 - Printed from Mimosa (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO-51.

Arg Glu Phe Ala Glu Arg Arg Leu Trp Gly Cys Asp Asp Leu Ser Trp 10 15 Arg Leu Asp Ala Glu Gly Cys Gly Pro Thr Pro Ser Asn Arg Ala Val 25 30 Lys His Arg Lys Pro Arg Pro Arg Ser Pro Ala Leu 35 40 (2) INFORMATION FOR SEQ ID NO:52 (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:52: Ser Asp His Ala Leu Gly Thr Asn Leu Arg Ser Asp Asn Ala Lys Glu 10 15 Pro Gly Asp Tyr Asn Cys Cys Gly Asn Gly Asn Ser Thr Gly Arg Lys 25 30 Val Phe Asn Arg Arg Arg Pro Ser Ala lie Pro Thr 35 40 (2) INFORMATION FOR SEQ ID NO:53- (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 44 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:53: Arg His lie Ser Glu Tyr Ser Phe Ala Asn Ser His Leu Met Gly Gly 10 15 Glu Ser Lys Arg Lys Gly Cys Gly lie Asn Gly Ser Phe Ser Pro Thr 20 25 30 Cys Pro Arg Ser Pro Thr Pro Ala Phe Arg Arg Thr 35 40 (2) INFORMATION FOR SEQ ID NO:54: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 38 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 54 : - 149 " Printed from Mimosa WO 98/51325 PCT/US98/10088 Ser Arg Glu Ser Gly Met Trp Gly Ser Trp Trp Arg Gly His Arg Leu l Asn Ser Thr Gly 20 Gly Asn Ala Asn Met 25 Asn Ala Ser Leu Pro Pro Asp 30 Pro Pro Val 35 Ser Thr Pro (2) INFORMATION FOR SEQ ID NO:55: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 39 amino acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:55: Ser Thr Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp 10 15 Ser Asp Ser Asp Thr Asn Ala Lys His Ser Ser His Asn Arg Arg Leu 25 30 Arg Thr Arg Ser Arg Pro Asn 35 (2) INFORMATION FOR SEQ ID NO:56: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid <C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:56: TCTCACTCCT CGAGATCCGG CGCTTATGAG AGTCCGGATG GTCGGGGGGG TCGGAGCTAT 60 GTGGGGGGCG GGGGTGGNTG TGGTAACATT GGTCGGAAGC ATAACCTGTG GGGGCTGCGT 120 ACCGCGTCGC CGGCCTGCTG GGACTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:57: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:57: TCTCACTCCT CGAGTCCTCG CTCTTTCTGG CCCGTTGTGT CCCGGCATGA GTCGTTTGGG 60 ATCTCTAACT ATTTGGGNTG TGGTTATCGT ACATGTATCT CCGGCACGAT GACTAAGTCT 120 AGCCCGATTT ACCCTCGGCA TTCGTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:58. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear Printed from Mimosa (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:58: TCTCACTCCT CGAGTAGTAG CTCCGATTGG GGTGGTGTGC CTGGGAAGGT GGTTAGGGAG 60 CGCTTTAAGG GGCGCGGTTG TGGTATTTCC ATCACCTCCG TGCTCACTGG GAAGCCCAAT 120 CCGTGTCCGG AGCCTAAGGC GGCCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO-59: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:59: TCTCACTCCT CGAGAGTTGG CCAGTGCACG GATTCTGATG TGCGGCGTCC TTGGGCCAGG 60 TCTTGCGCTC ATCAGGGTTG TGGTGCGGGC ACTCGCAACT CGCACGGCTG CATCACCCGT 120 CCTCTCCGCC AGGCTAGCGC TCATTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID N0:60.

(I) SEQUENCE CHARACTERISTICS: (A) LENGTH 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE. DNA <xi> SEQUENCE DESCRIPTION. SEQ ID N0:60: TCTCACTCCT CGAGCCACTC CGGTGGTATG AATAGGGCCT ACGGGGATGT GTTTAGGGAG 6 0 CTTCGTGATC GGTGGAACGC CACTTCCCAC CACACTCGCC CCACCCCTCA GCTCCCCCGT 120 GGGCCTAATT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:61- (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 16 8 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:61: TCTCACTCCT CGAGTCCGTG CGGGGGGTCG TGGGGGCGTT TTATGCAGGG TGGCCTTTTC 60 GGCGGTAGGA CTGATGGTTG TGGTGCCCAT AGAAACCGCA CTTCTGCGTC GTTAGAGCCC 120 CCGAGCAGCG ACTACTCTAG AATCGAAGGT CGCGCTAGAC CTTCGAGA 168 (2) INFORMATION FOR SEQ ID NO:62: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 135 base pairs (B) TYPE: nucleic acid 35 (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA Printed from Mimosa (xi) SEQUENCE DESCRIPTION: SEQ ID NO:62: TCTCACTCCT CGAGGGGCGC CGCCGATCAG CGGCGGGGGT GGTCCGAGAA CTTGGGGTTG 60 CCTAGGGTGG GGTGGGACGC CATCGCTCAC AATAGCTATA CGTTCACCTC GCGCCGCCCG 120 CGCCCCCCCT CTAGA 135 (2) INFORMATION FOR SEQ ID NO:63- (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS. single (D) TOPOLOGY, linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:63: TCTCACTCCT CGAGCGGTGG GGAGGTCAGC TCCTGGGGCC GCGTGAATGA CCTCTGCGCT 60 AGGGTGAGTT GGACTGGTTG TGGTACTGCT CGTTCCGCGC GTACCGACAA CAAAGGCTTT 120 CTTCCTAAGC ACTCGTCACT CCGCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO-64: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 177 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:64: TCTCACTCCT CGAGTGATAG TGACGGGGAT CATTATGGGC TTCGGGGGGG GGTGCGTTGT 60 TCGCTTCGTG ATAGGGGTTG TGGTCTGGCC CTGTCCACCG TCCATGCTGG TCCCCCCTCT 120 TTTTACCCCA AGCTCTCCAG CCCCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:65: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:65: TCTCACTCCT CGAGGAGCTT GGGTAATTAT GGCGTCACCG GGACTGTGGA CGTGACGGTT 60 TTGCCCATGC CTGGCCACGC CAACCACCTT GGTGTCTCCT CCGCCTCTAG CTCTGATCCT 120 CCGCGGCGCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:66: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:66: Printed from Mimosa TCTCACTCCT CGAGAACTAC GACGGCTAAG GGGTGTCTTC TCGGAAGCTT CGGCGTTCTT 60 AGTGGGTGCT CATTTACGCC AACCTCTCCA CCGCCCCACC TAGGATACCC CCCCCACTCC 120 GTCAATTCTA GAATCGAAGG TCGCGCTAGA CCTTCGAGA 159 (2) INFORMATION FOR SEQ ID NO:67: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:67: TCTCACTCCT CGAGCCCGAA GTTGTCCAGC GTGGGTGTTA TGACTAAGGT CACGGAGCTG 60 CCCACGGAGG GGCCTAACGC CATTAGTATT CCGATCTCCG CGACCCTCGG CCCGCGCAAC 120 CCGCTCCGCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:68: (1) SEQUENCE CHARACTERISTICS (A) LENGTH: 162 base pairs (B) TYPE, nucleic acid 15 (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:68: TCTCACTCCT CGAGGTGGTG CGGCGCTGAG CTGTGCAACT CGGTGACTAA GAAGTTTCGC 60 CCGGGCTGGC GGGATCACGC CAATCCCTCC ACCCATCATC GTACTCCCCC GCCCAGCCAG 120 20 TCCAGCCCTT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:69: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 176 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:69: TCTCACTCCT CGAGGTGGTG CGGCGCTGAT GACCCGTGTG GTGCCAGTCG TTGGCGGGGG 60 GGCAACAGCT TGTTTGGTTG TGGTCTTCGT TGTAGTGCGG CGCAGAGCAC CCCGAGTGGC 120 AGGATCCATT CCACTTCGAC CAGCTCTAGA ATCGAAGGTG CGCTAGACCT TCGAGA 176 (2) INFORMATION FOR SEQ ID NO:70. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:70: TCTCACTCCT CGAGTAAGTC CGGGGAGGGG GGTGACAGTA GCAGGGGCGA GACGGGCTGG 60 GCGAGGGTTC GGTCTCACGC CATGACTGCT GGCCGCTTTC GGTGGTACAA CCAGTTGCCC 120 Printed from Mimosa PCT /U S98/10088 TCTGATCGGT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:71: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (li) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION SEQ ID NO: 71 TCTCACTCCT CGAGGTCGAG CGCCAATAAT TGCGAGTGGA AGTCTGATTG GATGCGCAGG 60 GCCTGTATTG CTCGTTACGC CAACAGTTCG GGCCCCGCCC GCGCCGTCGA CACTAAGGCC 120 10 GCGCCCTCTA GAATCGAAGG TCGCGCTAGA CCTTCGAGA 159 (2) INFORMATION FOR SEQ ID NO:72 (I) SEQUENCE CHARACTERISTICS (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:72- TCTCACTCCT CGAGTAAGTG GTCGTGGAGT TCGAGGTGGG GCTCCCCGCA GGATAAGGTT 60 GAGAAGACCA GGGCGGGTTG TGGTGGTAGT CCCAGCAGCA CCAATTGTCA CCCCTACACC 120 TTTGCCCCCC CCCCGCAAGC CGGCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:73: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (li) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:73 TCTCACTCCT CGAGTGGGTT CTGGGAGTTT AGCAGGGGGC TTTGGGATGG GGAGAACCGT 6 0 AAGAGTGTCC GGTCGGGTTG TGGTTTTCGT GGCTCCTCTG CTCAGGGCCC GTGTCCGGTC 12 0 ACGCCTGCCA CCATTGACAA ACACTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:74: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:74: TCTCACTCCT CGAGTGAGAG CGGGCGGTGC CGTAGCGTGA GCCGGTGGAT GACGACGTGG 60 CAGACGCAGA AGGGCGGTTG TGGTTCCAAT GTTTCCCGCG GTTCGCCCCT CGACCCCTCT 120 CACCAGACCG GGCATGCCAC TACTTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 Printed from Mimosa (2) INFORMATION FOR SEQ ID NO:75: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY- linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:75.

TCTCACTCCT CGAGGGAGTG GAGGTTTGCC GGGCCGCCGT TGGACCTGTG GGCGGGTCCG 60 AGCTTGCCCT CTTTTAACGC CAGTTCCCAC CCTCGCGCCC TGCGCACCTA TTGGTCCCAG 120 CGGCCCCGCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO.76 (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO.76: TCTCACTCCT CGAGGATGGA GGACATCAAG AACTCGGGGT GGAGGGACTC TTGTAGGTGG 60 GGTGACCTGA GGCCTGGTTG TGGTAGCCGC CAGTGGTACC CCTCGAATAT GCGTTCTAGC 120 AGAGATTACC CCGCGGGGGG CCACTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:77: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 152 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: TCTCACTCCT CGAGTCATCC GTGGTACAGG GGCCAGTCAC GCCACACCCC GCCGGAGAGC TCTAGAATCG AAGGTCGCGC TAGACCTTCG SEQ ID NO.77: CATTGGAACC ATGGTGACTT CTCTGGTTCG 60 CCCCACCCCG GCCGCCCTAA TGCCACCATT 12 0 AG 152 (2) INFORMATION FOR SEQ ID NO:78: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:78: TCTCACTCCT CGAGATATAA GCACGATATC GGTTGCGATG CTGGGGTTGA CAAGAAGTCG 60 TCGTCTGTGC GTGGTGGTTG TGGTGCTCAT TNGTCGCCAC CCCGCGCCGG CCGTGGTCCT 120 CGCGGCACGA TGGTTAGCAG GCTTTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:79- Printed from Mimosa (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO.79: TCTCACTCCT CGAGTCAGGG CTCCAAGCAG TGTATGCAGT ACCGCACCGG TCGTTTGACG 60 GTGGGGTCTG AGTATGGTTG TGGTATGAAC CCCGCCCGCC ATGCCACGCC CGCTTATCCG 120 GCGCGCCTGC TGCCACGCTA TCGCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:80: I1) SEQUENCE CHARACTERISTICS (A) LENGTH. 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (il) MOLECULE TYPE DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:80: TCTCACTCCT CGAGTGGGCG GACTACTAGT GAGATTTCTG GGCTCTGGGG TTGGGGTGAC 60 GACCGGAGCG GTTATGGTTG GGGTAACACG CTCCGCCCCA ACTACATCCC TTATAGGCAG 120 GCGACGAACA GGCATCGTTA TACGTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:81- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:81: TCTCACTCCT CGAGGTGGAA TTGGACTGTC TTGCCCGCCA CTGGCGGCCA TTACTGGACG 60 CGTTCGACGG ACTATCACGC CATTAACAAT CACAGGCCGA GCATCCCCCA CCAGCATCCG 120 ACCCCTATCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:82: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single 30 (D) TOPOLOGY: linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:82: TCTCACTCCT CGAGTTGGTC GTCGTGGAAT TGGAGCTCTA AGACTACTCG TCTGGGCGAC 60 AGGGCGACTC GGGAGGGTTG TGGTCCCAGC CAGTCTGATG GCTGTCCTTA TAACGGCCGC 120 CTTACGACCG TCAAGCCTCG CACGTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:83: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 156 base pairs Printed from Mimosa (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ll) MOLECULE TYPE- DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:83: TCTCACTCCT CGAGTGGTAG TTTGAACGCA TGGCAACCGC GGTCATGGGT GGGGGGCGCG 60 TTCCGGTCAC ACGCCAACAA TAACTTGAAC CCCAAGCCCA CCATGGTTAC TNGTCACCCT 120 ACCTCTAGAA TCGAAGGTCG CGCTAGACCT TCGAGA 156 (2) INFORMATION FOR SEQ ID NO:84: (I) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 178 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:84.

TCTCACTCCT CGAGGTATTC GGGTTTGTCC CCGCGGGACA ACGGTCCCGC TTGTAGTCAG 6 0 GAGGCTACCT TGGAGGGTTG TGGTGCGCAG AGGCTGATGT CCACCCGTCG CAAGGGCCGC 120 AACTCCCGCC CCGGGTGGAC GCTCTCTAGA ATCGAAGGTC GCGCTAGACC CTTCGAGA 178 (2) INFORMATION FOR SEQ ID NO:85: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH- 162 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS: single 20 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:85- TCTCACTCCT CGAGCGTGGG GAATGATAAG ACTAGCAGGC CGGTTTCCTT CTACGGGCGC 6 0 GTTAGTGATC TGTGGAACGC CAGCTTGATG CCGAAGCGTA CTCCCAGCTC GAAGCGCCAC 120 GATGATGGCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:86: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (il) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:86: TCTCACTCCT CGAGTACTCC CCCCAGTAGG GAGGCGTATA GTAGGCCCTA TAGTGTCGAT 60 AGCGATTCGG ATACGAACGC CAAGCACAGC TCCCACAACC GCCGTNTGCG GACGCGCAGC 120 CGCCCGAACT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:87: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 159 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS single Printed from Mimosa WO 98/51325 PCT/US98/10088 (D) TOPOLOGY: linear (ii) MOLECULE TYPE. DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:87.

TCTCACTCCT CGAGATGGCC TAGTGTGGGT TACAAGGGTA ATGGCAGTGA CACTATTGAT 60 GTTCACAGCA ATGACGCCAG TACTAAGAGG TCCCTCATCT ATAACCACCG CCGCCCCNTC 12 0 TTTCCCTCTA GAATCGAAGG TCGCGCTAGA CCTTCGAGA 159 (2) INFORMATION FOR SEQ ID NO:88: (l) SEQUENCE CHARACTERISTICS- (A) LENGTH 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single 10 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:88 TCTCACTCCT CGAGAACGTT TGAGAACGAC GGGCTGGGCG TCGGCCGGTC TATTCAGAAG 60 AAGTCGGATA GGTGGTACGC CAGCCACAAC ATTCGTAGCC ATTTCGCGTC CATGTCTCCC 120 GCTGGTAAGT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:89: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 160 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS• single (D) TOPOLOGY- linear (ii) MOLECULE TYPE DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:89: TCTCACTCCT CGAGCTATTG TCGGGTTAAG GGTGGTGGGG AGGGGGGGCA TACGGATTCC 60 AATCTGGCTA GGTCGGGTTG TGGTAAGGTG GCCAGGACCA GCAGGCTTCA GCATATCAAC 120 CCGCGCGCTA CCCCCCCCTC CCGGTCTAGA ATCGAAGGTC 160 (2) INFORMATION FOR SEQ ID NO:90: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:90.

TCTCACTCCT CGAGTTGGAC TCGGTGGGGC AAGCACANTC ATGGGGGGTT TGTGAACAAG 60 TCTCCCCCTG GGAAGAACGC CACGAGCCCC TACACCGACG CCCAGCTGCC CAGTGATCAG 120 GGTCCTCCCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:91: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear Printed from Mimosa (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO.91: TCTCACTCCT CGAGTCAGGT TGATTCGTTT CGTAATAGCT TTCGGTGGTA TGAGCCGAGC 60 AGGGCTCTGT GCCATGGTTG TGGTAAGCGC GACACCTCCA CCACTCGTAT CCACAATAGC 120 CCCAGCGACT CCTATCCTAC ACGCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:92. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: TCTCACTCCT CGAGCTTTTT GCGGTTCCAG ATCTNTCGGT TGCGCAACGC CACGAACCCG GCCTTGGCCT CTAGAATCGA AGGTCGCGCT (2) INFORMATION FOR SEQ (I) SEQUENCE CHARACTERISTICS- (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY, linear (II) MOLECULE TYPE. DNA <xi) SEQUENCE DESCRIPTION: SEQ ID NO:93.

TCTCACTCCT CGAGGAGCAT CACCGACGGG GGCATCAATG AGGTGGACCT GAGTAGTGTG 60 TCGAACGTTC TTGAGAACGC CAACTCGCAT AGGGCCTACA GGAAGCATCG CCCGACCTTG 120 AAGCGTCCTT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:94: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (li) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:94: 3 0 TCTCACTCCT CGAGTTCGAA GGTGAGCAGC CCGAGGGATC CGACGGTCCC GCGGAAGGGC 60 GGCAATGTTG ATTATGGTTG TGGTCACAGG TCTTCCGCCC GGATGCCTAC CTCCGCTCTG 120 TCGTCGATCA CGAAGTGCTA CACTTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:95: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE, nucleic acid 35 (C) STRANDEDNESS- single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA SEQ ID NO:92: AGTCCGAGGT TCGAGGATTA CAGTAGGACG 60 AGTAATGTCT CCGATGCGCA CAATAACCGG 120 AGACCTTCGA GA 162 ID NO:93 Printed from Mimosa (xi) SEQUENCE DESCRIPTION: SEQ ID NO:95: TCTCACTCCT CGAGAGCCAG TANGCAGGGC GGCCGGGGTG TTGCCCCTGA GTTTGGGGCG 60 AGCGTTTTGG GTNGTGGTTG TGGTAGCGCC ACTTATTACA CGAACTCCAC CAGCTGCAAG 120 GATGCTATGG GCCACAACTA CTCGTCTAGA ATCGAAGGTC GCGNTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:96: (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:96: TCTCACTCCT CGAGATGGTG CGAGAAGCAC AAGTTTACGG CTGCGCGTTG CAGCGCGGGG 60 GCGGGTTTTG AGAGGGANGC CAGCCGTCCG CCCCAGCCTG CCCACCGGGA TAATACCAAC 120 CGTAATGCNT NTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:97: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- Single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO-97: TCTCACTCCT CGAGTTTTCA GGTGTACCCG GACCATGGTC TGGAGAGGCA TGCTTTGGAC 60 GGGACGGGTC CGCTTTACGC CATGCCCGGC CGCTGGATTA GGGCGCGTCC GCAGAACAGG 120 GACCGCCAGT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO-98: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 159 base pairs (B) TYPE: nucleic acid 25 (C) STRANDEDNESS- single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:98: TCTCACTCCT CGAGCAGGTG TACGGACAAC GAGCAGTGCC CCGATACCGG GANTAGGTCT 60 CGTTCCGTTA GTAACGCCAG GTACTTTTCG AGCAGGTTGC TCAAGACTCA CGCCCCCCAT 120 30 CGCCCTTCTA GAATCGAAGG TCGCGCTAGA CCTTCGAGA 159 (2) INFORMATION FOR SEQ ID NO:99: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:99: Printed from Mimosa WO 98/51325 PCT/US98/10088 TCTCACTCCT CGAGTGCCAG GGATAGCGGG CCTGCGGAGG ATGGGTCCCG CGCCGTCCGG 60 TTGAACGGGG TTGAGAACGC CAACACTAGG AAGTCCTCCC GCAGTAACCC GCGGGGTAGG 120 CGCCATCCCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO-100: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:100: TCTCACTCCT CGAGTTCCGC CGATGCGGAG AAGTGTGCGG GCAGTCTGTT GTGGTGGGGT 60 AGGCAGAACA ACTCCGGTTG TGGTTCGCCC ACGAAGAAGC ATCTGAAGCA CCGCAATCGC 120 AGTCAGACCT CCTCTTCGTC CCACTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:101: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH- 162 base pairs (B) TYPE nucleic acid 15 (C) STRANDEDNESS- single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:101: TCTCACTCCT CGAGACCGAA GAACGTGGCC GATGCTTATT CGTCTCAGGA CGGGGCGGCG 60 GCCGAGGAGA CGTCTCACGC CAGTAATGCC GCGCGGAAGT CCCCTAAGCA CAAGCCCTTG 120 20 AGGCGGCCTT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:102: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS• single (D) TOPOLOGY: linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:102: TCTCACTCCT CGAGAGGCAG TACGGGGACG GCCGGCGGCG AGCGTTCCGG GGTGCTCAAC 60 CTGCACACCA GGGATAACGC CAGCGGCAGC GGTTTCAAAC CGTGGTACCC TTCGAATCGG 120 GGTCACAAGT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:103: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH- 162 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:103: TCTCACTCCT CGAGGTGGGG GTGGGAGAGG AGTCCGTCCG ACTACGATTC TGATATGGAC 60 TTGGGGGCGA GGAGGTACGC CACCCGCACC CACCGCGCGC CCCCTCGCGT CTTGAAGGCT 120 Printed from Mimosa CCCCTGCCCT CTAGAATCGA AGGTCGCGCT AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID NO:104: (I) SEQUENCE CHARACTERISTICS.

(A) LENGTH. 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:104 TCTCACTCCT CGAGGCACTG GAAGTGCGAG GGCTCTCAGG CTGCCTACGG GGACAAGGAT 60 ATCGGGAGGT CCAGGGGTTG TGGTTCCATT ACAAAGAATA ACACTAATCA CGCCCATCCT 120 10 AGCCACGGCG CCGTTGCTAA GATCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:105: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 162 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:105 TCTCACTCCT CGAGCCGCGA GGAGGCGAAC CGGAGTCGCT TTTGGGACGC CACCCACCTG GACCCTAACT CTAGAATCGA AGGTCGCGCT TGGGACGGCT ATAAGAGGGA GATGAGCCAC 60 TCCCGCCCTC GCCGCCCCGC TAACTCTGGT 120 AGACCTTCGA GA 162 (2) INFORMATION FOR SEQ ID N0:106. (l) SEQUENCE CHARACTERISTICS: (A) LENGTH 177 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:106: TCTCACTCNT CGAGAGAGTT CGCGGAGAGG AGGTTGTGGG GGTGTGATGA CCTGAGTTGG 60 CGTCTCGACG CGGAGGGTTG TGGTCCCACT CCGAGCAATC GGGCCGTCAA GCATCGCAAG 120 CCCCGCCCAC GCTCCCCCGC ACTCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID NO:107: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS single (D) TOPOLOGY, linear (li) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:107.

TCTCACTCNT NGAGTGATCA CGCGTTGGGG ACGAATCTGA GGTCTGACAA TGCCAAGGAG 60 CCGGGTGATT ACAACTGTTG TGGTAACGGG AACTCTACCG GGCGAAAGGT TTTTAACCGT 120 AGGCGCCCCT CCGCCATCCC CANTTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 Printed from Mimosa (2) INFORMATION FOR SEQ ID NO:108. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 177 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO.108: TCTCACTCCT CGAGGCATAT TTCTGAGTAT AGCTTTGCGA ATTCCCACTT GATGGGTGGC 60 GAGTCCAAGC GGAAGGGTTG TGGTATTAAC GGCTCCTTTT CTCCCACTTG TCCCCGCTCC 120 CCCACCCCAG CCTTCCGCCG CACCTCTAGA ATCGAAGGTC GCGCTAGACC TTCGAGA 177 (2) INFORMATION FOR SEQ ID N0:109 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 158 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID N0:109: TCTCACTCCT CGAGCCGGGA GAGCGGGATG TGGGGTAGTT GGTGGCGTGG TCACAGGTTG 60 AATTCCACGG GGGGTAACGC CAACATGAAT GCTAGTCTGC CCCCCGACCC CCCTGTTTCC 120 ACTCCGTCTA GAATCGAAGG TCGCGCTAGA CCTTCGAG 158 (2) INFORMATION FOR SEQ ID NO.110: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 708 amino acids (B) TYPE: amino acid (C) STRANDEDNESS- (D) TOPOLOGY- unknown (il) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:110: Met Gly Met Ser Lys Ser His Ser Phe Phe Gly Tyr Pro Leu Ser lie 1 Phe Phe lie Val Val Asn Glu Phe Cys Glu Arg Phe Ser Tyr Tyr Gly Met Arg Ala 35 lie Leu lie Leu Tyr 40 Phe Thr Asn Phe lie 45 Ser Trp Asp Asp Asn 50 Leu Ser Thr Ala lie 55 Tyr His Thr Phe Val 60 Ala Leu Cys Tyr Leu Thr Pro lie Leu Gly Ala Leu lie Ala Asp Ser Trp Leu Gly Lys 65 70 75 80 Phe Lys Thr lie Val Ser Leu Ser lie Val Tyr Thr lie Gly Gin Ala 85 90 95 Val Thr Ser Val 100 Ser Ser lie Asn Asp 105 Leu Thr Asp His Asn 110 His Asp Gly Thr Pro 115 Asp Ser Leu Pro Val 120 His Val Val Leu Ser 125 Leu lie Gly Leu Ala 13 0 Leu lie Ala Leu Gly 135 Thr Gly Gly lie Lys 140 Pro Cys Val Ser Ala Phe Gly Gly Asp Gin Phe Glu Glu Gly Gin Glu Lys Gin Arg Asn 145 150 155 160 Arg Phe Phe Ser lie Phe Tyr Leu Ala lie Asn Ala Gly Ser Leu Leu 165 170 175 Printed from Mimosa Ser Thr lie lie Thr Pro Met Leu Arg Val Gin Gin Cys Gly lie His 180 185 190 Ser Lys Gin Ala Cys Tyr Pro Leu Ala Phe Gly Val Pro Ala Ala Leu 195 200 205 Met Ala Val Ala Leu lie Val Phe Val Leu Gly Ser Gly Met Tyr Lys 210 215 220 Lys Phe Lys Pro Gin Gly Asn lie Met Gly Lys Val Ala Lys Cys lie 5 225 230 235 240 Gly Phe Ala lie Lys Asn Arg Phe Arg His Arg Ser Lys Ala Phe Pro 245 250 255 Lys Arg Glu His Trp Leu Asp Trp Ala Lys Glu Lys Tyr Asp Glu Arg 260 265 270 Leu lie Ser Gin lie Lys Met Val Thr Arg Val Met Phe Leu Tyr lie 275 280 285 Pro Leu Pro Met Phe Trp Ala Leu Phe Asp Gin Gin Gly Ser Arg Trp 290 295 300 in Thr Leu Gin Ala Thr Thr Met Ser Gly Lys lie Gly Ala Leu Glu lie 305 310 315 320 Gin Pro Asp Gin Met Gin Thr Val Asn Ala lie Leu lie Val lie Met 325 330 335 Val Pro lie Phe Asp Ala Val Leu Tyr Pro Leu lie Ala Lys Cys Gly 340 345 350 Phe Asn Phe Thr Ser Leu Lys Lys Met Ala Val Gly Met Val Leu Ala 355 360 365 Ser Met Ala Phe Val Val Ala Ala lie Val Gin Val Glu lie Asp Lys 15 370 375 380 Thr Leu Pro Val Phe Pro Lys Gly Asn Glu Val Gin lie Lys Val Leu 385 390 395 400 Asn lie Gly Asn Asn Thr Met Asn lie Ser Leu Pro Gly Glu Met Val 405 410 415 Thr Leu Gly Pro Met Ser Gin Thr Asn Ala Phe Met Thr Phe Asp Val 420 425 430 Asn Lys Leu Thr Arg lie Asn lie Ser Ser Pro' Gly Ser Pro Val Thr 435 440 445 Ala Val Thr Asp Asp Phe Lys Gin Gly Gin Arg His Thr Leu Leu Val 450 455 460 Trp Ala Pro Asn His Tyr Gin Val Val Lys Asp Gly Leu Asn Gin Lys 465 470 475 480 Pro Glu Lys Gly Glu Asn Gly lie Arg Phe Val Asn Thr Phe Asn Glu 485 490 495 Leu lie Thr lie Thr Met Ser Gly Lys Val Tyr Ala Asn lie Ser Ser 500 505 510 Tyr Asn Ala Ser Thr Tyr Gin Phe Phe Pro Ser Gly lie Lys Gly Phe 25 515 520 525 Thr lie Ser Ser Thr Glu lie Pro Pro Gin Cys Gin Pro Asn Phe Asn 530 535 540 Thr Phe Tyr Leu Glu Phe Gly Ser Ala Tyr Thr Tyr lie Val Gin Arg 545 550 555 560 Lys Asn Asp Ser Cys Pro Glu Val Lys Val Phe Glu Asp lie Ser Ala 565 570 575 Asn Thr Val Asn Met Ala Leu Gin lie Pro Gin Tyr Phe Leu Leu Thr 580 585 590 Cys Gly Glu Val Val Phe Ser Val Thr Gly Leu Glu Phe Ser Tyr Ser 595 600 605 Gin Ala Pro Ser Asn Met Lys Ser Val Leu Gin Ala Gly Trp Leu Leu 610 615 620 Thr Val Ala Val Gly Asn lie lie Val Leu lie Val Ala Gly Ala Gly 625 630 635 640 Gin Phe Ser Lys Gin Trp Ala Glu Tyr lie Leu Phe Ala Ala Leu Leu 645 650 655 Leu Val Val Cys Val Val Phe Ala lie Met Ala Arg Phe Tyr Thr Tyr 35 660 665 670 lie Asn Pro Ala Glu lie Glu Ala Gin Phe Asp Glu Asp Glu Lys Lys 675 680 685 Asn Arg Leu Glu Lys Ser Asn Pro Tyr Phe Met Ser Gly Ala Asn Ser 690 695 700 Printed from Mimosa PCT /US98/10088 Gin Lys Gin Met 705 (2) INFORMATION FOR SEQ ID NO:111: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs 5 (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO.Ill: TCCGGACTCT CATAAGCGCC GG 22 (2) INFORMATION FOR SEQ ID NO-112: (l) SEQUENCE CHARACTERISTICS- (A) LENGTH 22 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (li) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION. SEQ ID NO.112: ACAACGGGCC AGAAAGAGCG AG 22 (2) INFORMATION FOR SEQ ID NO.113: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs .

(B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (il) MOLECULE TYPE- DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:113- (2) INFORMATION FOR SEQ ID NO:114■ (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS single (D) TOPOLOGY, linear ACACCACCCC AATCGGAGCT AC 22 (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:114: TCAGAATCCG TGCACTGGCC AA 22 (2) INFORMATION FOR SEQ ID NO:115: (i) SEQUENCE CHARACTERISTICS (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear 165 Printed from Mimosa (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:115: GCCCTATTCA TACCACCGGA GT (2) INFORMATION FOR SEQ ID NO:116: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS. single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA 10 (xi) SEQUENCE DESCRIPTION: SEQ ID NO:116: CATCAGTCCT ACCGCCGAAA AG (2) INFORMATION FOR SEQ ID NO:117: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid 15 (C) STRANDEDNESS- single (D) TOPOLOGY, linear (li) MOLECULE TYPE: DNA (Xi) SEQUENCE DESCRIPTION SEQ ID NO:117: CGTATAGCTA TTGTGAGCGA TG 20 (2) INFORMATION FOR SEQ ID NO.118 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: Single (D) TOPOLOGY: linear (il) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:118: ACGCGCGGAA CGAGCAGTAC CA (2) INFORMATION FOR SEQ ID NO.119: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:119: CCATAATGAT CCCCGTCACT AT (2) INFORMATION FOR SEQ ID NO:120- (i) SEQUENCE CHARACTERISTICS-(A) LENGTH. 22 base pairs Printed from Mimosa (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID N0:120.

AGACACCCCT TAGCCGTCGT AG 22 (2) INFORMATION FOR SEQ ID NO:121- (l) SEQUENCE CHARACTERISTICS (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single 10 (D) TOPOLOGY- linear (li) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:121 AGCTCCGTGA CCTTAGTCAT AA 22 (2) INFORMATION FOR SEQ ID NO.122: (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: Single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:122: TGCACAGCTC AGCGCCGCAC CA 22 (2) INFORMATION FOR SEQ ID NO-.123: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid 25 (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:123: ACGGGTCATC AGCGCCGCAC CA 22 (2) INFORMATION FOR SEQ ID NO:124: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid s (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO.124: TGTCACCCCC CTCCCCGGAC TT 22 167 Printed from Mimosa (2) INFORMATION FOR SEQ ID NO:125. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY: linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:125-ACTCGCAATT ATTGGCGCTC GA (2) INFORMATION FOR SEQ ID NO: 126 in (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE- DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO.126.

GTCTTCTCAA CCTTATCCTG CG (2) INFORMATION FOR SEQ ID NO.127: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single 20 (D) TOPOLOGY: linear (li) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO 127: AAAGCCCCCT GCTAAACTCC CA (2) INFORMATION FOR SEQ ID NO:128: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: Single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:128: CTGCGTCTGC CACGTCGTCA TC (2) INFORMATION FOR SEQ ID NO 12 9- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs (B) TYPE, nucleic acid 35 (C) STRANDEDNESS: single (D) TOPOLOGY, linear (n) MOLECULE TYPE: DNA Printed from Mimosa PCT /US98/10088 (xi) SEQUENCE DESCRIPTION: SEQ ID NO.129: GTTAAAAGAG GGCAAGCTCG GA 22 (2) INFORMATION FOR SEQ ID N0:130: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO.130-10 CCGAGTTCTT GATGTCCTCC AT 22 (2) INFORMATION FOR SEQ ID NO:131: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:131-TCCAATGCCT GTACCACGGA TG 22 (2) INFORMATION FOR SEQ ID NO:132: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:132: TCGCAACCGA TATCGTGCTT AT 22 (2) INFORMATION FOR SEQ ID NO:133: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single 30 (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:133: TGCATACACT GCTTGGAGCC CT 22 (2) INFORMATION FOR SEQ ID NO:134. (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single Printed from Mimosa (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:134 GAAATCTCAC TAGTAGTCCG CC (2) INFORMATION FOR SEQ ID NO 13 5- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE- DNA (Xl) SEQUENCE DESCRIPTION- SEQ ID NO 135: GCGGGCAAGA CAGTCCAATT CC (2) INFORMATION FOR SEQ ID NO:136- (I) SEQUENCE CHARACTERISTICS: 3_5 (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:136 20 GAGCTCCAAT TCCACGACGA CC (2) INFORMATION FOR SEQ ID NO:137: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO 137 GGTTGCCATG CGTTCAAACT AC (2) INFORMATION FOR SEQ ID NO:138 (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:138 TCCCGCGGGG ACAAACCCGA AT (2) INFORMATION FOR SEQ ID NO:139: Printed from Mimosa (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS. single (D) TOPOLOGY- linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:139: CTGCTAGTCT TATCATTCCC CA 22 (2) INFORMATION FOR SEQ ID NO.14 0- (I) SEQUENCE CHARACTERISTICS (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:140: CTATCGACAC TATAGGGCCT AC 22 (2) INFORMATION FOR SEQ ID NO-141: (l) SEQUENCE CHARACTERISTICS (A) LENGTH 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY- linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:141: TACCCTTGTA ACCCACACTA GG 22 (2) INFORMATION FOR SEQ ID NO 142: (i) SEQUENCE CHARACTERISTICS 25 (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:142: TTCTTCTGAA TAGACCGGCC GA 22 (2) INFORMATION FOR SEQ ID NO:143: (i) SEQUENCE CHARACTERISTICS (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:143: Printed from Mimosa CCACCACCCT TAACCCGACA AT (2) INFORMATION FOR SEQ ID NO:144: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 22 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO.144 AGGGGGAGAC TTGTTCACAA AC (2) INFORMATION FOR SEQ ID NO.145: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ll) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:145 CGGCTCATAC CACCGAAAGC TA (2) INFORMATION FOR SEQ ID NO.146- (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs 20 <B ) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:146 ATCGTCCTAC TGTAATCCTC GA (2) INFORMATION FOR SEQ ID NO:147: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS. single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO.147 GACACACTAC TCAGGTCCAC CT (2) INFORMATION FOR SEQ ID NO-148: (i) SEQUENCE CHARACTERISTICS: 35 (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA 22 22 22 22 Printed from Mimosa (xi) SEQUENCE DESCRIPTION: SEQ ID NO:148 CCATAATCAA CATTGCCGCC CT 22 (2) INFORMATION FOR SEQ ID NO:149- (I) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS- Single (D) TOPOLOGY linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO 149-10 CAAAACGCTC GCCCCAAACT CA 22 (2) INFORMATION FOR SEQ ID NO:150: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear , V (II) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO 150: GTAAACTTGT GCTTCTCGCA CC 22 (2) INFORMATION FOR SEQ ID NO:151- U) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (li) MOLECULE TYPE: DNA (xi) SEQXJENCE DESCRIPTION: SEQ ID NO:151.

CCATGGTCCG GGTACACCTG AA 22 (2) INFORMATION FOR SEQ ID NO:152: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single 30 (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:152: GTTACTAACG GAACGAGACC TA 22 (2) INFORMATION FOR SEQ ID NO.153: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE nucleic acid (C) STRANDEDNESS: single Printed from Mimosa (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:153: TGTTGGCGTT CTCAACCCCG TT (2) INFORMATION FOR SEQ ID NO:154: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (li) MOLECULE TYPE- DNA (xi) SEQUENCE DESCRIPTION SEQ ID NO:154: ACAACCGGAG TTGTTCTGCC TA (2) INFORMATION FOR SEQ ID NO.155: (I) SEQUENCE CHARACTERISTICS: 15 (A) LENGTH. 22 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS single (D) TOPOLOGY, linear (ii) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:155: 20 TAAGCATCGG CCACGTTCTT CG (2) INFORMATION FOR SEQ ID NO:156: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (II) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:156: TTATCCCTGG TGTGCAGGTT GA (2) INFORMATION FOR SEQ ID NO:157. (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:157: TATCAGAATC GTAGTCGGAC GG (2) INFORMATION FOR SEQ ID NO:158: Printed from Mimosa (l) SEQUENCE CHARACTERISTICS- (A) LENGTH. 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE- DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:158: CTTTGTAATG GAACCACAAC CC 22 (2) INFORMATION FOR SEQ ID NO:159 (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 22 base pairs TO (B) TYPE: nucleic acid (C) STRANDEDNESS single (D) TOPOLOGY, linear (ii) MOLECULE TYPE- DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:159: CGGTGGCTCA TCTCCCTCTT AT 22 (2) INFORMATION FOR SEQ ID NO:160. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 22 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS- single (D) TOPOLOGY linear (ll) MOLECULE TYPE- DNA (Xl) SEQUENCE DESCRIPTION- SEQ ID NO:16 0: ATCAGACTGG CTGGGACCAC AA 22 (2) INFORMATION FOR SEQ ID NO: 161. (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE. DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:161: CACAACCTCC TCTCCGCGAA CT 22 (2) INFORMATION FOR SEQ ID NO:162: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE, nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO. 162 Printed from Mimosa AGATTCGTCC CCAACGCGTG AT (2) INFORMATION FOR SEQ ID NO:163: (l) SEQUENCE CHARACTERISTICS (A) LENGTH: 22 base pairs (B)- TYPE: nucleic acid (C) STRANDEDNESS: Single (D) TOPOLOGY linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION: SEQ ID NO:163 GGGAATTCGC AAAGCTATAC TC !0 (2) INFORMATION FOR SEQ ID NO.164 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 22 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (il) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION. SEQ ID NO:164 CCCCGTGGAA TTCAACCTGT GA (2) INFORMATION FOR SEQ ID NO:165. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 17 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY linear (ii) MOLECULE TYPE: DNA (xi) SEQUENCE DESCRIPTION- SEQ ID NO:165 GTCGTCTTTC CAGACGT (2) INFORMATION FOR SEQ ID NO 166. (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 base pairs (B) TYPE- nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY, linear (ii) MOLECULE TYPE: DNA (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:166 CTTGCATGCC TGCAGGTCGA C (2) INFORMATION FOR SEQ ID NO:167: (l) SEQUENCE CHARACTERISTICS: 35 (A) LENGTH: 37 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown Printed from Mimosa WO 98/51325 PCT/US98/10088 (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO-167: Arg lie Ala Gly Leu Pro Trp Tyr Arg Cys Arg Thr Val Ala Phe Glu 10 15 Thr Gly Met Gin Asn Thr Gin Leu Cys Ser Thr lie Val Gin Leu Ser 25 30 Phe Thr Pro Glu Glu 35 (2) INFORMATION FOR SEQ ID NO:168- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.168: Arg Glu Phe Ala Glu Arg Arg Leu Trp Gly Cys Asp Asp Leu Ser Trp 10 15 Arg Leu Asp Ala Glu Gly Cys Gly Pro Thr Pro Ser Asn Arg Ala Val 25 30 Lys His Arg Lys Pro Arg Pro Arg Ser Pro Ala Leu 35 40 (2) INFORMATION FOR SEQ ID NO:169: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 41 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO 169: Ser Gly Ser His Ser Gly Gly Met Asn Arg Ala Tyr Gly Asp Val Phe 10 15 Arg Glu Leu Arg Asp Arg Trp Tyr Ala Thr Ser His His Thr Arg Pro 25 30 Thr Pro Gin Leu Pro Arg Gly Pro Asn 35 40 (2) INFORMATION FOR SEQ ID NO:170: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 20 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:170 Ser Thr Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp 10 15 Ser Asp Ser Asp 20 Printed from Mimosa (2) INFORMATION FOR SEQ ID NO:171: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 29 ammo acids (B) TYPE ammo acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO 171 Ser Thr Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp 10 15 Ser Asp Ser Asp Thr Asn Ala Lys His Ser Ser His Asn 25 (2) INFORMATION FOR SEQ ID NO:172. (i) SEQUENCE CHARACTERISTICS (A) LENGTH 19 ammo acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ll) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION- SEQ ID NO:172: Thr Asn Ala Lys His Ser Ser His Asn Arg Arg Leu Arg Thr Arg Ser 10 15 Arg Pro Asn (2) INFORMATION FOR SEQ ID NO:173: (I) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 9 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (II) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:173: Thr Asn Ala Lys His Ser Ser His Asn 1 5 (2) INFORMATION FOR SEQ ID NO:174: (i) SEQUENCE CHARACTERISTICS: 30 (A) LENGTH: 14 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:174.

Ser Ser His Asn Arg Arg Leu Arg Thr Arg Ser Arg Pro Asn 15 10 (2) INFORMATION FOR SEQ ID NO.175.

Printed from Mimosa (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:175: Arg Arg Leu Arg Thr Arg Ser Arg Pro Asn 15 10 (2) INFORMATION FOR SEQ ID NO-176: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 708 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO 176 Met Gly Met Ser Lys Ser His Ser Phe Phe Gly Tyr Pro Leu Ser lie 1 Phe Phe lie Val 20 Val Asn Glu Phe Cys 25 Glu Arg Phe Ser Tyr 30 Tyr Gly Met Arg Ala lie Leu lie Leu Tyr Phe Thr Asn Phe lie Ser Trp Asp 40 45 Asp Asn Leu Ser Thr Ala lie Tyr His Thr Phe Val Ala Leu Cys Tyr 50 55 60 Leu Thr Pro lie Leu Gly Ala Leu lie Ala Asp Ser Trp Leu Gly Lys 65 70 75 80 Phe Lys Thr lie Val 85 Ser Leu Ser lie Val 90 Tyr Thr lie Gly Gin 95 Ala Val Thr Ser Val Ser Ser lie Asn Asp Leu Thr Asp His Asn His Asp 100 105 110 Gly Thr Pro Asp Ser Leu Pro Val His Val Val Leu Ser Leu lie Gly 115 120 125 Leu Ala 130 Leu lie Ala Leu Gly 135 Thr Gly Gly lie Lys 140 Pro Cys Val Ser Ala Phe Gly Gly Asp Gin Phe Glu Glu Gly Gin Glu Lys Gin Arg Asn 145 150 155 160 Arg Phe Phe Ser lie 165 Phe Tyr Leu Ala lie 170 Asn Ala Gly Ser Leu 175 Leu Ser Thr lie lie 180 Thr Pro Met Leu Arg 185 Val Gin Gin Cys Gly 190 lie His Ser Lys Gin 195 Ala Cys Tyr Pro Leu 200 Ala Phe Gly Val Pro 205 Ala Ala Leu Met Ala 210 Val Ala Leu lie Val 215 Phe Val Leu Gly Ser 220 Gly Met Tyr Lys Lys Phe Lys Pro Gin Gly Asn lie Met Gly Lys Val Ala Lys Cys lie 225 230 235 240 Gly Phe Ala lie Lys Asn Arg Phe Arg His Arg Ser Lys Ala Phe Pro 245 250 255 Lys Arg Glu His 260 Trp Leu Asp Trp Ala 265 Lys Glu Lys Tyr Asp 270 Glu Arg Leu lie Ser Gin lie Lys Met Val Thr Arg Val Met Phe Leu Tyr lie 275 280 285 Pro Leu 290 Pro Met Phe Trp Ala 295 Leu Phe Asp Gin Gin 300 Gly Ser Arg Trp Thr Leu Gin Ala Thr Thr Met Ser Gly Lys lie Gly Ala Leu Glu lie 305 310 315 320 Gin Pro Asp Gin Met Gin Thr Val Asn Ala lie Leu lie Val lie Met Printed from Mimosa 325 330 335 Val Pro lie Phe 340 Asp Ala Val Leu Tyr 345 Pro Leu lie Ala Lys 350 Cys Gly Phe Asn Phe 355 Thr Ser Leu Lys Lys 360 Met Ala Val Gly Met 365 Val Leu Ala Ser Met 370 Ala Phe Val Val Ala 375 Ala lie Val Gin Val 380 Glu lie Asp Lys Thr Leu Pro Val Phe Pro Lys Gly Asn Glu Val Gin lie Lys Val Leu 385 390 395 400 Asn lie Gly Asn Asn Thr Met Asn lie Ser Leu Pro Gly Glu Met Val 405 410 415 Thr Leu Gly Pro Met Ser Gin Thr Asn Ala Phe Met Thr Phe Asp Val 420 425 430 Asn Lys Leu 435 Thr Arg lie Asn lie 440 Ser Ser Pro Gly Ser 445 Pro Val Thr Ala Val Thr Asp Asp Phe Lys Gin Gly Gin Arg His Thr Leu Leu Val 450 455 460 Trp Ala Pro Asn His Tyr Gin Val Val Lys Asp Gly Leu Asn Gin Lys 465 470 475 480 Pro Glu Lys Gly Glu Asn Gly lie Arg Phe Val Asn Thr Phe Asn Glu 485 490 495 Leu lie Thr lie Thr Met Ser Gly Lys Val Tyr Ala Asn lie Ser Ser 500 505 510 Tyr Asn Ala 515 Ser Thr Tyr Gin Phe 520 Phe Pro Ser Gly lie 525 Lys Gly Phe Thr lie 530 Ser Ser Thr Glu lie 535 Pro Pro Gin Cys Gin 540 Pro Asn Phe Asn Thr Phe Tyr Leu Glu Phe Gly Ser Ala Tyr Thr Tyr lie Val Gin Arg 545 550 555 560 Lys Asn Asp Ser Cys Pro Glu Val Lys Val Phe Glu Asp lie Ser Ala 565 570 575 Asn Thr Val Asn 580 Met Ala Leu Gin lie 585 Pro Gin Tyr Phe Leu 590 Leu Thr Cys Gly Glu Val Val Phe Ser Val Thr Gly Leu Glu Phe Ser Tyr Ser 595 600 605 Gin Ala 610 Pro Ser Asn Met Lys 615 Ser Val Leu Gin Ala 620 Gly Trp Leu Leu Thr Val Ala Val Gly Asn lie lie Val Leu lie Val Ala Gly Ala Gly 625 630 635 640 Gin Phe Ser Lys Gin 645 Trp Ala Glu Tyr lie 650 Leu Phe Ala Ala Leu 655 Leu Leu Val Val Cys 660 Val Val Phe Ala lie 665 Met Ala Arg Phe Tyr 670 Thr Tyr lie Asn Pro 675 Ala Glu lie Glu Ala 680 Gin Phe Asp Glu Asp 685 Glu Lys Lys Asn Arg 690 Leu Glu Lys Ser Asn 695 Pro Tyr Phe Met Ser 700 Gly Ala Asn Ser Gin Lys Gin Met 705 (2) INFORMATION FOR SEQ ID NO:177: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 3345 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ii) MOLECULE TYPE: DNA (ix) FEATURE: (A) NAME/KEY: Coding Sequence (B) LOCATION: 88...2583 (D) OTHER INFORMATION: Printed from Mimosa (xi) SEQUENCE DESCRIPTION: SEQ ID NO.177: GAATTCCGTC TCGACCACTG AATGGAAGAA AAGGACTTTT AACCACCATT TTGTGACTTA 60 CAGAAAGGAA TTTGAATAAA GAAAACT ATG ATA CTT CAG GCC CAT CTT CAC TCC 114 Met lie Leu Gin Ala His Leu His Ser 1 5 CTG TGT CTT CTT ATG CTT TAT TTG GCA ACT GGA TAT GGC CAA GAG GGG 162 Leu Cys Leu Leu Met Leu Tyr Leu Ala Thr Gly Tyr Gly Gin Glu Gly 10 15 20 25 AAG TTT AGT GGA CCC CTG AAA CCC ATG ACA TTT TCT ATT TAT GAA GGC 210 Lys Phe Ser Gly Pro Leu Lys Pro Met Thr Phe Ser lie Tyr Glu Gly 30 35 40 CAA GAA CCG AGT CAA ATT ATA TTC CAG TTT AAG GCC AAT CCT CCT GCT 258 ]_0 Gln Pro Ser I^e Ile p^e G^n p^e Lys Ala Asn Pro Pro Ala 45 50 55 GTG ACT TTT GAA CTA ACT GGG GAG ACA GAC AAC ATA TTT GTG ATA GAA 306 Val Thr Phe Glu Leu Thr Gly Glu Thr Asp Asn lie Phe Val lie Glu 60 65 70 CGG GAG GGA CTT CTG TAT TAC AAC AGA GCC TTG GAC AGG GAA ACA AGA 354 Arg Glu Gly Leu Leu Tyr Tyr Asn Arg Ala Leu Asp Arg Glu Thr Arg 15 75 80 85 TCT ACT CAC AAT CTC CAG GTT GCA GCC CTG GAC GCT AAT GGA ATT ATA 4 02 Ser Thr His Asn Leu Gin Val Ala Ala Leu Asp Ala Asn Gly lie lie 90 95 100 105 GTG GAG GGT CCA GTC CCT ATC ACC ATA GAA GTG AAG GAC ATC AAC GAC 450 Val Glu Gly Pro Val Pro lie Thr lie Glu Val Lys Asp lie Asn Asp 110 115 120 AAT CGA CCC ACG TTT CTC CAG TCA AAG TAC GAA GGC TCA GTA AGG CAG 498 Asn Arg Pro Thr Phe Leu Gin Ser Lys Tyr Glu Gly Ser Val Arg Gin 125 130 135 AAC TCT CGC CCA GGA AAG CCC TTC TTG TAT GTC AAT GCC ACA GAC CTG 546 Asn Ser Arg Pro Gly Lys Pro Phe Leu Tyr Val Asn Ala Thr Asp Leu 140 145 150 GAT GAT CCG GCC ACT CCC AAT GGC CAG CTT TAT TAC CAG ATT GTC ATC 5 94 Asp Asp Pro Ala Thr Pro Asn Gly Gin Leu Tyr Tyr Gin lie Val lie 155 160 165 CAG CTT CCC ATG ATC AAC AAT GTC ATG TAC TTT CAG ATC AAC AAC AAA 642 Gin Leu Pro Met lie Asn Asn Val Met Tyr Phe Gin lie Asn Asn Lys 170 175 180 185 ACG GGA GCC ATC TCT CTT ACC CGA GAG GGA TCT CAG GAA TTG AAT CCT 690 3 o Thr Gly Ala lie Ser Leu Thr Arg Glu Gly Ser Gin Glu Leu Asn Pro 190 195 200 GCT AAG AAT CCT TCC TAT AAT CTG GTG ATC TCA GTG AAG GAC ATG GGA 738 Ala Lys Asn Pro Ser Tyr Asn Leu Val lie Ser Val Lys Asp Met Gly 205 210 215 GGC CAG AGT GAG AAT TCC TTC AGT GAT ACC ACA TCT GTG GAT ATC ATA 786 Gly Gin Ser Glu Asn Ser Phe Ser Asp Thr Thr Ser Val Asp lie lie 35 220 225 230 GTG ACA GAG AAT ATT TGG AAA GCA CCA AAA CCT GTG GAG ATG GTG GAA 834 Val Thr Glu Asn lie Trp Lys Ala Pro Lys Pro Val Glu Met Val Glu 235 240 245 Printed from Mimosa AAC TCA ACT GAT CCT CAC CCC ATC AAA ATC ACT CAG GTG CGG TGG AAT 882 Asn Ser Thr Asp Pro His Pro lie Lys lie Thr Gin Val Arg Trp Asn 250 255 260 265 GAT CCC GGT GCA CAA TAT TCC TTA GTT GAC AAA GAG AAG CTG CCA AGA 930 Asp Pro Gly Ala Gin Tyr Ser Leu Val Asp Lys Glu Lys Leu Pro Arg 270 275 280 TTC CCA TTT TCA ATT GAC CAG GAA GGA GAT ATT TAC GTG ACT CAG CCC 978 Phe Pro Phe Ser lie Asp Gin Glu Gly Asp lie Tyr Val Thr Gin Pro 285 290 295 TTG GAC CGA GAA GAA AAG GAT GCA TAT GTT TTT TAT GCA GTT GCA AAG 1026 Leu Asp Arg Glu Glu Lys Asp Ala Tyr Val Phe Tyr Ala Val Ala Lys 300 305 310 GAT GAG TAC GGA AAA CCA CTT TCA TAT CCG CTG GAA ATT CAT GTA AAA 1074 Asp Glu Tyr Gly Lys Pro Leu Ser Tyr Pro Leu Glu lie His Val Lys 315 320 325 GTT AAA GAT ATT AAT GAT AAT CCA CCT ACA TGT CCG TCA CCA GTA ACC 1122 Val Lys Asp lie Asn Asp Asn Pro Pro Thr Cys Pro Ser Pro Val Thr 330 335 340 345 GTA TTT GAG GTC CAG GAG AAT GAA CGA CTG GGT AAC AGT ATC GGG ACC 1170 Val Phe Glu Val Gin Glu Asn Glu Arg Leu Gly Asn Ser lie Gly Thr 350 355 360 CTT ACT GCA CAT GAC AGG GAT GAA GAA AAT ACT GCC AAC AGT TTT CTA 1218 Leu Thr Ala His Asp Arg Asp Glu Glu Asn Thr Ala Asn Ser Phe Leu 365 370 375 AAC TAC AGG ATT GTG GAG CAA ACT CCC AAA CTT CCC ATG GAT GGA CTC 1266 Asn Tyr Arg lie Val Glu Gin Thr Pro Lys Leu Pro Met Asp Gly Leu 380 385 390 TTC CTA ATC CAA ACC TAT GCT GGA ATG TTA CAG TTA GCT AAA CAG TCC 1314 Phe Leu lie Gin Thr Tyr Ala Gly Met Leu Gin Leu Ala Lys Gin Ser 395 400 405 TTG AAG AAG CAA GAT ACT CCT CAG TAC AAC TTA ACG ATA GAG GTG TCT 1362 Leu Lys Lys Gin Asp Thr Pro Gin Tyr Asn Leu Thr lie Glu Val Ser 410 415 420 425 GAC AAA GAT TTC AAG ACC CTT TGT TTT GTG CAA ATC AAC GTT ATT GAT 1410 Asp Lys Asp Phe Lys Thr Leu Cys Phe Val Gin lie Asn Val lie Asp 430 435 440 ATC AAT GAT CAG ATC CCC ATC TTT GAA AAA TCA GAT TAT GGA AAC CTG 1458 lie Asn Asp Gin lie Pro lie Phe Glu Lys Ser Asp Tyr Gly Asn Leu 445 450 455 ACT CTT GCT GAA GAC ACA AAC ATT GGG TCC ACC ATC TTA ACC ATC CAG 1506 Thr Leu Ala Glu Asp Thr Asn lie Gly Ser Thr lie Leu Thr lie Gin 460 465 470 GCC ACT GAT GCT GAT GAG CCA TTT ACT GGG AGT TCT AAA ATT CTG TAT 1554 Ala Thr Asp Ala Asp Glu Pro Phe Thr Gly Ser Ser Lys lie Leu Tyr 475 480 485 CAT ATC ATA AAG GGA GAC AGT GAG GGA CGC CTG GGG GTT GAC ACA GAT 1602 His lie lie Lys Gly Asp Ser Glu Gly Arg Leu Gly Val Asp Thr Asp 490 495 500 505 CCC CAT ACC AAC ACC GGA TAT GTC ATA ATT AAA AAG CCT CTT GAT TTT 1650 Pro His Thr Asn Thr Gly Tyr Val lie lie Lys Lys Pro Leu Asp Phe Printed from Mimosa WO 98/51325 PCT/US98/10088 510 515 520 GAA ACA GCA GCT GTT TCC AAC ATT GTG TTC AAA GCA GAA AAT CCT GAG 1698 Glu Thr Ala Ala Val Ser Asn lie Val Phe Lys Ala Glu Asn Pro Glu 525 530 535 CCT CTA GTG TTT GGT GTG AAG TAC AAT GCA AGT TCT TTT GCC AAG TTC 1746 Pro Leu Val Phe Gly Val Lys Tyr Asn Ala Ser Ser Phe Ala Lys Phe 540 545 550 ACG CTT ATT GTG ACA GAT GTG AAT GAA GCA CCT CAA TTT TCC CAA CAC 17 94 Thr Leu lie Val Thr Asp Val Asn Glu Ala Pro Gin Phe Ser Gin His 555 560 565 GTA TTC CAA GCG AAA GTC AGT GAG GAT GTA GCT ATA GGC ACT AAA GTG 184 2 Val Phe Gin Ala Lys Val Ser Glu Asp Val Ala lie Gly Thr Lys Val 570 575 580 585 GGC AAT GTG ACT GCC AAG GAT CCA GAA GGT CTG GAC ATA AGC TAT TCA 1890 Gly Asn Val Thr Ala Lys Asp Pro Glu Gly Leu Asp lie Ser Tyr Ser 590 595 600 CTG AGG GGA GAC ACA AGA GGT TGG CTT AAA ATT GAC CAC GTG ACT GGT 1938 Leu Arg Gly Asp Thr Arg Gly Trp Leu Lys lie Asp His Val Thr Gly 605 610 615 GAG ATC TTT AGT GTG GCT CCA TTG GAC AGA GAA GCC GGA AGT CCA TAT 1986 Glu lie Phe Ser Val Ala Pro Leu Asp Arg Glu Ala Gly Ser Pro Tyr 620 625 630 CGG GTA CAA GTG GTG GCC ACA GAA GTA GGG GGG TCT TCC TTA AGC TCT 2 034 Arg Val Gin Val Val Ala Thr Glu Val Gly Gly Ser Ser Leu Ser Ser 635 640 645 GTG TCA GAG TTC CAC CTG ATC CTT ATG GAT GTG AAT GAC AAC CCT CCC 2082 Val Ser Glu Phe His Leu lie Leu Met Asp Val Asn Asp Asn Pro Pro 650 655 660 665 AGG CTA GCC AAG GAC TAC ACG GGC TTG TTC TTC TGC CAT CCC CTC AGT 213 0 Arg Leu Ala Lys Asp Tyr Thr Gly Leu Phe Phe Cys His Pro Leu Ser 670 675 680 GCA CCT GGA AGT CTC ATT TTC GAG GCT ACT GAT GAT GAT CAG CAC TTA 2178 Ala Pro Gly Ser Leu lie Phe Glu Ala Thr Asp Asp Asp Gin His Leu 685 690 695 TTT CGG GGT CCC CAT TTT ACA TTT TCC CTC GGC AGT GGA AGC TTA CAA 2226 Phe Arg Gly Pro His Phe Thr Phe Ser Leu Gly Ser Gly Ser Leu Gin 700 705 710 AAC GAC TGG GAA GTT TCC AAA ATC AAT GGT ACT CAT GCC CGA CTG TCT 2274 Asn Asp Trp Glu Val Ser Lys lie Asn Gly Thr His Ala Arq Leu Ser i 715 720 725 ACC AGG CAC ACA GAC TTT GAG GAG AGG GCG TAT GTC GTC TTG ATC CGC 2322 Thr Arg His Thr Asp Phe Glu Glu Arg Ala Tyr Val Val Leu lie Arg 730 735 740 745 ATC AAT GAT GGG GGT CGG CCA CCC TTG GAA GGC ATT GTT TCT TTA CCA 2370 lie Asn Asp Gly Gly Arg Pro Pro Leu Glu Gly lie Val Ser Leu Pro 750 755 760 GTT ACA TTC TGC AGT TGT GTG GAA GGA AGT TGT TTC CGG CCA GCA GGT 2418 Val Thr Phe Cys Ser Cys Val Glu Gly Ser Cys Phe Arg Pro Ala Gly 765 770 775 Printed from Mimosa CAC CAG ACT GGG ATA CCC ACT GTG GGC ATG GCA GTT GGT ATA CTG CTG 2466 His Gin Thr Gly lie Pro Thr Val Gly Met Ala Val Gly lie Leu Leu 780 785 790 ACC ACC CTT CTG GTG ATT GGT ATA ATT TTA GCA GTT GTG TTT ATC CGC 2514 Thr Thr Leu Leu Val lie Gly lie lie Leu Ala Val Val Phe lie Arg 795 800 805 ATA AAG AAG GAT AAA GGC AAA GAT AAT GTT GAA AGT GCT CAA GCA TCT 2562 lie Lys Lys Asp Lys Gly Lys Asp Asn Val Glu Ser Ala Gin Ala Ser 810 815 820 825 GAA GTC AAA CCT CTG AGA AGC TGAATTTGAA AAGGAATGTT TGAATTTATA TAGC 2617 Glu Val Lys Pro Leu Arg Ser 830 AAGTGCTATT TCAGCAACAA CCATCTCATC CTATTACTTT TCATCTAACG TGCATTATAA 2677 TTTTTTAAAC AGATATTCCC TCTTGTCCTT TAATATTTGC TAAATATTTC TTTTTTGAGG 2737 TGGAGTCTTG CTCTGTCGCC CAGGCTGGAG TACAGTGGTG TGATCCCAGC TCACTGCAAC 2797 CTCCGCCTCC TGGGTTCACA TGATTCTCCT GCCTCAGCTT CCTAAGTAGC TGGGTTTACA 2857 GGCACCCACC ACCATGCCCA GCTAATTTTT GTATTTTTAA TAGAGACGGG GTTTCGCCAT 2917 TTGGCCAGGC TGGTCTTGAA CTCCTGACGT CAAGTGATCT GCCTGCCTTG GTCTCCCAAT 2977 ACAGGCATGA ACCACTGCAC CCACCTACTT AGATATTTCA TGTGCTATAG ACATTAGAGA 3037 GATTTTTCAT TTTTCCATGA CATTTTTCCT CTCTGCAAAT GGCTTAGCTA CTTGTGTTTT 3097 TCCCTTTTGG GGCAAGACAG ACTCATTAAA TATTCTGTAC ATTTTTTCTT TATCAAGGAG 3157 ATATATCAGT GTTGTCTCAT AGAACTGCCT GGATTCCATT TATGTTTTTT CTGATTCCAT 3217 CCTGTGTCCC CTTCATCCTT GACTCCTTTG GTATTTCACT GAATTTCAAA CATTTGTCAG 32 77 AGAAGAAAAA AGTGAGGACT CAGGAAAAAT AAATAAATAA AAGAACAGCC TTTTGCGGCC 3337 GCGAATTC 334 5 (2) INFORMATION FOR SEQ ID NO:178: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 832 amino acids (B) TYPE. amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO.178: Met lie Leu Gin Ala His Leu His Ser Leu Cys Leu Leu Met Leu Tyr 1 Leu Ala Thr Gly 20 Tyr Gly Gin Glu Gly 25 Lys Phe Ser Gly Pro 30 Leu Lys Pro Met Thr 35 Phe Ser lie Tyr Glu 40 Gly Gin Glu Pro Ser 45 Gin lie lie Phe Gin 50 Phe Lys Ala Asn Pro 55 Pro Ala Val Thr Phe 60 Glu Leu Thr Gly Glu Thr Asp Asn lie Phe Val lie Glu Arg Glu Gly Leu Leu Tyr Tyr 65 70 75 80 Asn Arg Ala Leu Asp 85 Arg Glu Thr Arg Ser 90 Thr His Asn Leu Gin 95 Val Ala Ala Leu Asp 100 Ala Asn Gly lie lie 105 Val Glu Gly Pro Val 110 Pro lie Thr lie Glu 115 Val Lys Asp lie Asn 120 Asp Asn Arg Pro Thr 125 Phe Leu Gin Ser Lys 130 Tyr Glu Gly Ser Val 135 Arg Gin Asn Ser Arg 140 Pro Gly Lys Pro Phe Leu Tyr Val Asn Ala Thr Asp Leu Asp Asp Pro Ala Thr Pro Asn 145 150 155 160 Gly Gin Leu Tyr Tyr 165 Gin lie Val lie Gin 170 Leu Pro Met lie Asn 175 Asn Val Met Tyr Phe 180 Gin lie Asn Asn Lys 185 Thr Gly Ala lie Ser 190 Leu Thr Printed from Mimosa Arg Glu Gly Ser Gin Glu Leu Asn Pro Ala Lys Asn Pro Ser Tyr Asn 195 200 205 Leu Val lie Ser Val Lys Asp Met Gly Gly Gin Ser Glu Asn Ser Phe 210 215 220 Ser Asp Thr Thr Ser Val Asp lie lie Val Thr Glu Asn lie Trp Lys 225 230 235 240 Ala Pro Lys Pro Val Glu Met Val Glu Asn Ser Thr Asp Pro His Pro 245 250 255 He Lys lie Thr Gin Val Arg Trp Asn Asp Pro Gly Ala Gin Tyr Ser 260 265 270 Leu Val Asp Lys Glu Lys Leu Pro Arg Phe Pro Phe Ser lie Asp Gin 275 280 285 Glu Gly Asp lie Tyr Val Thr Gin Pro Leu Asp Arg Glu Glu Lys Asp 290 295 300 Ala Tyr Val Phe Tyr Ala Val Ala Lys Asp Glu Tyr Gly Lys Pro Leu 305 310 315 320 Ser Tyr Pro Leu Glu lie His Val Lys Val Lys Asp lie Asn Asp Asn 325 330 335 Pro Pro Thr Cys Pro Ser Pro Val Thr Val Phe Glu Val Gin Glu Asn 340 345 350 Glu Arg Leu Gly Asn Ser lie Gly Thr Leu Thr Ala His Asp Arg Asp 355 360 365 Glu Glu Asn Thr Ala Asn Ser Phe Leu Asn Tyr Arg lie Val Glu Gin 370 375 380 Thr Pro Lys Leu Pro Met Asp Gly Leu Phe Leu lie Gin Thr Tyr Ala 385 390 395 400 Gly Met Leu Gin Leu Ala Lys Gin Ser Leu Lys Lys Gin Asp Thr Pro 405 410 415 Gin Tyr Asn Leu Thr lie Glu Val Ser Asp Lys Asp Phe Lys Thr Leu 420 425 430 Cys Phe Val Gin lie Asn Val lie Asp lie Asn Asp Gin lie Pro lie 435 440 445 Phe Glu Lys Ser Asp Tyr Gly Asn Leu Thr Leu Ala Glu Asp Thr Asn 450 455 460 lie Gly Ser Thr lie Leu Thr He Gin Ala Thr Asp Ala Asp Glu Pro 465 470 475 480 Phe Thr Gly Ser Ser Lys lie Leu Tyr His lie lie Lys Gly Asp Ser 485 490 495 Glu Gly Arg Leu Gly Val Asp Thr Asp Pro His Thr Asn Thr Gly Tyr 500 505 510 Val lie lie Lys Lys Pro Leu Asp Phe Glu Thr Ala Ala Val Ser Asn 515 520 525 lie Val Phe Lys Ala Glu Asn Pro Glu Pro Leu Val Phe Gly Val Lys 530 535 54 0 Tyr Asn Ala Ser Ser Phe Ala Lys Phe Thr Leu lie Val Thr Asp Val 545 550 555 560 Asn Glu Ala Pro Gin Phe Ser Gin His Val Phe Gin Ala Lys Val Ser 565 570 575 Glu Asp Val Ala lie Gly Thr Lys Val Gly Asn Val Thr Ala Lys Asp 580 585 590 Pro Glu Gly Leu Asp lie Ser Tyr Ser Leu Arg Gly Asp Thr Arg Gly 595 600 605 Trp Leu Lys lie Asp His Val Thr Gly Glu lie Phe Ser Val Ala Pro 610 615 620 Leu Asp Arg Glu Ala Gly Ser Pro Tyr Arg Val Gin Val Val Ala Thr 625 630 635 640 Glu Val Gly Gly Ser Ser Leu Ser Ser Val Ser Glu Phe His Leu lie 645 650 655 Leu Met Asp Val Asn Asp Asn Pro Pro Arg Leu Ala Lys Asp Tyr Thr 660 665 670 Gly Leu Phe Phe Cys His Pro Leu Ser Ala Pro Gly Ser Leu lie Phe 675 680 685 Glu Ala Thr Asp Asp Asp Gin His Leu Phe Arg Gly Pro His Phe Thr S90 695 700 Phe Ser Leu Gly Ser Gly Ser Leu Gin Asn Asp Trp Glu Val Ser Lys 705 710 715 720 Printed from Mimosa lie Asn Gly Thr His 725 Ala Arg Leu Ser Thr 730 Arg His Thr Asp Phe 735 Glu Glu Arg Ala Tyr Val Val Leu lie Arg lie Asn Asp Gly Gly Arg Pro 740 745 750 Pro Leu Glu 755 Gly lie Val Ser Leu 760 Pro Val Thr Phe Cys 765 Ser Cys Val Glu Gly Ser Cys Phe Arg Pro Ala Gly His Gin Thr Gly lie Pro Thr 770 775 780 Val Gly Met Ala Val Gly lie Leu Leu Thr Thr Leu Leu Val lie Gly 785 790 795 800 lie lie Leu Ala Val Val Phe lie Arg lie Lys Lys Asp Lys Gly Lys 805 810 815 Asp Asn Val Glu 820 Ser Ala Gin Ala Ser 825 Glu Val Lys Pro Leu Arg 830 Ser (2) INFORMATION FOR SEQ ID NO:179: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 1827 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:179: Met Ala Arg Lys Lys Phe Ser Gly Leu Glu lie Ser Leu lie Val Leu 1 Phe Val lie Val 20 Thr lie lie Ala lie 25 Ala Leu lie Val Val 30 Leu Ala Thr Lys Thr 35 Pro Ala Val Asp Glu 40 lie Ser Asp Ser Thr 45 Ser Thr Pro Ala Thr Thr Arg Val Thr Thr Asn Pro Ser Asp Ser Gly Lys Cys Pro 50 > 55 60 Asn Val Leu Asn Asp Pro Val Asn Val Arg lie Asn Cys lie Pro Glu 65 70 75 80 Gin Phe Pro Thr Glu Gly lie Cys Ala Gin Arg Gly Cys Cys Trp Arg 85 90 95 Pro Trp Asn Asp 100 Ser Leu lie Pro Trp 105 Cys Phe Phe Val Asp 110 Asn His Gly Tyr Asn Val Gin Asp Met Thr Thr Thr Ser lie Gly Val Glu Ala 115 120 125 Lys Leu Asn Arg lie Pro Ser Pro Thr Leu Phe Gly Asn Asp lie Asn 130 135 140 Ser Val Leu Phe Thr Thr Gin Asn Gin Thr Pro Asn Arg Phe Arg Phe 145 150 155 160 Lys lie Thr Asp Pro 165 Asn Asn Arg Arg Tyr 170 Glu Val Pro His Gin 175 Tyr Val Lys Glu Phe 180 Thr Gly Pro Thr Val 185 Ser Asp Thr Leu Tyr 190 Asp Val Lys Val Ala Gin Asn Pro Phe Ser lie Gin Val lie Arg Lys Ser Asn 195 200 205 Gly Lys Thr Leu Phe Asp Thr Ser lie Gly Pro Leu Val Tyr Ser Asp 210 215 220 Gin Tyr Leu Gin lie Ser Ala Arg Leu Pro Ser Asp Tyr lie Tyr Gly 225 230 235 240 lie Gly Glu Gin Val 245 His Lys Arg Phe Arg 250 His Asp Leu Ser Trp 255 Lys Thr Trp Pro lie Phe Thr Arg Asp Gin Leu Pro Gly Asp Asn Asn Asn 260 265 270 Asn Leu Tyr 275 Gly His Gin Thr Phe 280 Phe Met Cys lie Glu 285 Asp Thr Ser Gly Lys Ser Phe Gly Val Phe Leu Met Asn Ser Asn Ala Met Glu lie 290 295 300 Phe lie Gin Pro Thr Pro lie Val Thr Tyr Arg Val Thr Gly Gly lie Printed from Mimosa 305 310 315 320 Leu Asp Phe Tyr lie Leu Leu Gly Asp Thr Pro Glu Gin Val Val Gin 325 330 335 Gin Tyr Gin Gin Leu Val Gly Leu Pro Ala Met Pro Ala Tyr Trp Asn 340 345 350 Leu Gly Phe Gin Leu Ser Arg Trp Asn Tyr Lys Ser Leu Asp Val Val 355 360 365 Lys Glu Val Val Arg Arg Asn Arg Glu Ala Gly lie Pro Phe Asp Thr 370 375 380 Gin Val Thr Asp lie Asp Tyr Met Glu Asp Lys Lys Asp Phe Thr Tyr 385 390 395 400 Asp Gin Val Ala Phe Asn Gly Leu Pro Gin Phe Val Gin Asp Leu His 405 410 415 Asp His Gly Gin Lys Tyr Val lie lie Leu Asp Pro Ala lie Ser lie 420 425 430 Gly Arg Arg Ala Asn Gly Thr Thr Tyr Ala Thr Tyr Glu Arg Gly Asn 435 440 445 Thr Gin His Val Trp lie Asn Glu Ser Asp Gly Ser Thr Pro lie lie 450 455 460 Gly Glu Val Trp Pro Gly Leu Thr Val Tyr Pro Asp Phe Thr Asn Pro 465 470 475 480 Asn Cys lie Asp Trp Trp Ala Asn Glu Cys Ser lie Phe His Gin Glu 485 490 495 Val Gin Tyr Asp Gly Leu Trp lie Asp Met Asn Glu Val Ser Ser Phe 500 505 510 lie Gin Gly Ser Thr Lys Gly Cys Asn Val Asn Lys Leu Asn Tyr Pro 515 520 525 Pro Phe Thr Pro Asp lie Leu Asp Lys Leu Met Tyr Ser Lys Thr lie 530 535 540 Cys Met Asp Ala Val Gin Asn Trp Gly Lys Gin Tyr Asp Val His Ser 545 550 555 560 Leu Tyr Gly Tyr Ser Met Ala lie Ala Thr Glu Gin Ala Val Gin Lys 565 570 575 Val Phe Pro Asn Lys Arg Ser Phe ,Ile Leu Thr Arg Ser Thr Phe Ala 580 585 590 Gly Ser Gly Arg His Ala Ala His Trp Leu Gly Asp Asn Thr Ala Ser 595 600 605 Trp Glu Gin Met Glu Trp Ser lie Thr Gly Met Leu Glu Phe Ser Leu 610 615 620 Phe Gly lie Pro Leu Val Gly Ala Asp lie Cys Gly Phe Val Ala Glu 625 630 635 640 Thr Thr Glu Glu Leu Cys Arg Arg Trp Met Gin Leu Gly Ala Phe Tyr 645 650 655 Pro Phe Ser Arg Asn His Asn Ser Asp Gly Tyr Glu His Gin Asp Pro 660 665 670 Ala Phe Phe Gly Gin Asn Ser Leu Leu Val Lys Ser Ser Arg Gin Tyr 675 680 685 Leu Thr lie Arg Tyr Thr Leu Leu Pro Phe Leu Tyr Thr Leu Phe Tyr 690 695 700 Lys Ala His Val Phe Gly Glu Thr Val Ala Arg Pro Val Leu His Glu 705 710 715 720 Phe Tyr Glu Asp Thr Asn Ser Trp lie Glu Asp Thr Glu Phe Leu Trp 725 730 735 Gly Pro Ala Leu Leu lie Thr Pro Val Leu Lys Gin Gly Ala Asp Thr 740 745 750 Val Ser Ala Tyr lie Pro Asp Ala lie Trp Tyr Asp Tyr Glu Ser Gly 755 760 765 Ala Lys Arg Pro Trp Arg Lys Gin Arg Val Asp Met Tyr Leu Pro Ala 770 775 780 Asp Lys lie Gly Leu His Leu Arg Gly Gly Tyr lie lie Pro lie Gin 785 790 795 800 Glu Pro Asp Val Thr Thr Thr Ala Ser Arg Lys Asn Pro Leu Gly Leu 805 810 815 lie Val Ala Leu Gly Glu Asn Asn Thr Ala Lys Gly Asp Phe Phe Trp 820 825 830 Asp Asp Gly Glu Thr Lys Asp Thr lie Gin Asn Gly Asn Tyr lie Leu Printed from Mimosa 835 840 845 Tyr Thr Phe Ser Val Ser Asn Asn Thr Leu Asp lie Val Cys Thr His 850 855 860 Ser Ser Tyr Gin Glu Gly Thr Thr Leu Ala Phe Gin Thr Val Lys lie 865 870 875 880 Leu Gly Leu Thr Asp 885 Ser Val Thr Glu Val 890 Arg Val Ala Glu Asn 895 Asn Gin Pro Met Asn 900 Ala His Ser Asn Phe 905 Thr Tyr Asp Ala Ser 910 Asn Gin Val Leu Leu 915 lie Ala Asp Leu Lys 920 Leu Asn Leu Gly Arg 925 Asn Phe Ser Val Gin 930 Trp Asn Gin lie Phe 935 Ser Glu Asn Glu Arg 94 0 Phe Asn Cys Tyr Pro Asp Ala Asp Leu Ala Thr Glu Gin Lys Cys Thr Gin Arg Gly Cys 945 950 955 960 Val Trp Arg Thr Gly 965 Ser Ser Leu Ser Lys 970 Ala Pro Glu Cys Tyr 975 Phe Pro Arg Gin Asp 980 Asn Ser Tyr Ser Val 985 Asn Ser Ala Arg Tyr 990 Ser Ser Met Gly lie Thr Ala Asp Leu Gin Leu Asn Thr Ala Asn Ala Arg lie 995 1000 1005 Lys Leu Pro Ser Asp Pro lie Ser Thr Leu Arg Val Glu Val Lys Tyr 1010 1015 1020 His Lys Asn Asp Met Leu Gin Phe Lys lie Tyr Asp Pro Gin Lys Lys 025 1030 1035 1040 Arg Tyr Glu Val Pro Val Pro Leu Asn lie Pro Thr Thr Pro lie Ser 1045 1050 1055 Thr Tyr Glu Asp Arg Leu Tyr Asp Val Glu lie Lys Glu Asn Pro Phe 1060 1065 1070 Gly lie Gin lie Arg Arg Arg Ser Ser Gly Arg Val lie Trp Asp Ser 1075 1080 1085 Trp Leu Pro Gly Phe Ala Phe Asn Asp Gin Phe lie Gin lie Ser Thr 1090 1095 1100 Arg Leu Pro Ser Glu Tyr lie Tyr Gly Phe Gly Glu Val Glu His Thr 105 1110 ' 1115 1120 Ala Phe Lys Arg Asp Leu Asn Trp Asn Thr Trp Gly Met Phe Thr Arg 1125 1130 1135 Asp Gin Pro Pro Gly Tyr Lys Leu Asn Ser Tyr Gly Phe His Pro Tyr 1140 1145 1150 Tyr Met Ala Leu Glu Glu Glu Gly Asn Ala His Gly Val Phe Leu Leu 1155 1160 1165 Asn Ser Asn Ala Met Asp Val Thr Phe Gin Pro Thr Pro Ala Leu Thr 1170 1175 1180 Tyr Arg Thr Val Gly Gly lie Leu Asp Phe Tyr Met Phe Leu Gly Pro 185 1190 1195 1200 Thr Pro Gin Val Ala Thr Lys Gin Tyr His Glu Val lie Gly His Pro 1205 1210 1215 Val Met Pro Ala Tyr Trp Ala Leu Gly Phe Gin Leu Cys Arg Tyr Gly 1220 1225 1230 Tyr Ala Asn Thr Ser Glu Val Arg Glu Leu Tyr Asp Ala Met Val Ala 1235 1240 1245 Ala Asn lie Pro Tyr Asp Val Gin Tyr Thr Asp lie Asp Tyr Met Glu 1250 1255 1260 Arg Gin Leu Asp Phe Thr lie Gly Glu Ala Phe Gin Asp Leu Pro Gin 265 1270 1275 1280 Phe Val Asp Lys lie Arg Gly Glu Gly Met Arg Tyr lie lie lie Leu 1285 1290 1295 Asp Pro Ala lie Ser Gly Asn Glu Thr Lys Thr Tyr Pro Ala Phe Glu 1300 1305 1310 Arg Gly Gin Gin Asn Asp Val Phe Val Lys Trp Pro Asn Thr Asn Asp 1315 1320 1325 lie Cys Trp Ala Lys Val Trp Pro Asp Leu Pro Asn lie Thr lie Asp 1330 1335 1340 Lys Thr Leu Thr Glu Asp Glu Ala Val Asn Ala Ser Arg Ala His Val 345 1350 1355 1360 Ala Phe Pro Asp Phe Phe Arg Thr Ser Thr Ala Glu Trp Trp Ala Arg Printed from Mimosa 1365 1370 1375 Glu lie Val Asp Phe Tyr Asn Glu Lys Met Lys Phe Asp Gly Leu Trp 1380 1385 1390 lie Asp Met Asn Glu Pro Ser Ser Phe Val Asn Gly Thr Thr Thr Asn 1395 1400 1405 Gin Cys Arg Asn Asp Glu Leu Asn Tyr Pro Pro Tyr Phe Pro Glu Leu 1410 1415 1420 Thr Lys Arg Thr Asp Gly Leu His Phe Arg Thr lie Cys Met Glu Ala 425 1430 1435 1440 Glu Gin lie Leu Ser Asp Gly Thr Ser Val Leu His Tyr Asp Val His 1445 1450 1455 Asn Leu Tyr Gly Trp Ser Gin Met Lys Pro Thr His Asp Ala Leu Gin 1460 1465 1470 Lys Thr Thr Gly Lys Arg Gly lie Val lie Ser Arg Ser Thr Tyr Pro 1475 1480 1485 Thr Ser Gly Arg Trp Gly Gly His Trp Leu Gly Asp Asn Tyr Ala Arg 1490 1495 1500 Trp Asp Asn Met Asp Lys Ser lie lie Gly Met Met Glu Phe Ser Leu 505 1510 1515 1520 Phe Gly lie Ser Tyr Thr Gly Ala Asp He Cys Gly Phe Phe Asn Asn 1525 1530 1535 Ser Glu Tyr His Leu Cys Thr Arg Trp Met Gin Leu Gly Ala Phe Tyr 1540 1545 1550 Pro Tyr Ser Arg Asn His Asn lie Ala Asn Thr Arg Arg Gin Asp Pro 1555 1560 1565 Ala Ser Trp Asn Glu Thr Phe Ala Glu Met Ser Arg Asn lie Leu Asn 1570 1575 1580 lie Arg Tyr Thr Leu Leu Pro Tyr Phe Tyr Thr Gin Met His Glu lie 585 1590 1595 1600 His Ala Asn Gly Gly Thr Val lie Arg Pro Leu Leu His Glu Phe Phe 1605 1610 1615 Asp Glu Lys Pro Thr Trp Asp lie Phe Lys Gin Phe Leu Trp Gly Pro 1620 1625 1630 Ala Phe Met Val Thr Pro Val Leu Glu Pro Tyr Val Gin Thr Val Asn 1635 1640 1645 Ala Tyr Val Pro Asn Ala Arg Trp Phe Asp Tyr His Thr Gly Lys Asp 1650 1655 1660 lie Gly Val Arg Gly Gin Phe Gin Thr Phe Asn Ala Ser Tyr Asp Thr 665 1670 1675 1680 lie Asn Leu His Val Arg Gly Gly His lie Leu Pro Cys Gin Glu Pro 1685 1690 1695 Ala Gin Asn Thr Phe Tyr Ser Arg Gin Lys His Met Lys Leu lie Val 1700 1705 1710 Ala Ala Asp Asp Asn Gin Met Ala Gin Gly Ser Leu Phe Trp Asp Asp 1715 1720 1725 Gly Glu Ser lie Asp Thr Tyr Glu Arg Asp Leu Tyr Leu Ser Val Gin 1730 1735 1740 Phe Asn Leu Asn Gin Thr Thr Leu Thr Ser Thr lie Leu Lys Arg Gly 745 1750 1755 1760 Tyr lie Asn Lys Ser Glu Thr Arg Leu Gly Ser Leu His Val Trp Gly 1765 1770 1775 Lys Gly Thr Thr Pro Val Asn Ala val Thr Leu Thr Tyr Asn Gly Asn 1780 1785 1790 Lys Asn Ser Leu Pro Phe Asn Glu Asp Thr Thr Asn Met lie Leu Arg 1795 1800 1805 lie Asp Leu Thr Thr His Asn Val Thr Leu Glu Glu Pro lie Glu lie 1B10 1815 1820 Asn Trp Ser 825 (2) INFORMATION FOR SEQ ID NO:180. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2284 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single Printed from Mimosa WO 98/51325 PCT/US98/10088 (D) TOPOLOGY: linear (il) MOLECULE TYPE: DNA (ix) FEATURE.

(A) NAME/KEY: Coding Sequence (B) LOCATION. 45...2099 (D) OTHER INFORMATION: (xi) SEQUENCE DESCRIPTION: SEQ ID NO:180" GCCTTACTGC AGGAAGGCAC TCCGAAGACA TAAGTCGGTG AGAC ATG GCT GAA GAT 56 Met Ala Glu Asp 1 AAA AGC AAG AGA GAC TCC ATC GAG ATG AGT ATG AAG GGA TGC CAG ACA 104 Lys Ser Lys Arg Asp Ser lie Glu Met Ser Met Lys Gly Cys Gin Thr 5 10 15 20 AAC AAC GGG TTT GTC CAT AAT GAA GAC ATT CTG GAG CAG ACC CCG GAT 152 Asn Asn Gly Phe Val His Asn Glu Asp lie Leu Glu Gin Thr Pro Asp 25 30 35 CCA GGC AGC TCA ACA GAC AAC CTG AAG CAC AGC ACC AGG GGC ATC CTT 2 00 Pro Gly Ser Ser Thr Asp Asn Leu Lys His Ser Thr Arg Gly lie Leu 40 45 50 GGC TCC CAG GAG CCC GAC TTC AAG GGC GTC CAG CCC TAT GCG GGG ATG 248 Gly Ser Gin Glu Pro Asp Phe Lys Gly Val Gin Pro Tyr Ala Gly Met 55 60 65 CCC AAG GAG GTG CTG TTC CAG TTC TCT GGC CAG GCC CGC TAC CGC ATA 296 Pro Lys Glu Val Leu Phe Gin Phe Ser Gly Gin Ala Arg Tyr Arg lie 20 7° 80 CCT CGG GAG ATC CTC TTC TGG CTC ACA GTG GCT TCT GTG CTG GTG CTC 344 Pro Arg Glu lie Leu Phe Trp Leu Thr Val Ala Ser Val Leu Val Leu 85 90 95 100 ATC GCG GCC ACC ATA GCC ATC ATT GCC CTC TCT CCA AAG TGC CTA GAC 392 lie Ala Ala Thr lie Ala lie lie Ala Leu Ser Pro Lys Cys Leu Asp 105 110 115 TGG TGG CAG GAG GGG CCC ATG TAC CAG ATC TAC CCA AGG TCT TTC AAG 440 Trp Trp Gin Glu Gly Pro Met Tyr Gin lie Tyr Pro Arg Ser Phe Lys 120 125 130 GAC AGT AAC AAG GAT GGG AAC GGA GAT CTG AAA GGT ATT CAA GAT AAA 488 Asp Ser Asn Lys Asp Gly Asn Gly Asp Leu Lys Gly lie Gin Asp Lys 135 140 145 CTG GAC TAC ATC ACA GCT TTA AAT ATA AAA ACT GTT TGG ATT ACT TCA 536 Leu Asp Tyr lie Thr Ala Leu Asn lie Lys Thr Val Trp lie Thr Ser 150 155 160 TTT TAT AAA TCG TCC CTT AAA GAT TTC AGA TAT GGT GTT GAA GAT TTC 584 Phe Tyr Lys Ser Ser Leu Lys Asp Phe Arg Tyr Gly Val Glu Asp Phe 165 170 175 180 CGG GAA GTT GAT CCC ATT TTT GGA ACG ATG GAA GAT TTT GAG AAT CTG 632 Arg Glu Val Asp Pro lie Phe Gly Thr Met Glu Asp Phe Glu Asn Leu 185 190 195 GTT GCA GCC ATA CAT GAT AAA GGT TTA AAA TTA ATC ATC GAT TTC ATA 680 Val Ala Ala lie His Asp Lys Gly Leu Lys Leu lie lie Asp Phe lie Printed from Mimosa 2 00 205 210 CCA AAC CAC ACG AGT GAT AAA CAT ATT TGG TTT CAA TTG AGT CGG ACA Pro Asn His Thr Ser Asp Lys His lie Trp Phe Gin Leu Ser Arg Thr 215 220 225 728 CGG ACA GGA AAA TAT ACT GAT TAT TAT ATC TGG CAT GAC TGT ACC CAT Arg Thr Gly Lys Tyr Thr Asp Tyr Tyr lie Trp His Asp Cys Thr His 230 235 240 776 GAA AAT GGC AAA ACC ATT CCA CCC AAC AAC TGG TTA AGT GTG TAT GGA Glu Asn Gly Lys Thr lie Pro Pro Asn Asn Trp Leu Ser Val Tyr Gly 245 250 255 260 824 AAC TCC AGT TGG CAC TTT GAC GAA GTG CGA AAC CAA TGT TAT TTT CAT Asn Ser Ser Trp His Phe Asp Glu Val Arg Asn Gin Cys Tyr Phe His 265 270 275 872 CAG TTT ATG AAA GAG CAA CCT GAT TTA AAT TTC CGC AAT CCT GAT GTT Gin Phe Met Lys Glu Gin Pro Asp Leu Asn Phe Arg Asn Pro Asp Val 280 285 290 920 CAA GAA GAA ATA AAA GAA ATT TTA CGG TTC TGG CTC ACA AAG GGT GTT Gin Glu Glu lie Lys Glu lie Leu Arg Phe Trp Leu Thr Lys Gly Val 295 300 305 968 GAT GGT TTT AGT TTG GAT GCT GTT AAA TTC CTC CTA GAA GCA AAG CAC Asp Gly Phe Ser Leu Asp Ala Val Lys Phe Leu Leu Glu Ala Lys His 310 315 320 1016 CTG AGA GAT GAG ATC CAA GTA AAT AAG ACC CAA ATC CCG GAC ACG GTC Leu Arg Asp Glu lie Gin Val Asn Lys Thr Gin lie Pro Asp Thr Val 325 330 335 340 1064 ACA CAA TAC TCG GAG CTG TAC CAT GAC TTC ACC ACC ACG CAG GTG GGA Thr Gin Tyr Ser Glu Leu Tyr His Asp Phe Thr Thr Thr Gin Val Gly 345 350 355 1112 ATG CAC GAC ATT GTC CGC AGC TTC CGG CAG ACC ATG GAC CAA TAC AGC Met His Asp lie Val Arg Ser Phe Arg Gin Thr Met Asp Gin Tyr Ser 360 365 370 1160 ACG GAG CCC GGC AGA TAC AGG TTC ATG GGG ACT GAA GCC TAT GCA GAG Thr Glu Pro Gly Arg Tyr Arg Phe Met Gly Thr Glu Ala Tyr Ala Glu 375 380 385 1208 AGT ATT GAC AGG ACC GTG ATG TAC TAT GGA TTG CCA TTT ATC CAA GAA Ser lie Asp Arg Thr Val Met Tyr Tyr Gly Leu Pro Phe lie Gin Glu 390 395 400 1256 GCT GAT TTT CCC TTC AAC AAT TAC CTC AGC ATG CTA GAC ACT GTT TCT Ala Asp Phe Pro Phe Asn Asn Tyr Leu Ser Met Leu Asp Thr Val Ser 405 410 415 420 1304 GGG AAC AGC GTG TAT GAG GTT ATC ACA TCC TGG ATG GAA AAC ATG CCA Gly Asn Ser Val Tyr Glu Val lie Thr Ser Trp Met Glu Asn Met Pro 425 430 435 1352 GAA GGA AAA TGG CCT AAC TGG ATG ATT GGT GGA CCA GAC AGT TCA CGG Glu Gly Lys Trp Pro Asn Trp Met lie Gly Gly Pro Asp Ser Ser Arg 440 445 450 1400 CTG ACT TCG CGT TTG GGG AAT CAG TAT GTC AAC GTG ATG AAC ATG CTT Leu Thr Ser Arg Leu Gly Asn Gin Tyr Val Asn Val Met Asn Met Leu 455 460 465 1448 Printed from Mimosa PCT /US98/10088 CTT TTC ACA CTC CCT GGA ACT CCT ATA ACT TAC TAT GGA GAA GAA ATT Leu Phe Thr Leu Pro Gly Thr Pro lie Thr Tyr Tyr Gly Glu Glu lie 470 475 480 1496 GGA ATG GGA AAT ATT GTA GCC GCA AAT CTC AAT GAA AGC TAT GAT ATT Gly Met Gly Asn lie Val Ala Ala Asn Leu Asn Glu Ser Tyr Asp lie 485 490 495 500 1544 AAT ACC CTT CGC TCA AAG TCA CCA ATG CAG TGG GAC AAT AGT TCA AAT Asn Thr Leu Arg Ser Lys Ser Pro Met Gin Trp Asp Asn Ser Ser Asn 505 510 515 1592 GCT GGT TTT TCT GAA GCT AGT AAC ACC TGG TTA CCT ACC AAT TCA GAT Ala Gly Phe Ser Glu Ala Ser Asn Thr Trp Leu Pro Thr Asn Ser Asp 520 525 530 1640 TAC CAC ACT GTG AAT GTT GAT GTC CAA AAG ACT CAG CCC AGA TCG GCT Tyr His Thr Val Asn Val Asp Val Gin Lys Thr Gin Pro Arg Ser Ala 535 540 545 1688 TTG AAG TTA TAT CAA GAT TTA AGT CTA CTT CAT GCC AAT GAG CTA CTC Leu Lys Leu Tyr Gin Asp Leu Ser Leu Leu His Ala Asn Glu Leu Leu 550 555 560 1736 CTC AAC AGG GGC TGG TTT TGC CAT TTG AGG AAT GAC AGC CAC TAT GTT Leu Asn Arg Gly Trp Phe Cys His Leu Arg Asn Asp Ser His Tyr Val 565 570 575 580 1784 GTG TAC ACA AGA GAG CTG GAT GGC ATC GAC AGA ATC TTT ATC GTG GTT Val Tyr Thr Arg Glu Leu Asp Gly lie Asp Arg lie Phe lie Val Val 585 590 595 1832 CTG AAT TTT GGA GAA TCA ACA CTG TTA AAT CTA CAT AAT ATG ATT TCG Leu Asn Phe Gly Glu Ser Thr Leu Leu Asn Leu His Asn Met lie Ser 600 605 610 1880 GGC CTT CCC GCT AAA ATA AGA ATA AGG TTA AGT ACC AAT TCT GCC GAC Gly Leu Pro Ala Lys lie Arg lie Arg Leu Ser Thr Asn Ser Ala Asp 615 620 625 1928 AAA GGC AGT AAA GTT GAT ACA AGT GGC ATT TTT CTG GAC AAG GGA GAG Lys Gly Ser Lys Val Asp Thr Ser Gly lie Phe Leu Asp Lys Gly Glu 630 635 640 1976 GGA CTC ATC TTT GAA CAC AAC ACG AAG AAT CTC CTT CAT CGC CAA ACA Gly Leu lie Phe Glu His Asn Thr Lys Asn Leu Leu His Arg Gin Thr 645 650 655 660 2024 GCT TTC AGA GAT AGA TGC TTT GTT TCC AAT CGA GCA TGC TAT TCC AGT Ala Phe Arg Asp Arg Cys Phe Val Ser Asn Arg Ala Cys Tyr Ser Ser 665 670 675 2072 GTA CTG AAC ATA CTG TAT ACC TCG TGT TAGGCACCTT TATGAAGAGA TGAAGAC Val Leu Asn lie Leu Tyr Thr Ser Cys 680 685 2126 ACTGGCATTT CAGTGGGATT GTAAGCATTT GTAATAGCTT CATGTACAGC ATGCTGCTTG 2186 GTGAACAATC ATTAATTCTT CGATATTTCT GTAGCTTGAA TGTAACCGCT TTAAGAAAGG 2246 TTCTCAAATG TTTTGAAAAA AATAAAATGT TTAAAAGT 2284 (2) INFORMATION FOR SEQ ID NO:181: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 685 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: 192 - Printed from Mimosa (D) TOPOLOGY unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:181: Met Ala Glu Asp Lys Ser Lys Arg Asp Ser lie Glu Met Ser Met Lys 1 Gly Cys Gin Thr Asn Asn Gly Phe Val His Asn Glu Asp lie Leu Glu Gin Thr Pro Asp Pro Gly Ser Ser Thr Asp Asn Leu Lys His Ser Thr 40 45 Arg Gly lie Leu Gly Ser Gin Glu Pro Asp Phe Lys Gly Val Gin Pro 50 55 60 Tyr Ala Gly Met Pro Lys Glu Val Leu Phe Gin Phe Ser Gly Gin Ala 65 70 75 80 Arg Tyr Arg lie Pro Arg Glu lie Leu Phe Trp Leu Thr Val Ala Ser 85 90 95 Val Leu Val Leu lie Ala Ala Thr lie Ala lie lie Ala Leu Ser Pro 100 105 110 Lys Cys Leu Asp Trp Trp Gin Glu Gly Pro Met Tyr Gin lie Tyr Pro 115 120 125 Arg Ser Phe Lys Asp Ser Asn Lys Asp Gly Asn Gly Asp Leu Lys Gly 130 135 140 lie Gin Asp Lys Leu Asp Tyr lie Thr Ala Leu Asn lie Lys Thr Val 145 150 155 160 Trp lie Thr Ser Phe Tyr Lys Ser Ser Leu Lys Asp Phe Arg Tyr Gly 165 170 175 Val Glu Asp Phe Arg Glu Val Asp Pro lie Phe Gly Thr Met Glu Asp 180 185 190 Phe Glu Asn Leu Val Ala Ala lie His Asp Lys Gly Leu Lys Leu lie 195 200 205 lie Asp Phe lie Pro Asn His Thr Ser Asp Lys His lie Trp Phe Gin 210 215 220 Leu Ser Arg Thr Arg Thr Gly Lys Tyr Thr Asp Tyr Tyr lie Trp His 225 230 235 240 Asp Cys Thr His Glu Asn Gly Lys Thr lie Pro Pro Asn Asn Trp Leu 245 250 255 Ser Val Tyr Gly Asn Ser Ser Trp His Phe Asp Glu Val Arg Asn Gin 260 265 270 Cys Tyr Phe His Gin Phe Met Lys Glu Gin Pro Asp Leu Asn Phe Arg 275 280 285 Asn Pro Asp Val Gin Glu Glu lie Lys Glu lie Leu Arg Phe Trp Leu 290 295 300 Thr Lys Gly Val Asp Gly Phe Ser Leu Asp Ala val Lys Phe Leu Leu 305 310 315 320 Glu Ala Lys His Leu Arg Asp Glu lie Gin Val Asn Lys Thr Gin lie 325 330 335 Pro Asp Thr Val Thr Gin Tyr Ser Glu Leu Tyr His Asp Phe Thr Thr 340 345 350 Thr Gin Val Gly Met His Asp lie Val Arg Ser Phe Arg Gin Thr Met 355 360 365 Asp Gin Tyr Ser Thr Glu Pro Gly Arg Tyr Arg Phe Met Gly Thr Glu 370 375 380 Ala Tyr Ala Glu Ser lie Asp Arg Thr Val Met Tyr Tyr Gly Leu Pro 385 390 395 400 Phe lie Gin Glu Ala Asp Phe Pro Phe Asn Asn Tyr Leu Ser Met Leu 405 410 415 Asp Thr Val Ser Gly Asn Ser Val Tyr Glu Val lie Thr Ser Trp Met 420 425 430 Glu Asn Met Pro Glu Gly Lys Trp Pro Asn Trp Met lie Gly Gly Pro 435 440 445 Asp Ser Ser Arg Leu Thr Ser Arg Leu Gly Asn Gin Tyr Val Asn Val 450 455 460 Met Asn Met Leu Leu Phe Thr Leu Pro Gly Thr Pro lie Thr Tyr Tyr 465 470 475 480 Printed from Mimosa Gly Glu Glu lie Gly 485 Met Gly Asn lie Val 490 Ala Ala Asn Leu Asn 495 Glu Ser Tyr Asp lie 500 Asn Thr Leu Arg Ser 505 Lys Ser Pro Met Gin 510 Trp Asp Asn Ser Ser 515 Asn Ala Gly Phe Ser 520 Glu Ala Ser Asn Thr 525 Trp Leu Pro Thr Asn 530 Ser Asp Tyr His Thr 535 Val Asn Val Asp Val 540 Gin Lys Thr Gin Pro Arg Ser Ala Leu Lys Leu Tyr Gin Asp Leu Ser Leu Leu His Ala 545 550 555 560 Asn Glu Leu Leu Leu 565 Asn Arg Gly Trp Phe 570 Cys His Leu Arg Asn 575 Asp Ser His Tyr Val 580 Val Tyr Thr Arg Glu 585 Leu Asp Gly lie Asp 590 Arg lie Phe lie val 595 Val Leu Asn Phe Gly 600 Glu Ser Thr Leu Leu 605 Asn Leu His Asn Met 610 lie Ser Gly Leu Pro 615 Ala Lys lie Arg lie 620 Arg Leu Ser Thr Asn Ser Ala Asp Lys Gly Ser Lys Val Asp Thr Ser Gly lie Phe Leu 625 630 635 640 Asp Lys Gly Glu Gly 645 Leu lie Phe Glu His 650 Asn Thr Lys Asn Leu 655 Leu His Arg Gin Thr 660 Ala Phe Arg Asp Arg 665 Cys Phe Val Ser Asn 670 Arg Ala Cys Tyr Ser 675 Ser Val Leu Asn lie 680 Leu Tyr Thr Ser Cys 685 (2) INFORMATION FOR SEQ ID NO:182- (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 54 amino acids (B) TYPE: amino acid (C) STRANDEDNESS- (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:182: Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Arg Val Gly Gin 10 15 Cys Thr Asp Ser Asp Val Arg Arg Pro Trp Ala Arg Ser Cys Ala His 25 30 Gin Gly Cys Gly Ala Gly Thr Arg Asn Ser His Gly Cys lie Thr Arg 40 45 Pro Leu Arg Gin Ala Ser 50 (2) INFORMATION FOR SEQ ID NO:183: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:183: Ser Ala Arg Asp Ser Gly Pro Ala Glu Asp Gly Ser Arg Ala Val Arg 10 15 Leu Asn Gly (2) INFORMATION FOR SEQ ID NO:184: Printed from Mimosa WO 98/51325 PCT/US98/10088 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY- unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:184 Asp Gly Ser Arg Ala Val Arg Leu Asn Gly Val Glu Asn Ala Asn Thr 10 15 Arg Lys Ser Ser Arg 20 (2) INFORMATION FOR SEQ ID NO:185. (i) SEQUENCE CHARACTERISTICS• (A) LENGTH: 19 amino acids (B) TYPE: amino acid (C) STRANDEDNESS (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.185: Glu Asn Ala Asn Thr Arg Lys Ser Ser Arg Ser Asn Pro Arg Gly Arg 10 15 Arg His Pro (2) INFORMATION FOR SEQ ID NO:186: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 ammo acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO.186 Thr Arg Lys Ser Ser Arg Ser Asn Pro Arg Gly 15 10 (2) INFORMATION FOR SEQ ID NO:187: (I) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 21 ammo acids (B) TYPE- ammo acid 30 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (II) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION- SEQ ID NO:187: Ser Arg Pro Tyr Ser Val Asp Ser Asp Ser Asp Thr Asn Ala Lys His 15 10 15 Ser Ser His Asn Arg 20 (2) INFORMATION FOR SEQ ID NO:188: Printed from Mimosa (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 19 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:188- Thr Asn Ala Lys His Ser Ser His Asn Arg Arg Leu Arg Thr Arg Ser 10 15 Arg Pro Asn (2) INFORMATION FOR SEQ ID NO 18 9. (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 24 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (il) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:189 Arg Tyr Lys His Asp lie Gly Cys Asp Ala Gly Val Asp Lys Lys Ser 10 15 Ser Ser Val Arg Gly Gly Cys Gly 20 (2) INFORMATION FOR SEQ ID NO:190: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 26 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY linear (il) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:190- Gly Cys Asp Ala Gly Val Asp Lys Lys Ser Ser Ser Val Arg Gly Gly 10 15 Cys Gly Ala His Ser Ser Pro Pro Arg Ala 20 25 (2) INFORMATION FOR SEQ ID NO:191: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 ammo acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO.191: Gly Ala His Ser Ser Pro Pro Arg Ala Gly Arg Gly Pro Arg Gly Thr 15 10 15 Met Val Ser Arg Leu 20 - 196 Printed from Mimosa (2) INFORMATION FOR SEQ ID NO:192: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ll) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:192: Glu Asn Ala Asn Thr Arg Lys Ser Ser Arg 15 10 (2) INFORMATION FOR SEQ ID NO-193: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH- 39 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY- unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:193.

Lys Lys Arg lie Ala Gly Leu Pro Trp Tyr Arg Cys Arg Thr Val Ala 10 15 Phe Glu Thr Gly Met Gin Asn Thr Gin Leu Cys Ser Thr lie Val Gin 25 30 Leu Ser Phe Thr Pro Glu Glu 35 (2) INFORMATION FOR SEQ ID NO:194: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:194.

Arg Lys Ser Ser Arg Ser Asn Pro Arg Gly 15 10 (2) INFORMATION FOR SEQ ID NO:195: (l) SEQUENCE CHARACTERISTICS: 30 (A) LENGTH: 9 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQXJENCE DESCRIPTION: SEQ ID NO: 195: Ser Asn Pro Arg Gly Arg Arg His Pro 1 5 (2) INFORMATION FOR SEQ ID NO:196: Printed from Mimosa WO 98/51325 PCT/US98/10088 (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 9 ammo acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO-196- Thr Asn Ala Lys His Ser Ser His Asn 1 5 (2) INFORMATION FOR SEQ ID NO.197: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH 10 ammo acids (B) TYPE- ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:197: Ser Ser His Asn Arg Arg Leu Arg Thr Arg 15 10 (2) INFORMATION FOR SEQ ID NO:198: (i) SEQUENCE CHARACTERISTICS (A) LENGTH: 10 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: TOPOLOGY- unknown (li) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:198: Arg Arg Leu Arg Thr Arg Ser Arg Pro Asn 15 10 (2) INFORMATION FOR SEQ ID NO-199: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH 19 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:199: Arg Val Gly Gin Cys Thr Asp Ser Asp Val Arg Arg Pro Trp Ala Arg 10 15 Ser Cys Ala (2) INFORMATION FOR SEQ ID NO:200: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 21 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: Printed from Mimosa (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO.200: Val Arg Arg Pro Trp Ala Arg Ser Cys Ala His Gin Gly Cys Gly Ala 5 1 5 10 15 Gly Thr Arg Asn Ser 20 (2) INFORMATION FOR SEQ ID N0:201. (l) SEQUENCE CHARACTERISTICS- (A) LENGTH. 19 amino acids (B) TYPE: amino acid in (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID N0:201 Gly Thr Arg Asn Ser His Gly Cys lie Thr Arg Pro Leu Arg Gin Ala 15 10 15 ]_5 Ser Gin His (2) INFORMATION FOR SEQ ID NO:202 (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH 40 amino acids (B) TYPE amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:202.

Ser Thr Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp 10 15 Ser Asp Ser Asp Thr Met Ala Lys His Ser Ser His Asn Arg Arg Leu 25 20 25 30 Arg Thr Arg Ser Arg Pro Asn Gly 35 40 (2) INFORMATION FOR SEQ ID N0:203. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE amino acid 30 (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:203- Tyr Ser Lys Val 1 (2) INFORMATION FOR SEQ ID NO:204- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids Printed from Mimosa (B) TYPE amino acid (C) STRANDEDNESS: (D) TOPOLOGY' unknown (li) MOLECULE TYPE peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID N0:204.

Phe Pro His Leu 1 (2) INFORMATION FOR SEQ ID NO 2 05: (I) SEQUENCE CHARACTERISTICS: (A) LENGTH. 4 amino acids (B) TYPE: amino acid 10 (C) STRANDEDNESS- (D) TOPOLOGY: unknown (II) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:205: Tyr Arg Gly Val 1 (2) INFORMATION FOR SEQ ID N0:206. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 4 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown <ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID N0:206: Tyr Gin Thr lie l Thr Glu Gin Phe 1 (2) INFORMATION FOR SEQ ID NO:208. (i) SEQUENCE CHARACTERISTICS- (A) LENGTH. 4 amino acids (B) TYPE: amino acid 35 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (2) INFORMATION FOR SEQ ID NO:207: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO.207: Printed from Mimosa WO 98/51325 PCT/US98/10088 (xi) SEQUENCE DESCRIPTION: SEQ ID N0:208: Thr Glu Val Met 1 (2) INFORMATION FOR SEQ ID NO:209: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 4 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (li) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION. SEQ ID N0:209: Thr Ser Ala Phe 1 (2) INFORMATION FOR SEQ ID N0:210- (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH. 4 ammo acids (B) TYPE, amino acid 15 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQXJENCE DESCRIPTION: SEQ ID NO:210: Tyr Thr Arg Phe 1 (2) INFORMATION FOR SEQ ID NO.211: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 717 base pairs (B) TYPE: nucleic acid (C) STRANDEDNESS: single (D) TOPOLOGY: linear (ll) MOLECULE TYPE: DNA (ix) FEATURE: (A) NAME/KEY. Coding Sequence (B) LOCATION: 1...714 (D) OTHER INFORMATION- (xi) SEQUENCE DESCRIPTION: SEQ ID NO:211: ATG TCC CCT ATA CTA GGT TAT TGG AAA ATT AAG GGC CTT GTG CAA CCC 48 Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 15 10 15 ACT CGA CTT CTT TTG GAA TAT CTT GAA GAA AAA TAT GAA GAG CAT TTG 96 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu 20 25 30 TAT GAG CGC GAT GAA GGT GAT AAA TGG CGA AAC AAA AAG TTT GAA TTG 144 Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 35 40 45 GGT TTG GAG TTT CCC AAT CTT CCT TAT TAT ATT GAT GGT GAT GTT AAA 192 Printed from Mimosa WO 98/51325 PCT/US98/10088 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 TTA ACA CAG TCT ATG GCC ATC ATA CGT TAT ATA GCT GAC AAG CAC AAC Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 ATG TTG GGT GGT TGT CCA AAA GAG CGT GCA GAG ATT TCA ATG CTT GAA Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 GGA GCG GTT TTG GAT ATT AGA TAC GGT GTT TCG AGA ATT GCA TAT AGT Gly Ala Val Leu Asp lie Arg Tyr Gly val Ser Arg lie Ala Tyr Ser 100 105 110 AAA GAC TTT GAA ACT CTC AAA GTT GAT TTT CTT AGC AAG CTA CCT GAA Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 ATG CTG AAA ATG TTC GAA GAT CGT TTA TGT CAT AAA ACA TAT TTA AAT Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 14 0 GGT GAT CAT GTA ACC CAT CCT GAC TTC ATG TTG TAT GAC GCT CTT GAT Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 GTT GTT TTA TAC ATG GAC CCA ATG TGC CTG GAT GCG TTC CCA AAA TTA Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 GTT TGT TTT AAA AAA CGT ATT GAA GCT ATC CCA CAA ATT GAT AAG TAC Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 TTG AAA TCC AGC AAG TAT ATA GCA TGG CCT TTG CAG GGC TGG CAA GCC Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 ACG TTT GGT GGT GGC GAC CAT CCT CCA AAA TCG GAT CTG GTT CCG CGT Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 GGA TCC CCA GGA ATT CCC GGG TCG ACT CGA GCG GCC GCA TCG TGA Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser 225 230 235 240 288 336 384 432 480 528 576 624 672 717 (2) INFORMATION FOR SEQ ID NO:212: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH1 238 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: protein (xi) SEQUENCE DESCRIPTION: SEQ ID NO:212: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys Phe 45 Glu Leu Printed from Mimosa Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser 225 230 235 (2) INFORMATION FOR SEQ ID NO:213: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 282 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:213 Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Gin Printed from Mimosa WO 98/51325 PCT/US98/10088 225 230 235 240 Gly Ser Lys Gin Cys 245 Met Gin Tyr Arg Thr 250 Gly Arg Leu Thr Val 255 Gly Ser Glu Tyr Gly 260 Cys Gly Met Asn Pro 265 Ala Arg His Ala Thr 270 Pro Ala Tyr Pro Ala 275 Arg Leu Leu Pro Arg 280 Tyr Arg (2) INFORMATION FOR SEQ ID NO:214 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH. 282 amino acids (B) TYPE: amino acid (C) STRANDEDNESS- (D) TOPOLOGY, unknown (ii) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:214.

Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Asp 225 230 235 240 His Ala Leu Gly Thr 245 Asn Leu Arg Ser Asp 250 Asn Ala Lys Glu Pro 255 Gly Asp Tyr Asn Cys Cys Gly Asn Gly Asn Ser Thr Gly Arg Lys Val Phe 260 265 270 Asn Arg Arg 275 Arg Pro Ser Ala lie 280 Pro Thr (2) INFORMATION FOR SEQ ID NO 215: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 279 ammo acids 35 (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide Printed from Mimosa (xi) SEQUENCE DESCRIPTION: SEQ ID NO:215: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Pro 225 230 235 240 Cys Gly Gly Ser Trp Gly Arg Phe Met Gin Gly Gly Leu Phe Gly Gly 245 250 255 Arg Thr Asp Gly 260 Cys Gly Ala His Arg 265 Asn Arg Thr Ser Ala 270 Ser Leu Glu Pro Pro Ser Ser Asp Tyr 275 (2) INFORMATION FOR SEQ ID NO:216: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 277 amino acids 25 (B) TYPE- ammo acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: : SEQ ID NO-216: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu Printed from Mimosa 115 120 125 Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 14 0 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Tip Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Gly 225 230 235 240 Ser Thr Gly Thr Ala 245 Gly Gly Glu Arg Ser 250 Gly Val Leu Asn Leu 255 His Thr Arg Asp Asn 260 Ala Ser Gly Ser Gly 265 Phe Lys Pro Trp Tyr 270 Pro Ser Asn Arg Gly His 275 Lys (2) INFORMATION FOR SEQ ID NO 217: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH. 277 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:217: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 14 0 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser His 225 230 235 240 Ser Gly Gly Met Asn Arg Ala Tyr Gly Asp Val Phe Arg Glu Leu Arg 245 250 255 Printed from Mimosa Asp Arg Trp Asn Ala Thr Ser His His Thr Arg Pro Thr Pro Gin Leu 260 265 270 Pro Arg Gly Pro Asn 275 (2) INFORMATION FOR SEQ ID NO:218: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (li) MOLECULE TYPE peptide (xi) SEQUENCE DESCRIPTION: : SEQ ID NO-218 Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser His 225 230 235 240 Ser Gly Gly Met Asn Arg Ala Tyr 245 (2) INFORMATION FOR SEQ ID NO:219: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:219: Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 10 15 Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu 20 25 30 Met Ser 1 Thr Arg Printed from Mimosa Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys Leu 125 Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys Lys 180 Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Asp 225 230 235 240 Val Phe Arg Glu Leu Arg Asp Arg 245 (2) INFORMATION FOR SEQ ID NO 220: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH 248 amino acids (B) TYPE: amino acid 20 (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE- peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:220: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 15 10 15 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu 20 25 30 Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 30 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala Printed from Mimosa 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Trp Asn 225 230 235 240 Ala Thr Ser His His Thr Arg Pro 245 (2) INFORMATION FOR SEQ ID NO:221: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 247 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION.

SEQ ID NO:221: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 18 5 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Thr Pro 225 230 235 240 Gin Leu Pro Arg Gly Pro Asn 245 (2) INFORMATION FOR SEQ ID NO:222: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 2 58 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS- (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:222: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro Printed from Mimosa 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp 60 Gly Asp Val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Asp 225 230 235 240 Val Phe Arg Glu Leu Arg Asp Arg Trp Asn Ala Thr Ser His His Thr 245 250 255 Arg Pro (2) INFORMATION FOR SEQ ID NO.223: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 257 amino acids (B) TYPE- amino acid (C) STRANDEDNESS (D) TOPOLOGY unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:223: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp 60 Gly Asp Val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Printed from Mimosa Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Trp Asn 225 230 235 240 Ala Thr Ser His His 245 Thr Arg Pro Thr Pro 250 Gin Leu Pro Arg Gly 255 Pro Asn (2) INFORMATION FOR SEQ ID NO.224: (1) SEQUENCE CHARACTERISTICS: (A) LENGTH: 267 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:224.

Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Asp 225 230 235 240 Val Phe Arg Glu Leu Arg Asp Arg Trp Asn Ala Thr Ser His His Thr 245 250 255 Arg Pro Thr Pro Gin Leu Pro Arg Gly Pro Asn 260 265 (2) INFORMATION FOR SEQ ID NO:225. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 277 amino acids Printed from Mimosa (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:225: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser His 225 230 235 240 Ser Gly Gly Met Asn Arg Ala Tyr Gly Asp Val Phe Arg Glu Leu Arg 245 250 255 Asp Arg Trp Asn 260 Ala Thr Ser Ala Ala 265 Thr Arg Pro Thr Pro 270 Gin Leu Pro Arg Gly Pro Asn 275 (2) INFORMATION FOR SEQ ID NO:226. (l) SEQUENCE CHARACTERISTICS- (A) LENGTH- 277 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:226: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn Printed from Mimosa 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Ala 225 230 235 240 Arg Asp Ser Gly Pro Ala Glu Asp Gly Ser Arg Ala Val Arg Leu Asn 245 250 255 Gly Val Glu Asn 260 Ala Asn Thr Arg Lys 265 Ser Ser Arg Ser Asn 270 Pro Arg Gly Arg Arg 275 His Pro (2) INFORMATION FOR SEQ ID NO:227: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 257 amino acids (B) TYPE: amino acid 20 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.227: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys Thr 140 Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala Printed from Mimosa 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Ala 225 230 235 240 Arg Asp Ser Gly Pro 245 Ala Glu Asp Gly Ser 250 Arg Ala Val Arg Leu 255 Asn G1v (2) INFORMATION FOR SEQ ID NO:228: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 259 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS- (D) TOPOLOGY• unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO 228: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Asp Gly 225 230 235 240 Ser Arg Ala Val Arg 245 Leu Asn Gly Val Glu 250 Asn Ala Asn Thr Arg 255 Lys Ser Ser Arg (2) INFORMATION FOR SEQ ID NO.229: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 257 ammo acids , 35 (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide Printed from Mimosa (xi) SEQUENCE DESCRIPTION' SEQ ID NO:229: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 14 0 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Glu Asn 225 230 235 240 Ala Asn Thr Arg Lys Ser Ser Arg Ser Asn Pro Arg Gly Arg Arg His 245 250 255 Pro (2) INFORMATION FOR SEQ ID NO.230: (l) SEQUENCE CHARACTERISTICS.

(A) LENGTH 248 anu.no acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION; : SEQ ID NO:230: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu cys His Lys Thr Tyr Leu Asn Printed from Mimosa 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Glu Asn 225 230 235 240 Ala Asn Thr Arg Lys 245 Ser Ser Arg (2) INFORMATION FOR SEQ ID NO:231: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO 231.

Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp Gly 60 Asp val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys Thr 140 Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp Leu 220 Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Lys 225 230 235 240 Ser Ser Arg Ser Asn 245 Pro Arg Gly (2) INFORMATION FOR SEQ ID NO:232: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 247 amino acids (B) TYPE: amino acid Printed from Mimosa PCT/U S98/10088 (C) STRANDEDNESS• (D) TOPOLOGY: unknown (11) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:232: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Asn 225 230 235 240 Pro Arg Gly Arg Arg His Pro 245 (2) INFORMATION FOR SEQ ID NO:233: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 24 9 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION' SEQ ID NO:233: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg He Ala Tyr Ser 100 105 110 Printed from Mimosa Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Thr Arg 225 230 235 240 Lys Ser Ser Arg Ser 245 Asn Pro Arg Gly (2) INFORMATION FOR SEQ ID NO:234: (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH. 277 amino acids (B) TYPE: amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:234: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Thr 225 230 235 240 Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp Ser Asp 245 250 255 Ser Asp Thr Asn Ala Lys His Ser Ser His Asn Arg Arg Leu Arg Thr 260 265 270 Arg Ser Arg Pro Asn Printed from Mimosa WO 98/51325 PCT/US98/10088 275 (2) INFORMATION FOR SEQ ID NO:235: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH- 258 amino acids (B) TYPE: amino acid (C) STRANDEDNESS- (D) TOPOLOGY, unknown (ii) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.235: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met .Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Thr 225 230 235 240 Pro Pro Ser Arg Glu Ala Tyr Ser Arg Pro Tyr Ser Val Asp Ser Asp 245 250 255 Ser Asp (2) INFORMATION FOR SEQ ID NO.236: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 259 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (li) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: : SEQ ID NO:236 Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 5 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu - 219 ■ Printed from Mimosa 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Arg 225 230 235 240 Pro Tyr Ser Val Asp Ser Asp Ser Asp Thr Asn Ala Lys His Ser Ser 245 250 255 His Asn Arg (2) INFORMATION FOR SEQ ID NO:2 3 7• (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 257 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION . SEQ ID NO:237: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr GlU Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Printed from Mimosa Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Thr Asn 225 230 235 240 Ala Lys His Ser Ser 245 His Asn Arg Arg Leu 250 Arg Thr Arg Ser Arg 255 Pro Asn (2) INFORMATION FOR SEQ ID NO:238: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 247 ammo acids (B) TYPE- amino acid (C) STRANDEDNESS- (D) TOPOLOGY: unknown (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.238.

Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Thr Asn 225 230 235 240 Ala Lys His Ser Ser 245 His Asn (2) INFORMATION FOR SEQ ID NO:239: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 amino acids (B) TYPE amino acid (C) STRANDEDNESS (D) TOPOLOGY, unknown (ii) MOLECULE TYPE, peptide Printed from Mimosa (xi) SEQUENCE DESCRIPTION. SEQ ID NO 23 9: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 10 15 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu 25 30 Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 5 35 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 in Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 -150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Ser Ser 225 230 235 240 His Asn Arg Arg Leu Arg Thr Arg 245 (2) INFORMATION FOR SEQ ID NO:240: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 248 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID N0:240: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 10 15 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu 25 30 Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 30 35 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Printed from Mimosa WO 98/51325 PCT/US98/10088 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Arg 225 230 235 240 Leu Arg Thr Arg Ser 245 Arg Pro Asn (2) INFORMATION FOR SEQ ID NO:241 (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 282 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO.241: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp 60 Gly Asp Val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Val 225 230 235 240 Gly Gin Cys Thr Asp 245 Ser Asp Val Arg Arg 250 Pro Trp Ala Arg Ser 255 Cys Ala His Gin Gly Cys Gly Ala Gly Thr Arg Asn Ser His Gly Cys lie 260 265 270 Thr Arg Pro 275 Leu Arg Gin Ala Ser 280 Ala His (2) INFORMATION FOR SEQ ID NO:242: (l) SEQUENCE CHARACTERISTICS: Printed from Mimosa (A) LENGTH. 257 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:242: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Val 225 230 235 240 Gly Gin Cys Thr Asp Ser Asp Val Arg Arg Pro Trp Ala Arg Ser Cys 245 250 255 Ala (2) INFORMATION FOR SEQ ID NO.243- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 259 ammo acids (B) TYPE ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:243- Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp GlU Gly Asp Lys 40 Trp Arg Asn Lys Lys Phe 45 Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys HIS Asn 65 70 75 80 Printed from Mimosa Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Val Arg 225 230 235 240 Arg Pro Trp Ala Arg 245 Ser Cys Ala His Gin 250 Gly Cys Gly Ala Gly 255 Thr Arg Asn Ser (2) INFORMATION FOR SEQ ID N0:244: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 257 amino acids (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (il) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:244: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro l Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys Val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Thr Printed from Mimosa WO 98/51325 PCT/US98/10088 225 230 235 240 Arg Asn Ser Hxs Gly Cys lie Thr Arg Pro Leu Arg Gin Ala Ser Gin 245 250 255 His (2) INFORMATION FOR SEQ ID NO:245: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH: 282 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ll) MOLECULE TYPE- peptide (Xl) SEQUENCE DESCRIPTION: SEQ ID NO:245: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Tyr 225 230 235 240 Lys His Asp lie Gly 245 Cys Asp Ala Gly Val 250 Asp Lys Lys Ser Ser 255 Ser Val Arg Gly Gly 260 Cys Gly Ala His Ser 265 Ser Pro Pro Arg Ala 270 Gly Arg Gly Pro Arg 275 Gly Thr Met Val Ser 280 Arg Leu (2) INFORMATION FOR SEQ ID NO-246. (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 262 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:246: Printed from Mimosa Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp 60 Gly Asp Val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 140 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys 175 Leu Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp 190 Lys Tyr Leu Lys Ser 195 Ser Lys Tyr lie Ala 200 Trp Pro Leu Gin Gly 205 Trp Gin Ala Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Arg Tyr 225 230 235 240 Lys His Asp lie Gly 245 Cys Asp Ala Gly Val 250 Asp Lys Lys Ser Ser 255 Ser Val Arg Gly Gly 260 Cys Gly (2) INFORMATION FOR SEQ ID NO.247: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 264 amino acids (B) TYPE, amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:247: Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu 20 Leu Glu Tyr Leu Glu 25 Glu Lys Tyr Glu Glu 30 His Leu Tyr Glu Arg 35 Asp Glu Gly Asp Lys 40 Trp Arg Asn Lys Lys 45 Phe Glu Leu Gly Leu 50 Glu Phe Pro Asn Leu 55 Pro Tyr Tyr lie Asp 60 Gly Asp Val Lys Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys 85 Pro Lys Glu Arg Ala 90 Glu lie Ser Met Leu 95 Glu Gly Ala Val Leu 100 Asp lie Arg Tyr Gly 105 Val Ser Arg lie Ala 110 Tyr Ser Lys Asp Phe 115 Glu Thr Leu Lys Val 120 Asp Phe Leu Ser Lys 125 Leu Pro Glu Met Leu 130 Lys Met Phe Glu Asp 135 Arg Leu Cys His Lys 14 0 Thr Tyr Leu Asn Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp Printed from Mimosa WO 98/51325 „ PCT/US98/10088 145 150 155 160 Val val Leu Tyr Met 165 Asp Pro Met Cys Leu 170 Asp Ala Phe Pro Lys Leu 175 Val Cys Phe Lys 180 Lys Arg lie Glu Ala 185 lie Pro Gin lie Asp Lys Tyr 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe 210 Gly Gly Gly Asp His 215 Pro Pro Lys Ser Asp 220 Leu Val Pro Arg Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Cys 225 230 235 240 Asp Ala Gly Val Asp 245 Lys Lys Ser Ser Ser 250 Val Arg Gly Gly Cys Gly 255 Ala His Ser Ser 260 Pro Pro Arg Ala (2) INFORMATION FOR SEQ ID NO:248- (l) SEQUENCE CHARACTERISTICS (A) LENGTH- 259 ammo acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (il) MOLECULE TYPE :: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO-248 : Met Ser Pro lie Leu Gly Tyr Trp Lys lie Lys Gly Leu Val Gin Pro 1 Thr Arg Leu Leu Leu Glu Tyr Leu Glu Glu Lys Tyr Glu Glu His Leu Tyr Glu Arg Asp Glu Gly Asp Lys Trp Arg Asn Lys Lys Phe Glu Leu 40 45 Gly Leu Glu Phe Pro Asn Leu Pro Tyr Tyr lie Asp Gly Asp Val Lys 50 55 60 Leu Thr Gin Ser Met Ala lie lie Arg Tyr lie Ala Asp Lys His Asn 65 70 75 80 Met Leu Gly Gly Cys Pro Lys Glu Arg Ala Glu lie Ser Met Leu Glu 85 90 95 Gly Ala Val Leu Asp lie Arg Tyr Gly Val Ser Arg lie Ala Tyr Ser 100 105 110 Lys Asp Phe Glu Thr Leu Lys val Asp Phe Leu Ser Lys Leu Pro Glu 115 120 125 Met Leu Lys Met Phe Glu Asp Arg Leu Cys His Lys Thr Tyr Leu Asn 130 135 140 Gly Asp His Val Thr His Pro Asp Phe Met Leu Tyr Asp Ala Leu Asp 145 150 155 160 Val Val Leu Tyr Met Asp Pro Met Cys Leu Asp Ala Phe Pro Lys Leu 165 170 175 Val Cys Phe Lys Lys Arg lie Glu Ala lie Pro Gin lie Asp Lys Tyr 180 185 190 Leu Lys Ser Ser Lys Tyr lie Ala Trp Pro Leu Gin Gly Trp Gin Ala 195 200 205 Thr Phe Gly Gly Gly Asp His Pro Pro Lys Ser Asp Leu Val Pro Arg 210 215 220 Gly Ser Pro Gly lie Pro Gly Ser Thr Arg Ala Ala Ala Ser Gly Ala 225 230 235 240 His Ser Ser Pro Pro Arg Ala Gly Arg Gly Pro Arg Gly Thr Met Val 245 250 255 Ser Arg Leu (2) INFORMATION FOR SEQ ID NO:249: (i) SEQUENCE CHARACTERISTICS: Printed from Mimosa (A) LENGTH: 44 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: : SEQ ID NO:249: Ser Gly Ser Pro Pro Cys Cys Cys Ser Trp Gly Arg Phe Met Gin Gly 1 5 Gly Leu Phe Gly Gly Arg Thr Asp Gly Cys Gly Ala His Arg Asn Arg Thr Ser Ala Ser Leu Glu Pro Pro Ser Ser Asp Tyr 40 (2) INFORMATION FOR SEQ ID NO 250: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH 40 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (n) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.250.

Ser His Ser Gly Gly Met Asn Arg Ala Tyr Gly Asp Val Phe Arg Glu 10 15 Leu Arg Asp Arg Trp Asn Ala Thr Ser His His Thr Arg Pro Thr Pro 25 30 Gin Leu Pro Arg Gly Pro Asn Ser 35 40 (2) INFORMATION FOR SEQ ID NO.251: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 21 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.251.

Asp Thr Asn Ala Lys His Ser Ser His Asn Arg Arg Leu Arg Thr Arg 10 15 Ser Arg Pro Asn Gly 20 (2) INFORMATION FOR SEQ ID NO:252: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH- 23 ammo acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:252: Cys Gly Ala Gly Thr Arg Asn Ser His Gly Cys lie Thr Arg Pro Leu 15 10 15 Printed from Mimosa WO 98/51325 PCT/US98/10088 Arg Gin Ala Ser Ala His Gly 20 (2) INFORMATION FOR SEQ ID NO:253- (i) SEQUENCE CHARACTERISTICS.

(A) LENGTH: 11 amino acids c (B) TYPE- amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (lx) FEATURE: (A) NAME/KEY. Modified Site (B) LOCATION: 1 (D) OTHER INFORMATION: "Xaa=Ser or Thr" (A) NAME/KEY- Modified Site (B) LOCATION: 3 (D) OTHER INFORMATION: "Xaa=Arg or Lys" (A) NAME/KEY Modified Site (B) LOCATION: 4 (D) OTHER INFORMATION. "Xaa=Lys or Arg" (A) NAME/KEY Modified Site (B) LOCATION: 6 (D) OTHER INFORMATION: "Xaa=Ser or Leu" (A) NAME/KEY Modified Site (B) LOCATION. 7 (D) OTHER INFORMATION "Xaa=Arg, lie, Val or Ser" (A) NAME/KEY: Modified Site (B) LOCATION: 8 (D) OTHER INFORMATION: "Xaa=Ser, Tyr, Phe or His" (A) NAME/KEY: Modified Site (B) LOCATION: 10 (D) OTHER INFORMATION "Xaa=Phe, His or Arg" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:253: Xaa Thr Xaa Xaa Ser Xaa Xaa Xaa Asn Xaa Arg 15 10 (2) INFORMATION FOR SEQ ID NO:254: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE- amino acid 30 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (ix) FEATURE: (A) NAME/KEY. Modified Site (B) LOCATION. 2 (D) OTHER INFORMATION: "Xaa=Ser, Ala or Gly" (A) NAME/KEY: Modified Site (B) LOCATION: 4 (D) OTHER INFORMATION: "Xaa=Val Or Gin" Printed from Mimosa (A) NAME/KEY: Modified Site (B) LOCATION 7 (D) OTHER INFORMATION: "Xaa=Pro, Gly or Ser" (A) NAME/KEY: Modified Site (B) LOCATION: 8 (D) OTHER INFORMATION. "Xaa=Trp or Tyr" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:254: Asp Xaa Asp Xaa Arg Arg Xaa Xaa 1 5 (2) INFORMATION FOR SEQ ID NO.255 (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 ammo acids (B) TYPE, ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (il) MOLECULE TYPE: peptide (ix) FEATURE: ]_5 (A) NAME/KEY: Modified Site (B) LOCATION: 7 (D) OTHER INFORMATION: "Xaa=Ala or Phe" (A) NAME/KEY. Modified Site (B) LOCATION: 8 (D) OTHER INFORMATION: "Xaa=Arg or His" (xi) SEQUENCE DESCRIPTION: SEQ ID NO:255: Val Arg Ser Gly Cys Gly Xaa Xaa Ser Ser 15 10 (2) INFORMATION FOR SEQ ID NO:256: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: amino acid 25 (C) STRANDEDNESS (D) TOPOLOGY, unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO-.256: Asn Thr Arg Lys Ser Ser Arg Ser Asn Pro Arg 15 10 (2) INFORMATION FOR SEQ ID NO:257: (i) SEQUENCE CHARACTERISTICS- (A) LENGTH- 11 amino acids (B) TYPE- amino acid (C) STRANDEDNESS.

(D) TOPOLOGY: unknown (n) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.257: Ser Thr Lys Arg Ser Leu lie Tyr Asn His Arg Printed from Mimosa 10 (2) INFORMATION FOR SEQ ID NO:258: (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 10 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (n) MOLECULE TYPE, peptide (xi) SEQUENCE DESCRIPTION. SEQ ID NO:258: Ser Thr Gly Arg Lys Val Phe Asn Arg Arg 15 10 (2) INFORMATION FOR SEQ ID NO:259: (l) SEQUENCE CHARACTERISTICS- (A) LENGTH: 11 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY, unknown (ll) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO.259: Thr Asn Ala Lys His Ser Ser His Asn Arg Arg 15 10 (2) INFORMATION FOR SEQ ID N0:260. (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE amino acid (C) STRANDEDNESS: (D) TOPOLOGY unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:260: Asp Ser Asp Val Arg Arg Pro Trp 1 5 (2) INFORMATION FOR SEQ ID NO:261: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids (B) TYPE: amino acid 30 (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:261.

Ala Ala Asp Gin Arg Arg Gly Trp 1 5 (2) INFORMATION FOR SEQ ID NO.262: (i) SEQUENCE CHARACTERISTICS: (A) LENGTH: 8 amino acids Printed from Mimosa WO 98/51325 PCT/US98/10088 (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (11) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION- SEQ ID NO:262: Asp Gly Arg Gly Gly Arg Ser Tyr 1 5 (2) INFORMATION FOR SEQ ID NO:263 (I) SEQUENCE CHARACTERISTICS- (A) LENGTH 4 ammo acids (B) TYPE: ammo acid TQ (C) STRANDEDNESS: (D) TOPOLOGY- unknown (II) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION SEQ ID NO:263: Arg Val Arg Ser 1 (2) INFORMATION FOR SEQ ID NO 264- (i) SEQUENCE CHARACTERISTICS: (A) LENGTH. 12 amino acids (B) TYPE: amino acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (il) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:264: Ser Val Arg Ser Gly Cys Gly Phe Arg Gly Ser Ser 15 10 (2) INFORMATION FOR SEQ ID NO.265 (l) SEQUENCE CHARACTERISTICS: (A) LENGTH: 11 amino acids (B) TYPE: ammo acid (C) STRANDEDNESS: (D) TOPOLOGY: unknown (ii) MOLECULE TYPE: peptide (xi) SEQUENCE DESCRIPTION: SEQ ID NO:265: Ser Val Arg Gly Gly Cys Gly Ala His Ser Ser 15 10 Printed from Mimosa

Claims (41)

WO 98/51325 PCT/US98/10088 WHAT IS CLAIMED IS:

1. A purified protein which specifically binds to a gastro-intestinal tract receptor selected from the group 5 consisting of HPTl, hPEPTl, D2H, and hSI.

2. A protein which binds specifically to a gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the 10 protein comprises an amino acid sequence selected from the group consisting of SEQ ID NOS:1-55 or a binding portion thereof.

3. A protein which binds specifically to a 15 gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the amino acid sequence of the protein is selected from the group consisting of SEQ ID NOS:1-55, or a binding portion thereof. 20 4. The protein of claim 2 which comprises the amino acid sequence substantially as set forth in: SEQ ID NO:

4, SEQ ID NO: 6, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 30, SEtf ID NO: 43, 'SEQ ID NO: 46, or SEQ ID NO: 52, or a binding portion thereof. 25

5. The protein of claim 3, the amino acid sequence of which consists of the amino acid sequence substantially as set forth in: SEQ ID NO: 4, SEQ ID NO: 6, SEQ ID NO: 22, SEQ ID NO: 23, SEQ ID NO: 30, SEQ ID NO: 43, 30 SEQ ID NO: 46, or SEQ ID NO: 52, or a binding portion thereof.

6. A protein of not more than 50 amino acids in length which specifically binds to a gastro-intestinal 35 transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the protein includes, positioned anywhere along its sequence, the contiguous amino - 234 - Printed from Mimosa WO 98/51325 PCT/US98/10088 acid sequence of: Xaa! Thr Xaa2 Xaa3 Ser Xaa4 Xaa5 Xaa6 Asn Xaa7 Arg (SEQ ID NO:253), where Xaax is Ser or Thr; Xaa2 is Arg or Lys; Xaa3 is Lys or Arg; Xaa4 is Ser or Leu; Xaas is Arg, lie, Val, or Ser; Xaa6 is Ser, Tyr, Phe, or His; and Xaa7 5 is Pro, His or Arg.

7. The protein of claim 6 which is not more than 40 amino acids in length. 10

8. The protein of claim 6 which is not more than 3 0 amino acids in length.

9. The protein of claim 6 which is not more than 20 amino acids in length. 15

10. A protein of not more than 50 amino acids in length which specifically binds to a gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the protein includes, 20 positioned anywhere along its sequence, the contiguous amino acid sequence of: Asp Xaa! Asp Xaa2 Arg Arg Xaa3 Xaa4 (SEQ ID N0:254) where Xaax is Ser, Ala, or Gly; Xaa2 is Val or Gin; Xaa3 is Pro, Gly, or Ser; and Xaa4 is Trp or Tyr. 25

11. The protein of claim 10 which is not more than 4 0 amino acids in length.

12. The protein of claim 10 which is not more than 3 0 amino acids in length. 30

13. The protein of claim 10 which is not more than 2 0 amino acids in length.

14. A protein of not more than 50 amino acids in 35 length which specifically binds to a gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the protein includes, - 235 - Printed from Mimosa WO 98/51325 PCT/US98/10088 positioned anywhere along its sequence, the contiguous amino acid sequence of: Val Arg Ser Gly Cys Gly Xaax Xaa2 Ser Ser (SEQ ID NO:255) , where Xaax is Ala or Phe; and Xaa2 is Arg or His. 5

15. The protein of claim 14 which is not more than 40 amino acids in length.

16. The protein of claim 14 which is not more than 10 30 amino acids in length.

17. The protein of claim 14 which is not more than 20 amino acids in length. 15

18. A protein of not more than 50 amino acids in length which specifically binds to a gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the protein includes, positioned anywhere along its sequence, the contiguous amino 20 acid sequence of: NTRKSSRSNPR (SEQ ID NO:256) or STKRSLIYNHR (SEQ ID NO:257) or STGRKVFNRR (SEQ ID NO:258) or TNAKHSSHNRR (SEQ ID NO:259).

19. A protein of not more than 50 amino acids in 25 length which specifically binds to a gastro-intestinal transport receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, in which the protein includes, positioned anywhere along its sequence, the contiguous amino acid sequence of: DSDVRRPW (SEQ ID NO:260) or AADQRRGW (SEQ 30 ID NO:261) or DGRGGRSY (SEQ ID NO:262).

20. A protein of not more than 50 amino acids in length which specifically binds to a gastro-intestinal transport receptor selected from the group consisting of 35 HPTl, hPEPTl, D2H, and hSI, in which the protein includes, positioned anywhere along its sequence, the contiguous amino - 236 - Printed from Mimosa Intellectual Property - . . j j A Office of NZ 501110 - 3 SEP 2001 WO 98/51325. received PCT/US98/10088~~~ acid sequence of: RVRS (SEQ ID NO:263) or SVRSGCGFRGSS (SEQ ID NO:264) or SVRGGCGAHSS (SEQ ID NO:265).

21. The protein of claim 1, 2, 3, 6, 10, 14, 18, 5 19, or 20 which is purified.

22. A composition comprising the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, or 20, bound to a material comprising an active agent, said active agent being of value 10 in the treatment of a mammalian disease or disorder.

23. The composition of claim 22 in which the active agent is a drug. 15

24. The composition of claim 22 in which the material is a particle containing the active agent.

25. The composition of claim 22 in which the material is a slow-release device containing the drug. 20

26. The composition of claim 22 in which the protein is covalently or noncovalently bound to the material.

27. A composition comprising a chimeric protein bound 25 to a material comprising an active agent, in which the chimeric protein comprises a sequence selected from the group consisting of SEQ ID NOS:1-55 or an HPT1-, hPEPTl-, D2H- or hSI- binding portion thereof fused via a covalent bond to an amino acid sequence of a second protein, m which the active agent is of 30 value in the treatment of a mammalian disease or disorder.

28. A composition comprising the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, or 20 covalently bound to a particle containing a drug. 35

29. A composition comprising the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, or 20 covalently bound to a drug. - 237 - Intellectual Property Office of NZ R^™y£D50 liJiOL WO 98/51325 RECtW fc 9 PCT/9S98/lVo88

30. The composition of claim 22 which facilitates the transport of the active agent through human or animal gastrointestinal tissue.

31. The use of a protein of any one of claims 1, 2, 3, 6, 10, 14, 18, 19 or 20 bound to a material comprising an active agent, said active agent being of value in the treatment of a mammalian disease or disorder, for the manufacture of a purified composition for delivering said active agent in vivo to a subj ect.

32. The use of a protein of any one of claims 1, 2, 3, 6, 10, 14, 18, 19 or 20 bound to a material comprising an active agent, said active agent being of value in the treatment of a mammalian disease or disorder, for the manufacture of a purified composition, which facilitates the transport of the active agent through human or animal gastro-intestinal tissue, for delivering a drug to a subject.

33. The use of a protein of any one of claims 1, 2, 3, 6, 10, 14, 18, 19 or 20 bound to a material comprising an active agent, said active agent being of value in the treatment of a mammalian disease or disorder, for the manufacture of a purified composition for delivering a drug to a subject.

34. The use according to claim 31 in which the composition is for oral administration.

35. The use according to claim 31 in which the active agent is a drug.

36. The use according to claim 31 m which the subject is a human. - 238 - 501110 WO 98/51325 PCT/US98/10088

37. The use according to claim 35 in which the subject is a human.

38. The use according to claim 31 m which said composition facilitates the transport of the .active agent through human or animal gastro-intestinal tissue.

39. The use according to claim 33 m which the composition is for oral administration. Intellectual Property Office of NZ - 3 SEP 2001 received - 238A - WO 98/5132S

40. A pharmaceutical composition comprising the composition of claim 22 in a pharmaceutically acceptable carrier suitable for use in humans m vivo.

41. A chimeric protein comprising at least 6 contiguous amino acids of an ammo acid sequence selected from the group consisting of SEQ ID NOS:1-55, that specifically bind to a gastro-intestinal tract receptor selected from the group consisting of HPTl, hPEPTl, D2H or hSI, fused via a covalent bond to an amino acid sequence of a second protein.

42. An antibody which is capable of immunospecifically binding the protein of claim 2, 3, 6, 10, 14, 18, 19 or 20. 15

43. A molecule comprising a fragment of the antibody of claim 42, which fragment is capable of immunospecifically binding said protein. 20 44. A purified derivative of the protein of claim 1 or 2, which displays one or more functional activities of said protein.

45. The derivative of claim 44 which is able to be 25 bound by an antibody directed against said protein.

46. A fragment of the protein of claim 2 comprising a domain of said protein. 30 47. A fragment of the protein of claim 3 comprising a domain of said protein.

48. A nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID 35 NOS:111-164, except as found in nature. Intellectual Property Office of NZ - 239 - - 3 SEP 2001 RECEIVED WO 98/51325 PCT/US98/10088

49. A nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS:56-109, except as found m nature.

50. An isolated nucleic acid comprising a nucleotide sequence encoding the protein of claim 1.

51. A nucleic acid comprising a nucleotide sequence encoding the protein of claim 2, 3, 6, 10, 14, 18, 19 or 20.

53. An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of SEQ ID NOS:111-164 .

54. An isolated nucleic acid comprising a nucleotide sequence selected from the group consisting of seq id nos:56-1Q9.

56. An isolated nucleic acid comprising a nucleotide sequence complementary to the nucleotide sequence of claim 51.

57. An isolated nucleic acid comprising a nucleotide sequence encoding a fragment of the protein of claim 1, 2, or 3, which fragments bind to said gastro-intestinal tract receptor.

58. A nucleic acid comprising a nucleotide sequence encoding the chimeric protein of claim 41.

52. The nucleic acid of claim 51 which is a DNA.

55. The nucleic acid of claim 51 which is isolated.

59. A nucleic acid comprising a nucleotide sequence encoding the fragment of claim 41. Intellectual Property Office of NZ 240 - 3 SEP 2001 RECEIVED h'i 1110 » s. WO 98/51325 PCT/US98/10088

60. The nucleic acid of claim 58 which is isolated.

61. The nucleic acid of claim 59 which is isolated.

62. A recombinant cell containing -the nucleic acid of claim 48, 49 or 50.

63. A recombinant cell containing the nucleic acid of claim 51.

64. A recombinant cell containing the nucleic acid of claim 58.

65. A method of producing a protein comprising growing a recombinant cell containing the nucleic acid of claim 48, 49 or 50 such that the encoded protein is expressed by the cell, and recovering the expressed protein.

66. A method of producing a protein comprising growing a recombinant cell containing the nucleic acid of claim 51 such that the encoded protein is expressed by the cell, and recovering the expressed protein.

67. A method of producing a protein comprising growing a recombinant cell containing the nucleic acid of claim 58 such that the encoded protein is expressed by the cell, and recovering the expressed protein.

68. The product of the process of claim 65.

69. The product of the process of claim 66.

70. The product of the process of claim dTitetlectual Property Office of NZ - 241 - -3 SEP 2001 RECEIVED 501110 WO 98/51325 PCT/US98/10088

71. A pharmaceutical composition comprising a therapeutically effective amount of a composition comprising the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, or 20; and a pharmaceutically acceptable carrier.

72. The chimeric protein of claim 41 in which said second protein is a drug.

73. A nucleic acid comprising a nucleotide sequence encoding the protein of claim 72.

74. A pharmaceutical composition comprising a therapeutically effective amount of the protein of claim 72, and a pharmaceutically acceptable carrier.

75. A pharmaceutical composition comprising a therapeutically effective amount of the nucleic acid of claim 73.

76. The use of a nucleic acid according to claim 73 for the manufacture of a pharmaceutical composition, for use in a method of delivering a drug to a subject.

77. The use of the protein of claim 1, 2, 3, 6, 10, 14, 18, 19 or 20, bound to a material comprising a drug, said drug being of value in the treatment of a mammalian disease or disorder, for the manufacture of a composition for use in a method of treating or preventing a disease in a subject.

78. The use of the protein of claim 1, 2, 3, 6, 10, 14, 18, 19 or 20 covalently bound to a particle containing a drug for the manufacture of a composition, for use in a method of treating or preventing a disease in a subject. Intellectual Property _ 242 _ 0ffice of NZ - 3 SEP 2001 received 50 1110 WO 98/51325 PCT/US98/10088

79. The use of the protein of claim 1, 2, 3, 6, 10, 14, 18, 19 or 20 covalently bound to a drug for the manufacture of a composition, for use in a method of treating or preventing a disease in a subject.*

80. The use according to claim 77 in which the disease or disorder is selected from the group consisting of: hypertension, diabetes, osteoporosis, hemophilia, anemia, cancer, migrains, and angina pectoris.

81. The use according to claim 77 m which the subject is a human.

82. A composition comprising the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, 20, or 46 wherein the protein is coated onto or absorbed onto or covalently bonded to the surface of a nano- or microparticle.

83. A nano- or microparticle formed from the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, 20, or 46.

84. The composition of claim 82, wherein the nano- or microparticle is a drug-loaded or drug-encapsulating nano- or microparticle.

85. A method of detecting or measuring the level of a gastro-intestinal tract receptor in a sample, comprising contacting a sample suspected of containing a gastro-intestinal tract receptor with the protein of claim 1, 2, 3, 6, 10, 14, 18, 19, 20, or 46 under conditions conducive to binding between the protein and any of said receptor in said sample, and detecting or measuring any of said binding that occurs, m which the detected intellectual Property Office of NZ - 243 - - 3 SEP 2001 received 50 \ A WO 98/51325 PCT/US98/10088 or measured amount of binding indicates the presence or amount of the receptor in the sample.

86. A method of identifying a molecule that specifically binds to a ligand selected from the group consisting of the protein of claim 1, 2, 3, 6, 10, 14, 18, or 19, a fragment of said protein comprising a domain of the protein, and a nucleic acid encoding said protein or fragment, comprising (a) contacting said ligand with a plurality of molecules under conditions conducive to binding between said ligand and the molecules; and (b) identifying a molecule within said plurality that specifically binds to said ligand.

87. An isolated nucleic acid encoding a fragment of a gastro-intestinal tract receptor selected from the group consisting of HPTl, hPEPTl, D2H, and HSI, or encoding a chimeric protein comprising said fragment, said fragment consisting essentially of the extracellular domain of the receptor.

88. A cell containing and capable of expressing a recombinant nucleic acid encoding a fragment of a gastrointestinal tract receptor selected from the group consisting of HPTl, hPEPTl, D2H, and hSI, or encoding a chimeric protein comprising said fragment, said fragment consisting essentially of the extracellular domain of the receptor.

89. The cell of claim 88 which contains an expression vector comprising a nucleotide sequence encoding said fragment operably linked to a heterologous promoter. Intellectual Property Office of NZ _ 244 - " 3 SEP 2001 received 501110 WO 98/51325 ~ PCT/US98/10088 '

90. A method for identifying a molecule that specifically binds to a gastro-intestinal tract receptor selected from the group consisting of HPTl, hPEPTl, D2H or hSI, comprising contacting a fragment o*f the receptor, or a chimeric protein comprising said fragment, with a plurality of -test molecules under conditions conducive to binding between said fragment or protein and the molecules, and identifying a molecule within said plurality that specifically binds to said fragment or protein, in which the fragment consists essentially of the extracellular domain of the receptor.

91. The composition of claim 22 for use as a medicament.

92. The composition of claim 28 for use as a medicament.

93. The composition of claim 29 for use as a medicament.

94. The composition of claim 82 for use as a medicament.

95. The composition of claim 23 in which the drug is insulin or leuprolide.

96. The composition of claim 24 in which the active agent is insulin or leuprolide.

97. The composition of claim 25 m which the drug is insulin or leuprolide. Intellectual Property Office of NZ 245 - ~ 3 SEP 2001 received WO 98/51325 PCT/US98/10088

98. The composition of claim 28 in which the drug is insulin or leuprolide.

99. A purified protein according to claim 1 substantially as herein described or exemplified.

100. A protein according to claim 2, 3, 6, 10, 14, 18, 19 or 20 substantially as herein described or exemplified.

101. A composition according to claim 22 substantially as herein described or exemplified.

102. A composition according to claim 27, 28 or 29 substantially as herein described or exemplified.

103. A use according to claim 31, 32 or 33 substantially as herein described or exemplified.

104. A pharmaceutical composition according to claim 40 substantially as herein described or exemplified.

105. A chimeric protein according to claim 41 substantially as herein described or exemplified.

106. A antibody according to claim 42 substantially as herein described or exemplified.

107. A molecule according to claim 43 substantially as herein described or exemplified.

108. A purified derivative according to claim 44 substantially as herein described or exemplified. 246 Intellectual Property Office of NZ - 3 SEP 2001 RECEIVED WO 98/51325 PCT/US98/10088

109. A fragment according to claim 46 or 47 substantially as herein described or exemplified.

110. A nuclerc acid according to claim 48, 49, 50, 51, 56, 57, 58 or 59 substantially as herein described or exemplified.

111. A recombinant cell according to claim 62, 63 or 64 substantially as herein described or exemplified.

112. A method according to claim 65, 66 or 67 substantially as herein described or exemplified.

113. A product according to claim 68, 69 or 70 substantially as herein described or exemplified.

114. A pharmaceutical composition according to claim 71, 74 or 75 substantially as herein described or exemplified.

115. A nucleic acid according to claim 73 substantially as herein described or exemplified.

116. A method according to claim 75 substantially as herein described or exemplified.

117. A use according to claim 77, 78 or 79 substantially as herein described or exemplified.

118. A composition according to claim 82 substantially as herein described or exemplified.

119. A nano- or microparticle according to claim 84 substantially as herein described or exemplified. Intellectual Property 247 Office of NZ - 3 SEP 2001 RECEIVED 5011 "*0 WO 98/51325* PCT/US98/10088

120. A method according to claim 85 substantially as herein described or exemplified.

121. A method according to claim 86 substantially as herein described or exemplified. #

122. An isolated nucleic acid according to claim 87 substantially as herein described or exemplified.

123. A cell according to claim 88 substantially as herein described or exemplified.

124. A method according to claim 90 substantially as herein described or exemplified. Intellectual Property Office of NZ - 3 SEP 20Q1 RECEIVE0 - 248 -