CA2175579A1

CA2175579A1 - Structured synthetic antigen libraries as diagnostics, vaccines and therapeutics

Info

Publication number: CA2175579A1
Application number: CA002175579A
Authority: CA
Inventors: Chang Yi Wang; Timothy J. Zamb; Jacek Kowalski; Alan M. Walfield; Wayne C. Koff; John Ye; Barbara Hosein; Douglas F. Nixon; Stephen M. Kaminsky
Original assignee: Individual
Current assignee: United Biomedical Inc
Priority date: 1993-10-26
Filing date: 1994-10-26
Publication date: 1995-05-04
Also published as: WO1995011998A1; EP0725838A4; IL111417A0; EP0725838A1; AU8091694A

Abstract

The present invention relates to "structured synthetic antigen libraries" (SSAL) composed of related peptides synthesized simultaneously in a single peptide synthesis. This "stuctured" library contrasts to those libraries previously described as "random peptide libraries" in that the order or structure within a synthetic antigen is provided by invariant amino acid residues that define the framework sequence of the synthetic antigen. The specific amino acids and their frequency of appearance at a variant locus within aligned peptide sequences is defined by the primary sequences of the several variants that make up the alignment used to construct the antigen peptide library. A method of constructing an open diagnostic, vaccine or therapeutic for a mutational infectious agent is also provided. The invention further provides the SSAL in diagnostic methods, kits vaccination methods, vaccine compositions and pharmaceutical compositions. The libraries are prepared from variable domains in proteins and provide improved vaccines, diagnostics and therapeutics for infectious agents, etc., from such proteins.

Description

~ WO9S111998 217~7,g PCT/IJS94112268 :~l~UelU~:Il SYNTHETIC ANTIGEN T,TFIR1~T~TT.`.C
AS DIAGNOSTICS, VACCINES AND THERAPEUTICS
FT~T~n OF THE INVT`NTION
The present invention relates to "structured synthetic antigen libraries" (SSAL) composed of related peptides synthesized simultaneously in a single peptide synthesis.
This "structured" library contrasts to those libraries previously described as "random peptide libraries" in that the order or structure within a synthetic antigen is 15 provided by invariant amino acid residues that def ine the rL ~Lk sequence of the synthetic antigen. The specific amino acids and their CL e~uc ... y of appearance at a variant locus within aligned peptide se~u~..ces is defined by the primary sequences of the several variants that make up the alignment used to construct the antigen peptide library. A
method of constructing an open diagnostic, vaccine or therapeutic for a mutational infectious agent is also provided. The invention further provides the SSAL in diagnostic methods, kits, vaccination methods, vaccine 25 compositions and rh;lrr~~~utical compositions. The libraries are prepared from variable domains in proteins and provide improved vaccines, diagnostics and therapeutics for infectious agents, etc., from such proteins.
BACKGROUND OF TTTF TNV~NTION
Synthetic peptides have been used increasingly to map antigenic or 1 -, ; c sites on the surface of proteins.
These approaches have used: (l) nested sets of peptides as in "site-directed serology" to identify diagnostically useful epitopes; (2) overlapping sets of peptides to scan the length of a protein antigen to locate antigenic sites (a technique colloquially known as pepscan), and (3) totally random peptide libraries which can be systematically analyzed to characterize particular antigenic sites.

Wo 95/11998 PCr/USs~/l2268 217~9 In this regard, synthetic peptides provide a powerful tool for the diagnostic field, particularly in the areas of diagnostic virology, blood screening and others. For example, using site-directed serology, sensitive and specif ic tests have been developed f or the envelope and core proteins of human i -'-ficiency virus (HIV) [see, e.g., U . S . Pat . Nos . 4 , 7 3 5 , 8 9 6 and 4 , 87 9 , 2 12 ], f or the gag protein of human T-ly ~Ll~ic viruses I and II (HTLV I/II) [see, e.g., U.S. Pat. No. 4,833,071], and the structural and n.", ~LLU~:LUL~1 proteins of the hepatitis C virus (HCV) [see, e.g., U.S. Pat. No. 5,106,726].
In certain cases, peptide-based assays are unmatched in their ability to differentiate between or among closely related viruses, a problem often associated with other common diagnostic tests based on either viral lysates or ~' inAntly-produced proteins. While having many advantages, there are still pitfalls to peptide-based immunodiagnostics, as well as the other formats for qnnstic tests, when dealing with infectious agents that vary significantly over time or may have many serotypes . Peptide-based and other i i ~nr ~tics are static by design and can not ~ e such variation.
As is well-known, many proteins from infectious agents, or sites on those proteins, can vary in sPT~Pnre by strain.
At a given time or in a particular geographic locale, a particular antigen may contain one cr multiple point mutations relative to an arbitrary prototype strain.
Dynamic variation thereby creates an extremely complex antigenic profile for a given site for which sensitive or specific detection may be increasingly difficult as the site "drifts" further from the prototype. I hPlPRR, each antigen maintains a ~r, Lell motif" or IILLLU-_LU~l rL I~ Lh" that is representative of the prototype sequence from which the variants arise. The structural LL . ~Lh ~ WO95111998 21 7~7~ PCTiUS94/12268 , consists of generally invariant residues which can be exploited in con~unction with the inherently variable residues to design improved diagnostic tests that overcome the limitations of the present tests.
Accordingly, the present invention provides a means to ~U-I LLU-,~ a synthetic peptide library termed a 'I~LLU- ~UL'd synthetic antigen library" (SSAL) which addresses the need for "open" diagnostics, i.e. diagnostic kits which can expre6sly ~ t;e antigenic variation.
Site-directed synthetic peptides also find use as vaccines. Vaccine development and delivery has been highly ~u~-_ess rul as a public health strategy in the control of acute viral and bacterial infectious diseases. However, several major globally i ~L Lal~L infectious diseases has proven elusive to attempts at designing effective vaccines.
A r~ ULL~.lL theme among certain pathogens, contributing to llncllrr c.ccful or limited success of vaccines, is antigenic variation. Extensive antigenic variation, for example, is a hallmark for example of HIV (AIDS), rhinovirus (the common cold), influenza virus (flu), Borrelia hllr~dorferi (Lyme disease), (Ihl~mydia (pelvic inflammatory disease), pl~ i~1m falci~ar-~m (malaria), ~rrYr~~~ S~L (Chagas' disease). While biotechnology may enable production of large ~uantities of purif ied antigens containing protective epitopes, antigenic variation has thus far proved to be a significant obstacle for vaccine development in certain pathogens .
Effcctive vaccines generally must induce long term memory I er~ollses SO that a s-~hceq~ t protective response is elicited immediately upon exposure to a pathogen. A single recombinant protein or a single peptide confers only limited protective immunity to pathogens that exhibit extensive antigenic variation because of their limited capacity to induce heterotypic immune responses. Synthetic peptide W0 95/1 1998 PCI IUS9~/12268 technology using SSALs offers a powerful tool to develop the next generation vaccines because they ~nc -c a broad range of sequences required to overcome problems associated with antigenic variation a3 previously ~liRr~lcs_<l for diagnostic markers. L
~herefore, a novel strategy for vaccine-induced protection against pathogens having multiple strains, forms, serovars, serotypes, etc., is required. The "structured synthetic antigen library" (SSAL) provides a solution to the problem of developing vaccines for such pathogens for the following reasons. Pirst, SSALs can be synthesized to address serological variants of each major neutralizing determinant for a pathogen, then combined for effective multi ~ ~ vaccines. For example, current strategies for HIV may focus on the use of a mixture of 5-lO synthetic peptides representing variant c-q11_n-_- within a clade for a single principal neutralizing domain (V3 loop). An antigenic peptide library can be cu~ ed to address not only the 5-lO major l~-yL_S ..~ative se~-e1~es, but also the 20 th~-llc:~n(lc of variants within each clade that the virus is capable of producing escape from neutralizing antibodies.
Second, SSALs can be synthesized to provide T-helper epitopes for a pathogen, thus ~Ve~L~ in~ the problem of -t; c population diversity ( i . e . L e~ ,..ders vs .
non-responders) which has thus far limited the effioacy of experimental synthetic peptide vaccines. Third, SSALs can also be synthesized to include cytotoxic T cell (CTL) epitopes, again to overcome the problem of i , ic population diversity. Such SSALs can thereby stimulate CTL
responses in broad population~-., which in conjunction with a complementary serological response, is required for successful vaccines.
The "structured synthetic antigen library" differs from the random peptide libraries known in the art (see, e.g., ~ WO 95/11958 2 i 7 S 5 7 9 PCT/US94/12268 Jung et al., 1992, ~ng~. Chem. 1~- ~a- ~,. 31:367).
Fully random peptide libraries, produced both recombinantly and 6ynthetically, have been used to identify receptor ligands, to determine 6ubstrate specificity, to identify epitopes. ~oreover, recombinantly pLod.lced antibody librarie6, typically containing millions of randomly generated antibody Fab chains, have been de6cribed to select for high affinity antibodies.
Among the previous report6 was the construction and screening of va6t libraries of peptides and proteins for the ability to bind a variety of receptor molecules. Thi6 wa6 conducted by genetic techniques fu6ing DNA encoding a peptide to a filamentou6 bacteriophage phage coat protein, to another protein for di6play on E. coli cell6, or to a DNA
binding protein 6uch as a lac repre660r. In those approaches, peptides with the desired binding properties were isolated by affinity selection (panning) on an immobilized receptor. After several rounds of panning, individual clones from an enriched clonal population were analyzed to rl~t~rmin.o the binding properties and seguences of the peptides that they contained.
Further, there are reports of synthetic peptide combinatorial libraries composed of mixtures of free peptides in quantities sufficient for use in ligand-binding assays. In one instance, the screening of a positional scanning synthetic library combined with an iterative selection and synthesis process, permitted the systematic identification of optimal peptide ligands from a "random peptide library" composed of 34 million hexa-peptides (Dooley et al., 1993, ~ ~i- ~2:1509).
~ ohren et al. (1993, J. ~. ~. 177: 1713) reported a site-specific positional scanning synthetic peptide library (certain residues were fixed (anchors) and other positions had a random a~ ,L i ~ of residues) . This peptide library WO 95/11998 6 PCrll~S9~112268 was used to identiy critical residues in peptide binding in the "pockets of NHC class I Rb molecules. Blake et 81.
(1992, Bioconiuq Chem. 3:510) described random tetrapeptides (a set of 50,62S peptides) and hexapeptides (a set of 16,777,216 peptides) to identify residues important in the reactivity of the peptides with antibodies. The library was highly variable at each position.
.~` ^hf.-- et ~l. (1992, Eur. J. Immunol. 22: 1405) reported a "limited complexity" peptide library to analyze the peptide preference of MHC class I molecules. This study taught that a singly L~:~Le:selll.ed T cell epitope could be detected and further suggested using the library to identify new T cell epitopes. The peptide library consisted of 8-mer or 9-mer libraries/mixtures with equimolar amounts of the variable amino aoids at the multiply substituted positions and was used for epitope mapping. The peptides were based on the 9-mer Sendai virus NP peptide and the VSV-N peptide (specific for the mouse Kb locus) with the variable amino ~cids from vLL~vlldingly known Kb epitopes.
E~ortin et ~l. (1992, Biochem. Int. Z6: 731) reported a series of S synthetic peptide libraries of 7 amino acids in length wherein the last two carboxyl reæidues were totally random, i . e. included all twenty amino acids in equal proportion. One of these mixtures inhibited platelet adhesion.
Gras-Nasse et al. (1992, Pe~tide ~ç~. 5: 211) described a "mixotope" synthetic peptide library from the hypervariable gpl20 V3 loop of }IIV-1 for use as a vaccine.
The mixotope was 22-25 amino acids in length with invariant 30 and variant positions. The variant-position residues (also termed degenerate positions) consisted only of those amino acids which exceeded 7% Ahlln~l~n~-e at a particular position and were incorporated in equimolar amounts.

~WO9~/11998 1 7Ss7~ PCrlUSs4/12268 The peptide library approaches described above with libraries of, ~ i 70~1 complexity generally provide a research tool for the selection of a particular ligand or antibody to facilitate further drug design. The present invention provides ''~LLu~LuL.l synthetic antigen libraries"
SSAL as the key bioactive ingredient in vaccines, diagnostics and therapeutics. Such libraries provide the broad range of seqUon~oc necessitated by the strain or antigenic variation by anticipating temporal and geographic variation in antigens. The resulting SSALs are collections of peptides that maintain the character of the antigen and can simultaneously provide cross-reactivity to multiple strains of an infectious agent.
STIMMARY OF THE INVT NrrToN
The present invention provides a library of peptides comprising a known antigenic site, epitope or ligand on a protein molecule, wherein (1) said library is optionally linked to a substantially invariant peptide domain or a core branched sequence; (2) said library consists of an ensemble domain; (3) the overall peptide length is about 8 to about 100 amino acids; (4) the sequence of said onC- I lo domain is repre6ented by a cnnConc~c formula; (5) said onco~hle is Jcnic, diagnostic for the epitope or is a therapeutic;
and (6) the cnnConc~l~ formula provides that each sequence position in the ensemble contain either a single amino acid or multiple amino acids, and that when a position contains multiple amino acids, (a) the identity and ratio of those amino acids being determined by the relative prevalence of amino acids in a consensus of known variant sequences for that epitope or ligand or (b) the identity of those amino acids being determined by the amino acids present in a c~nConcl~c of known variant sequences for that epitope or ig~ nd the rat1o of ~no aoldG 1~ eq~li~olar.

WO 95111998 PCT/Us9~/12268 2i~5~ 9 Furthermore the antigenic sites, epitopes or ligands of the library are from a virus, bacterium, paracite, tumor antigen, allergen or other protein antigen, from a diagnostic marker site of an infectiouc agent or disease, 5 from a therapeutically valuable ligand or from a helper T
cell epitope or a cytotoxic T lymphocyte tCTL) epitope.
More particula~ly, these antigenic sites, epitopes or ligands can be from an envelope, core, NS1, NS3, NS4 or NS5 proteins of HCV; a gpl20 V3 loop, gp41 envelope protein or the gp40 envelope protein of HIV; an envelope protein of HTLV I/II; an HA protein or a mutant HA domain of influenza A virus; an major outer membrane protein of rhlamvdia trachomtis; a neur~mini~Ace~ SAPA or CRA site of :~. cruzi;
an envelope B cell slte or helper T cell site of type 2 I)engue virus; an M protein of streptococcus; a carbohydrate recognition site of selectin; a y~ ; C~ llR helper T cell sites; a CTL epitope from HIV gag specific for HLA-B27; or an OspA, OsaB, OspC or flagellin protein of Lyme disease.
SSAL libraries of the present invention are also shown in Figs. 2-11, 13, and 15-34.
Another aspect of the invention i6 directed to peptide compositions containing the above libraries, including ; c peptide compositions.
still another aspect of the invention provides a method of detecting antibodies associated with an infectious agent or a disease state which comprises using an effective amount of the subject peptide compositions in an 1 --ccny uu~.lu~. Also provided is a method of detecting infection or a disease state which comprises contacting an effective amount of the sub~ect peptide , jt;,rnc with a body fluid, tissue or tissue extract in an; --cc~y ~u~e-lu~:~
for a time sufficient to form a complex between said peptide composition and any antibody in said fluid, said tissue, or said tissue extract, and subjecting said complex to a ~WO95/11998 17~S79 PC~/USg~/12268 detecting means. Preferably, the i CCAy procedure is an ELISA or a PHA procedure. Similarly, the invention provides a kit for detection or diagnosis of an infectious agent or a disease state comprising a f irst container 5 adapted to contain the subject peptide composition. In one L, the kit is an ELISA or PHA test kit.
A still further aspect of the invention relates to a method of treating a disease which comprises administering an effective amount of the subject peptide compositions to a patient for a time sufficient to elicit an efficacious result. In this respect, the invention provides a pharmaceutical composition comprising the subject library, or a rhAr~-^~utically acceptable salt thereof, and a rhArr ~~ltically acceptable carrier.
Yet a further aspect of the invention is directed to a method of vaccination which comprises administering to a mammal an amount of the subject library effective to produce a protective immune response against an infectious agent which provided the basis of the library as well as a vaccine composition therefor. Such vaccine composition contains the subject library, or an acceptable salt thereof, formulated with immunologically acceptable carrier or adjuvant.
Another aspect of the invention is directed to a method of making an open diagnostic, vaccine or therapeutic composition for a mutatable infectious agent or for a protein cite of known diversity which comprises (l) aligning a collection of primary amino acid sPq~ n- ~C for a related family of antigens, epitopes, diagnostic markers or therapeutic sites from said mutatable infectious agent or said protein site; (2) identi~ying invariant and variant amino acid positions in said A1 i~ L; (3) calculating a ronc~.ncl-e formula for the sequence of a DLLu-;LuLed synthetic antigen library (SSAL); (4) preparing the SSAL from the ~ Qnc,~ncl1c formula. The cnnc~ncllc formula is ~9Ot~rm1n~ by WO95/11998 217 ~ 9 ` PcrluS9J/12268 from ~YAm;n;n~ each sequence position in the alignment to ascertain the presence of a single amino acid or multiple amino acid6. When a position contains multiple amino acids, then (a) the identity and ratio of the multiple amino acids 5 at that position in the SSAL i6 ~l~t~rm;n~ by the relative prevalence of amino acids in the alignment, (b) the identity of the multiple amino acids is determined by the amino acids present in the ~lignment and the ratio of the multiple amino acids is equimolar, or (c) the identity of the multiple amino acids is determined by the amino acids present in the Al il3 L and the ratio of the multiple amino acids is weighted towards the pL~ ',' ;ni~nt amino acid at said position. The process is repeated for each position of the sequence which can be r~Lesc ..Led by multiple amino acids .
This method for making a vaccine, diagnostic or therapeutio SSAL can be used with mutatable infectious agents from a virus, bacterium or parasite or with a protein site from a tumor antigen, allergen or other protein antigen, from a helper T cell epitope or a CTL epitope.
Similarly, the family of antigens, epitopes, diagnostic markers or therapeutic sites can be selected from an envelope, core, NSl, NS3, NS4 or NS5 proteins of HCV; a gpl20 V3 loop, gp41 envelope protein or the gp40 envelope protein of HIV; an envelope protein of HTLV I/II; an HA
protein or a mutant HA domain of influenza A virus; an major outer membrane protein of t'hl~mvdia trachomtis: a neuraminidase, SAPA or CRA site of ~. ~; an envelope 1~
cell site or helper T cell site of type 2 Dengue virus; an M
protein of streptococcus; a ca.l,u~lydLc.te recognition site of selectin; a pL~ lC helper T cell site; a CTL epitope from HIV gag specific for HLA-B27; or an OspA, OspB, OspC or ~1~ge111n prot~1n of Lyme ~se~e.

W095111998 5~79 j PCT/U594/12268 BRIEF DECrRTPTT(~` OF TTTF ~RAWTNGS
Fig. l illustrates a generalized mathematical and chemical representation of a structured synthetic antigen library (SSAL).
,. 5 Fig. 2 illustrates a primary amino acid sequence alignment (SEQ ID NO:l) of an antigenic NS4 peptide (47-mer~
from eight representative strains of HCV. rqore weight (i.e.
three times the listing frequency) is given to the prototype 6equence used in the construction of an SSAL HCV NS4 to allow easy calculation of the ratio for each amino acid at the specif ied position .
Fig. 3 illustrates a primary amino acid scquence A1 1,; t (SEQ ID NO:2) of an antigenic core peptide (61-mer) from eight IepLese..tative strains of ~CV. An SSAL HCV
15 core is similarly cu.. sLLù- ~ed based on the given antigen ~Llu-;Lu~e and its source sequences.
Fig. 4 illustrates a primary amino acid s~q~l~n~ e ~lil; L (SEQ ID NO:3) of an antigenic NS3 peptide (80-mer) from eight Ley~eSe:llLatiVe strains of HCV.
An SSAL HCV NS3 is similarly ~un~Llu.Led based on the given antigen ~LLU-,LUL~: and its source S~-q~lenr~c.
Fig. 5 illustrates a primary amino acid sequence A1i; -- L (SEQ ID NO:4) of an antigenic NS5 peptide (44-mer, Sequence ~l) from eight representative strains of HCV. An SSAL HCV NS5 (#l) is similarly constructed based on the given antigen structure and its source sequences.
Fig. 6 illustrates a primary amino acid sequence A1 i- ~nt (SEQ ID NO:5) of an antigenic NS5 peptide (41-mer, Sequence ~2) from eight representative strains of E~CV. An SSAL HCV NS5 (#2) is similarly constructed based on the given antigen ~LluuLure and its source c~q~ n~ c.
Fig. 7 illustrates a primary amino acid sequence - alignment (SEQ ID No: 6) of an antigenic Env peptide (55-mer, Sequence #l) from sixteen representative strains of HCV. An Wo95/11998 21~ ~S~ ~ PCTIU59~112268 SSAL HCV ENV (~l) is similarly cull,LLu-,~ed ba6ed on the given antigen l,LLu.,Lu~ and it6 60urce sequence6.
Fig. 8 ilLustrates a primary amino acid sequence alignment (SEQ ID N0:7) of an antigenic Env peptide (44-mer, Sequence #2) from sixteen L~uL.se11~ative 6trains of HCV. An SSAL HCV ENV (#2) is constructed ba6ed on the given antigen 6tructure and its source sequences.
Fig. 9 illustrates a primary amino acid sequence alignment (SEQ ID N0:8) of an antiyenic Env peptide (40-mer, Sequence #3) from sixteen ~,uLes~~ ative strains of HCV. An SSAL HCV ENV (#3 ) is constructed based on the given antigen structure and its 60urce sPql1~n~ c~
Fig. lO illustrates a primary amino acid SPq~PnrP
alignment (SEQ ID N0:9) of an antigenic Env peptide (35 mer, Sequence #4) from ten r~uLes~l,tative 6trains of HCV. An SSAL HCV ENV ( ~4 ) i6 constructed based on the given antigen structure and it6 60urce s~ .. c.
Fig. ll illustrates a primary amino acid s~q~ e ~li; L (SEQ ID NO:lO) of an antigenic NSl peptide from eight representative strains of HCV. An SSAL HCV NSl is constructed based on the given antigen structure and its source 6equences.
Fig. 12 illu6trates four types of heteromeric synthetic carriers used in the production of branched SSALs: dendritic (A), linear (i3), tree-like (C), and tetrameric (D).
Attached onto the core carriers is the SSAL 6ynthesized according to a 6pecif ied mathematical and chemical Lt:pr~s~1.Lation. Each dash shown within a peptide represents a particular amino acid ~ULL- ~IJ.. ~l;n~ to a conserved 30 position in the original protein. Each 0 represents a position where peptides in the library contain any one of a f ixed number of amino acids determined from the composition o 1 the amino acid pool used in that cycle of synthesis . The J~Wo95111998 SS79 PCTIU59J/12268 variable positions can be at any position in the peptide except branch points.
Fig. 13A illustrates a primary amino acid sequence aliynment (SEQ ID N0:11, top Fig. 13B) of an antigenic gpl20 Env peptide (33 mer), .e~Le~ellLing the principal neutralizing ll_t_rminAnt (PND), the V3 domain, of the HIV-l.
An SSAL HIV-l gpl20 V3 is constructed according to a previously optimized V3 rL ~Lk (Fig. 13B; SEQ ID N0: 11) .
Sixteen V3 6equences representative of geographically distinct HIV field isolates were used with more weight given to the r-^nC~nC~ S s-quonre to allow easier calculation of ratios for the various amino acids. An SSAL is constructed based on the given antigen structure and its source sequences. The SSAL HIV-l gpl20 V3 of 109 complexity is shown in Fig. 13C (SEQ ID N0:41). Fig. 13D depicts the neutralization titers found in the serum derived from the guinea pigs i i 701 with the SSALs of the indicated complexities and neutralization titers evoked by monovalent octameric V3; - V3MN and V39l_330 Fig. 14 illustrates a primary amino acid sequence A~ t (SEQ ID No:12) of a highly antigenic gp41 Env peptide (35 mer) from 50 ~e~LéSel~l atiVe strains of HIV-l.
The source of the S_q-lon^_C is derived from Human Retroviruses and AIDS, 1993, ed. by G. Myers et al.
published by Los Alamos National Laboratory, USA. An SSAL
is c~,l.DLLu~Led based on the given antigen structure and its source sequences.
Fig. 15 illustrates a primary amino acid Dequence Ali; -- L (SEQ ID No:13) of an antigenic gp40 Env peptide (36 mer) from 2^ representative strains of Type 2 ~uman T -~^ficiency Virus. The source of the sequon~^~-c is from Human Retroviruses and AIDS, 1993, ed. by G. Myers et al.
pub l~he~ by Los Al~a N.tion~l L:boratory, II:A. An S5AL

2 ~ 7 5 ~ ~ 9 14 PCrlUss4/l2268 is constructed based on the given antigen structure and its source sequences.
Fig. 16 illustrates a primary amino acid sequence alignment of three antigenic Env peptides from HTLV-I/II
(SEQ ID NO:14, Fig. 16A; SEQ ID NO:15, Fig. 16B; SEQ ID
NO:16, Fig. 16C). An SSAL i5 cu,,,LLuuLed based on the given antigen YLLUULUL~ and its source s~ e~:.
Fig. 17 illustrate8 a primary amino acid 6eu,u~ _e (SEQ
ID NO:17) of the critical domaln from Influenza A virus hemagglutinin (HA), a surface glycoprotein capable of eliciting virus-neutralizing antibodies.
t of the complete amino acid sequence of this domain, from residues 91-164, is shown for the H3N2 strains of Inf luenza virus A . The numbering of the amino acids begins with the f irst amino acid of the mature HA sequence (i.e. following cleavage of the signal sequence) as the number one . The amino acid sequences were extracted f rom the EMBL and the SWISSPROT Databases.
Fig. 18A illustrates a primary amino acid sequence alignment (SEQ ID NO:18) of the critical HA domain from the HlN1 strains of Inf luenza A virus capable of eliciting virus neutralizing antibodies.
The structural organizatlon of the antigenic sites and the sequence sources are the same as described in Fig. 17.
WIL and FPR834 strains each harbor a single amino acid deletion within this domain. The location of the deletion is marked by X. Therefore, a separate library was _o~.~LLu-~ed to r~:uL~st.lL these c~qu~-n~ (SEQ ID NO:40; Fig.
18B) .
Fig. 19 illustrates a primary amino acid s~ql-~n~
ill;, L (SEQ ID NO:19) of the critical HA domain from two H2N2 strains of Inf luenza A virus capable of eliciting virus neutralizing antibodies.

~W095/11998 2 ~ 75~ 7 1 PcrluS94112268 Fig. 20 illustrates an SSAL for a Nutant Form of the Critical Influenza Virus HA Neutralizing Domain. Synthetic peptides were made from the Cpqllon,~Pc that immediately f lank the loop structure present in the HA neutralizing domain which is capable of P~ it~n~ antibody that partially neutralizes virus infectivity. Therefore, these sequences are likely to form a conformationally-~PpPn~lPnt epitope important f or eliciting a protective immune response .
Antigen libraries have been constructed to mimic this antigenic IjL~U~,LU~C! by linking the amino acid sequences present on either side of the loop. These libraries represent deletions of the amino acid residues pre6ent between Cys residues at positions 97 and 139 in the original HA domain structure for the H3N2 type virus. The sequence of the deletion libraries wa6 extended at the amino-terminus to include all amino acids I~,L~_.-Led between the first Cysteine residue of the loop structure and the next Cys residue on the amino-terminal side of the loop. Structured libraries representing this mutant form of the HA
neutralizing domain are presented for H3N2, HlN1 and H2N2 influenza A viruses (SEQ ID NO:20, Fig. 20A; SEQ ID NO:21, Fig. 20B; SEQ ID No:22, Fig. 20C; respectively).
Fig. 21 illustrates the amino acid sequence of the first variable domain ~rom the major outer membrane protein (MOMP) of Chlamydia trachomatis. The primary amino acid sequence for the ~Lru-:LuL~:d synthetic antigen library for VDI" and VDIb is shown in Figs. 21A and 21B respectively (SEQ ID No:23 and SEQ ID NO:24, respectively). The chlamydia VDI domain sequences were extracted from the EMBL
database and from Raltenboeck, et al. (1993, J. Bacteriol.
175: 487) .
Fig. 22 illustrates the amino acid sequence of the fourth variable domain (VDIV) from the major outer membrane protein (MOMP) of Chlamydia L~ ~-,I u~ Lis. The primary amino Wo 95111998 21~ S 5 ~ ~ PCT/~1591ll2268 ~
acid sequence for the structured synthetic antigen library for VDIV,~ and VDIVb is shown in Figs. 22A and 22B
respectively (SEQ ID NO:25 and SEQ ID N0:26, respectively).
The chlamydia VDIV domain sequences were extracted from the EMBL database and from Kaltenboeck, et al. (1993).
Fig. 23 illustrates a primary amino acid sequence alignment (SEQ ID NO:27) derived from a 3eries of 43 repeat6 present as a contiguous sequence of amino acid residues (596-1110) in the T. erlzzi neur~-nini~ protein (NA) and a series of 14 repeats present as a contiguous sequence in the T . cruz i SAPA protein .
Fig. 24 illustrates a primary amino acid sequence alignment (SEQ ID NO:28) of a series of 21 repeats present as a contiguous sequence in T. eruzi cytoplasmic repetitive antigen (CRA).
Fig. 25A illustrates a primary amino acid sequence alignment (SEQ ID NO:29) of a 65 mer peptide L~ple6t~ ative of the critieal neutralizing determinant of the Env protein from 33 antigenic variants of type 2 Dengue virus.
Fig. 25B illustrates a primary amino acid sequence (SEQ
ID N0:30) of a 17-mer peptide Le~LèScll~atiVe of a helper T
cell epitope present in the Env protein from 33 antigenic variants of type 2 Dengue virus.
Fig. 26 illustrates a primary amino acid seq~ nr~e ~1;; L (SEQ ID N0:31) of three ~y~ ecific sequences from the N-terminus of the streptococcal M protein. This N-terminus portion of the protein has been shown to be the site that elicits opsonie, or anti-phagocytic, antibodies.
Fig. 27 illustrates a primary amino acid sequence Al;~3 ~ (SEQ ID N0:32) of a T cell helper epitope from the collagen type I/II proteins.
Fig. 2~3 illustrates a primary amino aeid sequenee i~li; ~ (SEQ ID NO:33) of a ~_~LbollydLclte recognition site ~rom a group of selectin molecules.

~WO95/11998 Figs. 29A and B illustrates SSALs from two helper T
cell determinants (Th), SEQ ID NO:34 and SBQ ID NO:35, respectively. The general feature of such SSALs is that the degenerate Th sequence is separated from the LHRH sequence, 5 EHWSYGLRPG, by two Gly residues. The Gly residues act as a spacer between the Th epitope and the LHRH sequence.
Fig. 30 illustrates a primary amino acid 5~q~ nl e alignment (SEQ ID NO: 36) of an HLA-B27 restricted self peptide antigen. The antigen sequences are derived from histone H3, HSP89a, HSP89B, HEF 2, Helicase, rihos protein and L28 (ribosomal protein).
Fig. 31 illustrates a primary amino acid sequence alignment (SEQ ID NO:37) of a HLA B-27 restricted HIV GAG
peptide CTL antigen. The GAG CTL antigen sequences represent those derived from HIV-l MN, HIV-l ELI, HIV-2 ROD
and multiple HIV patient isolates.
Fig. 32 illustrates two SSALo5pA antigens by ~-h~mic~l and mathematical representations #1 and #2 (SEQ ID NO: 38 and SEQ ID NO:39, respectively). The libraries were IlF~cignFrl according to the primary sequence ~1 i j L provided for 12 isolates of European and North American origin by Wallich et al; 1992 (ref. cited in Example 16). Thcse included five isolates of g nnSp~ciF8 B. burgdorferi sensu stricto (ospA
serotype 1), five isolates of B. garinii (one of ospA
serotype 4, three of ospA serotype 6, and one of an unexpressed ospA), and ~.F ~ FC from two isolates of the V461 g~l~o7.~ecies (ospA serotype 2). The amino acid ratios for variable positions were adjusted to give the B.
burgdorferi sensu stricto sequences a weight of 50%, so as to account for the greater than 90% prevalence of this g~nncre~ies in North America. B. garinii and V46l isolates were weighted to approximately 25%, to account for the - distributions of these gF nnsp~ni E~c in Europe.

W095/11998 2~S57~ PcrluS9~112268 ~
SSALonpA #1 includes ospA amino acids 175-234 and t2 is an overlapping 6equence of residues 210-273. The overl2p in the two libraries is to ensure that each contains B and T
cell antigenic determinants, both being required for protective immunity.
Fig. 33 illustrates an SSAL OspC antigen by chemical and mathematical representations (SEQ ID NO:42).
Fig . 34 illustrates an SSAL f lagellin antigen by chemical and mathematical Le~es~:,.Lations (SEQ ID NO:43).
DETAILED DESCRIPTION OF TIIF INVENTION
In this invention, a "structured 6ynthetic antigen library" or SSAL ~ ~JLL~=alJ~lldS to the antigenic region of a protein from an infectious agent or from another biologically important system, a diagnostic marker or a variable therapeutic domain of a protein. The design of ~n SSAL covers the broad range of s:~qll~nn "~ embraced by geographical or population differences and temporal variation in these molecules.
A 'laLLU~;LuL-:d 6ynthetic antigen library" or SSAL is -' of an ordered set of from at least 3 to several million different but related peptides having sequences imposed upon an invariant :~LLu~Lul~ll rL ~ILk capable of maintaining the antigenicity, diagnostic value or therapeutic bioactivity of that site. Fig. 1 provides a mathematical and chemical formula for the SSA~, which f ormulation describes the relative ratio of amino acids at each position in the SSAL. AAl to AAi represent the amino acid sequence from N- to C-terminus of the library, j varies from 1 to n where n Lcpr~se.. L:. the number of pos6ible different amino acids known at the ith amino acid position.
Following this formula, a SSAL can be prepared in a single 6ynthe6is .

W095/11998 7Ss7,g PCTiUS94/12268 The sequence of the SSAL is determined by aligning the primary amino acid sequences of a related family of ~ntigens, markers or diagnostics and identifying the invariant and variant loci within the Al il, L. The invariant loci generally represent the structural framework of the SSAL. The degeneracy within the SSAL is determined by the loci within the Al i~ L that harbor different amino acid residues relative to an arbitrary prototype sequence.
After def~rminin~ which amino acids are to be at each position, the degree of degeneracy for the multiresidue containing positions in the SSAL library is determined from the number of variants each individual amino acid re~Les~,.Ls. The SSAL is then synthesized with single amino acids at the invariant positions and with the requisite degeneracies at variant positions. Thus in a simple manner, the specific amino acids and their LL~U~ Y of appearance at each position within the SSAL is defined by the primary sequences of the different antigens or molecules that make up the A 1 i; -t .
2 0 In another : ' i L, amino acids not known to exist at a variant locus within aligned epitopes can also be in_~LI~UL.~ed into the library. For example, all residue~; of a specific class of amino acids (e.g. hydrophobic, charged, neutral or polar) can be i~ .L~-~L~ted at a variable locus.
The class of amino acids i1l~oL~JL lted at a specific locus can be ~ t~rmin~d by the ~L~ " ;nAnt type of amino ~cid found at that variant locus within the antigen alignment.
In broad application, the sequence of any SSAL can be def ined by homologous portions of existing primary amino acid ~ from a family of proteins or peptides of known biological significance. The sequences encoding the ~amily of proteins or peptides encoding the target antigenic - site, diagnostic marker or variable therapeutic domain of a protein or peptide may represent: l) alternate alleles of W0 95111998 2 ~ g Pcrlu59~112268 the same gene; 2) genes that are evolutionarily related a6 deduced from sequence homology, but isolated from different strains or 6pecies of similar organisms; 3) genes encoding ~LLU~:LU~11Y similar proteins with similar function but i601ated from very different organisms, or; 4) repeated sequences within the same gene, or repeats from different genes within the same organism or repeats within homologous genes from different, but related organisms. The order within any SSAL is provided by invariant amino acid residues that are defined as the framework sequence of the 6elected protein or peptide.
The SSAL can be fron about 8 to About 100 amino acids in length ~L.rF~n~l;n~ on the particular site and preferably from about 10 to about 100. The overall length of the SSAL
can be minimized to provide the fewe6t re6idue6 nece66ary to elicit the de6ired biological re6pon6e. Similarly, extra re6idue6 can be added to the ends of the SSAL. For example, }~KK can be added at the amino terminu6 to increa6e peptide solubility, cysteine can be added to facilitate directed coupling to carrier molecules, and methionine can be added for cyanogen bromide cleavage if nece6sary.
Moreover, the SSAL can be a domain within a peptide or can have other antigenic, diagno6tic or therapeutic 6ite6 ~ttached to it. The SSAL can be attached to a core 6equence for fac$1e delivery. Theæe core sequences include dendritically branched cores, linear array type branched cores or randomly branched cores (e.g. poly-L-lysine). The branched cores can be composed of an amino acid or an amino Acid analog having two amino groups and one carboxyl group, each group capable of forming a peptide bond linkage.
Preferably such amino acids are lysine or a lysine analog such as ornithine. The amino acid analog can be an ~-amino acid, a B-~mino acid, or any other either n~tural or non-natural amino acid with two amino groups and one carboxyl ~ wo 9S111998 2 1 7 ~ S 7 ~ Pcr/uss4/12268 group available for forming peptide bond~. Preferred branched peptides of the invention are dimers, teLL c and octamers, ~p~ 11 y those having a branching core structure composed of lysine. Similarly, the branched cores can contain other residues int~L~ Lsed among the branching residues a6 depicted, for example, in Fig. 12.
If the variant sequences comprise insertions and/or deletions of amino acids relative to the prototype sequence, separate SSALs may need to be prepared to fully ~ te the various strains. Such SSALs preserve the immunoreactivity or bioactivity of the desired antigen, marker or therapeutic.
The subject SSALs can also be used to form conjugates, i.e., the SSAL, either in branched or linear ~orm can be coupled directly or indirectly, by methods known in the art, to carrier proteins such as bovine serum albumin (BSA), human serum albumin (HSA), or to red blood cells or latex particles .
As used herein, natural amino acids are the 20 amino acids commonly found in proteins (i.e. alanine, aspartic acid, asparagine, arginine, cysteine, glycine, glutamine, glutamic acid, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tyrosine, tryptophan and valine). The natural amino acids also include the D- and L- forms o~ such amino acids.
As used herein "unnatural amino acids" include both D-and L- forms of any other amino acids whether found in a protein, whether found in nature or whether synthetically produced. Unnatural amino acids can include, but are not limited to, B-alanine, ornithine, norleucine, norvaline, hydroxyproline, thyroxine, ~mm~-amino butyric acid, homoserine, citrulline and the like.
- The SSALs of the present invention are thus from antigenic sites of viruses, bacteria, parasites, tumor W09~11998 ~ 1 PCr/US94112268 antigens, allergens or other protein antigens as well as the relevant diagnostic marker sites of these infectious agents or diseases. Llkewise the SSALs include helper T cell epitopes, CTL epitopes or other var$able immune epitopes.
By way of example, SSALs can be prepared from the envelope, core, NSl, NS3, NS4 or NS5 proteins of HCV; the gpl20 V3 loop, gp41 envelope protein or the gp40 envelope protein of HIV; the envelope protein of HTLV I/II; the HA protein and a mutant HA domain of influenza A virus (in~ lin~ the H3N2, H2N2 and HlNl strains); the major outer membrane protein of ChlamYdia trachomitis (inrll-~l;nq the VDI and VDIV sites);
the neur~m;n;~ e (NA), SAPA and CRA sites of ~C- saa~.; the envelope B cell site and helper T cell site of type 2 Dengue virus (or other types); the M protein of streptococcus; the caLl,~,y~Lc.te recognition site of selectln; yLI ;F:r~ lC
helper T cell site6; the CTL epitope from HIV gag ~r~rif;r for HLA B-27; and the OspA, OspB, OspC and flagellin proteins of Lyme disease (from Borrelia 1~11rqdorferi and ~.
qarinii. The SSALs of this invention further include but, are not limited to, those specifically provided in the Examples, 6 and Sequence Listings.
The SSALs are prepared by chemical synthesis using standard t~ hniqll~ well known in the art such as the solid-phase synthetic route pioneered by Merrif ield. The coupling of multiple amino acids at a given position i8 accomplished by providing a mixture of the desired amino acids at the appropriate ratios. If necessary the ratio of amino acids in the mixture can be varied to account for different coupling efficiency of those amino acids. The SSALs can also be produced by standard L~ in~nt DNA technology, pooled in batch, isolated if ne~ec~ lLy and used as provided in accordance with the invention. Preferably, SSALs are yLeyaLed by chemical synthesis.

W095/11998 2I 7~7.~1 PCrllJS94/12268 The peptide compositions of the present invention can be composed of one or more SSAL. Preferably such compositions contain from one to about 30 SSALs ~p~n~lin~ on the application.
SSALs in accordance with this invention are synthesized and tested to determine immunoreactivity, elicitation of neutralizing antibodies, diagnostic utility, therapeutic value and the like of the SSAL as described in the Examples, by ELISA, by PHA, by bioassay or any other technique appropriate to the source of the SSAL. For example, the efficacy of the HCV SSALs in detecting and diagnosing NANBH
and HCV infection is rl~t~rm;n~ by testing it for reactivity with serum Fp~-Ci with known immunoreactivity for HCV.
Such scrum panels are commercially available. The strategy for serological validation, naturally, depends on the ~Yr~cted characteristics of the target epitopes. For example, universal i ~ inF-nt epitopes, such as the gp41 trA- ` ~ne peptide of HIV-l, can be screened by a single representative serum sample from a patient known to be infected with the virus. Epitopes which are not recognized by all infected individuals, or those for which antibody is produced late or only transiently, and ~Cp~ciAl ly epitopes which give rise to neutralizing antibodies, may need to be screened by large panels of sera. Both methods of screening can be employed in the present invention to as6ess the SSALs for selectivity and sensitivity.
Based on the i ~activities of the SSALs, they are useful in a vaccine composition to treat or prevent the infection caused by the infectious agent from which they are derived. These vaccine compositions containing one or more SSALs, alone or when coupled to a carrier or polymerized to homo- or hetero-dimers or higher oligomers by cysteine - oxidation, by induced disulfide cross-linking, or by use of homo- or hetero-functional multivalent cross-linking W095/11998 PCT/IJS9~/12268 reagents, can be introduced into normal subjects to stimulate production of antibodies. Similarly the subject SSALs can be formulated in a vaccine composition using adjuvants, pharmaceutically-acceptable carriers or other 5 ingredients routinely provided in vaccine compositions.
Such f ormulations are readily determined by one of ordinary 6kill in the art and include formulations for immediate release and for 6u6tained release, e.g., mi.:L..~ Ation.
The pre6ent vaccines can be administered by any convenient route including subcutaneous, oral, intramuscular, illLL~V~ uus, or other parenteral or enteral route.
Similarly the vaccine6 can be admini6tered as a single dose or divided into multiple doses for administration.
The vaocine composition6 o~ the instant invention contain an i --~eective amount of the SSAL to treat or prevent the target infection. Such compositions in dosage unit form can contain about 0. l ,ug to about l mg of the peptide (or mixture o~ peptides) per kg body weight. When delivered in multiple doses, the dosage unit f orm is conveniently divided into the appropriate amounts per dosage .
The SSAL peptide compositions prepared in accordance with the present invention can be used to detect or diagnose a target infection by using them as the te6t reagent in an enzyme-linked i --~c ~ I,e--~ assay (ELISA), an enzyme -' L as6ay, a passive hemagglutination assay (e.g., PHA
test) or other well-known; CCAyS. In accordance with the present invention, any suitable ~ --ccAy can be u6ed with the SSALs. Such technique6 are well known to the ordinarily 6killed arti6an and have been de6cribed in many 6tandard immunology manuals and text6, see for example, by Harlow et 311. (1988, Antibodies: A Labo~atorY Manual, Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, 726 pp. In a preferred e~mhO~;- L, the; ~~~ y is an ELISA

~W095/11998 ,?~ 7S~7~ . Pcrlu594/12268 using a solid phase coated with the pcptide compositions of the present invention . ELISA ~ hn i q~c are well known in the art. In another preferred ~ a; L the i ~Accay is a PHA assay.
The i --cAAys of the present invention are used to screen body fluids and tissues for the ~LeS~ e of the target inf cctious agent or diagnostic marker . The body f luids which can be subjected to such screening include blood and blood fractions (e.g. 6erum), saliva, or any other fluid which contains antibodies specific for the target infectious agent or pathogen.
The example5 serve to illustrate the present invention and are not to be used to limit the scope of the invention.

WO95/11998 21~ ~ ~7 ~ PCT/USg~/12268 , . ~ . .

E le l V~; DETECTION OF HCV ANTIBODIES BY NS--4 SSAL IN
PATIENTS FROr5 GEOGRAPHICALLY DISTINCT REGIONS

An HCV NS4 antigen library was designed according to the sequence listing shown in Fig. 2. The HCV NS4 SSAL was synthesized as linear peptides using the solid phase peptide synthesis strategy employing Fmoc chemistry with standard side chain protecting groups. Protected amino acids were added sequentially during the synthesi6 process from C- to N-terminus according to the mathematical and chemical ~eu~èse.lLation of the library as shown in Fig. 2B. The protected amino acid reagent added at each of these cycles consists of a collection of amino acid types, where the total cul~cellLL~tion of amino acids i6 identical to that of non-variable positions, but the ratio of each type was set by an algorithm specific to the application of the individual SSAL . The deprotection and cleavage yL uceduL ~s were carried out by standard y~uceduL~s applicable to the Fmoc chemistry of the synthesis. As controls, two NS4 peptides with CuLL~ . Sin~ HCV-l and HCV J-8 sequences as listed in Fig. 2 were also synthesized as linear peptides for comparison of their relative HCV immunoreactivity.
Wells of a microtiter plate were coated with NS-4 HCV-l peptide, the J-8 variant analogue, or the NS-4 SSAL at 5 ,ug/mL. The NS--4 SSAL peptide was selectively substituted in 27/47 (57%) of its amino acids. In five of these substituted positions, the substituted amino acid ~ ULL-~y" ~in~ to the J-8 sequence represents only 10% of the total. Specimens from two qroups were tested: Japanese h a;~lysis patients, a group at high risk for hepatitis C
~HCV) infection, for whom r~c ' ;n:~nt C-l00 assay data was available, and plasma donors confirmed seropositive for HCV
by recombinant immunoblot (RIBA-II). Two samples that were non-reactive on the HCV-l peptide reacted with both the J-8 W095111998 7SS79 ~ PCTIUS94/12268 peptide and the SSAL peptide (Table la), despite the minority I~Lese.-~ation of the ~-8 sequence in the SSAL
peptide. Both 6amples were non-reactive with the HCV-l ,~ -inAnt C-100 protein, indicating that additional antigenic regions that may exist in the C-100 protein do not contribute toward recognition of the Japanese patient antibody. The SSAL NS-4 peptide also detected samples that reacted with the HCV-l peptide and recombinant C-100 protein but not with the J-8 peptide (Table lb). In five samples in which both HCV-l and J-8 peptides were reactive (Table lc), the ~e~ .,.e to the SSAL peptide was greater than to either of the single strain peptides derived from HCV-l or J-8. In the case of Japanese dialysis patient No. 97, the ab:.uLl a~-ce on the SSAL peptide was twice as high a6 on either HCV-l or J-8 peptide.
The results obtained from the above evaluation of critical serological sample6 with HCV i ~ SAy5 .1~ LL~ted a clear advantage in using the SSAL antigen design approach for the detection of antibodies to infectious agents, particularly those with high variability such as HCV.

WO9~;/11998 2t'l S~ ~ PCTIUS9.~112268 ~ ` O
U N ~ N O 1 0 111 _I ~r u ~r u) o o ,i ,~ ~ ~ ~ ~ o J~
o o o Lrl o Iq O
i O O O O O _i ~1 ~ ~1 ~ , 1 ~1 U~:> O O N O 1-- U~ fi ~ - h C O O .-1 ~i N O ~ ~
tq ~-1 O ~`I Q ~ r O Q ~r ~`1 . U
Q
Z ~
S 1:1, H _ U
,~1 ~ C~ O Q O ~r N ~ a ~ ~ ~
2 -I a o rl ~ ~ a R' U ~ o ~ Cl Cl O O O O O O
OW:~Z ~D~
a t~ U~
~Cu~ r' C
o o r ~ Cl r D Cl, D D ~C r . ul C C 3 z o O O
U) O U-- ON N

Woss/11998 7SS79 PCr/uss4ll2268 ....

r- le 2 SSAL HCV PEPTIDES CAN BE EFFECTIVELY FnT~M~TT A~T~n INTO AN HCV
TMMTT~TnA~AY WITH IMPROVED :ikNnlllvlll AND SPECIFICITY
OVER ASSAYS USING INDIVIDUAL HCV-l PEPTIDES
Five HCV antigen [NS4, core, N53, N55(#1) and N55t#2) ]
libraries were designed according to the sPqn~n- ~ li6ting shown in Figs. 2, 3, 4, 5 and 6 respectively. All five HCV
antigen SSALs were synthesized as linear peptides using the solid phase peptide synthesi6 strategy employing Fmoc chemistry with standard side chain protecting groups according to the mathematical and chemical ~ .,Lation of each of the libraries as illustrated in Figs. 2, 3, 4, 5 and 6. All deprotection and cleavage methods were identical to standard p~ dUL e:S applicable to the Fmoc chemistry of the synthesis .
SSAL peptides NS4, core, N55(~1), N55(#2), and NS3 were coated onto wells of a microtiter plate at a ~_O~ Lr ation of 2, 1, O . 5, O . 5 and 5 ~Lg/Ml, respectively. The N53 SSAL
peptide was employed as a conjugate to bovine serum albumin (BSA). The reactivity of the mixed SSAL peptides was compared to the standard HCV peptides under the same formulation. Sensitivity was evaluated using commercially available low titer and mixed titer HCV panels from Boston Biomedica, Inc. In five samples from panels 102 and 202, the SSAL mixture of peptides provided greater sensitivity than did the mixture of individual HCV-l peptides (Table 2).
According to RIBA-II analysis, the reactivity of the five samples that had higher absorbance on the SSAL mixture of peptides varied: patterns of core only, NS-3 only and mixtures of NS-3/NS-4 or NS-3/core were all represented.
specificity of the two formats tested over 264 random plasma samples was equivalent. One well- characterized ~alse positive sample, detected by the HCV-l mixture but not by WO95111998 217S~7~ PcrluS9~/12268 RIBA-II, was negative on the SSAL peptide mixture. The resultg rl ~L~ed improved HCV i ~cs~y performance using SSAL-based HCV peptides.
E le 3 BRANCHED SSALS K~KS~kl~LlN~i THE HIGHLY VARIABLE YET
ANTIGENIC
REGIONS OF THE HCV ENV AND NSl PROTEINS ARE USED AS THE KEY
INGREDIENTS IN A POLYVALENT VACCINE FOR THF: ~K~;V~1~ 1 lON
OF INFECTION BY HIGHLY DIVERGENT GLOBAL STRAINS OF HCV
HCV has been associated with liver disease, both acute and chronic, and with the development of llver cancer.
Chronic liver disease occurs in at least 50% of infected individuals, and up to 20~ oithese go on to develop cirrhosis. Variation in immune responsiveness to HCV may account for the variation in outcome associated with HCV
inf ection .
In this HCV vaccine development approach, branched SSALs derived from highly variable yet antigenic regions of the HCV Env and NSl proteins are used in a polyvalent vaccine. More specifically, five HCV SSALs derived from both the Env and NSl protein regions are designed according to the sequence listing shown in Figs. 7, 8, 9, lO and ll.
Analogues of the four Env peptides and one NSl peptide with 8~qllOnt~C ~.ULL ~IJ-~.- 11n~ to the HCV ~-l strain were previously found to be antigenic, in that they reacted with sera from HCV- infected individuals. In this HCV vaccine design, all five HCV Env and NSl SSALs are synthesized as branched peptides onto a heteromeric dendritic branched carrier as shown in Fig. 12a, using the solid phase peptide synthesis strategy employing Fmoc chemistry with ~ WO 95/11998 21 7~ 79 `~ PCT/US94/12268 ,~
Vl 2 0 2 H 1~ 1-( C4 1-1 O I -~ I I + I
V
U
I ~ + + ~ ,~
O V~
C ~ O I ~
c u ~ u . _ .
t 7 L
,~
.C o -.( r~ ~ ~ Z O
t J o 0~ N .r t~l V ~ 13 - g N
~ ~ ~ ~J O ~ 0 5 t - V~ ~ t ~ , L
~ ~ ~ X ~ t` N ~t t~ ,~ Q C
U~ V~ t~ V) ~ ~ ~ e~
Vi ~ W r~
:> ~i ` ~ O ~` O N r~ 3 ~ 3 ~ x rl v N 111 N ~1 (U C1 C) p,, ,~ o o O ,~ O , h~
Ql o ~:) O ~ C~l ~
~ x ~ X .,, r4 ~rl ~?4 r~
o ~ o u~ o c~

WO9~/11998 32 PcrNS9~/12268 standard side chain protecting groups. Protected amino acids are added sequentially during the 6ynthesis process from C-toN-terminus according to the mathematical and chemical representation of each of the libraries. All deprotection and cleavage methods are carried by standard ~L~ eduL~s applicable to the Fmoc chemistry of the 6ynthesis.
After the synthesis of five HCV Env/NSl SSALs as branched peptide6, they are formulated with appropriate vaccine vehicles and ad~uvants and tested for their capacity to induce HCV neutralizing antibodies in laboratory animals which will block the growth of HCV isolates in vitro. Following d LL~lLion of neutr~lizing antibody production in vivo, the SSALs can be formulated into microparticles or other appropriate vaccine delivery systems for efficacy testing in human volunteers at risk for HCV infection.
Exam~le 4 SYNTHETIC AIDS VACCINE FOR THE PREVENTION OF HIV-l INFECTION
USING SSAL Rh~ hl~ATIVE OF UNIQUE AMINO ACID SEQUENCE
FRAME DERIVED FROM THE GPl20 V3 REGION
The third hypervariable region (V3 ) of the human ~- ficiency virus-l (~IV-l) external envelope 25 glycoprotein (gpl20) contains a cysteine loop, the V3 loop, that constitutes the principal neutralizing determinant (PND).
To enhance the ; o~ i city of PND peptides, the V3-derived peptides have been presented as radial eight-branched octamers based on an optimized amino acid sequence frame. A
30 heptalysyl core bearing eight reactive NH2-termini served as a carrier onto which eight identical V3 peptides were attached by solid-phase synthesis . Signif icant titers of V3 reactive neutrallzing antibodies were observed after immunization with this branched peptide vaccine (Wang et al. l99l, Sclepce 25q:
35 285). This V3 peptide ~ J thus serves as a promising approach for an AIDS vaccine. However, two major problems ~Wo95/11998 ~3 PcT/Uss~/12268 were encountered subsequently with; -; irity during the development of such a vaccine. The sources of the problems, the hypervariability of HIV-l V3 and the genetic restriction of peptide vaccines by the human hosts, served to render a ; 5 simple peptide insufficiently i J nir for application in a peptide-based vaccine.
First, the hypervariability of the HIV-l gpl20 V3 PND
presented a major challenge. Although immunizations with a mixture of the branched octameric V3 peptides carefully lO 6elected from the HIV-l V3 CPql~pncec shown in Figure 13A
resulted in broadly crossreactive anti-V3 neutralizing antibodies capable of reacting with and neutralizing isolates from each major clade of HIV-l (e.g. Clades A, B, C, D, E~, intra-clade variation, e.g., variation within clade A (e.g. A-15 l. A-2..., A-n) still presented a sirJnific~nt obstacle to an ef f ect ive vaccine .
The second major problem, the genetic restriction of the cellular immune response to a peptide i -, , became evident during a phase I clinical trial using the monovalent 20 branched octameric V3~5N peptide in a prototype vaccine. Only a s~hrop~ tion of the vaccinees L ~ ." v.-~ed to the V3 i J
after a six-month three-dose protocol. This narrow range in the responsiveness of humans to the peptide-based vaccine was a manifestation of the simplicity of the i , design:
25 The monovalent peptide, comprised of eight identical V3MN
peptides attached to heptalysine core, had an insufficient number of T cell epitopes for broad responsiveness in the genetically diverse human population.
More diversity associated with the V3 CPqllPnrPc is needed 30 to allow the synthetic i j to simultaneously induce neutralizing antibodies to the globally diverse HIV-l V3 spectrum (e . g . inter-clade and intra-clade variation) and to - elicit responses in the majority of genetically variable human subj ects .

W095/11998 ~ PCr~S9~112268 ~1 ?~5~9 Because of the large numbers of different viruses expressing different V3 loop primary 6equences, the synthesis of individual c~ql~n~Pc~ either by synthetic or by Lec l~n~nt DNA means becomes impractical, if not impossible. Clearly, 5 the production of an array of V3 i ,_.1S as an SSAL, by a single peptide synthesis, provides a useful solution to this problem. Furthermore, a V3 SSAL can impart an additional advantage to a peptide-based vaccine. By the insertion of amino acid mixtures at the variable loci within the V3 epitope lO array, a V3 SSAL i -, - can elicit protective immune responses specific to HIV-l types that have not yet been produced by the readily mutable virus, thereby anticipating viral evolution and preventing evasion of the immune Lea~v1-se through escape mutation.
A branched SSAL with sequences derived from the previously characterized V3 region of the HIV-l gpl20 protein can be synthesized onto a standard heptalysyl core or any one of the four types of heteromeric branched core resins shown in Fig. 12 using the solid phase Fmoc chemistry with standard 20 side chain protecting groups. For example, a V3 SSAL was synthesized onto a heptalysyl core resin according to the mathematical and chemical Ie~Le:s~ tion of the library as 6hown in Figs. 13A and B. The protected amino acid reagent added at each of the5e cycles (re~sase~.Ling a variant 25 position), consists of a collection of amino acid types, where the total ~ ull~en~L~tion of amino acids at the variant position is identical to that o~ non-variable positions, but the ratio of each type is set by an algorithm specif ic to the application of the individual SSAL. All deprotection and 30 cleavage methods were carried out identically to standard procedures for the Fmoc chemistry of the synthe6is.
The SSAL HIV-l V3 i , -~ was evaluated by immunizations followed by assays for serological and cellular immune activities; and, in r' LLc,tion of the value and -095/11998 7$~79 PCTIUS94/12268 flexibility for the complexity of the V3 SSAL, complexity was varied and its inf luence on ; ,_.-icity was observed .
a~ S~ oloaY of res~onse to the qC~T ElTV-al~120 V3 5 i - 1. Following the synthesis of the SSAL ~IV-gpl20 V3, the peptide library was formulated in 0 . 5~6 alum and in~ected into four guinea pigs at 100 ,ug per dose. Three and six weeks after the first injection, sera from the animals were tested by a panel of anti-peptide ELISAs (enzyme-linked i - ~orbent 10 assays) for their immune reactivities against the i i7in~
V3 peptide SSAL as well as for reactivities to other individual V3 peptides whose sequences tFigure 13A) contributed to the construction of the V3 SSAL. As shown in Table 3, the SSAL HIV-gp 120 V3 i -J_., induced significant 15 antibody responses (Logl0 titer > 4 . 0) to itself, the "V3 library", and more significantly"n LL~ted broad reactivity to monovalent V3 peptide8 with 6equences derived from geographically distinct worldwide isolatcs. In this experiment, the branched SSAL IIIV-1 gpl20 V3 d~ ~Lated 20 potent i ~ icity and elicited highly crossreactive V3 antibodies.
Another aspect of the serological reactivity evoked by the V3 SSAL i -~- is neutralization activity, i.e., the capability of the anti-sera to neutralize or inactivate infectious virus. The ability to elicit viral neutralization 25 by antibodies i5 an i .. L~-L ~ L of a protective vaccine against HIV infection. Neutralization activity was determined by the Rapid Microplaque Assay (MT-2 assay) decribed by Han60n et al. (1990, J. Clin. Microbiol. 28:2030) in which the infectivity of HIV-1 (preincubated with 30 neutralizing serum) is determined on HIV-sensitive MT-2 cells.
The particular rqT-2 assay was a stringent assay in which the endpoint was the antibody dilution at which 90~ of input - virus, North American HIV-l isolate MN of Clade B, is neutralized. This endpoint is e.L~Lecsed as the ~IMT-2MNgoll.

W095/11998 2i~ S~ ~ PcrllJS9l/12268 The pooled antiserum from the V3 SSAL-immunized guinea pigs had an MT-2MN90 of 1:2430. This titer was comparable to the MT-2mn90 of 21:2430 achieved by guinea pigs administered comparable doses of an analogous monovalent peptide; ~g~n 5 representing the V3 sequence for EiIV-1 MN only. These titers were comparable dispite the minute proportion of V3~N sequence present in the SSAL; In ~n (1o-11%) compared to the proportion of V3~N in the monovalent formulation (lOOS).
Neutralization activity for the anti-SSAL V3 antibodies 10 in the pooled guinea pig anti-sera was also ~Pt~m;n~d by a stringent assay, the infectivity reduction assay (IRA), described in White-Scharf et al. (1993, Virology L92:197), which measures neutralization activity against primary f ield isolates. IRA neutralization activity, expressed as 15 infectious units (IU) blocked lOOS by 1:10 dilution of serum, was 10 IU against the Zambian HIV-1 isolate Zam200370, a field isolate of Clade C, which was not contained in the V3 SSAL.
This cross-neutralization activity illustrates the library ' s capacity for broad i , ~ ity.
b) CTL res~onse ts the ~C~T ~}IV-qP120 V3 i ....
Cell-mediated immunity, including cytotoxic T lymphocyte L~ se (CTL) is an important ~ ~.en~ of the protective immune response to HIV-1. CTL response results in the 25 destruction of HIV-infected cells. The CTL response to a V3 SSAL; ~, is useful both in a prophylactic vaccine by preventing the tri~n-~; Rcinn of infected cells, and in a therapeutic vaccine by reducing the viral load in HIV-infected individuals. CTL responsiveness to a peptide i , in 30 humans i8 determined by how well the i , i8 recognized by the Class I HLA antigens of the genetically diverse human population. Antigens of greater complexity are more likely to be recognized by the Class I antigens of a greater part of the population. 'rhe capability of the V3 SSAL to be WOgS/11998 1 7SS7 PCT/US94112268 ~ 3 `.

recognized by a Cla5s I major histocompatibility complex and evoke CTL wa6 ' LL~Led in a mouse model system.
The immunizations were accomplished by the following protocol: The V3 SSAL branched peptide was formulated into . 5 polylactide-co-glycolide microparticles that are suspended in phosphate-buffered saline (O'Hagan et al. 1991, Vaccine, 9:768) and injected intraperitoneally into a group of female Balb/c mice (6-~ weeks old), three mice per group, at 100 ~g of peptide in 0. 5 ml of microparticulate vaccine per dose.
Booster immunizations were administered on days 12 and 21.
Animals were sacrificed on day 27 and splenocyte6 collected and restimulated with a monovalent linear peptide of s~qU~nr-~
I-G-P-G--R-A-F-Y-T-T, cuLL~ n~lin~ to a portion of the V3 domain of HIV-l ~ that was present in the SSAL as an individual sequence. The same peptide was also used to sensitize target cell6. The cultured splenocytes were restimulated on days 7 and 14 after sacrifice and assayed for CTL 6 day6 later by SlCr-release assay (Hioe et al. 1990, J.
Virol . 64: 6246) . Target cells were SlCr-labeled A20 .1-11 (H-2d) ceLls pulsed with the above monovalent linear peptide or irrelevant peptide and no peptide as controls. A highly significant level of CTL immune reactivity specific ~or the V3MN peptide was observed, as ~e~L~E_.ILed by per cent Slcr released of (% specific lysis) 51%, 42~, and 21% for respective Effector:Target ratios of 150, 50, and 17. By comparison, CTL results with unsensitized target cells and for target cells coated with an irrelevant peptide were less than

3~. Splenocytes cultured from placebo-i i 7~ mice displayed only background levels of SlCr release.
c) Ol:!tim; 7~tiQn of cc~r, l~Y;tv. The V3 SSAL
evaluated above in parts a and b, with a complexity of 10l3, - was evaluated for; ,_.liCity relative to that of a series of V3 libraries of ~l;mini~::hin~ complexities. Complexity is a WO 95/11998 PcrluS9~112268 ~
2~5~9 highly significant parameter for; ,~nicity as it determines not only the number of different peptides that is delivered by an SSAL YaCCine, but also the dose of any individual peptide in the SSAL.
Four V3 SSAL i , -- of increasingly simpler compositions were synthesized, of complexities 1012, lO9, lo6, and 103 . These were used to immunize groups of f ive guinea pigs, as described in part a of this Example. The composition of the V3 SSAL of 109 complexity is illustrated in Figure 13c. Neutralization activities (against HIV-1 MN) elicited by those libraries were ~ t~ n~d for the pooled anti-sera of each group at week 6 as MT--2MNgo values and results were compared to the neutralization titer of the anti-sera to the original V3 SSAL (Figure 13B) and to neutralization titers evoked by monovalent branched V3 peptide - J -- Neutralization activities are plotted in Figure 13D and show a strongly 6ignificant neutralization titer for the SSAL of 109 complexity (MT-2MNgo 1:9081) and the other SSALs elicited titers comparable to the results obtained from immunization with monovalent V3MN i ,~.l. Immunization with a monovalent V3 i , other than MN that was also present in the libraries evoked poorer cross-neutralizing activity against HIV-l MN than was evoked by any of the SSAL i regardless of library complexity.
The V3 SSAL i , - of complexities 10l3 and 103 were compared for i -, i city by CTL assays against V3MN-pulsed cells. Mice were immunized with these two libraries and evaluated for CTL by the protocols described in part b. At Effector:Target cell ratio of 150, results were 5396 specific release for the CTL elicited by SSAL of 1013 sequences and 79%
specific release for the SSAL of 103 sequences, strongly suggestive of the inf luence of complexity on the ~ W095/11998 21 7S~7~ ; . PCTIUS94112268 3g 0 o~ o O ~ 0 ~
~ $ =~ U
115 H o O c O o ,., ~o ~ . , O
o O
0 o o 2 o o ~ ~ O n 0 o v~ u ,_, 1 ,o _ N o~ ,5 R
. __ o~ ~J 0 _ r~
-o ~
O > O .. ' .~ o ~ U
2 ~
H o < ~ ~ W
O O
~ ~ 'n ~ ~ ~ ~
~ ~ Y O ' ~
Il~ O Il'~ O

WO 95/11998 PCT/IJS94~12268 21~S~9 ~ ~

3 i city of HIV-1 gpl20 V3 SSAL i 3 - , and a clear illu6tration of the potent i n~.ni city of a V3 SSAL.
r le 5 HIGHLY ANTIGENIC REGION OF THE GP41 ~RAN~M~MRRANF

The gp41 trA clne envelope protein of the human ff$ciency virus, Type 1 (HIV-l) contains an i "~ 1n Int antigenic region useful for detection of antibodies to HIV-1. Although this antigenic region is highly ~_ul.s~L~,~d, variant amino acid substitutions have been observed in f ield isolates.
An ~IV-1 gp41 antigen (35 mer) library was ~ ;qn~-l according to the sequence listing of 50 field isolates shown in Pig. 14. The HIV-1 gp41 SSAL was synthesized as linear peptides using solid phase Fmoc chemistry. Wells of a microtiter plate were coated with the SSAL HIV-1 gp41 at 1 ~g/mL. Confirmed HIV-1 positive spoci ~ from geographical diverse areas were tested with the HIV-1 gp41 SSAL and found to have strong seroreactivities.
Due to the inherent design advantages associated with the SSAL, such a gp41 antigen preparation is useful for comprehensive HIV-1 antibody detection.
r le 6 A HIGHLY ANTIGENIC REGION OF THE GP40 ~RAN~MFMRRANF

The gp40 transmembrane envelope protein of the human ~r~ficiency virus, Type 2 (HIV-2~ contains an ~n:~nt antigenic region useful for detection of antibodies to HIV-2. Although this antigenic region is ~WO95/11998 ~ 7S~7~ ` PCT/US94/1226 highly conserved, variant amino acid substitutions have been observed in f ield isolates .
An HIV-2 gp40 antigen (36mer) library is de6igned according to the sequence listing of 20 f ield isolates shown in Fig. 15. The HIV-2 gp40 SSAL i6 synthesized as linear peptides using the solid phase Fmoc chemistry. Wells of a microtiter plate can be coated with the SSAL HIV-2 gp40 at l~Lg/mL. Confirmed HIV-2 positive specimens, mostly from West Africa are tested against these S5ALs.
Due to the inherent design advantages associated with the SSAL, such an HIV-2 gp40 SSAL antigen pLe~ ion is thus u6eful for comprehen6ive HIV-2 antibody detection in the event of antigenic drift.
R-- le 7 SIMULTANEOUS DETECTION OF HTLV I/II ANTIBODIES WITH SSALS
DERIVED FROM THE HIGHLY ANTIGENIC REGIONS OF THE
RY~RRrl~T PORTION (GP46) AND rrR~N~MRMRR~NR
PORTION (GP21) OF THE HTLV ENVELOPE PROTEIN
HTLV-I and HTLV-II are closely related viruses with extensive serologic cross-reactivity. HTLV-I is the causative agent of adult T-cell leukemia and a declel~e~--tive neurological disorder, HTLV-I-associated myelopathy (HAM) or tropical spastic paraparesis. Disease association with HTLV-II is less well-es~hl i Ch~l, but HTLV-II virus has been identified in patients with a neurological disease similar to HAM.
HTLV-I and HTLV-II can be transmitted by blood transfusion. T -'. in~nt epitopes have been identified in the gp46 external portion and gp21 tr~l ~ille portion of the HTLV-I envelope that are useful for detection of antibodies to HTLV-I.
Three HTLV I/II antigen (gp46, gp21) libraries are designed according to the sequence listing shown in 16A, B
and C. All three HTLV-I/II antigen SSALs are synthesized as Wo9Stll998 ? ~5~ 42 PC~IUS9~11n68 linear peptides using the solid phase peptide synthesis strategy employing Fmoc chemistry. SSAL peptides 16A
(gp46), 16B (gp46) and 16C (gp21) can be individually coated onto wells of a microplate at a ~ LLction of 2 ILg/mL.
These SSAL peptides are tested with sera from patients infected with either HTLV-I or HTLV-II. SSAL HTLV-I/II
peptides thus d LL~ted their cost-ef~ective feature in screening for HTLV-I/II antibodies when compared with the use of individual HTLV-I or ~TLV-II peptide antigens.
E le 8 SSAL BASED UNIVERSAL FLU VACCINE FOR THE PROTECTION
AGAINST DONINANT HUI~AN FLU VIRUS TYPBS
(e.g. H3N2, hlN1 AND H2N2) Inf luenza A viruses belong to the Orth' y,~-,viridae family and cause febrile respiratory disease on a frequent and worldwide ba6is. High morbidity and mortality are typically associated with epidemics caused by inf luenza A
viruses. Young children, the elderly and patients with chronic illn~cF~c are ~crec;Ally susceptible to r ~;~
resulting from infection. Antigenic variation occurs with exceptionally high frequency in influenza A virus.
Mutations in the genes encoding the viral surface glycoproteins, namely the hemagglutinin (HA) and neurAm1nitllCc proteins (NA) are rapidly selected for by immune responses mounted against the virus. ThQse mutations a 1 Ate around the regions containing antigenic site6 such that the mutants are capable of escaping i-m~munity~
thereby causing 1e~ULL_I-L epidemics of respiratory disease on nearly an annual basis. Because the existence of large numbers of different virus strains and their capacity to escape immune surveillance through mutation, new vaccines structured to block infection by the current circulating virus strain(s) must be formulated on an annual basis.

-~, W095/11998 21 7~S 7~ PCT/I~S94/12268 Herein is described a realistic alternative to this approach by using l~LLU.LULed synthetic antigen libraries (SSALs).
The libraries are desiyned to mimic the major neutralizing 6ites present on the HA molecule, which is the dominant l ,~., of this virus. Therefore, a single vaccine d of a simple mixture of structured libraries, based upon the dominant HA antigenic sites which are capable of eliciting virus neutralizing antibodies, should eliminate and replace the need to prepare "customized" Flu virus vaccines in the conventional manner.
The first step in the de6ign of an effective Flu vaccine i6 the identif ication of the primary amino acid 6equence of the critical domain from Influenza A viru6 hemagglutinin (HA) surface glycoprotein capable of eliciting virus neutralizing antibodies.
A synthetic peptide representing the amino-terminus of this domain (residues 91-108), when used as a vaccine ,_., was found capable of protecting mice from virulent virus challenge (Muller, et al. 1982, Proc NAtl Acad 5ci USA
79: 569). Its carboxyl-terminus (residues 138-164), was also found to confer partial protection to mice (Shapira et al, 1984, Proc Natl Acad Sci TT.':A 81: 2461) . A7 1~, t of the complete amino acid sequence of this domain, from re6idue 91-164, i6 hereby de6cribed for the H3N2 strains of Influenza virus A. The major ~LLu~ LUL~I1 feature of this domain is a loop formed by a disulfide bond between the Cysteine (Cy6) re6idue6 at positions 97 and 139. The numbering of the amino acids begins with the f irst amino acid of the mature HA sequence (i.e. following cleavage of the signal sequence) a6 the fir6t residue. The primary srquDnl-o of the ..LLu~LuL2d synthetic library for the H3N2 viruses ( i . e . SSAL1 Inf A-HA-H3N2 ) is described in Fig . 17 .
Extending the amino acid sequence framework information obtained from H3N2 strains of influenza virus A to the Hl~l WO 9S/11998 PCr/lJS9l/12268 2~5~g ' ' strains, a separate SSAL (i.e. SSAL2 Inf A-HA-HlN1) for HlN1 was 6imilarly constructed a6 shown in Fig. 18A. However, there were two 6train6 in the HlN1 family, namely WIL and -FPR 834 that were found to harbor a 6ingle amino acid deletion within thi6 domain. A third library wa6 therefore ~w~la~LueLed (SSAL3 Inf A--HA--HlNl) to re~Lè6ell~ the two 6equence6 (Fig. 18B).
The "f. JLk l information of the amino acid 6equences representing the riA critical neutralizing site of two H2N2 strains was further extended, and u6ed to C~ LU :~ a fourth SSAL (i.e. SSAL 4 Inf A-HA-H2N2) (Fig. 19).
Since synthetic peptides made ba6ed upon the sequences that immediately flank the loop aLLul_LuLe present in the HA
neutralizing domain were found capable of eliciting antibody that partially neutralizes virus infectivity, these flanl~ing sequences are likely to form a conformationally ~orPnflont epitope important for eliciting a protective immune re6pon6e. Three more SSAL6 leyLè6el.~ing a mutant form of the HA critical neutralizing domain ba6ed on the sequences of H3N2, HlN1 and H2N2 were thus con6tructed to mimic thi6 "Base" aLLu~LuLe by 6imply linking the amino acid sequence6 pre6ent on either side of the loop as shown in Fig. 20.
These libraries lel . eSell~ ~ll.ti~nF: of the amino acid residues present between Cys residue6 at po6ition6 97 and 139 in the original critical HA domain structure de3cribed above . The sequence of the deletion libraries ( i . e. SSAL5 Inf A-HA-H3N2, SSAL6 Inf A-HA-HlN1, SSAL7 Inf A-HA-H2N2 ) was extended at the amino-terminus to include all residues represented between the f irst Cysteine residue of the loop aLLU- ~uLe and the next Cys residue on the amino-terminal side of the loop.
Based on the above iLLUl_LULe design, seven SSAL6 are synthesized as branched peptides. The6e seven SSALs are tested alone and in combinations for the induction of virus-WO 95/11998 1 7S~ 79 PCT/I~S94/12268 specific antibody responses and virus n~uLL~lizing antibody responses in laboratory animals. A response capable of neutralizing the broad-spectrum of human influenza viruses is anticipated . Following d LL aLion of virus S neutralizing L~ Jo11ses in vitro, mixtures of the these SSALs are formulated with appropriate vaccine vehicles and adjuvants and used to immunize mice. Following LLation of an immune response to the influenza A HA
SSALs in mice, the animals are rhA11.on~ed with the different types of human viruses to determine the breadth of protection conferred by this novel i , . Following mouse protection studies and ~1 LLaLion of safety in laboratory animals, the SSAL-based vaccine is tested for efficacy in human volunteers.
r le 9 NEUl'RAT T~T`-- DOMAIN OF THE MAJOR OUTER M~MRR~NF~ PROTEIN
(MOMP) FOR PROTECTION FROM C~LANYDIA INFECTION

Chlamydia trachomatis is an obligate intrA- ~l 1111Ar bacterial pathogen that is the leading cause of sexually transmitted disease (in~ in~ cervicitis, epidldymitis, urethritis and pelvic infli tVLy disease) in industrialized countries, and the dominant pathogen associated with preventable hl in~n~'~ (i.e. trachoma) in developing countries. The many different isolates of this pathogen have been divided into three biovars ba6ed upon their nucleic acid composition, antigenicity and pathogenicity. Isolates that produce disease in humans come from the ly ~ _ alluloma venereum and trachoma biovars.
Within these two biovars, the different bacterial strains s are further subdivided into at least 15 different serovars based upon serologic responses. These serovars are ~ollected into three ~:l UU,L uu~s based largely upon antigenic WO95/11998 2~5~7~ PcrluS9~/12268 '~1 responses to the major outer membrane protein (MOMP): the B
serogroup contains the B, Ba, D, E, L1 and L2 serovars; the intermediate group is composed of the F, G, K and L3 -serovars; while, the C ~ LOULUU~U covers the A, C, H, I, and J serovars. The MOMP protein contains major antigenic structures that cause the production of serovar-, 8~:LuyLuu~
and species-specif ic antibody responses . Since it is the serovar- ~nd S~:LU~UU,U ~-~uecific antibody L~Duul.ses, particularly the anti-MOMP L-:~uUII=ie8 that are capable of neutralizing chlamydial infectivity, MOMP has been the focus of subunit vaccine research and development. MOMP ~LUULUL~:
is conserved between serovars and consists of four variable domains separated by constant region6. The variable domains I and IV (i.e. VDI and VDIV) contain ~ntigenic sites which elicit protective neutralizing antibody (Zhang et al. 1989, Infect. Immun. 57:636; Yuan et al. 1989, Infect. Immun.
57: 1040) Alignment of the amino acid sequence of the f irst variable domain (VDI) from the 15 known serovars of C.
I LC~ JO~ tiS has est~hl; Ci1P~1 two primary amino acid seuu~n~_e patterns, designated VDI" and VDIb. The prototype VDIA
pattern is defined by the VDI domain from C. trachomatis serovar A (two alternate s~uuel - es for the VDI domain of serovar A exist, designated Al and A2 above) and r~=~LesenLs MOMP residues 81-106. Serovars A, C, H, I, J, R and L3 are representatives of the VDI~, pattern. The C. l.L ' tis serovar B VDI domain is the prototype VDIb pattern and represents MOMP residues 81-104. Serovars B, Ba, D, E, L1 and L2 are L~pr~r_~.Latives of the VDIb pattern. The primary amino acid sequences for the SSALs for VDIA and VDIb are described in Figs. 21A and 21B.
Alignment of the amino acid sequences of the f ourth variable domain (VDIV) from the 15 known C. /.L ' . tls 2 serovars has est~hl; ChPd two primary amino acid sequence W095111998 ~I 7~7~ PCT/US94/122~i8 patterns, designated VDIVa and VDIVb. The prototype VDIVa pattern is defined by the VDIV domain from C. trachomatis serovar A and I~L~__.IL8 MOMP residues 312-341. Serovars A, B, Ba, D, E, I, Ll and L2 are representatives of the VDIVa pattern. The C. trachomatis serovar C VDIV domain i8 the prototype VDIVb pattern and ~.cs~:..Ls MOMP residues 312-342. Serovars C, F, G, H, J, K and L3 are representatives of the VDIVb pattern. The primary amino acid s~q~ nr~ for the structured synthetic epitope library for VDIVa and VDIVb is described in Figs. 22A and 22B.
Based on the above vaccine design, four SSALs were synthesized as branched peptides using Fmoc chemistry.
Since the VDI and VDIV domains do not contain potent helper T cell epitopes one can be provided to both libraries.
Addition of the helper epitope is required to potentiate antibody-producing B cell re~ ,.,ses to the chlamydial SSALs.
A T cell helper epitope, named A8, ha6 been located between MOMP variable domains I and II (Su et al. 1990, J pYn Med 172:203). This helper epitope sequence was inr]l~ at the amino-terminus of the VDI and VDIV libraries in an unmodified and non-degenerate form to elicit the T cell r~Cpnnc~c required for the production of neutralizing antibodies to the degenerate B cell domains present within the SSALs.
Following synthesis of the VDI and VDIV SSALs, they were formulated into eYperimental vaccines with Freund's Complete tFCA) and Incomplete Adjuvants (IFA) and used to immunize groups of five guinea pigs, at 100 ,ug of peptide per dose with FCA on week O and with IFA f or booster i i 75~tions on weeks 3 and 10, intraperitoneally. Blood was collected from the guinea pigs on weeks O, 5, 8, and 12, octssed into serum, then stored until evaluated by anti-pept1cle E1IEA on hoEologou- pept1de~ . Ev~ry group of f lve s ~
WO 9511 1998 i PC rruss4rl226s 2~75~7~
animals that had received Chlamydia MOMP SSAL displayed strong anti-peptide reactivity at weeks 5, 8, and 12, with ELISA reactivities of >0.5 A492r"l, at serum dilution of 1:100,000. Serum samples from week 5 or 8 were pooled for each group, serially diluted and tested by dot i -- .,y blot assay (Zhang et al. 1989) for cross-reactivity to the elementary bodies (EB) of a selection of Chlamydia trachomatis serovars LepLesell~ing each of the three se~v~Lvu~,=. (EBs of serovarfi B,Ba,D,E,F,G,A,C,H,I,J,and K).
The anti-serum to the VDIa, VDIb, VDIVa, and VDIV}, SSAL
vaccinefi were cross-reactive to all of the EBs at a 1:200 dilution. The reactivity of the VDI, SSAL anti-serum was compared to guinea pig anti-serum made against an ;~n:l- 1 o~o--c but monovalent VDI peptide i -, - L ~y~ es~ ing serovar A .
The antibodies in the SSAL anti-serum bound to the E8s of each serovar approximately equally while the antibodies in the anti-VDl A monovalent antiserum bound preferentially to the serovar A EBs with reduced reactivities f or the heteL~,ye~ ,u~ EBs, in a clear ~1 ~Lcltion of the improved ; ,_.,icity of ~ered by the SSAL. The SSAL guinea pig anti-sera can be further tested to ~1 ~LCIte the capacity of the SSAL; -, ~~ to induce antibodies that block C.
trachomatis infections to all relevant serovars in vitro;
prevent vaginal infections in mice; and prevent infections of the eyê and sexually transmitted disease in primate6.
The VDI and VDIV libraries were initially administered by parenteral immunization. Following ~' ~L,-tion of antibody production specif ic to C . trachomatis elementary bodies that is capable of blocking infection in vitro, the SSALs can be incorporated into microparticles, or other appropriate biode~radable delivery 6ystems, for oral, vaginal or ocular delivery. Since chlamydial infections occur at mucosal surfaces, antigen delivery to these W09~/ll998 PcrluS94/12268 s$,~9 surfaces is considered critical for successful immunization to prevent these infections.
r le 10 SSALs FROM Trypanosoma cruzi ANTIGENS AS EXAMPLES OF
- PEPTIDE r TRR~RT~q DERIVED FROM REPEATS WITHIN THE SAME GENE
AND USED AS DIAGNOSTIC REAGENTS FOR THE
DETECTION OF T. cruzi INFECTION
Chagas ' disease, caused by the parasite T. cruzi, is a chronic disease affecting about Z0 million people in South and Central America. Transmission occurs through the bite of a hematophagous insect carrying infective Lly~ ctigotes in its excreta. The acute phase of the infection i5 symptomless, with 90% of infected individuals progressing to the chronic phase. This pha6e is often characterized by myocarditis, and since no infective organisms ~re detectible at this stage, it is speculated that an autoimmune proces6 is involved. An effective diagnostic tefit for Chagas ' disease is urgently needed, both for early detection of acutely infected individuals, and for prevention of transmission by blood transfu6ion.
Several T. cr;~zi antigens have been found to be proteins characterized by regions of highly repeated sequence. In the few cases that have been investigated, the repeats constitute antigenic sites. For example, shed acute-phase antigen (SAPA) is a lipid-anchored family of proteins shed from T. cruzi during the acute phase of the infection.
Some of the members of this family possess neurAm1n~ ce activity but all members contain variable numbers of a specific repeated sequ~-nre. The s^quF~nr~ of SEQ ID NO:27 has been found to be an i ~ inAnt B-cell epitope (Prioli et al., 1992, Mol. Birrh~r. P~rasitol 52:85) which is found in as many as 43 imperfect tandem repeats in SAPA
prote~ns. This is a very good candidate for the design of a W095/11998 PCrNSs~/l2268 2~ ~ 5S~

diagnostic test because it is potentially reactive with all members of the family; in addition it offers the possibility of early detection during the acute phase of Chagas ' disease .
The contiguous sequ~nre of residues 596-1110 of one member of this family (lined up in rows of twelve residues) is shown in Fig. 23. An ~n~lo~ollc region from another member of the SAPA family is shown in Fig. 23. The two proteins combined contain 57 imperfect repeats. An SSAL
designed to consist of tandem repeats of several such partially degenerate ~r~r~r-rs is also shown in Fig. 23.
Another T. crzlzi antigen useful for design of a diagnostic test is known as cytoplasmic repetitive antigen (CRA). It has been ~ LL~I~ed that the CF~A antigen is r~cr,r,ni 7F.rl by all of a panel of 6 chagasic sera, and none of a panel of 9 sera from patients with infections that produce cross-reacting antibodies (Krieger et al., 1992, Am J Tro Med Hva 46:427). By analogy with other ~. cruzi antigens, repeated sequences are likely to constitute antigenic sites in this protein as well. Computer analysis of the sequence of the CRA gene with a program that detects potential epitopes in proteins shows that 21 imperfect repeats of a 14 residue sequence are highly likely to be antigenic. The 21 repeats are shown in FLg. 24. A diagnostic SSAL consists of numerous tandem repeats of this sequence. An SSAL based on tandem repeats of C~A E~l~l,.n~c is also Le~LesellLed in Fig.
24 .
By analogy with the previous example, it can also contain equal amounts of the two residues at any given variant position.
Diagnostic reagents based on the SSALs as designed used either individually or in combination would constitute a significant i . ~,v~ L over peptide reagents derived from a WO 9S/11998 SS~9 PCT/U594/12268 single sequence because they have the potential to react with a broader spectrum of antibodies.
. Based on the above design, two SSALs are synthesized as linear peptides using Fmoc chemistry. Both SSALs are employed a6 solid phase antigens and formulated into an -cc~y, both individually and in combination, for the detection of antibodies to !l. cruzi.
r le 11 SSALDE~GUE VIRUS-BASED VACCINE FOR IISMUNIZATION AGAINST
VARIANTS OF DENGUE VIRUSES

Dengue viru6es are members of the Flavivirus f amily and cause }~ i r fever and shock syndrome which result in significant morbidity/mortality in Asia, and Central/South America. Dengue viruses are divided principally into four lo major antigenic groups (Dengue subtypes 1-4), wlth each group subdivided into several antigenic variants. Vaccine development for Dengue virus has thus far proven difficult, since enhancing epitopes from one or more subtypes primes individuals for Pnh~n~PA disease complications.
s~LuvLu.~ d Synthetic Antigen Libraries (SSAL) provide signif icant advantages f or the design of Dengue vaccines, since variable protective neutralizing B cell antibody epitopes can be covered through this approach, along with T
cell helper epitopes.
As an example, an SSAL derived from the critical neutralizing domain (aa 121-185) of the Env protein of the type-2 Dengue virus is constructed as shown in Fig. 25A, based on the primary amino acid sequence i.l i; t from 33 antigenic variants. Three other SSALs derived from the cvL.. :~v.. ding Env regions, on the primary amino acid sequence i-11; Ls, from antigenic variants of Dengue subtypes 1, 3 and 4 can be ~;u..:,~, u~ Led similarly.
These ~our SSALs ( i . e . Dengue Env 1, 2, 3 and 4 ) can be combined with a library derived from amino acids 352-368 of WO 95111998 ~ PCT/[~S9-11122G8 the Dengue 2 Env protein representing a T helper epitope capable of conferring T helper effects to heterologous flaviviruse6, thus l~t;ng the requisite T-B requirements for immunization against variants of four subtypes of Dengue viruses.
The combined four SSALs can be formulated w$th appropriate vaccine vehicles and adjuvants and tested for their capacity to induce neutralizing antibodies in laboratory animals to block the replication of all relevant antigenic variants of Dengue virus in vitro. Following ~' L.~.tion of neutralizing antibody production in vivo, the SSALs are then inc." ~,Le.ted into microparticles or other appropriate vaccine delivery systems f or ef f icacy testing in human volunteers at risk for infection by Dengue viruse6.
Exam~le 12 STREPTOCOCCAL M PROTEIN E~ASED VACCINE USING THE SSAL
APPROACH
The efforts to date in producing an effective vaccine against Group A streptococcal infection have been thwarted by the serological diversity of the antiphagocytic and major protective antigen designated streptococcal M protein. Only type 6pecific ~ ".ses have been elicited using M-protein based vaccines. M protein sequences are characterized by a seven amino acid periodicity (Hosein, B., McCarty, M., and Fischetti, V.A., Proc. Natl. Acad. Sci. USA, 76, 3765, 1979) that allows them to a6sume an alpha-helical coli structure that is rare among bacterial surface molecules but is common in mammalian proteins. The extreme variability at the amino terminus has formed the basis for serological typing of M
proteins, with over l00 distinct M types being identif ied to date .

WOgS/11998 1 7~79 PCT~S94/12268 The N-terminus of M proteins from across a number of types has been shown to produce opsonic, or antiphagocytic, - antibodie6. In humans and experimental animals, systemic immunity to group A streptococcal infections is due primarily to these y~ ecif ic opsonic antibodies directed against the variable amino-terminal half of M proteins. An SSAL based on an antigenic peptide ~.LL~ ULe derived from the N-terminus of the str~rtrrorr~ 1 M protein was constructed. The N-terminus of Type 5 M protein was aligned as a prototype with ~.ULL~ in~ regions of M proteins from Types l and 6 in such a way as to preserve the register across types of the seven-amino acid periodicity. Despite the wide diversity of sequences across types, 1eucine is completely conserved in positions 28 and 35, thus providing an anchor structure for the SSAL. CULL ~ J~ n J regions of M protein from Types l9 and 24 are also aligned for s~q~nre comparison, although the extreme diversity of these two 5~qU~nrl,c has prevented us from inrlll~inq them into the construction of an SSAL streptococcal M.
A 35 mer SSAL beginning at the invariant C-terminal Leucine position designed according to the Ill~t` Lical and chemical representation of the library (Fig. 26), is synthesized onto a heteromeric (Lysine and Alanine copolymer core) octa-branched carrier employing Fmoc chemistry.
Following synthesis of the ~-CAt,~ , this SSAL is formulated with appropriate vaccine vehicles and ad~uvants and tested for its capacity to elicit protective opsonic, or anti-phagocytic, antibodies in laboratory animals.
Following ~ LLt:tion of opsonic antibody production directed against multiple strains of streptococcus pyogenes, the SSAL is then incorporated into microparticles or other appropriate vaccine delivery systems for efficacy testing in human volunteers at risk for streptococcal infection.

~,q PCr/USslll226R
F le 13 S~T~rr~r~r~c~ Type I/II (~h)-BASED TREATMENT OF RHEUMATOID
ARTHRITIS

Rheumatoid arthritis (RA) is a common autoimmune disease in which the joints become painful and swollen under immune system attack. The symptoms of RA can be relieved somewhat by treatment with steroids or anti-cell proliferatlon drugs, but these therapies have serious side effects. Recently, Trentham et al. (1993, Science 261:172~) have described a treatment of RA in human volunteers by oral tolerization with collagen.
In a related study in a mouse model of collagen-induced arthritis (/qyers et al. 1993, J. Immunol. 150;4652), a peptide from collagen that was an effective competitor with collagen for MHC Clafis II moleculeG was identified. When mice were co-~ i 7ed with collagen and thi6 ~C class II
binding collagen peptide, the inrjrl~nre and severity of arthritis were reduced. In another related study (ICu et al.
1993, Eur. J. Immunol. 23:591), a collagen peptide was found to prevent the development of arthritis in the rat model.
Resistance to collagen-induced arthritis in rats and rhesus monkeys was found following an immunization protocol with attenuated type-II collagen aimed at maintaining T cell epitopes, thus implying that collagen-related T cell epitopes are i _ L~-IL in inducing a tolerized T cell state, thus preventing the development or induction of the autoimmune disease (Hart et al. 1993, Fllr. J. T -1.
23: 1588) .
From the above observations, we theref ore adopted the use of a collagen derived T cell epitope SSAL, an approach to broaden the responder population f or the treatment of patients with RA.

~W095111998 ~ 7~ PC~IUS94112268 More specifically, a collagen T cell epitope SSAL
(12mer in size) as shown in Fig. 27 can be used to i i ~e patients with RA with a total of from l00 to 500 ~g per dose, emulsified in incomplete Freund's adjuvant at multiple sites and administered intradermally. The patients are monitored at entry, l month, 2 months and 3 months after immunization for various clinical parameters and symptoms.
It is predicted that S~AT.--~TT~ ~ Type I/II (~h)--based ; zation will induce T cell tolerization to the collagen, thus relieving the symptoms of RA.
r le 14 SSAL~eleCtLn5--BASED TREATNENT OF INFLAMNATION
Developing effective anti-inflammatory _ ~~ is a major goal for medical 6ciences. In a recent report, pertussis toxin was found to bind target cells through thc carbohydrate recognition properties of two subunits, 52 and 53, which share amino acid sequence similarity with the lectin domains of the eukaryotic "s~lertin" family.
Selectins are a group of proteins that play an important role in promoting the reversible rolling of leukocytes at sites of tissue inflammation. To date, three selectins have been characterized and all shared a terminal lectin domain. P-selectin and E-selectin appear on activated endothelial cells and mediate adherence of leukocytes by recognition of sialylated and non-sialylated determinants.
L-selectin (human leukocyte homing receptor, hLHR) is present on lymphocytes and mediates homing to lymphoid tissue that bears sialyated glycoconjugates.
More i Lc~ ly, this report found that pertus6is toxin 52 and S3 subunits, and synthetic peptides representing their caLbo1lyllL~l~e recognition domain6, competitively inhibit adherence of neutrophils to selectin-coated 6urface6 and to endothelial cell6.

Wo 95111998 ~ , PcrluS9~112268 Based on the above observation, an SSAL representing the carbohydrate recognition domains is useful to treat inflammation. A careful eYamination of peptide ,cP~ r derived from multiple selectins and pertussis toxin subunits that block neutrophil adherence has allowed construction of an SSAL as shown in Fig. 28, based on sPlP~;nc or peptides from prokaryotic organisms with related sequences, for the treatment of inf lammation .
~ore specifically, a selectin SSAL is synthesized as linear peptide6 according to the mathematical and chemical representation shown in Fig. 28. Following the synthesi6, the SSAL cPler~inc can be formulated with an appropriate delivery 6ystem for application at site of inflammation.

r le 15 HELPER T CELL (Th) EPITOPE BASED SSALs ENHANCE ANTIBODY
RESPONSE TO LE~H IN A GENETICALLY DIVERSE POPULATION

B cell Lt ci~,..6es to a foreign antigen, resulting in the production of antibodies to that antigen, are modulated by T
cell Ie,,~u1.ses. During antigen processing, domains or epitopes that are composed of 6pecif ic linear segments of the antigen are recognized by and are presented in the context of MHC class II molecules on the surfaces of antigen presenting cells. The peptide segments presented in this fashion are recognized by CD4+ T cells. These activated T
cells stimulate [termed helper T cells (Th) ] or suppress [termed .'U~J~JL~SS~L T cell (Ts) ] antibody production to B
cell epitopes present on the foreign molecule in an antigen specific manner. Often, synthetic peptide antigens are poor - because they do not effectively stimulate Th e~ .s~6. This lack of immune reactivity can be overcome Wo 95/11998 1 7~S 71q PCT/US94112268 5~
by the addition of known Th epitopes to the synthetic antigen .
~ A number of Th epitopes have been characterized to date. They range in size from approximately 15-30 amino acid re6idues in length and often share common structural features and may contain specific landmark sequences. For example, a common feature iG amphipathic helices, which are alpha-helical structures with hydrophobic amino acid residueæ dominating one face of the helix and with charged and polar re6idues dominating the 6urrounding faces. Th epitopes frequently contain additional primary amino acid patterns such as a Gly or charged residue followed by two to three l~ydLu~ Obic residues, followed in turn by a charged or polar residue. This pattern defines what are called Rothbard s~q~lenc~-c. Also, Th epitopes often obey the 1, 4, 5, 8 rule, where a positively charged residue is followed by lly.lLuL~hobic residues at the fourth, fifth and eighth positions after the charged residue. Since all of these structures arc composed of common ll~dLuuhobic, charged and polar amino acids each ~LLuuLuL~ can exist simultaneously within a single Th epitope.
A further class of Th epitopcs have been defined as being universal, in that they are capable of stimulating B
cell responses in most members of a population that ~k~uL esses diverse HLA haplotypes . A representative of this class of epitope is more appropriately described as ~1 icr.l.,us, because a majority, but not all, individuals expressing different HLA antigens are capable of r~cronAin~
to it. Most, if not all, of the pL~ ir-ll,nlc T cell epitopes contain at least one and usually more than one of the Th epitope features described above.
The design and synthesis of two SSALs that act as artif icial Th epitopes and can elicit immune responses to WO 95/11998 PCT/U594~12268 s8 luteinizing hormone releasing hormone (LHRH) in all members of a genetically diverse population are described below.
The general feature of such SSALG i8 that the degenerate Th sequence is separated from the LHRH sequence, positions 19-28 of SEQ ID N0:34, by two Gly residues. The Gly residues act as a spacer between the Th epitope and the LHRH sequence. The Gly spacer and the LHRH sequence are invariAnt in the library sequences.
The degenerate helper T cell epitope present in SSAL1 THlLHRH is modeled after a universal epitope identified from the F protein of measles virus (Partidos et al. l990, J Gen Virol :Zl: 2099). This epitope is .~kLe~e,l-ed by residues 288-302 of the F protein. Charged residues Glu/Asp are added at position 1 to increase the charge surrounding the hydrophobic face of the epitope. This face is made up of residues at positions 2, 5, 8, 9, 10, 13 and 16. The hydrophobic residues commonly associated with p~ ; cr~ c epitopes are added at these positions. A Rothbard sequence is indicated by the residues in bold type in the prototype ~C-~Il.,nce. Sequences obeying the 1, 4, 5, 8 rule are underlined in the prototype sequence.
The degenerate helper epitope present in SSAL2 TH2LHRH
is modeled after the universal epitope identified from the hepatitis B virus surface antigen (Greenstein et al. 1992, J
Immunol 48: 3970). This epitope is l_l.L~serlLed by residues 19-33 of HBsAg. Positively charged Lys and Arg residues are added at posltions 1, 2, 3 and 5 to increa6e the charge surrounding the hydrophobic face of the helical structure.
The charged amino acids at position 3 also contributes the required residue to generate a sequence obeying the 1, 4, 5, 8 rule (residues underlined in the prototype sequence).
Hydrophobic residues at positions 4, 6, 7, 10, 11, 13, 15 and 17 make up the lly.l- o~l,ol,ic f ace of an amphipathic helix .

~ W095/11ss8 217SS79 PCr~Ss~/12268 A ~othbard sequence is indicated by the residues in bold type in the prototype sequence.
. The SsALs described above and as shown in Flgs. 29 A
and B are therefore the first artificial members of the , 5 ~ c~n~c class of helper epitopes. The synthetic Th:
LHRH library can be used as a therapy for the LL~ L of androgen driven diseases including prostate cancer, prostatic hyperplasia, breast cancer and ~ LLiosis. In addition, this same synthetic antigen can be used as a contraceptive in both males and females, with a focus on companion animals. It will also be used to prevent the ty11dL - commonly known as "boar taint", the offensive aroma and f lavor of meat caused by the presence of testosterone in intact male food animals. Thc synthetic peptide libraries can be formulated in an appropriate carrier, which may include but is not limited to saline, alum, mineral or vegetable oil emulsions, microparticles, other biodegradable vehicles, and the like. Additional ~ ' can also be added as adjuvants to further potentiate the immune response to the SSAL formulations including, but not limited to, pluronic polymers, lipid amines, saponin, muramyl dipeptides or their ~n~l og~lPc~ cytokines, derivatives of cytokines, and lipids which may be either mixed with or covalently linked to the synthetic peptide libraries. The final formulation can be administered by immunization, oral delivery or delivery to other mucosal surfaces.
The production of ~L.u- Lu._d synthetic Th: LHRH
libraries provides an ideal model for testing the efficacy of the synthetic universal helper epitopes, since LHRH is a self molecule and therefore not antigenic, i.e. antibody is not ~ duced to unmodif ied LHR}~ when exogenously ~dmin1stere~ .

W0 95/11998 2 ~ ~ ~ 5 ~ ~ PCTIUS9~12268 r le 16 = - -SsALHu B27 self~~P~RdEeLATED ARTHRITIS
The MHC molecule HLA-B27 is very strongly linked with ankylosing spondylitis and related arthropathies, including reactive arthritis. It i8 thought that these autoimmune diseases are caused by cytotoxic T cells reacting against an arthritogenic peptide of cellular or mlcrobial origin, which is selectively presented by HLA-B27. The stability of HLA
molecules on the surface of a cell is dc~_~d~ll~ on the binding of a peptide, and in an uninfected cell HLA
molecule6 are stabilized by self-derived peptides. When an individual with HLA-B27 becomes infected with a microorganism which contains an arthritogenic peptide, cytotoxic T cells recogni2e the HLA-B27 peptide complex and ilttack the cell leading to autoimmune disease. An SSAL
based on the seqll~n~pc of naturally produced self peptides can be used to competitively occupy the peptide binding sites and prevent the ~les~..L~tion of the arthritogenic peptide by HLA-B27.
SSAL peptides based on the s~qUpnrpc of naturally ~L~.~ essed self-peptides from HLA-B27 molecules can be used as competitive blockers to prevent presentation of arthritogenic peptides. An SSAL HLA-B27 self-peptides is synthe6ized as linear peptides according to the mathematical and chemical r~L~se -tation shown in Fig. 30. Following the synthesis, the SSAL HLA-B27 self-peptides can be formulated in an appropriate delivery system for application at the site of inf lammation .

WO95/11998 7~S7~ PC~IUS94/12268 Exam~le 17 SSAL -BASED VACCINE THERAPY
CTL epltgPe D ATMENT OF DISEASE

- Cytotoxic T ly ' yLes (CTL) are a vital part of the immune response in many diseases, including infectious diseases such as influenza and AIDS, and in some r-l iqnAnt diseases such as malignant melanoma. A strong cellular response is an; L~--L L of a prophylactic vaccine and, in addition stimulation o~ the CTL response, can be used therapeutically to improve 6elected CTL rPcroncpc~ The stimulation of this cellular response can be achieved by the use of peptides representing target epitopes within a protein, such as the gag protein of HIV. The advantage of the SSALCTL epltope based vaccine therapy is that sPq--PnrPC
from a large number of related but not identical sequences in a virus can be covered. CTL epitopes that are restricted by the HLA-B27 molecule, from virus variants of HIV-l, HIV-2 and variants found from sequencing patients viruses can be incorporated into a SSAL-based vaccine. This gives a set of peptides which can stimulate a broadly-reactive CTL response and uve~ . possible escape mutant seq~PnrPc The SSALCTL
epitope vaccine can be PnrAr5-11 Ated with a microparticle to be administered parenterally. In this example (see Fig.
31), the SSALCTL ep~tope vaccine therapy is designed on an epitope within the gag protein of HIV that has been shown to be restricted by HLA-B27. The SSAL can be administered to HIV-infected patients with HLA-B27 to stimulate an HIV
specific CTL Lc~yul~3e. Patients can be monitored at entry and each month following administration for; u.. L in immune function and clinical status.

W095111998 ` PCTIUS9~llZ268 ~
2~rl55~

r le 18 DIAGNOSTIC MAR~ERS AND VACCINE FOR LY~E DISEASE
Lyme disease or Lyme borreliosis is an inflammatory illness in humans and other maDals with varied involvements of the skin, heart, liver, kidneys, 1oclcel-~tal system, and the central and peripheral nervous systems (Steere.
1989, N Enql J Med 321:S86). The etiological agents are a group of spirochetes which are collectively known as Borrelia burgdorferl, and which are transmitted by ticks of the genus Ixodes. The disease is of global distribution and is the most prevalent tick-borne disease in the U. S .
Prevention i8 by tick-control methods, since there is no effective vaccine available.
The organisms responsible for Lyme disease have been subdivided by recent phyl ogon~t; r. analysis into at least three g-,nnSp~ c.c referred to as B. burgdorferl sensu strictu, Borrelia garinii, and Borrelia a~zelii (formerly known as VS461). B. burgdorferi sensu strictu is common in Europe and North America. It accounts for over 90~ of isolates in the United States. It tends to be correlated with arthritis. Borrelia garinii is found in Europe and tends to be associated with neuroborreliosis. B. afzelii isolates have been found in Europe and Japan and are linked to acrodermatitis chronica atrophicans (Cann et al, 1992 339 :1598) . Antigenic variation can be correlated to these subgroups by serotypic and sequence differences in major outer surface proteins OspA and OspB. This variation occurs at both inter- and intra-species levels. The inf luence of antigenic variation and strain specif icity can be observed in the limited cross-protection between isolates in animal vaccine trials with Osp subunit antigens (Marconi W09S/11998 S$~g . . PCT/U594/12268 et al. 1993 Tnfect T ' ~ 2611) and in a lack of natural cross-immunity in humans.
Recombinant proteins vo,,~ lin~ to variable major outer surface proteins (osp) of B. burgdorferii, OspA and , 5 OspB have been proven in mouse models to elicit protective neutralizing antibodies and to have potential as protective vaccines (Telford et al. 1993 J EYn Med 178:755). OspA ix the more c~,..,,eL v~d of the two among North American isolates but was found to display considerable variability (Wallich et al. 1992 Infect I 60: 4856). OspA and OspB vaccines formulated to ~ VL~C:D~JVlld to single strains do not clicit strongly _LV.-S ~LvLective immunity. SSALs can provide significant advantages for the design of Lyme disease vaccine, since variable protective epitopes can be effectively included for wide efficacy.
OspA antigenicity h2s been extensivcly analyzed.
Studies with monoclonal antibodies and patient antibodies have shown that the C-terminal half, re6idues 108 to 273, is exposed to the human immune system (s~hllhArh et al. 1991 Infect J 59:1911-1515). This half of OspA is highly variable between strains and bears one or more crucial non-contiguous antigenic determinants between residues 133 to 273 which are responsible for strain-specific protective immunity. Class II T cell determinants have been identified at residues 221-235, 258-273, and 248 to 263 and B cell epitopes are recognized by human serum samples on the terminal 60 residues, 211-273 (Shanafelt et al. 1992 J
1~3:218).
Two useful SSALs depicted in Figs. 32A and B can be 3 o constructed as OspA SSAL ; ~ -, based on the primary amino acid sequence ~1;; 1. from 12 antigenically variant icolates and antigenic mapping. They are useful for the detection of antibodies to 80rrelia in sera and body fluids when f ormulated into i CSAys, and they are key WO95/11998 ~2 ~ 55~ 9 - Pcrlu~i9~/12268 components of a protective vaccine for humans and animals when incorporated into microparticles or other appropriate vaccine delivery system. .
ospc is another heterogeous outer surf ace protein S common to the three gc~n~sp~-ies of the Borrella burgdorferl group. OspC elicits a pr~ l n~nt antibody respon6e early in infection making it a useful diagnostic marker and studies with animal models have shown it to be a candidate for an effective vaccine (Jauris-Heipke et al, 1993). It is somewhat more variable in sequence and antigenicity than OspA, a heterogeneity that suggests that it can be effectively applied to diagnostics and vaccines as an SSAL
antigen. An SSAL (Fig. 33) useful for those applications can be .:c,l.aLLu~;Led based on an alignment of primary amino acid s~-q~ n~ n from OspC of 15 Borrelia isolates. These sequences were published: 13 in the NCBI CD/ROM database "Entrez" as NCBI seq ID nos. 311392, 311394, 313272, 313274, 313276, 313278, 313280, 434658, 434662, 434664, 434666, 4437z5, and 495736; and two in GenBank as accession nos.
U08284 and X62162. The SSAL i , as shown in Fig. 33 is incorporated into vaccine and diagnostic compositions.
A central region of the primary sequence for the f lagellin protein of B . hurgdorferi comprises a cluster of antigenic determinants that cause the production of ~ntibodies specif ically associated with Lyme disease .
Serological detection and mea~SuL. ~ of these anti3~odies is useful for the diagnosis of Lyme disease and for the prognosis of clinical outcome (.~:r~hnF~ r et al, 1992 Infect ImmYn 60:316). This segment is e~l ,~~ ~'9 by a region spanning f lagellin residues 175-235 that is described by the SSAL of Fig. 34. This SSAL was designed in accordance with amino acid se.luences deduced for 12 isolates from North America and Europe (Collins and Peltz, 1991 Tn~ec~ J

I wo 9S/11998 21 7S$ PCTiUSg~/l2268 59:514; Picken, 1992 J ol~n rl;crobiol 30:99; Jauris-Heipke et al, 1993 r~ed M;crobiol T -1 ~:37). It includes a con6erved framework sequence that is species-specific and common to the isolates of B. burgdorl'eri, and ten positions . 5 within the framework sequence that vary among the isolates.
Though flagellin sequence is more c~,selv~:d than OspA
6equence f or the relevant g~n--spe~; ~c of Borrelia, the pattern of i801ate to isolate variation f or f l ~ l l; n does display some correlation with the heterogeneity of OspA
(Jauris-Heipke et al, 1993; Picken, 1992).
Antibodies to regions of flagellin outside of the segment described by the SSAL of Fig. 34 share homologies and cross-reactivities to the fl~llinc of other bacteria.
Thereby, a reduced specificity for the detection of Lyme disease is imparted to i -;~cæ /ys which are sensitized with f lagellin antigens that include the cross-reactive domains (C~hn~ et al, 1992). Conversely, the isolate-specif ic variation of the 6pecies-specif ic central domain suggeæts reduced antigenicity for; ccays which are sensitized to flagellin by single-strain central region peptides. Thus, a specific and comprehensive SSAL such as that of Fig. 34 is advantageously employed in; ---e~ys for the di~7nr~æic and prognosis of Lyme disease. A peptide composition comprised o~ the SSAL depicted in Fig. 34 is useful for the detection of antibodies to the Borreli~
burqdorl'eri group of bacteria in sera and body fluids in North America and Europe, and contribute6 to the diagnosis and prognosis of ~yme disease.
;

wo 95~11998 ~ ,5~ ~ Pcrrl~S9~/12268 SEQUENCE LISTING
( 1~ GENERAL INFORMATION:
(i~ APPLICANT: United Rtl '1ri:1, Inc (ii1 TITLE OF INVENTION: Structured Synthetic ATltige~ Llbrarle~
(SSAL~ ~or Diagnostic~, Vaccines and Th~r~Plltics (iii~ NUMBER OF ~tl~;UU~ : 43 ( iV~ ~:u~E~ J - ~ ; ADDRESS:
(Al ~nnR~crT.~: Maria C.H. Lin, Esq.
(B STREET: 345 Park Avenue (C CITY: New York (D l STATE: New York ( E COUNTRY: U . S . A .
(F~ ZIP: 10154 (v~ COM UTER RT~AnART T FORM:
A MEDIUM TYPE: Floppy diRk B` COMPUTER: IBM PC ca~npatible C OPERATING SYSTEM: PC-DOS/MS-DOS
~ D I SOFTWARE: WordPer~ect 5 .1 vi~ CURRENT APPLICATION DATA:
(A~ APPT T DTION NUMBER:
(B) FILING DATE: 26-OCT-1994 ( C ~ CLASS IFICATION:
(vii) PRIOR APPLICATION DATA:
(A~ APPLICATION N[~MBER: US 08/143, 412 (B~ FILING DATE: 26-OCT-1993 (viii~ ATTORNEY/AGENT INFORMATION:
(A) NAME: Lln, Maria C.H.
(B) REGISTRATION NUMBER: 29, 323 (C~ K~KL.._~;/DOCKET N[~MBER: 1151-4120PCT
(ix) TELECOMMUNICATION INFORMATION:
(A~ TELEPHONE: 212-758-4800 (B~ TELEFAX: 212-751-6849 (2) INFORMATIûN FOR SEQ ID NO:l:

~W095/11998 ~79 PCT/US94/12268 (i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 47 amino acids (B) TYPE: amino acid . (D) TOPOLoGY: linear (ii) Mr)T~rTJT.T TYPE: peptide .
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: l ( D ) OTHER INFORMATION: / note= " S8; N2 "
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 (D) OTHER INFORMATION: /note= "G8;Dl;Ql"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "K6;R4"
(ix) FEATTIRE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "P8;Vl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "A8;V2"
( ix) FEATURE:
(A) NAT~E/KEY: Modified-site ~B) LOCATION: 6 (D) OTHER INFORMATION: /note= "I6;V4"
(ix) FEATURE:
(A) NA~ME/KEY: Modif ied-site ( B ) LOCATION: 7 (D) OTHER INFORMATION: /note= "I6;V2;A2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: lO
(D) OTHER INFORMATION: /note= "R8 ;K2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 W0 9s/ll998 ~ iS~ ~ PCTIU59~112268 (D) OTHER INFORMATION: /note= "V9;Il"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "R3;Q5;E2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "B8;A2"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 24 (D) OTHER INFORMATION: /note= "S4;A6"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOQTION: 25 (D) OTHER INFORMATION: /note= "Q4 ;S6"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "H8;Kl;Rl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 27 (D) OTHER INFORMATION: /note= "L8;A2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "P8;A2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 29 (D) OTHER INFO}~MATION: /note= "Y8;L2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 30 (D) OTHER INFORNATION: /note= "I9;Fl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOQTION: 32 (D) OTHER INFORMATION: /note= "Q8;E2"

WO95/11998 1 7~79 PCTIUS94112268 ( ix ) FEATURE:
(A) NAME/KEY: Modified-site t B ) LOCATI ON: 3 4 . (D) OTHER INFORMATION: /note= "M8;Q2"
( ix) FEATURE ~
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 35 (D) OTHER INFOR~MATION: /note= "M4 ;Q4 ;R2"
(ix) FEATURE:
(A) NAME/KEY: Modified-fiite (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "L8;Ml;Il"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 9 (D) OTHER INFORMATION: /note= "Q8;M2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 40 (D) OTHER INFORMATION: /note= "F8;L2"
(ix) FEATURE:
(A) NAME/KEY: Mn~l 1 f i Pd-site (B) LOCATION: 42 (D) OTHER INFORMATION: /note= "Q8 jS2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 44 (D) OTHER INFORMATION: /note= "A8;I2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "L8;Q2"
(Xi) SEQUENCE L~E~:Kl}~LloN: SEQ ID NO:l:
Ser Gly Lys Pro Ala Ile Ile Pro Asp Arg Glu Val Leu Tyr Arg Glu l0 15 Phe Asp Glu Met Glu Glu Cys Ser Gln His Leu Pro T
20 25 yr Ile Glu Gln Gly Met Met Leu Ala Glu Gln Phe Lys Gln Ly~ Ala Leu Gly Leu W095111998 a~ ss~ PCTIIJS9~112268 . . ..

(2) INFORMATION FOR SEQ ID NO:2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTa: 61 amino acids .
(B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ( ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "P9 jG
(ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "K3 jR7 ( i x ) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "N3;T7"
(ix) FEATURE:
(A) NANE/KEY: Nodif ied-site (B) LOCATION: 21 (D) OTHER INFORNATION: /note= "V9;I1"
( ix) FEATURE:
(A) NANE/KEY: Modified-site ( B ) LO CATI ON: 4 8 (D) OTHER INFORMATION: /note= "T9;Pl"
( ix ) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 52 (D) OTaER INFORMATION: /note= "S9;W1"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:2:
Ser Thr Asn Pro Lys Pro Gln Lys Lys Asn Lys Arg Asn Thr Asn Arg Arg Pro Gln Asp Val Lys Phe Pro Gly Gly Gly Gln Ile Val Gly Gly Val Tyr Leu Leu Pro Arg Arg Gly Pro Arg Leu Gly Val Arg Ala Thr ~WO95/11998 1 7SS7g PC~/US9~/1226 Arg Lys Thr Ser Glu Arg Ser Gln Pro Arg Gly Arg Ar . (2) INFORMATION FOR SEQ ID NO:3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 80 amino acids (B) TYPE- amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ix) FEATURE:
(A) NAME/KEY: M~ ~1 i f i ed-site (B) LOCATION: l ( D ) OTHER INFORMATION: / note= "K9; Rl "
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "L6;I4"
( ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "E8;Al;Sl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 7 (D) OTHER INFORMA~ION: /note= "V4;A3;Tl;Fl;Yl"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "K9;Rl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION 2 9 (D) OTHER INFORMATION: /note= "K8 ;A2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "V4;S3;Tl;R2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 32 WO95/11998 2i~5~9 PCrlU594/12268 ~D) OTHER INFORMATION: /note= "A5;G5"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 33 (D) OTHER INFORMATION: /note= "L8;M2"
(ix) F
(A) NAME/KEY: Modified-site (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "I6 jL3 jVl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "N9 jHl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 51 ( D ) OTHER INFORMATI ON: / note= " T 9; Al "
( ix) FEATURE -(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 5 2 (D) OTHER INFORMATION: /note= "57;Il;Q2"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 54 (D) OTHER INFORMATION: /note= "D9 jNl"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 59 (D) OTHER INFORMATION: /note= "A9 jSl"
( ix ) FEATURE -(A) NAME/KEY: Modified-Gite (B) LOCATION: 67 ( D ) OTHER INFORMAT ION: / note= " Y 5; FS "
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 79 (D) OTHER INFORMATION: /note= "T8;V2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 80 (D) OTHER INFORMATION: /note= "C8;A2"

~ WO 95111998 2 I 7 ~ 5 7 9 PCTI~JS94/12268 (xi) SEQUENCE DESCRIPTION: SEQ ID NO: 3:
. Lys Ala Ile Pro Leu Glu Val Ile Lys Gly Gly Arg His Leu Ile Phe Cy6 His Ser Lys Lys Lys Cy~ Asp Glu Leu Ala Ala Lys Leu Val Ala Leu Gly Ile A6n Ala Val Ala Tyr Tyr Arg Gly Leu Asp Val Ser Val Ile Pro Thr Ser Gly Asp Val Val Val Val Ala Thr Asp Ala Leu Met Thr Gly Tyr Thr Gly Asp Phe Asp Ser Val Ile Asp Cys Asn Thr Cys (2) INFORMATION FOR SEQ ID NO:4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 44 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) M~-T.T~rTTT.T. TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "R9 ;P1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "P8;Ll;V1"
(ix) FEATURE-(A) NAME/KEY: Modified-site ( B ) LOCATION: 10 (D) OTHER INFORMATION: /note= "V5;L4;I1"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "T5;54;P1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site 2 ~ ~ 55 PCT/IIS94/12268 (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "K4 jD4;R2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "D9 jGl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "E5 jV4;Ql"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "P9 jAl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (8) LOCATION: 22 (D) OTHER INFORMATION: /note= "V8 jT2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 24 (D) OTEIER INFORMATION: /note= "H8 jLl;Al"
( ix) FEATURE:
(A~ NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "P8;A2"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 30 (D) OTHER INFORMATION: /note= "P9;Sl"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "P5;T3;Vl;Il"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite ( B ) LOCATION: 3 2 (D) OTHER INFORMATION: /note= "K8;Pl;Rl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 33 WO95/11998 1 7~79 PCrlll594/12268 (D) OTHER INFORNATION: /note= "54 jA4;Kl jQl"
( ix) FEATURE:
. (A) NAME/KEY: Modified-site (B) LOCATION: 34 (D) OTHER INFORNATION: /note= "P8;T2"
( i x ) FEATURE:
(A) NANE/KEY: Nodified-site (B) LOCATION: 36 (D) OTHER INFORNATION: /note= "V5;I4;Tl"
( ix) FEATURE:
(A) NANE/KEY: Modif ied-site ( B ) LOCAT I ON: 4 2 (D) OTHER INFORNATION: /note= "~7;R3"
(ix) FEATURE:
(A) NANE/KEY: Nodified-site (B) LOCATION: 44 (D) OTHER INFORNATION: /note= "T9;Al"
( ix) FEATURE:
(A) NANE/KEY: Modified-site ( B ) LOCAT I ON: 4 l (D) OTHER INFORNATION: /note= "K4 ;R6"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:4:
Ala Arg Pro Asp Tyr A~;n Pro Pro Leu Val Glu Thr Trp Lys Lys Pro lO 15 Asp Tyr Glu Pro Pro Val Val His Gly Cys Pro Leu Pro Pro Pro Lys Ser Pro Pro Val Pro Pro Pro Arg Lys Lys Arg Thr (2) INFORNATION FOR SEQ ID NO:5:
( i ) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 36 amino acids (B) TYPE: amino acid (D ) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ;

( ix) FEATURE:
(A) NANE/KEY: Modified-site Wo 9~111998 PCTIUS94/12268 ~
2~55~ 9 ( B ) LOCATION: 2 (D) OTHER INFORMATION: /note= "E8;D2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: ~
(D) OTHER INFORMATION: /note= "V8 jT2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "A8;El;Dl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l1 (D) OTHER INFORMATION: /note= "T9;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 4 (D) OTHER INFORMATION: /note= "A6;T4"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 6 (D) OTHER INFORMATION: /note= "G5;K3;A2"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:5:
Cys Glu Pro Glu Pro Asp Val Ala Val Leu Thr Ser Met Leu Thr Asp 5 l0 15 Pro Ser His Ile Thr Ala Glu Ala Ala Gly Arg Arg Leu Ala Arg Gly ser Pro Pro Ser (2) INFORMATION FOR SEQ ID N0:6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 55 amino acid6 (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) M~T.T'~'TJT.T~` TYPE: peptide ( ix) FEATURE:

~W095111998 7Ss79 PCT/US94~12268 (A) NAME/KEY: Modif ied- ite ( B ) LOCATION: 2 (D) OTHER INFORMATION: /note= "Ll2 jI2 jV2"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= 1'V7 jT4 jI5"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Al2 jV4"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "Yl2;V2;A2"
(ix) FEATURE:
(A) NAME/KEY: Nodified-site (B) LOCATION: lO
(D) OTHER INFORMATION: /note= "Q5 jEll"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Rll jH3 jK2"
(ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "S5 jV7 jI4"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l5 (D) OTHER INFORMATION: /note= "T3 jSl3"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: l 6 (D) OTHER INFORMATION: /note= "Gl2 jS2 jT2"
( ix) FEATURE:
(A) NANE/KEY: Nodified-site ( B ) LOCATION: l 7 (D) OTHER INFORMATION: /note= "L5 jI7 jS2 jG2"
(ix) FEATURE:
(A) NANE/KEY: Modified-site Wo 95/11998 PCT/US91/12268 $ 7 (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "H12 jY2;M2"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 20 (D) OTHER INFORMATION: /note= "V14;A2"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "P5;sg;T21' ( i x ) F EATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "S10;A2;N2;D2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LO CAT ION: 2 9 ( D ) OTHER INFORMATION: / note= " I 15; Vl "
( ix ) FEATURE:
(A) NAME/KEY: Modified--site ( B ) LO CATI ON: 3 0 (D) OTHER INFORMATION: /note= "V12;T4"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "Y12;W4"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 32 (D) OTHER INFORMATION: /note= "E12;Q4"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 3 (D) OTHER INFORMATION: /note= "A12;L4"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 3 4 (D) OTHER INFORMATION: /note= "All;Hl;T2;Q2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 35 W095/11998 21 7~7,~ PCT/IIS94/12268 (D) OTHER INFORMATION: /note= "D13 ;A2 ;Nl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 6 (D) OTHER INFORMATION: /note= "A9;L2;M4;Vl"
- (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "I12;V4"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 8 tD) OTHER INFORMATION: /note= "L9;M7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 40 (D) OTHER INFORMATION: /note= "Tll;L2;V2;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION 47 (D) OTHER INFORMATION: /note= "V12;E4"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 48 (D) OTHER INFORMATION: /note= "R12;N2;K2"
( i x ) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 49 (D) OTHER INFORMATION: /note= "E12;D2;V2"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site ( B ) LOCATION: 5 0 (D) OTHER INFORMATION: /note= "G9;S2;N4;Dl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 51 ( D ) OTHER INFORMATION: / note= " N 14; G2 "
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 53 (D) OTHER INFORMATION: /note= "S14;L2"

WO95/11998 ~5~,~ 80 PCr/U~94112268 ( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "A4;S5 jF2;T4;Vl"
(ix) FEATURE:
(A) NAME/XEY: Modified-site (B) LOCATION: 54 (D) OTHER INFORMATION: /note= "Rl5;Hl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:6:
ys Leu Thr Val Pro Ala Ser Ala Tyr Gln Val Arg Asn Ser Thr Gly 5 l0 15 eu Tyr His Val Thr A6n Asp Cys Pro Asn Ser Ser Ile Val Tyr Glu la Ala Asp Ala Ile Leu His Thr Pro Gly Cys Val Pro Cys Val Arg Glu Gly Asn Ala Ser Arg Cys (2) INFORMATION FOR SEQ ID NO:7:
(i) SEQUENCE '~AT~Ar'l'F~T.~TICS:
(A) LENGTH: 44 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: l inear (ii) M ~T.T~t TTT.T~ TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 (D) OTHER INFORMATION: /note= "Vl2;E4"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "Rl2;K2;N2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "El2;V2;D2"
( ix) FEATURE:

~ WO 95/11998 `t 7~5 7 ~ PcrltJS94/12268 (A) NAME/KEY: Modif ied-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "G9 jS2;N4 jDl"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "N14;G2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-æite (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "A4;55;F2;T4;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "514;L2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "R15;Hl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION- 12 (D) OTHER INFORMATION: /note= "V12;I4"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTEIER INFORMATION: /note= "A12;Q2;P2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 14 ( D ) OTHER INFORMATION: / note= "M3; V6; L7 "
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFoRMATIoN: /note= "T14;52"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (El) LOCATION: 17 (D) OTHER INFORMATION: /note= "T12;N4"
(ix) FEATURE:
(A) NAMB/KEY: Modified-site ~ 55~ PCIIUS9~/12268 (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "V9 jL7"
( ix~ FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 20 (D) OTHER INFORMATION: /note= "T5;A7;V4"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "R12;Q2;K2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 22 (D) OTHER INFORMATION: /note= "D5 jN7 jQ2 jH2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 23 (D) OTHER INFORMATION: /note= "G5;A2 jS2;V2;P2;R2;Nl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 24 (D) OTHER INFORMATION: /note= "R5;55;T2;G4"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 2 5 (D) OTHER INFORMATION: /note= "L5;V3;I4;A4"
( ix) FEATURE:
(A) NAME/REY: Modified-site ( B ) LOCATI ON: 2 6 (D) OTHER INFORMATION: /note= "P12;L4"
ix) FEATURE:
(A) NAME/REY: Modified-site ( B ) LOCATION: 2 7 (D) OTHER INFORMATION: /note= "A3 ;T13"
(ix) FEATURE:
(A) NAME/REY: Modif ied- t (B) LOCATION: 28 Sl e (D) OTHER INFORMATION: /note= "Tll;Q2;Hl;Al;Rl"
( ix ) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 29 WO9s/ll998 1 7S~79 PCT/US9V12268 tD) OTHER INFORMATION: /note= "Q5;T7;G2;Nl;S1"
(ix) FEATURE:
. (A) NAMB/KEY: Modified-site ( B ) LO CATI ON: 3 O
(D) OTHBR INFORMATION: /notc= "LlO;I6"
( ix) FEATURE:
(A) NAMB/KEY: Modified-site (B) LOCATION: 3 2 (D) OTHER INFORMATION: /note= "R12;T4"
ix) FEATURE:
(A) NAME/REY: Modified-site ( B ) LOCATION: 3 4 (D) OTHER INFORMATION: /note= "I7;V9"
(ix) FEATURE:
(A) NAME/KEY: Modified-6ite ( B ) LOCATION: 3 6 (D) OTHER INFORMATION: /note= "L12;M4"
( ix ) FEATURE:
(A) NAME/KEY: Mnrl i f i ~rl-site (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "L12 jI2;V2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 9 (D) OTHER INFORMATION: /note= "G12;M4"
(ix) FBATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 40 (D) OTHER INFORMATION: /note= "S7;A8;T1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 42 (D) OTHE~ INFOR~ATION: /note= "T9;A7"
( ix ) FEATURB:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 4 3 (D) OTHER INFORMATION: /note= "L9;F5;Vl;A1"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:7:
Cys Val Arg Glu Gly Asn Ala Ser Arg Cys Trp Val Ala Met Thr Pro Wo 95/11998 ~ 8 4 PCr~Ss~/12268 5 l0 15 Thr Val Ala Thr Arg Asp Gly Lys Leu Pro Ala Thr Gln Leu Arg Arg His Ile Asp Leu Leu Val Gly Ser Ala Thr Leu Cys (2) INFORMATION FOR SEQ ID NO:8:
(i) SBQUENCE r~ARAl~lrT~RT!~TIcs (A) LENGTH: 40 amino acids (B) TYPE: amino acid (D ) TOPO~OGY: linear (ii) MnT.T.`~ lTT.T. TYPE: peptide ( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "Tl2;I3;Ml"
( ix) FEATURE:
(A) NAMI:/KEY: Modified-site ( B ) LOCATION: 3 (D) OTHER INFORMATION: /note= "Fl2;V3;Il"
( ix) FEATURE:
(A) NAME:/KEY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Rl2;Q3;El"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "Rl3;H3"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "Hl3;Y3"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 9 (D) OTHER INFORM~ATION: /note= "W7;E5;V2;N2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-slte (B) LOCATION: l0 W095/11998 17~79 PcrNS94/12268 (D) OTHER INFORMATION: /note= "T12 jF4"
( ix) FEATURE:
- (A) NAME/KEY: Modif ied-site (B) LOCATION: ll (D) OTHER INFORMATION: /note= "T7 jV~ jL2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION- 13 (D) OTHER INFORMATION: /note= "G3 jDlljE2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 0 (D) OTHER INFORMATION: /note= "Pl4 jQ2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 2 2 (D) OTHER INFORMATION: /note= "Hl5 jTl"
( ix~ FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 3 (D) OTHER INFORMATION: /note= "I9 jLl jV6"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 24 (D) OTHER INFORMATION: /note= "TlO jS6"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B) LOCATION 3 l (D) OTHER INFORMATION: /note= "Dl5 jNl"
( ix ) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 34 (D) OTHER INFORMATION: /note= "Ml4 jL2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-Gite (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "Nl5 jSl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 40 (D) OTHER INFORMATION: /note= "T8 jA6 jL2"

WOgS/1199X 2~ 5~ 9 PCr/l~S94112268 ~

(xi) SEQUENCE DESCRIPTION: SEQ ID NO:8:
he Thr Phe Ser Pro Arg Arg His Trp Thr Thr Gln Gly Cys Asn Cys er Ile Tyr Pro Gly His Ile Thr Gly His Arg Met Ala Trp Asp Met Met Met Asn Trp Ser Pro Thr Thr (2) INFORMATION FOR SEQ ID NO:9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 35 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "A5;G5"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-6it (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "E3;H2;D2;51;Tl;Ql"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 ~D) OTHER INFORMATION: /note= "H7;I2;Yl"
( ix) FBATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "V7;Sl;Tl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "T6;S3;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modified-sit (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "G9;Ql"

WO9511 1998 PcrluS94~12268 2~ 7~S7~
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 11 . (D) OTHER INFORMATION: /note= "S2 jQl;R2;A2 jNl;Tl;El (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "H2 ;R4 ;52 ;K2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "T6;S2;Nl;Al"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 16 (D) OTHER INFORMATION: /note= "V2;T6;Ml;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-6ite ( B ) LOCATION: 17 (D) OTHER INFORMATION: /note= "52;A2;N2;Q2;Hl;Rl"
(ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "G4;53;R2;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "F2 ;L8"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 o (D) OTHER INFORMATION: /note= "V7;A2;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "57;G3"
( ix ) FEATURE:
(A) NAME/KlEY: Modif ied-site (B) LOCATION: 22 ( D ) OTHER INFORMATION: / note= "L5; M3; W2 "
( ix) FEATURE:

Wo 95/11998 PCrlUS9~/12268 ~
5~19 (A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 2 4 (D) OTE~ER INFORMATION: /note= "A3 ;53 jT4"
( ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 25 (D) OTIIER INFORMATION: /note= "P3 jQ3 jS2 jTl jLl"
( ix) FEATURE -(A) NAME/KEY: Modif ied-site (B) LOCATION: 27 (D) OTE~ER INFORNATION: /note= "A6;P4"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "K4 jS5;Rl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 0 (D) OTIIER INFORMATION: /note= "N4 ;K6"
(ix) FEATURE:
(A) NAME/KEY: M~rl i f i ed-site ( B ) LOCATION: 3 2 (D) OTEIER INFORMATION: /note= "Q9;Yl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 4 (D) OTIIER INFORMATION: /note= "I8 ;V2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 ( D ) OTHER INFORMATION: / note= "A4; Q2; V3; Tl "
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "G4;A6"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 23 (D) OTHER INFORMATION: /note= "L5;F5"
(ix) FEATURE:
(A) NAME/KEY: Modified-site Wo 95/11998 PCT/US94/122(;8 ~17sS7~9 ' ' ~9 (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "V4;I5;Ll"
., (xi) SEQUENCE DESCRIPTION: SEQ ID NO:9:
Val Asp Ala Glu Thr His Val Thr Gly Gly Ser Ala Gly His Thr Val Ser Gly Phe Val Ser Leu Leu Ala Pro Gly Ala Lys Gln Asn Val Gln Leu Ile Asn t2) INFORMATION FOR SEQ ID NO:10:
(i) SEQUENCE CHARACTERISTICS:
tA) LENGTH: 44 amino acids (B) TYPE: amino a~id (D) TOPOLOGY: linear (ii) r~ToT~r~lcrTrlr~ TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION:
(D) OTHER INFORMATION: /note= "Q8;Il;V1"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "P8;Tl;A1"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "I8 jL2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "55;T3 jEl jQ1"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "Y9 jH1"
~ix) FEATURE:

WO 95/11998 21 7 5 ~ 7 9 PCT/USg~/12268 ~
~ . ' (A) NAME/KEY: Modified-site (B) LOCATION: g (D) OTHER INFORMATION: /note= "A6;Dl jTl;E2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "N5;E2;Ml;Tl;Dl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "*8;N2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "*8 ;V2"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "G5;Sl;Pl;Al;T2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "56;El;Dl;N2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site tB) LOCATION: 15 (D) OTHER INFORMATION: /note= "G5;Rl;Sl;Il;Dl;Pl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "P4 jLl;S2;Ql;Gl;El"
( iX ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Q7;El;M2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "P8;A2"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site Wo 95/11998 . PCr/US94/12268 ~S7~
( B ) LOCATION: 2 8 ( D ) OTHER INFORMATION: / note= "K4; R5; P l "
. (ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 29 (D) OTHER INFOR~ATION: /note= "P8;Q2"
( ix) FEATURE:
(A) NAME/KEY: Mn~lifi-~rl--site (B) LOCATION: 3l (D) OTHER INFORMATION: /note= "G9;Tl"
(ix) FEATURE:
(A) NAME/KEY: Nodified-site ( B ) LOCATION: 3 2 (D) OTHER INFORMATION: /note= "I9;Vl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATI ON: 3 4 ( D ) OTHER INFORM~TION: / note= "P9; S l "
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 36 ( D ) OTHER INFORMATION: / note= "K5; S4; Rl "
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "S6;El;Q2;Tl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l0:
Gln Gly Trp Gly Pro Ile Ser Tyr Ala Asn Xaa Xaa Gly Ser Gly Pro Asp Gln Arg Pro Tyr Cys Trp HiG Tyr Pro Pro Lys Pro Cys Gly Ile Val Pro Ala Lys Ser Val Cys Gly Pro Val Tyr Cys ( 2 ) INFORMATION FOR SEQ ID NO: ll:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 32 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear .

Wo 95/1 1998 PCTIITS9~/12268 21~55~9 (ii) Mt LT.rTTT r TYPE: peptide ix ) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "El2;K4"
( ix) FEATURE:
(A) NANE/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORNATION: /note= "Sll jP2 ;A2 jTl"
( ix ) FEATURE
(A) NAME/KEY: Mt rli ~ site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "E9;R2;K2;Ql;Tl;Vl"
( ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Nl5;Vl"
( ix) FEATURE:
(A) NANE/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "Tl5;Al"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: l 0 (D) OTHER INFORMATION: /note= "Pl4;Ll;Hl"
( ix ) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: ll (D) OTHER INFORMATION: /note= "Nl2;Sl;Y3"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l2 (D) OTHER INFORMATION: /note= "Nl3;Yl;Kl;Sl"
( ix) FEATURE:
(A) NAME/KEY: Nodified-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "Nl5;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 14 WO 95111998 PcrluS94/12268 ~17Ss,~9 , ~,`, . I ~ ~

(D) OTHER INFORMATION: /note= "Tl3 jKl;Vl;Il"
(ix) FEATURE:
. (A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "Rl4 ;Kl;Tl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: l 6 ( D ) OTHER INFORMATION: / note= "K9; R3; Tl; El; Q2 "
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: l 7 (D) OTHER INFORMATION: /note= "Sll jR2 jG3"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 18 (D) OTHER INFORMATION: /note= "IlO jV3;Ll;T2"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l9 (D) OTHER INFORMATION: /note= "H8 jP4 jR2 jGljSl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 O
(D) OTHER INFORMATION: /note= "Il2 jL2 jV2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 22 (D) OTHER INFORMATION: /note= "Pl4 jWl jRl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 4 (D) OTHER INFORMATION: /note= "Q9 jR6 jKl"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-6ite ( B ) LOCATION: 2 5 (D) OTHER INFORMATION: /note= "All jT3 jVl jSl"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "FlO;W2;L3;Yl"

$~9 ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "Yl2 jF2 jSl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 8 (D) OTHER INFORMATION: /note= "A6;T7;R2 jKl"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION 29 (D) OTHER INFORMATION: /note= "Tl4;Rl;Sl"
( ix ) FEATrJRE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 0 (D) OTHER INFORMATION: /note= "Gl3;Kl;R2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "D7;E3;N3;Ql;Yl;Rl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:ll:
lu Ser Val GlU Ile Asn Cys Thr Arg Pro Asn Asn Asn Thr Arg Lys 5 l0 15 er Ile His Ile Gly Pro Gly Gln Ala Phe Tyr Ala Thr Gly Asp Met (2) INFORMATION FOR SEQ ID NO:12:
(i) SEQUENCE r~r~RAI''rRRT~TICS
(A) LENGTH: 35 amino ~cids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MtlT.T.rrrTR TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "I9;V38;L3"
(ix) FEATURE:

WO 95111998 ~S5;~9 PCTIUS94/12268 (A) NAME/KEY: Modif ied-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "L49;Ql"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "V41;L5;I4"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "R47;Sl;T2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "Y47;Fl;L2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "L49;Il"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "K27;Q6;Gl;R16"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "D47;El;N2"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFOR?lATION: /note= "Q49;Rl"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "L48;Rl;Il"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-5ite (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "L49;Ml"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site Wo 95/11998 - PCT/U591112268 5~

(B) LOCATION: 17 ~D) OTHER INFORMATION: /note= "I42 jF2 jM2 jGl jL3"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LO CAT ION: 1 8 (D) OTHER INFORMATION: /note= "W49 ;Ll"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l9 (D) OTHER INFORMATION: /note= "G49;Wl"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site ( B ) LOCATION: 2 0 (D) OTHER INFORMATION: /note= "C49;Rl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "S48;K2"
( ix) FEATURE:
(A) NAME/~EY: Modif ied-site ( B ) LOCAT ION: 2 3 (D) OTHER INFORMATION: /note= "K49;Rl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 4 ( D ) OTHER INFORMATION: / note= "L4 3; H4; I 2; Al "
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "I47;V3"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "T46;Y3;Pl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 9 (D) OTHER INFORMATION: /note= "A22;T20 jN5;Fl;52"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 31 WO95/11998 PCT~US94/12268 ~17.~S~

(D) OTHER INFORMATION: /note= "P48;K2"
(ix) FEATURE:
-~ (A) NAME/KEY: Nodified-site (B) LOCATION: 34 (D) OTHER INFOR15ATION: /note= "A26;T8;S14;Rl;Nl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 35 ( D ) OTHER INFORMATION: / note= " 54 8; T2 "
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-sitc (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "G46;El;Rl;Nl;Sl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:12:
Arg Ile Leu Ala Val Glu Arg Tyr Leu Ly5 A5p Gln Gln Leu Leu Gly Ile Trp Gly Cys Ser Gly Lys Leu Ile Cys Thr Thr Ala Val Pro Tr Asn Ala Ser (2) INFORMATION FOR SEQ ID NO:13:
(i) SEQUENCE t'TTAR~'rR~T.qTICS:
(A) LENGTH: 36 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear ( ii ) MnTlRr'uT~F~ TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "Tl9;S1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 10 (D) OTHER INFORMATION: /note= "Ql;K18;A1"
(ix) FEATURE~
(A) NAME/KEY: Modi~ied-site WO95/11998 ~ 51$ 1 ~ PCT/US9~112268 1--( B ) LOCATION: l l (D) OTHER INFORMATION: /note= "Dl9 jHl"
(ix) FEATURE: ^' (A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "R2;Ql6;Ll;Xl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "Ll9 ;Xl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-slte (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "Sl4;A6"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "Tl8 jSl jXl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 2 (D) OTHER INFORMATION: /note= "Wl9 jRl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 3 (D) OTHER INFORMATION: /note= "V7;Al;Pl2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 5 (D) OTHER INFORMATION: /note= "Dl0;E4;A6"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "Sll;T8;Nl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 13:
Arg Val Thr Ala Ile Glu Lys Tyr Leu Gln Asp Gln Ala Arg Leu Asn 5 lO 15 Ser Trp Gly Cys Ala Phe Arg Gln Val Cys His Thr Thr Val Pro Tr I wo 95/1 1998 PC~/U594/12268 ~ 99 Val Asn Asp Ser (2) INFORMATION FOR SEQ ID NO:14:
(i) SEQUENCE ~'T-TAT~A(''l'T~T2T.STICS:
(A) LENGTH: 4 5 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) l-ltlT.T~CTTT.T~ TYPE: peptide (ix) FEATURE:
(A) NAMEIKEY: Modi~ied-site (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "Ll;I1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "Nl;T1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "Tl;S1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Sl;T1"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "Ll;P1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Al;S1"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 16 ( D ) OTHER INFORMATI ON: / note= "L1; V1 "
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site t (B) LOCATION: 17 WO 95/11998 2 1~ 5 ~ 7 9 PCT/US9 1/12268 1--(D) OTHER INFORMATION: /note= "Pl jHl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ..
(B) LOCATION- 18 (D) OTHER INFORMATION: /note= "Hl;Dl"
(ix~ FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 20 (D) OTHER INFORMATION: /note= "Wl;Dl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) ~OCATION: 22 (D) OTHER INFORMATION: /note= "Dl;El"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LO CATI ON: 2 4 (D) OTHER INFORMATION: /note= "Il;Vl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 2 6 (D) OTHER INFORMATION: /note= "El;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 2 9 (D) OTHER INFORMATION: /note= "Il;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 30 (D) OTHER INFORMATION: /note= "Pl;Sl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 32 (D) OTHER INFOR15ATION: /note= "Kl;Tl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 33 (D) OTHER INFORMATION: /note= "Sl;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "Ll;Il"

! Wo 9sl11998 ~ 7$S~ PcrluS94112268 (ix) FEATURE:
(A) NAME/KEY: Nodif ied-site ( B ) LOCATION: 3 7 (D) OTHER INFORMATION: /note= "Tl;Kl"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 38 (D) OTHER }NFORMATION: /note= "Ll;Fl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 39 (D) OTHER INFORNATION: /note= "Vl;Il"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 14:
Leu Asn Thr Glu Pro Ser Gln Leu Pro Pro Thr Ala Pro Pro Leu Leu l0 15 Pro His Ser Trp Leu Asp His Ile Leu Glu Pro Ser Ile Pro Trp Lys Ser Lys Leu Leu Thr Leu Val Gln Leu Thr Leu Gln Ser (2) INFORMATION FOR SEQ ID NO:15:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGT}~: 37 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) M ~T.FrTTT.F TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 2 (D) OTHER INFOFMATION: /note= "Fl;Yl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "Dl;Ql"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "Ql;Rl"

~ ~ ~5~ ~ PCTluS9~112268 1--(ix) FEATURE:
(A) NAME/KEY: ModifiQd-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Il jLl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: lO
(D) OTHER INFORMATION: /note= "Vl;Tl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: ll (D) OTHER INFORMATION: /note= "Sl;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Sl;Dl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "Pl;Nl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "Ll;Il"
( i x ) F EATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 5 (D) OTHER INFORMATION: /note= "Sl;Al"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "Tl;Pl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 l (D) OTHER INFORMATION: /note= "Gl;Al"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT ION: 3 2 (D) OTHER INFORMATION: /note= "Sl;Tl"
(ix) FEATURE:

WO95/11998 2~ 7SS7~ PCr/US94/12268 (A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 4 (D) OTHER INFORMATION: /note= "Sl;Rl"
. .
(xi) SEQUENCE l~:s~:K~ ON: SEQ ID NO:15:
Cy6 Phe Asp Pro Gln Ile Gln Ala Ile Val Ser Ser Pro Cys His Asn l0 15 Ser Leu Ile Leu Pro Pro Ser Leu Ser Pro Val Pro Thr Leu Gly Ser Ar Ser Arg Arg Ala g 35 (2) INFORMATION FOR SEQ ID NO:16:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 71 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: linear ( i i ) M ~ TTr-F TYPE: peptide ( ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 7 (D) OTHEK INFORMATION: /note= "Ql jHl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "Kl jQl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l9 (D) OTHER INFORMATION: /note= "Ll jIl"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "Kl jRl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LQCATION: 2 2 (D) OTHER INFORMATION: /note= "IljVl"

WO 95/11998 PcrluS9~112268 ~
'19 ( ix) FEATURE:
(A) NAMB/KEY: Modified-site (B) LOCATION: 24 (D) OTHER INFORMATION: /note= "Nl jQl" .-ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 47 (D) OTHER INFORMATION: /note= "Ll;Il"
ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "Rl;Cl"
ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 54 (D) OTHER INFORMATION: /note= "Pl;Ll"
ix ) FEATURE:
(A) NAME/KEY: M~-l;fiPcl-site (B) LOCATION: 57 (D) OTHER INFORMATION: /note= "Tl;Sl"
ix) ~EATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 59 ( D ) OTHER I NFORMATI ON: / note= " S l; T l "
ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 62 (D) OT~ER INFORMATION: /note= "Pl;Sl"
ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 63 (D) OTHER INFORMATION: /note= "Il;Vl"
xi) SEQUENCE DESCRIPTION: SEQ ID NO:16:
al Asp Lys Asp Ile Ser Gln Leu Thr Gln Ala Ile Val Lys His His 5 l0 15 ys His Leu Leu Lys Ile Ala Asn Tyr Ala Ala Gln His Arg Arg Gly eu A~p Leu Leu Phe Trp Glu Gln Gly Gly Leu Cys Lyo Ala Leu Gln WO95/11998 1 7S~7~ PCT/US94/12268 Glu Gln Cys Arg Phe Pro His Ile Thr His Ser His Val Pro Ile Leu Gln Glu Arg Pro Pro Leu Glu ( Z ) INFORMATION FOR SEQ ID NO :17:
( i ) SEQUENCE t'~TAR~t'TF.RT ~TICS:
(A) LENGTH: 74 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULB TYPE: peptide iX ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "595;G5"
(iX) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "K82;Nl8"
( iX) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 4 (D) OTHER INFORMATION: /note= "Y86;Fl4"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 0 (D) OTHER INFORMATION: /note= "S95;Y5"
(iX) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 30 ( D ) OTHER INFORMATION: / note= " F9 5; C5 "
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 32 (D) OTHER INFORMATION: /note= "N50;T45;55"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 34 (D) OTHER INFORMATION: /note= "G90;N5;D5"

WO 95/11998 PCrlUSs~112268 , 21755~ lo~
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 36 (D) OTHER INFORMATION: /note= "T60;N40" .
( ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "W9O;L10"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 39 (D) OTHER INFORMATION: /note= "G95 jE5"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 43 (D) OTHER INFORMATION: /note= "N77 jS23"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 47 (D) OTHER INFORMATION: /note= "560 jY30 jS10"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "G9O jE10"
( ix) FEATURE:
(A) NAME/KEY: Nodified-site (B) LOCATION: 53 (D) OTHER INFORMATION: /note= "P72;S27"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 54 (D) OTHER INFORMATION: /note= "D46jV18;A18;G18"
( iX) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 55 (D) OTHER INFOR~ATION: /note= "N82;S14;K4"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 56 (D) OTHER INFORMATION: /note= "G68 jS32"
( ix ) FEATURE:

WO 951~1998 ~1 7S~ 7~ PCT/US94~12268 (A) NAME/KEY: Modif ied-sit (B) LOCATION: 65 (D) OTHER INFORMATION: /note= "Y55;T45"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 6 6 (D) OTHER INFORMATION: /note= "R82;E18"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 68 (D) OTHER INFORMATION: /note= "G64;E36"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 69 (D) OTHER INFORMATION: /note= "S86;Y14"
( i x ) FEATURE:
~A) NAME/REY: Modified-site (B) LOCATION: 70 (D) OTHER INFORMATION: /note= "T72;K27"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 73 (D) OTHER INFORMATION: /note= "V86;A14"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 74 (D) OTHER INFORMATION: /note= "L91;Q9"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 6 (D) OTHER INFORMATION: /note= "S91;N9"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:17:
Ser Lys Ala Tyr Ser Ser Cys Tyr Pro Tyr Asp Val Pro Asp Tyr Ala Ser Leu Arg Ser Leu Val Ala Ser Ser Gly Thr Leu Glu Phe Ile Asn Glu Gly Phe Thr Trp Thr Gly Val Thr Gln Asn Gly Gly Ser Ser Ala WO9~/11998 PcrnlS94/12268 ~
2i~S57~ 108 Cys Lys Arg Gly Pro Asp Asn Gly Phe Phe Ser Arg Leu A6n Trp Leu Tyr Lys Ser Gly Ser Thr Tyr Pro Val Leu ~.

(2) INFORMA~ION FOR SEQ ID NO:18:
(i) SEQUENCE rT-Tl~T~r'l'T~T.STICS:
(A) LENGTE~: 78 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: linear (ii) M-)T.r~lTr.F TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "T85;I15"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Y85;D15"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "A85;I15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 39 (D) OTHER INFORMATION: /note= "S85;F15"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 40 (D) OTHER INFORMATION: /note= "W84;N8;T8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 41 (D) OTHER INFORMATION: /note= "P84;W8;L8"
( ix ) FEATT~RE:
(Al NAME/KEY: Modified-site (B) LOCATION: 42 (D) OTHER INFORMATION: /note= "N62;K23;T15"

WO95/11998 PCr/US94/12268 , =~
1~9 2175573 ( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 43 (D) OTHER INFORMATION: /note= "H85;G15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 44 (D) OTHER INFORMATION: /note= "T69 ;N31"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATI ON: 4 5 (D) OTHER INFORMATION: /note= "V70;T15;Q15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 47 (D) OTHER INFORMATION: /note= "K62;R23;N15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite ( B ) LOCATION: 4 9 (D) OTHER INFORMATION: /note= IlV85;Gl5'' ( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATI ON: 5 0 (D) OTHER INFORMATION: /note= "T85;S15"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 51 ~D) OTHER INFORMATION: /note= "A84;S8;N50"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "S69 ;A31"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 54 (D) OTHER INFORMATION: /note= "S85;K15"
( ix) FEATURE:
(A) NAMI~/KEY: Modified-site (B) LOCATION: 55 (D) OTHER INFORMATION: /note= "H85;R15"
(ix) FEATURE:

PCr/U59~112268 (A) NAME/KEY: Modified-site (B) LOCATION: 56 (D) OTHER INFORMATION: /note= "K55 jA15 jN15 jG15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 57 (D) OTHER INFORMATION: /note= "G84 jP8 jR8"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 58 (D) OTHER INFORMATION: /note= "K70;D15;R15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 6 0 (D) OTHER INFORMATION: /note= "585;G15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 62 (D) OTHER INFORMATION: /note= "Y85;F15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 63 (D) OTHER INFORMATION: /note= "R85;S15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 64 (D) OTHER INFORMATION: /note= "N85;R15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 66 (D) OTHER INFORMATION: /note= "L85;N15"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 69 (D) OTHER INFORMATION: /note= "T85;Y15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite ( B ) LOCATION: 7 0 (D) OTHER INFORMATION: /note= "E70 jG15;K15"
(ix) FEATURE:
(A) NAME/I~EY: Modified-site ~ Wo 95/1 1998 PCT/USgl/12268 17~
(B) LOCATION: 71 (D) OTHER INFORMATION: /note= "K85;S15"
( ix) FEATURE ~
(A) NAME/KEY: Modif ied-site (B) LOCATION: 72 (D) OTHER INFORMATION: /note= "N77;G15;E8"
( ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 73 (D) OTHER INFORMATION: /note= "G77;515;E8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION~ 74 (D) OTHER INFORMATION: /note= "L62 ;S23 ;T15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 77 (D) OTHER INFORMATION: /note= "N70;K15;V15"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 7 8 (D) OTHER INFORMATION: /note= "L92;Q8"
(xi) SEQUENCE l~ Kl~ : SEQ ID NO:18:
Pro Asn Ser Glu Asn Gly Thr Cys Tyr Pro Gly Tyr Phe Ala Asp Tyr Glu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile Phe Pro Lys Ser Ser Trp Pro Asn His Thr Val Thr Ly6 Gl Val Thr Ala Ser cy5 Ser His Lys Gly Lys Ser Ser Phe Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Asn Gly Lcu Tyr Pro Asn Leu (2) INFORMATION FOR SEQ ID NO:l9:
(i) SEQUENCE ~ARA~rF:~T.cTICS:
(A) LENGTH: 77 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear WO9S/11998 2~rl55 PCTIUS94/12268 ( i i ) MnT T~ rrrT.r~ TYPE: peptide ( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site ( B ) LOCATION: 5 (D) OTHER INFORMATION: /note= "Dl;Yl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Gl;Sl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Sl;Nl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 50 ( D ) O$HER INFORMATION: / note= "Rl; Kl "
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 57 (D) OTHER INFORMATION: /note= "Nl;Kl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 7 0 (D) OTHER INFORMATION: /note= "El;Kl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION 7 2 (D) OTHER INFORMATION: /note= "Sl;Pl"
(ix) FEATURE:
(A) NAME/ICEY: Modif ied-site (B) LOCATION: 73 (D) OTHER INFORMATION: /note= "Dl;Nl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:l9:
Glu Asn Pro Arg Asp Gly LeU Cys Tyr Pro Gly Ser Phe Asn Asp Tyr 5 l0 15 Glu Glu Leu Lys His Leu Leu Ser Ser Val Lys His Phe GlU Lys Val ~ WO 95/11998 PCT/US94/12268 ~7$~79 Ly6 Ile Leu Pro Lys Asp Arg Trp Thr Gln ~is Thr Thr Thr Gly Gl .. Ser Arg Ala Cys Ala Val Ser Gly Asn Pro Ser Phe Phe Arg Asn Met Val Trp Leu Thr Lys Glu Gly Ser Asp Tyr Pro Val Ala (2) INFORMATION FOR SEQ ID NO:20:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 47 amino acids (B) TYPE: ar~ino acid (D) TOPOLOGY: linear (ii) M~)T.T~.l'TJT.T~ TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "G64 jV36"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "N77;D23"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "T59;K41"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "V95;I5"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "S95;G5"
( ix ) FEATURE ~
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "K82 ;N18"
( ix) FEATURE:
(A) NAME/KEY: Modified-site Wo 95/1 1998 ; ~ PC rlUS94112268 (B) LOCATION: 18 (D) OTHER INFOR~ATION: /note= "Y86;F14"
( ix) FEATURE: ..
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 2 o (D) OTHER INFORMATION: /note= "S90;N10"
( ix) FEATURE:
~A) NAME/KEY: Modif ied-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "G90;E10"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "P73 ;S27"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "D46;V18;A18;G18"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "N82 jS14 ;K4"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 2 9 (D) OTHER INFORMATION: /note= "G68;S32"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 38 (D) OTHER INFORMATION: /note= "Y55;T45"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 39 (D) OTHER INFORMATION: /note= "K82;E18"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 41 (D) OTHER rNFORMATION: /note= "G64;E36"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 42 WO95/11998 ~-7SjS PCTIUS9J/12268 (D) OTHER INFORMATION: /note= "586 jYl4"
( ix) FEATURE:
.~ (A) NAME/KEY: Modified-site (B) LOQTION: 43 (D) OTHER INFORMATION: /note= "T73 ;K27"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 46 (D) OTHER INFORMATION: /note= "V86;Al4"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 47 (D) OTHER INFORMATION: /note= "L90;Ql0"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:20:
Asp Gly Phe Gln Asn Glu Thr Trp Asp Leu Phe Val Glu Arg Ser Lys l0 15 Ala Tyr Ser Ser Cys Cys Lys Arg Gly Pro Asp Asn Gly Phe Phe Ser Arg Leu Asn Trp Leu Tyr Lys Ser Gly Ser Thr Tyr Pro Val Leu (2) INFORMATION FOR SEQ ID NO:21:
(i) SEQUENCE r~ARArTRRT~TIcs:
(A) LENGTH: 50 amino acids (B) TYPE: amino acid ( D) TOPOLOGY: linear (ii) M-T.RrlTr.R TYPE: peptide ( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "E85;Dl5"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 (D) OTHER INFORMATION: /note= "S85;Pl5"
(ix) ~EATURE:
(A) NAME/KEY: Modif ied-6ite WO 95111998 ~ PcrlllS94/12268 (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "F77;L15 jV8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "S85;P15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "K70;V15;Q15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 7 (D) OTHER INFORMATION: /note= "K70;E15;R15"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOQTION: 8 (D) OTHER INFORMATION: /note= "592;E8"
ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "A85;V15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 2 2 (D) OTHER INFORMATTON: /note= "T85;I15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 5 (D) OTHER INFORMATION: /note= "S85;K15"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 26 (D) OTHER INFORMATION: /note= "S85;K15"
( iX ) FEAT~RE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "H85;R15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 28 WO95/11998 7Ss7g PCr/USs4/12268 ~D) OTHER INFORMATION: /note= "K55;A15;N15;G15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 9 (D) OTHER INFORMATION: /note= "G84;P8;R8"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 30 (D) OTHER INFORMATION: /note= "K70;D15;R15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 2 (D) O~HER INFORMATION: /note= "585;G15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 34 (D) OTHER INFORMATION: /note= "Y85;F15"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "R85;S15"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 6 (D) OTHER INFORMATION: /note= "N85;R15"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 8 (D) OTHER INFORMATION: /note= "L85;N15"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 41 (D) OTHER INFORMATION: /note= "T85 jY15"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 4 2 (D) OTHER INFORMATION: /note= "E70;G15;K15"
( ix ) FEATURE:
- (A) NAME/KEY: Modified-site (B) LOCATION: 43 (D) OTHER INFORMATION: /note= "K85;S15"

WO95/11998 21r~s5~9 PCr/USg~/i2268 ` 11 8 ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 44 (D) OTHER INFORMATION: /note= "N77;G15;E8" .
ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "G77;515 jE8"
ix) FEATURE:
(A) NAME/KEY: Modif ied-slte (B) LOCATION: 46 (D) OTHER INFORMATION: /note= "L62;S23;T15"
ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 4 9 (D) OTHER INFORMATION: /note= "N70;K15;V15"
ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 50 (D) OTHER INFORMATION: /note= "L92;Q8"
xi) SEQUENCE DESCRIPTION: SEQ ID NO:21:
Glu Ser LeU Phe Ser Lys Lys Ser Trp Ser Tyr Ile Ala Glu Thr Pro sn Ser Glu Asn Gly Thr Cys Cys Ser Ser His Lys Gly Lys Ser Ser he Tyr Arg Asn Leu Leu Trp Leu Thr Glu Lys Asn Gly Leu Tyr Pro Asn Leu (2) INFORMATION FOR SEQ ID NO:22:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 49 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: linear (ii) M-lT.T~'f'TTT.T~' TYPE: peptide (ix) FEATURE:

WO 9~/11998 7Ss7 ~ PCT/US94/12268 (A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "Sl;Rl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 0 (D) OTHER INFORMATION: /note= "Dl;Yl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 l (D) OTHER INFORMATION: /note= "Gl;Sl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "Nl;Kl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 42 (D) OTHER INFORMATION: /note= "El;Kl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 44 (D) OTHER INFORMATION: /note= "Sl;Pl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "Dl;Nl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:22:
Asp Arg Leu Leu Ser Val Pro Glu Trp Ser Tyr Ile Met Glu 5 l0 Lys Glu Asn Pro Arg Asp Gly Leu Cys Cyc: Ala Val Ser Gly Asn Pro Ser Phe Phe Arg Asn Met Val Trp Leu Thr LYG Glu Gly Ser Asp Tyr Pro Val Ala (2) INFORMATION FOR SEQ ID NO:23:

WO9~/11998 2~ ~ 557 9 PCT/USg~/12268 ~

(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 26 amino acLds (B) TYPE: amino acid ( D ) TOPOLOGY: 1 inear (ii) M~lT.T~CTTT.T~ TYPE: peptide ix) FEATURE:
(A) NAMEIKEY: Modified-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "A7 jEl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "55;Nl;Kl;Rl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFOR~TATION: /note= "V6;Al;Tl"
( iX) FEATURE -(A) NAME/KEY: Modified-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "A7;El"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "G7 ;Dl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "E3 ;Q4 ;Sl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "K2;N6"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "V2;T5;Kl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 22 WO95/11998 ~17~S PCI/US94/12268 tD) OTHER INFORUATION: /note= "Al;T6 jVl"
. (xi) SEQUENCE DESCRIPTION: SEQ ID NO:23:
Glu Phe Gln Met Gly Ala Ala Pro Thr Thr Ser Asp Val Ala Gly Leu 5 l0 15 Glu Lys Asp Pro Val Ala Asn Val Ala Ar ( 2 ) INFORMATION FOR SEQ ID NO: 2 4:
(i) SEQUENCE rT~T~AcTT~T?T~cTI
(A) LENGTH: 24 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) Mt)T.T~CrTT.T~ TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "A5;Dl"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: l0 (D) OTHER INFORMATION: /note= "T3;A2;51"
( ix ) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: ll (D) OTHER INFORMATION: /note= "T4;Dl;Al"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-site ( B ) LOCATION: l 5 (D) OTHER INFORMATION: /note= "A5;Sl"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "Vl;A3;T2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site - (B) LOCATION: l9 (D) OTHER INFORMATION: /note= "S5;Tl"

Wo 95/11998 PCI/I~S94/12268 ~~-$`5~19 1~

( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "L4;C2"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 24:
Glu Phe Gln Met Gly Ala Lys Pro Thr Thr Thr Thr Gly Asn Ala Va Ala Pro Ser Thr Leu Thr Ala Arg (2) INFORMATION FOR SEQ ID NO:25:
(L) SEQUENCE ~ R~ TRRT~TIC5 (A) LENGTII: 30 amino acid (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: l (D) OTHER INFORMATION: /note= "L2;56"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 3 (D) OTllER INFORMATION: /note= "Kl;T4;E3"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION- 4 (D) OTHER INFORMATION: /note= "Pl;A4 ;T3"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-sit (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "V2;I6"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "L2;F6"
( ix ) FEATURE:
(A) NAME/KEY: Modi~ied-site ~W095/11998 ~17~ PC~IUS94/12268 (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "T4 jV4"
( ix ) FEATURE:
(A~ NAME/KEY: Modif ied-site (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "K2 jA6"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 20 (D) OTHER INFORMATION: /note= "T2 jD5 jE1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 22 (D) OTHER IN~ORMATION: /note= "V2 jK6"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 23 (D) OTHER INFORMATION: /note= "52 jT3;A3"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 24 (D) OTHER INFORMATION: /note= "S6 jGl;Nl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "N2 ;G6"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) 10CATION: 28 (D) OTHER INFORMATION: /note= "E3 ;Q5"
( ix) FEATURB:
(A) NAME/KEY: Modif ied-site ( B ) LO CATI ON: 3 0 (D) OTHER INFORMATION: /note= "A2 ;G6"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:25:
Leu Ala Lys Pro Val Leu Asp Thr Thr Thr Leu Asn Pro Thr Ile Ala Gly Lys Gly Thr Val Val Ser Ser Ala Glu Asn Glu Leu Ala W095/11998 2~7 ~ 9 PC~IIISg~llZZ68 (2) INFORMATION FOR SEQ ID NO:26:
(i) SEQUENCE C~iARA('`'l'T~'RT,STIC5 (A) LENGTH: 31 amino acids ~.
(B) TYPE: amino acid ( D ) TOPOLOGY: 1 i near (ii) M -T.RC IT.T. TYPE: peptide ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "A6;Vl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "E5;Tl;Kl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "A6;Pl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 ~ D ) OTHER INFORMATION: / note= " I 5; V2 "
( i x ) FEATURE:
(A) NAME/KEY: Modified--site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "L6;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modified--site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "V6;Il"
(ix) FEATURE:
(A) NAME/KEY: Modified-fiite (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "A6;Tl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "K5;C2"
(ix) FEATURE:

W09S/11998 2~ PcrluS94/12268 (A) NAME/REY: Modified-site ( B ) LOCATION: 2 O
(D) OTHER INFORMATION: /note= "S5;Tl;Al"
ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 22 (D) OTHER INFORMATION: /note= "V6;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 23 (D) OTHER INFORMATION: /note= "52;A4;Gl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 4 (D) OTHER INFORMATION: /note= "A3;S4"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "G5;N2"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 6 (D) OTHER INFORMATION: /note= "T2;S5"
( ix) FEATURE:
(A) NAMB/KEY: Modified-site ( B ) LOCAT I ON: 2 7 (D) OTHER INFORNATION: /note= "D3;E4"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 8 (D) OTHER INFORMATION: /note= "N5;G2"
( ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "E3 ;D2 ;Q2"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site ( B ) LOCATI ON: 3 0 (D) OTHER INFORMATION: /note= "L5;I2"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site WO 95ll 1998 2 ~ 7 5 5 7 PCTNS94/12268 ( B ) LOCATION: 31 (D) OTHER INFORMATION: /note= "A5;S2"
(xi) SEQIJENCE DESCRIPTION: SEQ ID NO:26:
eu Ala Glu Ala Ile Leu Asp Val Thr Thr Leu Asn Pro Thr Ile Ala ly Lys Gly Ser Val Val Ser Ala Gly Thr Asp Asn Glu Leu Ala (2) INFORMATION FOR SEQ ID NO:27:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGT~: 12 amino acids (B) TYPE: amino acid ( D) TOPOLOGY: linear (ii) l.ltlT.FCl,lT.F TYPE: peptide ix) FEATURE:
(A) NANE/KEY: Nodif ied-site (B) LOCATION: 3 (D) OT}~ER INFORMATION: /note= "S55;T2"
ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 6 (D) OT}IER INFORNATION: /note= "537;G20"
ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "T56;A1"
ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 12 (D) OTEIER INFORMATION: /note= "A33 jV24"
xi) SEQUENCE DESCRIPTION: SEQ ID NO:27:
sp Ser Ser Ala His Ser Thr Pro Ser Thr Pro Ala (2) INFORMATION FOR SEQ ID NO:28:
(i) SEQUENCE ~IiARArlrT~RT~sTIc5 ~Wo 95/11998 ~ PCr/US94/12268 (A) LENGTH: 14 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: l inear (ii) Mr TTCTTT.T TYPE: peptide ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "Vl9;A2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "Al9 ;V2"
( ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "Al9;T2"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORT~ATION: /note= "R3 ;Kl8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "Tl5;M4;A2"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:28:
Val Ala Glu Ala Glu Lys Gln Arg Ala Ala Glu Ala Thr Lys (2) INFORMATION FOR SEQ ID NO:29:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 65 amino zlcids (B) TYPE: amino acid ( D) TOPOLOGY: linear ( i i ) MrlT.T~ ~'TJT.T' TYPE: peptide - (ix) FEATURE:
(A) NAME/KEY: Modif ied-~ite ( B ) LOCATION: 2 WOg~/11998 ~ 9 ` PCTIUS94111268 (D) OTHER INFORMATION: /note= "R32 jQl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "N31jS2"
~ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 6 (D) OTHER INFORMATION: /note= "K3;E30"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-fiite (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "V16;I16;Fl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-slte (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "Q30;L2 jHl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 19 (D) OTHER INFORD5ATION: /note= "I29;V4"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (8) LOCATION: 21 (D) OTHER INFORMATION: /note= "I28;V5"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LO CAT ION: 2 9 (D) OTHER INFORMATION: /note= "H22;N10;~31"
(iX) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 4 0 (D) OTHER INFORMATION: /note= "K32 ;El"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 42 (D) OTHER INFORMATION: /note= "I27;Ll;V5"
( i x ) F EATURE:
( A) NAME/ KEY: Modif ied-site (B) LOCATION: 43 (D) OTHER INFORMATION: /note= "K30;V3"

~ WO 95/11998 1 7 ~ S 7 3 PCT/US94/12268 . . .

i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 44 A, (D) OTHER INFORMATION: /note= "I24;V9"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 50 (D) OTHER INFORMATION: /note= "I30;Ml;Tl;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 61 (D) OTHER INFORMATION: /note= "V32;Il"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 64 (D) OTHER INFORMATION: /note= "E31;Dl;Ql"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:29:
Cys Lys Lys Asn Met Lys Gly Lys Val Val Gln Pro Glu Asn Leu Glu Tyr Thr Ile Val Ile Thr Pro His Ser Gly Glu Glu His Ala Val Gl Asn Asp Thr Gly Lys His Gly Lys Glu Ile Lys Ile Thr Pro Gln Ser Ser Ile Thr Glu Ala Glu Leu Thr Gly Tyr Gly Thr Val Thr Met Glu Cys (2) INFORMATrON FOR SEQ ID NO:30:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 17 amino acids (B) TYPE: amino acid ( D) TOPOLOGY: linear (ii) M~lT.TrTTT.T` TYPE: peptide (ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 1 WO 95/1 1998 PC rluS9~112268 3 i`

(D) OTHER INFORMATION: /note= "I32 jTl"
(ix~ FEATURE:
(A) NAME/KEY: Modif ied-6ite .
(B) LOCATION: 4 (D) OTHER INFORMATION: /note= "N32 ;Vl"
( ix ) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "V31;Al;Gl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOQTION: 12 (D) OTHER INFORMATION: /note= "S32 ;Rl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "N32;Kl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "I31;V2"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOQTION: 17 (D) OTHER INFORMATION: /note= "E32;Dl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:30:
Ile Thr Val Asn Pro Ile Val Thr Glu Lys Asp Ser Pro Val Asn Ile Glu ( 2 ) INFORMATION FOR SEQ ID NO: 31:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 42 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOT.T~`CTTT.T~ TYPE: peptide W095/11998 7SS7g PCTIUS94112268 (ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) ~OCATION: 2 . (D) OTHER INFORMATION: /note= "TljVl;*"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "VljFl;*"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "Tl;Nl;Pl"
( ix) FEATURE-(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "R2;Gl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "G2;Dl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) I,OCATION: 7 (D) OTHER INFORMATION: /note= "T2 ;Gl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-~ite (B) ~OCATION: 8 (D) OTHER INFORMATION: /note= "Il;Nl;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 9 ~D) OTHER INFORMATION: /note= "Sl;Pl;El"
(ix~ FEATURE:
(A) NAME/KEY: Modified-site ( B ) ~OCATION: l 0 (D) O~HER INFORMATION: /note= "Dl;Rl;Nl"
(ix) FEATURE-(A) NAME/KEY: Modif ied--site ( B ) LOCATION: l l (D) OTHER INFORMATION: /note= "P2;El"
( ix) FEATURE:

WO 95111998 i t PCTI[~594/12268 ~
~17~9 (A) NAME/REY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Ql;Vl jDl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "Rl;Il;Kl"
) FEATURE-lX (A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "A2;El"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "Kl;Dl;Rl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "E2 ;Ll"
1X) FEATURE -(A) NAME/KEY: Modified- t (B) LOCATION: 17 81 e ( D ) OTHER INFORMATION: / note= "A2; Ll "
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 18 (D) OTHER INFOFMATION: /note= "L2 ;Al"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: l 9 (D) OTHER INFORMATION: /note= "Dl;N2"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 20 (D) OTHER INFORMATION: /note= "K2 ;Nl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 l (D) OTHER INFORMATION: /note= "YZ;Pl"
( 1X ) FEATURE :
(A) NAME/KEY: Modif ied-site Wo 9~111998 17~ 7 PCTIUS94112268 . ! "~ 1,, (B) LOCATION: 22 (D) OTHER INFORMATION: /note= "El;Al;Dl"
. ( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 3 (D) OTHER INFORMATION: /note= "Ll;Il;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 4 (D) OTHER INFORMATION: /note= "E2;Ql"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 2 6 (D) OTHER INFORMATION: /note= "Hl;Il;Sl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "Dl;Rl;Ml"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 29 ( D ) OTHER INFORMATION: / note= "Kl; Rl; Ql "
(ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 30 (D) OTHER INFORMATION: /note= "Tl;Hl;Al"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "Kl;El;Nl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 33 (D) OTHER INFORMATION: /note= "El;Kl;Dl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 4 (D) OTHER INFORMATION: /note= "Gl;Dl;Kl"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 36 -2175~7~ 8 ~

(D) OTHER INFORMATION: /note= "K2 jTl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site .
( B ) LOCAT ION: 3 7 (D) OTHER INFORMATION: /note= "T2 ;A1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( 3 ) LOCATION: 3 8 (D) OTHER INFORMATION: /note= "E2 ;R1"
( ix ) FEATUE'~E:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 39 (D~ OTHER INFORNATION: /note= "N2;L1"
( iX) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 4 0 (D) OTHER INFORMATION: /note= "E2;N1"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 41 (D) OTHER INFORMATION: /note= "Gl;Nl;L1"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite ( B ) LOCAT ION: 4 2 (D) OTHER INFORMATION: /note= "L2;Al"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:31:
Arg Thr Val Thr Arg Gly Thr Ile Ser Asp Pro Gln Arg Ala Lys Glu Ala Leu Asp Lys Tyr Glu Leu Glu Asn His Asp Leu Lys Thr Lys Asn Glu Gly Leu Lys Thr Glu Asn Glu Gly Leu (2) INFORMATION FOR SEQ ID NO:32:
(i) SEQUENCE CEaFACTERISTICS:
(A) LENGTH: 12 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: linear WO 95111998 ~ ~ PCTNS94/12268 (ii) MnT~Fl ~TT.T~ TYPE: pept$de ( ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOQTION: 2 (D) OTHER INFORNATION: /note= "E3;Il"
( ix) FEATURE:
(A) NANE/KEY: Modified-6ite (B) LOCATION: 4 (D) OTHER INFORNATION: /note= "A3 ;Pl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 5 (D) OTHER INFORNATION: /note= "Q3;Pl"
(ix) FEATURE:
(A) NANE/KEY: Modified-6ite ( B ) LOCATION: 7 (D) OTHER INFORMATION: /note= "P3 ;Sl"
(ix) FEATURE:
(A) NANE/KEY: Nodified-site (B) LOCATION: lO
(D) OTHER INFORMATION: /note= "E3 ;Rl"
(ix) FEATURE:
(A) NANE/KEY: Modified-site ( B ) LO QTI ON: l l (D) OTHER INFORMATION: /note= "P3;51"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 32:
Pro Glu Gly Ala Gln Gly Pro Arg Gly Glu Pro Gly (2) INFORMATION FOR SEQ ID NO:33:
( i ) SEQUENCE ~TARA-'l'T.'RT .~:TICS
(A) LENGTH: 14 amino acids (B) TYPE: amino ~cid ( D) TOPOLOGY: linear (ii) MnT.RCTTT.T~ TYPE: peptide (ix) FEATURE:
(A) NANE/KEY: Modif ied-site Wo 95/11998 ~ PCrlUS94112268 2~

(B) LOCATION: 2 (D) OTHER INFORMATION: /note= "El;Al;Nl"
( ix) FEATURE: ,~
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "E2 ;Dl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "N2;El"
( i x ) F EATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 9 tD) OTHER INFORMATION: /note= "Sl;K2"
( ix) FEATURE ~
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: l 0 (D) OTHER INFORMATION: /note= "Il;Tl;Vl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "Sl;P2"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "Y2 ;Fl"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCI~TION: 14 (D) OTHER INFORMATION: /note= "S2;Yl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 33:
Lys Glu Glu Ile Glu Tyr Leu Asn Ser Ile LeU Ser Tyr Ser (2) INFORMATION FOR SEQ ID NO:34:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 27 i~mino acids (B) TYPE: amino acid (D) TOPOLOGY: linear ~W095/11998 SS7,~ PCTIU594112268 (ii) M~TFcrTTF TYPE: peptide ~ (ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "D0.50;E0.50"
( ix) FEATTJRE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 2 ( D ) OTHER INFORMATION: /note= "L0 . 2 5; I0 . 25; V0 . 2 5; F0 . 2 5 "
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "E0.50;DO.50"
( ix ) FEATT~RE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 ( D ) OTHER INFORMATION: /note= "L0 . 25; I 0 . 2 5; V0 . 2 5; F0 . 2 5 "
(ix) FEATtlRE:
(A) NAME/KEY: Nodified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "K0.50;R0.50"
(ix) FEATTJRE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATTJRE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATTJRE:
(A) NAME/KEY: Modified-site (B) LOCATION: g (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
( ix) FEATTjRE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "K50;R50"
(ix) FEATT~RE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 Wo 95111998 ~¢~T~ 9 ~ ~ ~ PCrlUS94/12~68 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
( ix) FEATURE:
(A) NAME/REY: Modified-6ite .
(B) LOC~TION: 13 (D) OTHER INFORMATION: /note= "D50;E50"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:34:
Asp Leu Glu Leu Lys Gly Leu Leu Leu His Lys Leu Asp Gly Leu Gly Gly Glu His Trp Ser Tyr Gly Leu Arg Pro Gly (2) INFORMATION FOR SEQ ID NO:35:
(i) SEQUENCE t'TTARAcTRRT-cTIcs (A) LENGTH: 29 ~mino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MnT.F:ClTT.F TYPE: peptide ( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "K50;R50"
( ix) FEATURE:
(A) NAME/KEY: Modified-E;ite (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "K50;R50"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 3 (D) OTHER INFORMATION: /note= "K50;R50"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"

~ WO 95/11998 2 1 7 5 S 7 ~ ~ PCr/U594/12268 ( ix) FEATURE:
~A) NAMB/KEY: Modified-site tB) LOCATION: 5 (D) OTHER INFORMATION: /note= "F34;R33;R33"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "R50;R50"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "Q20;L20;I20;F20;V20"
(ix) FEATURE:
(A) NAME/R!EY: Modified-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "L25;I25;V25;F25"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:35:
Ly6 Lys Lys Leu Phe Leu Leu Thr LyE~ Leu Leu Thr Leu Pro Gln Ser W0 95/11998 ?,~ 9 PCr/llS94~12268 Leu Gly Gly Glu His Trp Ser Tyr Gly Leu Arg Pro Gly ( 2 ) INFORMATION FOR SEQ ID NO: 3 6: ~, ( i ) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 9 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "R5;Gl;Fl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "Y2;I2;Vl;Wl;Sl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "Ql;K3;Ll;Dl;Nl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "K2;E3;Pl;Gl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "Sl;I2;Vl;Al;Pl;Ll"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "T2;V2;I2;Gl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "El;K2;Dl;Vl;Ll;Hl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 9 ~ W095/11998 2~ 7SS7g PCr/1~594/12268 (D) OTHER INFORMATION: /note= "Ll;K3;R2;Al"
,~ (xi) SEQUENCE DESCRIPTION: SEQ ID NO:36:
Arg Arg Tyr Gln Lys Ser Thr Glu Leu (2) INFORMATION FOR SEQ ID NO:37:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: lO amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) M lT.FCrTTT~ TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l (D) OTHER INFORNATION: /note= "K8;Rl"
( ix) FEATURE:
(A) NAME/KEY: Nodified-site (8) LOCATION: 3 (D) OTHER INFORMATION: /note= "W8;Vl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "I8;Vl"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "I5;V3;Ql"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "L5;I2;M2"
( ix) FEATURE:
(A) NAME/KEY: Modi~ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "G8;Rl"
( ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 9 WO95/11998 PCr/US9~/12268 5~

(D) OTHER INFORMATION: /note= "N8;Ql"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ,~
(B) LOCATION: l0 (D) OTHER INFORMATION: /note= "K8 ;Rl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:37:
Lys Arg Trp Ile Ile Leu Gly Leu Asn Lys (2) INFORMATION FOR SEQ ID NO:38:
(i) SEQUENCE ~TTAR~ RT~cTIcs (A) LENGTEI: 58 amino acids (B) TYPE: amino acid ( D ) TOPOLOGY: l inear (ii) M lr.T~-'TTT.T~ TYPE: peptide ( ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOQTION: 2 (D) OTHER INFORMATION: /note= "T76;Xl6;N8"
(ix) FEATURE:
(A) NAME/REY: Nodified-site (B) LOCATION: 3 (D) OTHER INFORM~TION: /note= "L84;Vl6"
( ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "A58;T42"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "A67;N25;S8"
(ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "D50;E42;G8"
(ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 9 ~ Wo 95/11998 21 7~S 79 PCr/US94/12268 (D) OTHER INFORMATION: /note= "T67;V25;A8"
( ix ) FEATURE:
(A) NAME/KEY: Nodified-site (B) LOCATION: l0 (D) OTHER INFOR/5ATION: /note= "T92;A8"
- (ix) FEATURE:
(A) NANE/KEY: Nodified-slte (B) LOCATION: ll (D) OTHER INFORNATION: /note= "L92;V8"
( ix) FEATURE:
(A) NANE/KEY: Nodi~ied-site (B) LOCATION: 12 (D) OTHER INFORNATION: /note= "V34;E33;K33"
(ix) FEATURE:
(A) NANE/KEY: Nodif ied-site ( B ) LOCATION: l 3 (D) OTHER INFORNATION: /note= "V92;I8"
( i x ) FEATURE:
(A) NAME/REY: N~r1 i f i e~l-site ( B ) LOCATION: 14 (D) OTHER INFORMATION: /note= "K75;T25"
(ix) FEATURE:
(A) NANE/KEY: Nodif ied-site (B) LOCATION: l9 (D) OTHER INFORMATION: /note= "T75;V25"
(ix) FEATURE:
(A) NANE/KEY: Nodif ied-6ite (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "592;K8"
( i x ) FEATURE:
(A) NANE/REY: Modified-site ( B ) LOCATrON: 2 2 (D) OTHER INFORMATION: /note= "K92;R8"
( ix ) FEATURE:
(A) NANE/KEY: Nodified-site (B) LOCATION: 23 (D) OTHER INFORXATION: /note= "N50;E34;Hl6"
(ix) FEATURE:
(A) NAME/KEY: Nodified-site ( B ) LOCATION: 2 5 (D) OTHER INFORNATION: /note= "S42;A25;Ll7;P8;D8"

WO9~/11998 ~rl5$rl9 PCrlUSs4/l2268 (lx) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 26 (D) OTHE~ INFORMATION: /note= "K92 jN8"
(ix) FEATURE:
(A) NAME/KEY: Modified-~ite ( B ) LOCAT ION: 2 7 (D) OTHER INFORMATION: /note= "592 jN8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (8) LOCATION: 29 (D) OTHER INFORMATION: /note= "E92;K8"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT I ON: 3 O
(D) OTHER INFORMATION: /note= "V75;I25"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "T67;533"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 3 2 (D) OTHER INFORMATION: /note= "V92 ;A8"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 33 (D) OTHER INFORMATION: /note= "E50;A42;S8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "N75;D25"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 3 7 (D) OTHER INFORMATION: /note= "T73 ;527"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 9 (D) OTHER INFORMATION: /note= "T58;S42"
( ix) FEATURE:

WO 95/11998 PcrllJS94112268 (A) NAME/KEY: Modified-site ( B ) LOCATION: 4 O
(D) OTHER INFORMATION: /note= "T58;534;A8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 41 (D) OTHER INFORMATION: /note= "Q58;A34;58"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCAT ION: 4 2 (D) OTHER INFORMATION: /note= "A92;G8"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 43 (D) OTHER INFORNATION: /note= "T92;S8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 47 (D) OTHER INFORMATION: /note= "G58;A42"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 48 (D) OTHER INFORMATIoN: /note= "A58;K26;58;N8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATI ON: 5 O
(D) OTHER INFORMATION: /note= "D58;N34;Q8"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOQTION: 51 (D) OTHER INFORMATION: /note= "S84;A8;E8"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOQTION: 52 (D) OTHER INFORMATION: /note= "K42;G42;S8;N8"
(ix) FEATURE:
(A) NAME/KEY: Modiried-site (B) LOCATION: 53 (D) OTHER INFORMATION: /note= "T92;S8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site WO95111998 ~ Pcr~S94/12268 ( B ) LOCATION: 3 8 ( D ) OTHER INFoRMATIoN: / note= " D7 3; N2 7 "
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:38:
Gly Thr Leu Ala Ala Asp Gly Ly6 Thr Thr Leu Val Val Lys Glu Gl 5 l0 15 Thr Val Thr Leu Ser Lys Asn Ile Ser Lys Ser Gly Glu Val Thr Val Glu Leu Asn Asp Thr Asp Thr Thr Gln Ala Thr Lys Lys Thr Gly Ala Trp Asp Ser Lys Thr Ser Thr Leu Thr I le (2) INFORMATION FOR SEQ ID NO:39:
(i) SEQUENCE ~ RAr~lrFRT-~TICS:
(A) LENGTH: 71 amino 2Icids (B) TYPE: a~nino acid ( D ) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "N75;D25"
ix) FEATURE:
(A) NAME/REY: Modified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= 1~T73 ;S27"
(ix) FEATURE:
(A) NAME/REY: Modif ied-site (B) LOCATION: 4 (D) OTHER INFORMATION: /note= "D73;N23"
( iX) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "T58;S42"
(ix) FEATURE:
(A) NAME/KEY: Modified-site WO95/11998 ~79 PCI/IJS94/12268 (B) LOCATION: 6 tD) OTHER INFORNATION: /note= "T58;533;A9"
( ix) FEATURE:
(A) NAME/KEY: Nodified-site (B~ LOCATION: 7 (D) OTHER INFORNATION: /note= "Q58;A33;S9"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "A92;G8"
( i x ) FEATIJRE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 9 (D) OTHER INFORNATION: /note= "T92;S8"
( ix) FEATURE -(A) NAME/REY: Modified-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "G58;A42"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "A58;K25;S8;N9"
(ix) FEATURE-(A) NAME/KEY: Modif ied-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "D58;N33;Q9"
(ix) FEATURE:
(A) NAME/KEY: Modi~ied-~iite (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "583;A8;E9"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 18 (D) OTHER INFORMATION: /note= "K42;G42;S8;N8"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "T92;58"
(ix~ FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 24 Wo 95111998 ~ 9 PCrlUS94/12268 (D) OTHER INFORMATION: /note= "558;T42"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "V92 jA8"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "S92 ;N8"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "K75;Q17;R8"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "K75;T25"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 32 (D) OTHER INFORMATION: /note= "D42;N25;Q25;A8"
( i x ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 33 (D) OTHER INFORMATION: /note= "L92;I8"
( ix) FEATURE:
(A) NAME/KEY: Modified- t (B~ T.--~'ATT~ 36 51 e (D) OTHER INFORMATION: /note= "T92 ;L8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 7 (D) OTHER INFORMATION: /note= "K92;T8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 38 (D) OTHER INFORMATION: /note= "E58;Q33;N9"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 39 (D) OTHER INFORMATION: /note= "D58;N33;G9"

217~$7~

( ix) FEATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 4 3 (D) OTHER INFORMATION: /note= "V92 jS8"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 45 ( D ) OTHER INFORMATION: / note= "K5 8; Q3 3; N9 "
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 49 (D) OTHER INFORMATION: /note= "A75;N25"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "N58;K42"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 56 (D) OTHER INFORMATION: /note= "S42;T33;K17;N8"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 58 (D) OTHER INFORMATION: /note= "V92 ;A8"
( ix) FEATURE:
(A) NAME/KEY: Mn~ i f i ~--site (B) LOCATION: 61 (D) OTHER INFORMATION: /note= "K58;T42"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 62 (D) OTHER INFORMATION: /note= "T58;K42"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 64 (D) OTHER INFORMATION: /note= "D83 ;K17"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-iite (B) LOCATION: 66 (D) OTHER INFORMATION: /note= "L67;I33"
( ix ) F EATURE:

Wo95111998 ~ 55~ PCT/US9J/12268 ~

(A) NAME/KEY: Modified-site (B) LOQTION: 68 (D) OTHER INFORMATION: /note= "N75 jD1~ jA8"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 71 (D) OTHER INFORMATION: /note= "R25;K75"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:39:
sn Asp Thr Asp Thr Thr Gln Ala Thr LYB Lys Thr Gly Ala Trp Asp er Lys Thr Ser Thr Leu Ile Ser Val Asn Ser Lys Lys Thr Lys Asp Leu Val Phe Thr Lys Glu Asp Thr Ile Thr Val Gln Lys Tyr Asp Ser Ala Gly Thr Asn Leu Glu Gly Ser Ala Val Glu Ile Lys Thr Leu As Glu Leu Lys Asn Ala Leu Arg (2) INFORMATION FOR SEQ ID NO:40:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 78 amino ~Icids (B) TYPE: amino acid (D) TOPOLOGY: linear ( i i ) M~l}.RCTTT .R TYP E: pepti de ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "Al jIl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 46 (D) OTHER INFORMATION: /note= "Fl jTl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ~ WO95/11998 21 7~S79 PCT/US94/12268 (B) LOCATION: 5 (D) OTHER INFORMATION: /note= "Vl;Al"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 52 (D) OTHER INFORMATION: /note= "Sl;Al"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 56 (D) OTHER INFOR~ATION: /note= "Rl;El"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 70 (D) OTHER INFORMATION: /note= "Kl;El"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 72 (D) OTHER INFORMATION: /note= "Gl;El"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite ( B ) LOCATION: 7 3 (D) OTHER INFORMATION: /note= "Dl;Gl"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite (B) LOCATION: 74 (D) OTHER INPORMATION: /note= "Ll jSl"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:40:
ro A6n Ser Glu A6n Gly Ala Cys Tyr Tyr Gly Asp Phe Ile A6p Tyr l0 15 lu Glu Leu Arg Glu Gln Leu Ser Ser Val Ser Ser Phe Glu Arg Phe Glu Ile Phe Pro Lys Glu Ser Ser Trp Pro Asn His Thr Phe Asn Gly Val Thr Val ser Cys Ser His Arg Gly Lys 8er Ser Phe Tyr Arg Asn Leu Leu Trp Leu Thr Lys Lys Gly Asp Leu Tyr Pro Lys Leu 6~ 70 75 W095/11998 ~9 . PCr/US94/12268 ( 2 ) INFORMATION FOR SBQ ID NO: 4 l:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 32 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) M lT.RCrJT.F: TYPE: peptide ( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: l (D) OTHER INFORMATION: /note= "E6;K3;Tl"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 2 (D) OTHER INFORMATION: /note= "56;P3;Tl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 4 (D) OTHER INFORNATION: /note= "E7;Ll;Ql;Vl"
( i x ) FEATURE:
(A) NAME/KEY: Modified-3ite (B) LOCATION- 8 (D) OTHER INFORMATION: /note= "T5.5;I2.5;Sl;Pl"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: ll (D) OTHER INFORMATION: /note= "N9;Sl"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "N9 ;Kl"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "N9;Yl"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 14 ( D ) OTHER INFORMATION: / note= " -8; I l; Rl "
(ix) FEATURE:

09~/11998 ~$7~ PCr/US94/lZ268 (A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "T9 ;Q1"
( ix ) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 16 (D) OTHER INFORMATION: /note= "R8;K1;51"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "K9;R1"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite (B) LOCATION: 18 (D) OTHER INFORMATION: /note= "55.5 jG2.5;I1;R1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "I9 ;V1"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "I6;L3;M1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 23 (D) OTHER INFORNATION: /note= "P9;W1"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 2 5 (D) OTHER INFORMATION: /note= "R6.5;Q3.5"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION- 27 (D) OTHER INFORMATION: /note= "F6.5;W3.5"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "Y8;V1;H1"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site ~ 5~ PCr/l~S94/12268 (B) LOCATION: 29 ( D ) OTHER INFORMATION: / note= "A6 . 5; T3 . 5 "
(ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 30 (D) OTHER INFORMATION: /note= "T9;I1"
( ix) FEATURE:
(A) NANE/KEY: Modified-6ite ( B ) LOCATI ON: 3 2 (D) OTHER INFORMATION: /note= "D3 jK2.5;Q2.5 jEl;G1"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:41:
lu Ser Val Glu Ile Asn Cys Thr Arg Pro Asn Asn Asn Xaa Thr Ar ys Ser Ile His Ile Gly Pro Gly Arg Ala Phe Tyr Ala Thr Gly Asp (2) INFORMATION FOR SEQ ID NO:42:
(i) SEQUENCE ('T~ARA~'TT~RT~TICS:
(A) LENGTH: 45 amino acids (Il) TYPE: amino acid ( D ) TOPOLOGY: 1 inear (ii) lllOT.T~'TTT.T~' TYPE: peptide (ix) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 1 (D) OTHER INFORMATION: /note= "T60;K26;D7;A7"
(ix) FEATURE:
(A) NANE/KEY: Nodified-site (B) LOCATION: 3 (D) OTHER INFORMATION: /note= "I93;N7"
( ix ) FEATURE:
(A) NANE/KEY: Modified-site (B) LOCATION: 4 (D) OTHER INFORNATION: /note= "T67;K20;A13"
( ix) FEATURE:
(A) NANE/KEY: Modified-site ( B ) LOCATION: 5 ~ WO9S111998 21 7~$79 PCrlU594112268 (D) OTHER INFORMATION: /note= "Q46 jE40;T7;K7"
( i x ) F EATURE:
(A) NAME/KEY: Modif ied-s ite (B) LOCATION: 7 (D) OTHER INFORMATION: /note= "L86;T7;I7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 8 (D) OTHER INFORMATION: /note= "553;D33;N7;G7"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 9 (D) OTHER INFORMATION: /note= "G40;K26;V20;A7;S7"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 10 (D) OTHER INFORMATION: /note= "L87;I13"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 11 (D) OTHER INFORMATION: /note= "K85;N15"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 12 (D) OTHER INFORMATION: /note= "N60;G20;D20"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 13 (D) OTHER INFORMATION: /note= "S67;L33"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 14 (D) OTHER INFORMATION: /note= "E86;G14"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 15 (D) OTHER INFORMATION: /note= "E66;G27;K~"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "K93;S7"

WO 95111998 217 ~ 9 PCTIIJS94/12268 ~
. ` !
., . ~
1!~6 ( ix ) FBATURE:
(A) NAME/KEY: Modif ied-site ( B ) LOCATION: 18 (D) OTHER INFORMATION: /note= "E67 jT13;A13;K7"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 19 (D) OTHER INFORMATION: /note= "R67;E33"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 21 (D) OTHER INFORMATION: /note= "E40;A34;D13 jK13"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 22 (D) OTHER INFORMATION: /note= "K40 jA26 jE13;N7;D7;T7"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 23 (D) OTHER INFORMATION: /note= "A87;V13"
( ix) FEATURE:
(A) NAME/KEY: Modified-6ite ( B ) LO CATI ON: 2 5 ( D ) OTHER INFORMATION: / note= " K6 6; D13; E7; N7; Q7 "
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 2 6 (D) OTHER INFORMATION: /note= "C93 ;A7"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 27 (D) OTHER INFORMATION: /note= "S93;N7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 28 (D) OTHER INFORMATION: /note= "E60;D12;K7;Q7;A7;T7"
( ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 2 9 (D) OTHER INFORMATION: /note= "E40;K20;A13;D13;T7;S7"
( ix ) FEATURE:

WO 95/11998 1 7~$ 79 PCr/U594/12268 (A) NAME/KEY: Modif ied-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "T93 ;57"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 32 (D) OTHER INFORMATION: /note= "N60;K26;T20;D7;A7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 35 (D) OTHER INFORMATION: /note= "K72;E7;T7;Q7;57"
( ix ) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 3 6 (D) OTHER INFORMATION: /note= "540;D33;G13;E7;N7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 37 (D) OTHER INFORMATION: /note= "547;E20;G13;N13;K7"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 38 (D) OTHER INFORMATION: /note= "H93 ;Q7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 39 (D) OTHER INFORMATION: /note= "A80;T20"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCATION: 4 0 (D) OTHER INFORMATION: /note= "E46;D27;Q20;V7"
(ix) FEATURE:
(A) NAME/KEY: Modified-site ( B ) LOCAT I ON: 4 3 (D) OTHER INFORMATION: /note= "I40;K33;V13;A7;L7"
(ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 44 (D) OTHER INFORMATION: /note= "Q40;E26;A20;V7;D7"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-6ite WO95111998 2~55 PCrlUS94112268 (B) LOCATION: 45 (D) OTHER INFORMATION: /note= "G40;N34;D13;513"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:42:
hr Leu Ile Thr Gln Lys Leu Ser Gly Leu Lys Asn Ser Glu Glu Leu y6 Glu Lys Ile Glu Lys Ala Lys Lys Cys Ser Glu Glu Phe Thr Asn ys Leu Lys Ser Ser His Ala Glu Leu Gly Ile Gln Gly (2) INFORMATION FOR SEQ ID NO:43:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 61 amino acids (B) TYPE: amino acid (D) TOPOLOGY: linear (ii) MOLECULE TYPE: peptide ( ix) FEATURE:
(A) NAME/KEY: Nodif ied-site (B) LOCATION: 6 (D) OTHER INFORMATION: /note= "N83 ;T17"
( ix) FEATURE:
(A) NAME/REY: Modif ied-6ite (B) LOCATION: 17 (D) OTHER INFORMATION: /note= "A75;S25"
(ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 25 (D) OTHER INFORMATION: /note= "S83 ;A17"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 29 (D) OTHER INFORMATION: /note= "A92;S8"
( i x ) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 31 (D) OTHER INFORMATION: /note= "A50;T50"
( ix ) FEATURE:

~ WO95/11998 217SS7~ PCr/uss4ll2268 lS9 (A) NAME/KEY: Modif ied-site (B) LOCATION: 34 (D) OTHER INFORMATION: /note= "A75;T25"
( ix) FEATURE:
(A) NAME/KEY: Modif ied-site (B) LOCATION: 41 (D) OTHER INFORMATION: /note= "V67 jA33"
( ix) FEATURE:
(A) NAME/KEY: Modified-site (B) LOCATION: 43 (D) OTHER INFORMATION: /note= "Q83;E17"
(ix) FEATURE:
(A) NAMEIREY: Mn~lifi~cl-site (B) LOCATION- 50 (D) OTHER INFORMATION: /note= "A67 ;T33"
( ix ) FEATURE:
(A) NAME/KEY: Mr~rl i f i e-l-s ite (B) LOCATION: 56 (D) OTHER INFORMATION: /note= "583 jT17"
(xi) SEQUENCE DESCRIPTION: SEQ ID NO: 43:
al His Val Gly Ala Asn Gln Asp Glu Ala Ile Ala Val Asn Ile Tyr la Ala Asn Val Ala Asn Leu Phe Ser Gly Glu Gly Ala Gln Al A
20 25 30 a la Gln Ala Ala Pro Val Gln Glu Gly Val Gln Gln Glu Gly Ala Gln Gln Pro Ala Pro Ala Thr Ala Pro Ser Gln Gly Gly Val Asn

Claims

We Claim:

1. A library of peptides comprising a known antigenic site, epitope or ligand on a protein molecule, wherein (1) said library is optionally linked to a substantially invariant peptide domain or a core branched sequence;
(2) said library consists of an ensemble domain;
(3) the overall peptide length is about 8 to about 100 amino acids;
(4) the sequence of said ensemble domain is represented by a consensus formula;
(5) said ensemble is immunogenic, diagnostic for the epitope or is a therapeutic; and (6) the consensus formula provides that each sequence position in the ensemble contain either a single amino acid or multiple amino acids, and that when a position contains multiple amino acids, (a) the identity and ratio of those amino acids being determined by the relative prevalence of amino acids in a consensus of known variant sequences for that epitope or ligand or (b) the identity of those amino acids being determined by the amino acids present in a consensus of known variant sequences for that epitope or ligand and the ratio of amino acids is equimolar.

2. The library of Claim 1, wherein said antigenic site, epitope or ligand is from a virus, bacterium, parasite, tumor antigen, allergen or other protein antigen, from a diagnostic marker site of an infectious agent or disease, from a therapeutically valuable ligand or from a helper T cell epitope or a cytotoxic T lymphocyte (CTL) epitope.

3. The library of Claim 1, wherein said antigenic site, epitope or ligand is from an envelope, core, NS1, N53, NS4 or NS5 proteins of HCV; a gp120 V3 loop, gp41 envelope protein or the gp40 envelope protein of HIV; an envelope protein of HTLV
I/II; an HA protein or a mutant HA domain of influenza A
virus; an major outer membrane protein of Chlamydia trachomtis; a neuraminidase, SAPA or CRA site of T. cruzi;
an envelope B cell site or helper T cell site of type 2 Dengue virus; an M protein of streptococcus; a carbohydrate recognition site of selectin; a promiscuous helper T cell sites; a CTL epitope from HIV gag specific for HLA-B27; or an OspA, OspB, OspC or flagellin protein of Lyme disease.

4. The library of Claim 1 wherein said ensemble domain is represented by an SSAL library sequence provided in any one of Figs. 2-11, 13-34 (SEQ ID NO:1-43).

5. A peptide composition comprising the library of Claim 1.

6. The peptide composition of Claim 5 which is immunogenic.

7 A method of detecting antibodies associated with an infectious agent or a disease state which comprises contacting an effective amount of a peptide composition according to Claim 5 with a sample in an immunoassay procedure for a time sufficient to form a complex between said peptide composition and any antibody in said sample, and detecting the presence of the complex.

8. A method of detecting infection or a disease state which comprises contacting an effective amount of a peptide composition of, Claim 5 with a body fluid, tissue or tissue extract in an immunoassay procedure for a time sufficient to form a complex between said peptide composition and any antibody in said fluid, said tissue, or said tissue extract, and detecting the presence of said complex.

9. The method of Claim 7 wherein said immunoassay procedure is an ELISA or a PHA procedure.

10. The method of Claim 8 wherein said immunoassay procedure is an ELISA or a PHA procedure.

11. A kit for detection or diagnosis of an infectious agent or a disease state comprising a first container adapted to contain the peptide composition of Claim 5.

12. The kit of Claim 11 wherein said kit is an ELISA
or PHA test kit.

13. A method of treating a disease which comprises administering an effective amount of a composition of Claim 5 to a patient for a time sufficient to elicit an efficacious result.

14. A pharmaceutical composition comprising the library of Claim 1, or a pharmaceutically acceptable salt thereof, and a pharmaceutically acceptable carrier.

15. A method of vaccination which comprises administering an amount of the library of Claim 1 to a mammal effective to produce a protective immune response against an infectious agent which provided the basis of the library.

16. A vaccine composition comprising a library of Claim 1, or a salt thereof, and immunologically acceptable carrier.

17. A method of making an open diagnostic, vaccine or therapeutic composition for a mutatable infectious agent or for a protein site of known diversity which comprises (1) aligning a collection of primary amino acid sequences for a related family of antigens, epitopes, diagnostic markers or therapeutic sites from said mutatable infectious agent or said protein site;
(2) identifying invariant and variant amino acid positions in said alignment;
(3) calculating a consensus formula for the sequence of a structured synthetic antigen library (SSAL);
(4) preparing said SSAL from said consensus formula.

18. The method of Claim 17 wherein said consensus formula provides that each sequence position in the alignment contains either a single amino acid or multiple amino acids, and that when a position contains multiple amino acids, (a) the identity and ratio of said multiple amino acids being determined by the relative prevalence of amino acids in said alignment, (b) the identity of said multiple amino acids being determined by the amino acids present in said alignment and the ratio of said multiple amino acids is equimolar, or (c) the identity of said multiple amino acids being determined by the amino acids present in said alignment and the ratio of said multiple amino acids is weighted towards the predominant amino acid at said position.

19. The method of Claim 17 wherein said mutatable infectious agent is from a virus, bacterium or parasite.

20. The method of Claim 17 wherein said protein site is from a tumor antigen, allergen or other protein antigen, from a helper T cell epitope or a cytotoxic T lymphocyte (CTL) epitope.

21. The method of Claim 17, wherein said family of antigens, epitopes, diagnostic markers or therapeutic sites is from an envelope, core, NS1, NS3, NS4 or NS5 proteins of HCV;
a gp120 V3 loop, gp41 envelope protein or the gp40 envelope protein of HIV; an envelope protein of HTLV I/II; an HA
protein or a mutant HA domain of influenza A virus; an major outer membrane protein of Chlamydia trachomtis; a neuraminidase, SAPA or CRA site of T. cruzi; an envelope B
cell site or helper T cell site of type 2 Dengue virus; an M
protein of streptococcus; a carbohydrate recognition site of selectin; a promiscuous helper T cell sites; a CTL epitope from HIV gag specific for HLA-B27; or an OspA, OspB, OspC or flagellin protein of Lyme disease .