EP1250426A2 - Secreted and transmembrane polypeptides and nucleic acids encoding same - Google Patents

Secreted and transmembrane polypeptides and nucleic acids encoding same

Info

Publication number
EP1250426A2
EP1250426A2 EP00983846A EP00983846A EP1250426A2 EP 1250426 A2 EP1250426 A2 EP 1250426A2 EP 00983846 A EP00983846 A EP 00983846A EP 00983846 A EP00983846 A EP 00983846A EP 1250426 A2 EP1250426 A2 EP 1250426A2
Authority
EP
European Patent Office
Prior art keywords
seq
polypeptide
acid sequence
pro
amino acid
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Ceased
Application number
EP00983846A
Other languages
German (de)
English (en)
French (fr)
Inventor
Kevin P. Baker
Maureen Beresini
Laura Deforge
Luc Desnoyers
Ellen Filvaroff
Wei-Qiang Gao
Mary E. Gerritsen
Audrey Goddard
Paul J. Godowski
Austin L. Gurney
Steven Sherwood
Victoria Smith
Timothy A. Stewart
Daniel Tumas
Colin K. Watanabe
William L. Wood
Zemin Zhang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Genentech Inc
Original Assignee
Genentech Inc
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Priority claimed from PCT/US1999/028301 external-priority patent/WO2000032776A2/en
Priority claimed from PCT/US1999/028634 external-priority patent/WO2000036102A2/en
Priority claimed from PCT/US1999/028564 external-priority patent/WO2000055319A1/en
Priority claimed from PCT/US1999/028551 external-priority patent/WO2000053750A1/en
Priority claimed from PCT/US1999/028565 external-priority patent/WO2000037638A2/en
Priority claimed from PCT/US1999/030095 external-priority patent/WO2000037640A2/en
Priority claimed from PCT/US1999/030999 external-priority patent/WO2001005836A1/en
Priority claimed from PCT/US1999/030911 external-priority patent/WO2000075316A1/en
Priority claimed from PCT/US1999/031274 external-priority patent/WO2000053752A2/en
Priority claimed from PCT/US1999/031243 external-priority patent/WO2000053751A1/en
Priority claimed from PCT/US2000/000219 external-priority patent/WO2000053753A2/en
Priority claimed from PCT/US2000/000376 external-priority patent/WO2000053755A2/en
Priority claimed from PCT/US2000/000277 external-priority patent/WO2000053754A1/en
Priority claimed from PCT/US2000/003565 external-priority patent/WO2001053486A1/en
Priority claimed from PCT/US2000/004341 external-priority patent/WO2000053756A2/en
Priority claimed from PCT/US2000/004342 external-priority patent/WO2000078961A1/en
Priority claimed from PCT/US2000/004414 external-priority patent/WO2001004311A1/en
Priority claimed from PCT/US2000/004914 external-priority patent/WO2000075327A1/en
Priority claimed from PCT/US2000/005004 external-priority patent/WO2000053757A2/en
Priority claimed from PCT/US2000/005601 external-priority patent/WO2000056889A2/en
Priority claimed from PCT/US2000/005841 external-priority patent/WO2000053758A2/en
Priority claimed from PCT/US2000/006319 external-priority patent/WO2000053760A2/en
Priority claimed from PCT/US2000/006884 external-priority patent/WO2001005972A1/en
Priority claimed from PCT/US2000/007377 external-priority patent/WO2001019991A1/en
Priority claimed from PCT/US2000/007532 external-priority patent/WO2000070050A1/en
Priority claimed from PCT/US2000/008439 external-priority patent/WO2000073454A1/en
Priority claimed from PCT/US2000/013705 external-priority patent/WO2000073445A2/en
Priority claimed from PCT/US2000/014042 external-priority patent/WO2000077037A2/en
Priority claimed from PCT/US2000/014941 external-priority patent/WO2000073348A2/en
Priority claimed from PCT/US2000/015264 external-priority patent/WO2000073452A2/en
Priority claimed from PCT/US2000/020710 external-priority patent/WO2001009327A2/en
Priority claimed from PCT/US2000/022031 external-priority patent/WO2001040464A1/en
Priority claimed from PCT/US2000/023522 external-priority patent/WO2001016319A2/en
Priority claimed from PCT/US2000/023328 external-priority patent/WO2001016318A2/en
Priority claimed from PCT/US2000/030952 external-priority patent/WO2001049715A2/en
Priority claimed from PCT/US2000/030873 external-priority patent/WO2001040465A2/en
Priority to EP06000587A priority Critical patent/EP1690872A3/en
Priority to EP06000589A priority patent/EP1661997A1/en
Priority to EP06000581A priority patent/EP1666494A1/en
Priority to EP06000582A priority patent/EP1666495A1/en
Priority to EP06000585A priority patent/EP1661996A1/en
Priority to EP06000583A priority patent/EP1686134A3/en
Priority to EP06000588A priority patent/EP1690873A3/en
Priority to EP10005292A priority patent/EP2228446A1/en
Priority to EP06000584A priority patent/EP1669371A3/en
Priority to EP05025102A priority patent/EP1672070A3/en
Application filed by Genentech Inc filed Critical Genentech Inc
Priority to EP06000586A priority patent/EP1688497A1/en
Publication of EP1250426A2 publication Critical patent/EP1250426A2/en
Ceased legal-status Critical Current

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • C07K14/4701—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals not used
    • C07K14/4748—Tumour specific antigens; Tumour rejection antigen precursors [TRAP], e.g. MAGE
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P19/00—Drugs for skeletal disorders
    • A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P27/00—Drugs for disorders of the senses
    • A61P27/16—Otologicals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P43/00—Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P7/00—Drugs for disorders of the blood or the extracellular fluid
    • A61P7/02—Antithrombotic agents; Anticoagulants; Platelet aggregation inhibitors
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/46—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates
    • C07K14/47—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans from vertebrates from mammals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00—Medicinal preparations containing peptides

Definitions

  • the present invention relates generally to the identification and isolation of novel DNA and to the recombinant production of novel polypeptides.
  • Extracellular proteins play important roles in, among other things, the formation, differentiation and maintenance of multicellular organisms.
  • secreted polypeptides or signaling molecules normally pass through the cellular secretory pathway to reach their site of action in the extracellular environment.
  • Secreted proteins have various industrial applications, including as pharmaceuticals, diagnostics, biosensors and bioreactors.
  • Most protein drugs available at present, such as thrombolytic agents, interferons, interleukins, erythropoietins, colony stimulating factors, and various other cytokines, are secretory proteins.
  • Their receptors, which are membrane proteins, also have potential as therapeutic or diagnostic agents.
  • Efforts are being undertaken by both industry and proficient to identify new, native secreted proteins. Many efforts are focused on the screening of mammalian recombinant DNA libraries to identify the coding sequences for novel secreted proteins. Examples of screening methods and techniques are described in the literature [see, for example, Klein et al., Proc. Natl. Acad. Sci. 93:7108-7113 (1996); U.S. Patent No. 5,536,637)].
  • Membrane-bound proteins and receptors can play important roles in, among other things, the formation, differentiation and maintenance of multicellular organisms.
  • membrane-bound proteins and cell receptors include, but are not limited to, cytokine receptors, receptor kinases, receptor phosphatases, receptors involved in cell-cell interactions, and cellular adhesin molecules like selectins and integrins. For instance, transduction of signals that regulate cell growth and differentiation is regulated in part by phosphorylation of various cellular proteins. Protein tyrosine kinases, enzymes that catalyze that process, can also act as growth factor receptors. Examples include fibroblast growth factor receptor and nerve growth factor receptor.
  • Membrane-bound proteins and receptor molecules have various industrial applications, including as pharmaceutical and diagnostic agents.
  • Receptor immunoadhesins for instance, can be employed as therapeutic agents to block receptor-ligand interactions.
  • the membrane-bound proteins can also be employed for screening of potential peptide or small molecule inhibitors of the relevant receptor/ligand interaction.
  • Efforts are being undertaken by both industry and proficient to identify new, native receptor or membrane-bound proteins. Many efforts are focused on the screening of mammalian recombinant DNA libraries to identify the coding sequences for novel receptor or membrane-bound proteins.
  • the invention provides an isolated nucleic acid molecule comprising a nucleotide sequence that encodes a PRO polypeptide.
  • the isolated nucleic acid molecule comprises a nucleotide sequence having at least about 80% nucleic acid sequence identity, alternatively at least about 81 % nucleic acid sequence identity, alternatively at least about 82% nucleic acid sequence identity, alternatively at least about 83 % nucleic acid sequence identity, alternatively at least about 84% nucleic acid sequence identity, alternatively at least about 85% nucleic acid sequence identity, alternatively at least about 86% nucleic acid sequence identity, alternatively at least about 87 % nucleic acid sequence identity, alternatively at least about 88% nucleic acid sequence identity, alternatively at least about 89% nucleic acid sequence identity, alternatively at least about 90% nucleic acid sequence identity, alternatively at least about 91 % nucleic acid sequence identity, alternatively at least about 92% nucleic acid sequence identity, alternatively at least about 93 % nucleic acid sequence identity, alternatively at least about 94 % nucleic acid sequence identity, alternatively at least about 95% nucleic acid sequence identity,
  • the isolated nucleic acid molecule comprises a nucleotide sequence having at least about 80% nucleic acid sequence identity, alternatively at least about 81 % nucleic acid sequence identity, alternatively at least about 82 % nucleic acid sequence identity, alternatively at least about 83 % nucleic acid sequence identity, alternatively at least about 84% nucleic acid sequence identity, alternatively at least about 85% nucleic acid sequence identity , alternatively at least about 86 % nucleic acid sequence identity , alternatively at least about 87 % nucleic acid sequence identity, alternatively at least about 88% nucleic acid sequence identity, alternatively at least about 89% nucleic acid sequence identity, alternatively at least about 90% nucleic acid sequence identity, alternatively at least about 91 % nucleic acid sequence identity, alternatively at least about 92% nucleic acid sequence identity, alternatively at least about 93 % nucleic acid sequence identity, alternatively at least about 94 % nucleic acid sequence identity, alternatively at least about 95% nucleic acid
  • the invention concerns an isolated nucleic acid molecule comprising a nucleotide sequence having at least about 80% nucleic acid sequence identity, alternatively at least about 81 % nucleic acid sequence identity , alternatively at least about 82 % nucleic acid sequence identity , alternatively at least about 83 % nucleic acid sequence identity, alternatively at least about 84% nucleic acid sequence identity, alternatively at least about 85% nucleic acid sequence identity, alternatively at least about 86% nucleic acid sequence identity, alternatively at least about 87% nucleic acid sequence identity, alternatively at least about 88% nucleic acid sequence identity, alternatively at least about 89% nucleic acid sequence identity, alternatively at least about 90 % nucleic acid sequence identity, alternatively at least about 91 % nucleic acid sequence identity, alternatively at least about 92% nucleic acid sequence identity, alternatively at least about 93% nucleic acid sequence identity, alternatively at least about 94% nucleic acid sequence identity, alternatively at least about 95% nucleic acid sequence identity
  • Another aspect the invention provides an isolated nucleic acid molecule comprising a nucleotide sequence encoding a PRO polypeptide which is either transmembrane domain-deleted or transmembrane domain- inactivated, or is complementary to such encoding nucleotide sequence, wherein the transmembrane domain(s) of such polypeptide are disclosed herein. Therefore, soluble extracellular domains of the herein described PRO polypeptides are contemplated.
  • Another embodiment is directed to fragments of a PRO polypeptide coding sequence, or the complement thereof, that may find use as, for example, hybridization probes, for encoding fragments of a PRO polypeptide that may optionally encode a polypeptide comprising a binding site for an anti-PRO antibody or as antisense oligonucleotide probes.
  • nucleic acid fragments are usually at least about 10 nucleotides in length, alternatively at least about 15 nucleotides in length, alternatively at least about 20 nucleotides in length, alternatively at least about 30 nucleotides in length, alternatively at least about 40 nucleotides in length, alternatively at least about 50 nucleotides in length, alternatively at least about 60 nucleotides in length, alternatively at least about 70 nucleotides in length, alternatively at least about 80 nucleotides in length, alternatively at least about 90 nucleotides in length, alternatively at least about 100 nucleotides in length, alternatively at least about 110 nucleotides in length, alternatively at least about 120 nucleotides in length, alternatively at least about 130 nucleotides in length, alternatively at least about 140 nucleotides in length, alternatively at least about 150 nucleotides in length, alternatively at least about 160 nucleotides in length, alternatively at least about 170 nucleo
  • novel fragments of a PRO polypeptide-encoding nucleotide sequence may be determined in a routine manner by aligning the PRO polypeptide-encoding nucleotide sequence with other known nucleotide sequences using any of a number of well known sequence alignment programs and determining which PRO polypeptide-encoding nucleotide sequence fragment(s) are novel. All of such PRO polypeptide-encoding nucleotide sequences are contemplated herein. Also contemplated are the PRO polypeptide fragments encoded by these nucleotide molecule fragments, preferably those PRO polypeptide fragments that comprise a binding site for an anti-PRO antibody.
  • the invention provides isolated PRO polypeptide encoded by any of the isolated nucleic acid sequences hereinabove identified.
  • the invention concerns an isolated PRO polypeptide, comprising an amino acid sequence having at least about 80% amino acid sequence identity, alternatively at least about 81 % amino acid sequence identity, alternatively at least about 82% amino acid sequence identity, alternatively at least about 83 % amino acid sequence identity, alternatively at least about 84% amino acid sequence identity, alternatively at least about 85% amino acid sequence identity, alternatively at least about 86% amino acid sequence identity, alternatively at least about 87% amino acid sequence identity, alternatively at least about 88% amino acid sequence identity, alternatively at least about 89% amino acid sequence identity, alternatively at least about 90% amino acid sequence identity, alternatively at least about 91 % amino acid sequence identity, alternatively at least about 92% amino acid sequence identity, alternatively at least about 93% amino acid sequence identity, alternatively at least about 94% amino acid sequence identity, alternatively at least about 95 % amino acid sequence identity, alternatively at least about
  • the invention concerns an isolated PRO polypeptide comprising an amino acid sequence having at least about 80% amino acid sequence identity, alternatively at least about 81 % amino acid sequence identity, alternatively at least about 82% amino acid sequence identity, alternatively at least about 83 % amino acid sequence identity, alternatively at least about 84% amino acid sequence identity, alternatively at least about 85 % amino acid sequence identity, alternatively at least about 86% amino acid sequence identity, alternatively at least about 87% amino acid sequence identity, alternatively at least about 88% amino acid sequence identity, alternatively at least about 89% amino acid sequence identity, alternatively at least about 90% amino acid sequence identity, alternatively at least about 91 % amino acid sequence identity, alternatively at least about 92% amino acid sequence identity, alternatively at least about 93% amino acid sequence identity, alternatively at least about 94% amino acid sequence identity, alternatively at least about 95% amino acid sequence identity, alternatively at least about 96% amino acid sequence identity, alternatively at least about 97% amino acid sequence identity, alternatively at least about 98% amino acid sequence identity and alternative
  • the invention provides an isolated PRO polypeptide without the N-terminal signal sequence and/or the initiating methionine and is encoded by a nucleotide sequence that encodes such an amino acid sequence as hereinbefore described.
  • Processes for producing the same are also herein described, wherein those processes comprise culturing a host cell comprising a vector which comprises the appropriate encoding nucleic acid molecule under conditions suitable for expression of the PRO polypeptide and recovering the PRO polypeptide from the cell culture.
  • Another aspect the invention provides an isolated PRO polypeptide which is either transmembrane domain-deleted or transmembrane domain-inactivated.
  • Processes for producing the same are also herein described, wherein those processes comprise culturing a host cell comprising a vector which comprises the appropriate encoding nucleic acid molecule under conditions suitable for expression of the PRO polypeptide and recovering the PRO polypeptide from the cell culture.
  • the invention concerns agonists and antagonists of a native PRO polypeptide as defined herein.
  • the agonist or antagonist is an anti-PRO antibody or a small molecule.
  • the invention concerns a method of identifying agonists or antagonists to a PRO polypeptide which comprise contacting the PRO polypeptide with a candidate molecule and monitoring a biological activity mediated by said PRO polypeptide.
  • the PRO polypeptide is a native PRO polypeptide.
  • the invention concerns a composition of matter comprising a PRO polypeptide, or an agonist or antagonist of a PRO polypeptide as herein described, or an anti-PRO antibody, in combination with a carrier.
  • the carrier is a pharmaceutically acceptable carrier.
  • Another embodiment of the present invention is directed to the use of a PRO polypeptide, or an agonist or antagonist thereof as hereinbefore described, or an anti-PRO antibody, for the preparation of a medicament useful in the treatment of a condition which is responsive to the PRO polypeptide, an agonist or antagonist thereof or an anti-PRO antibody.
  • the invention provides vectors comprising DNA encoding any of the herein described polypeptides.
  • Host cell comprising any such vector are also provided.
  • the host cells may be CHO cells, E. coli, or yeast.
  • a process for producing any of the herein described polypeptides is further provided and comprises culturing host cells under conditions suitable for expression of the desired polypeptide and recovering the desired polypeptide from the cell culture.
  • the invention provides chimeric molecules comprising any of the herein described polypeptides fused to a heterologous polypeptide or amino acid sequence.
  • Example of such chimeric molecules comprise any of the herein described polypeptides fused to an epitope tag sequence or a Fc region of an immunoglobulin.
  • the invention provides an antibody which binds, preferably specifically, to any of the above or below described polypeptides.
  • the antibody is a monoclonal antibody, humanized antibody, antibody fragment or single-chain antibody.
  • the invention provides oligonucleotide probes which may be useful for isolating genomic and cDNA nucleotide sequences, measuring or detecting expression of an associated gene or as antisense probes, wherein those probes may be derived from any of the above or below described nucleotide sequences. Preferred probe lengths are described above.
  • the present invention is directed to methods of using the PRO polypeptides of the present invention for a variety of uses based upon the functional biological assay data presented in the Examples below.
  • Figure 1 shows a nucleotide sequence (SEQ ID NO:l) of a native sequence PR0177 cDNA, wherein SEQ ID NO:l is a clone designated herein as "DNA16438-1387".
  • Figure 2 shows the amino acid sequence (SEQ ID NO: 2) derived from the coding sequence of SEQ ID NO:l shown in Figure 1.
  • Figure 3 shows a nucleotide sequence (SEQ ID NO:3) of a native sequence PR03574 cDNA, wherein
  • SEQ ID NO:3 is a clone designated herein as "DNA 19360-2552".
  • Figure 4 shows the amino acid sequence (SEQ ID NO: 4) derived from the coding sequence of SEQ ID NO: 3 shown in Figure 3.
  • Figure 5 shows a nucleotide sequence (SEQ ID NO:5) of a native sequence PRO1280 cDNA, wherein SEQ ID NO:5 is a clone designated herein as "DNA33455-1548" .
  • Figure 6 shows the amino acid sequence (SEQ ID NO: 6) derived from the coding sequence of SEQ ID NO: 5 shown in Figure 5.
  • Figure 7 shows a nucleotide sequence (SEQ ID NO:7) of a native sequence PR04984 cDNA, wherein SEQ ID NO:7 is a clone designated herein as "DNA37155-2651".
  • Figure 8 shows the amino acid sequence (SEQ ID NO:8) derived from the coding sequence of SEQ ID NO:
  • Figure 9 shows a nucleotide sequence (SEQ ID NO:9) of a native sequence PR04988 cDNA, wherein SEQ ID NO:9 is a clone designated herein as "DNA38269-2654".
  • Figure 10 shows the amino acid sequence (SEQ ID NO: 10) derived from the coding sequence of SEQ ID NO: 9 shown in Figure 9.
  • Figure 11 shows a nucleotide sequence (SEQ ID NO: 11) of a native sequence PRO305 cDNA, wherein SEQ ID NO: 11 is a clone designated herein as "DNA40619-1220" .
  • Figure 12 shows the amino acid sequence (SEQ ID NO: 12) derived from the coding sequence of SEQ ID NO: 11 shown in Figure 11.
  • Figure 13 shows a nucleotide sequence (SEQ ID NO: 13)ofa native sequence PRO 1866 cDN A , wherein SEQ ID NO:13 is a clone designated herein as "DNA44174-2513".
  • Figure 14 shows the amino acid sequence (SEQ ID NO: 14) derived from the coding sequence of SEQ
  • Figure 15 shows a nucleotide sequence (SEQ ID NO: 15) of a native sequence PR04996 cDNA, wherein SEQ ID NO: 15 is a clone designated herein as "DNA44675-2662".
  • Figure 16 shows the amino acid sequence (SEQ ID NO: 16) derived from the coding sequence of SEQ ID NO: 15 shown in Figure 15.
  • Figure 17 shows a nucleotide sequence (SEQ ID NO: 17) of a native sequence PRO4406 cDNA, wherein SEQ ID NO: 17 is a clone designated herein as "DNA45408-2615".
  • Figure 18 shows the amino acid sequence (SEQ ID NO: 18) derived from the coding sequence of SEQ ID NO: 17 shown in Figure 17.
  • Figure 19 shows a nucleotide sequence (SEQ ID NO: 19) of a native sequence PROl 120 cDNA, wherein
  • SEQ ID NO: 19 is a clone designated herein as "DNA48606-1479".
  • Figure 20 shows the amino acid sequence (SEQ ID NO:20) derived from the coding sequence of SEQ ID NO: 19 shown in Figure 19.
  • Figure 21 shows a nucleotide sequence (SEQ ID NO:21) of a native sequence PRO4990 cDNA, wherein SEQ ID NO:21 is a clone designated herein as "DNA52753-2656" .
  • Figure 22 shows the amino acid sequence (SEQ ID NO: 22) derived from the coding sequence of SEQ ID NO:21 shown in Figure 21.
  • Figure 23 shows a nucleotide sequence (SEQ ID NO:23) of a native sequence PR0738 cDNA, wherein SEQ ID NO:23 is a clone designated herein as "DNA53915-1258".
  • Figure 24 shows the amino acid sequence (SEQ ID NO:24) derived from the coding sequence of SEQ
  • Figure 25 shows a nucleotide sequence (SEQ ID NO:25) of a native sequence PR03577 cDNA, wherein SEQ ID NO:25 is a clone designated herein as "DNA53991-2553".
  • Figure 26 shows the amino acid sequence (SEQ ID NO: 26) derived from the coding sequence of SEQ ID NO: 25 shown in Figure 25.
  • Figure 27 shows a nucleotide sequence (SEQ ID NO: 27) of a native sequence PRO 1879 cDNA, wherein SEQ ID NO:27 is a clone designated herein as "DNA54009-2517".
  • Figure 28 shows the amino acid sequence (SEQ ID NO:28) derived from the coding sequence of SEQ ID NO:27 shown in Figure 27.
  • Figure 29 shows a nucleotide sequence (SEQ ID NO : 29) of a native sequence PRO 1471 cDN A , wherein SEQ ID NO:29 is a clone designated herein as "DNA56055-1643" .
  • Figure 30 shows the amino acid sequence (SEQ ID NO: 30) derived from the coding sequence of SEQ ID NO:29 shown in Figure 29.
  • Figure 31 shows a nucleotide sequence (SEQ ID NO:31) of a native sequence PROl 114 cDNA, wherein SEQ ID NO:31 is a clone designated herein as "DNA57033-1403".
  • Figure 32 shows the amino acid sequence (SEQ ID NO: 32) derived from the coding sequence of SEQ ID NO: 31 shown in Figure 31.
  • Figure 33 shows a nucleotide sequence (SEQ ID NO:33) of a native sequence PRO1076 cDNA, wherein
  • SEQ ID NO:33 is a clone designated herein as "DNA57252-1453".
  • Figure 34 shows the amino acid sequence (SEQ ID NO:34) derived from the coding sequence of SEQ ID NO:33 shown in Figure 33.
  • Figure 35 shows a nucleotide sequence (SEQ ID NO:35) of a native sequence PR01483 cDNA, wherein SEQ ID NO:35 is a clone designated herein as "DNA58799-1652".
  • Figure 36 shows the amino acid sequence (SEQ ID NO: 36) derived from the coding sequence of SEQ ID NO:35 shown in Figure 35.
  • Figure 37 shows a nucleotide sequence (SEQ ID NO:37) of a native sequence PR04985 cDNA, wherein SEQ ID NO:37 is a clone designated herein as "DNA59770-2652".
  • Figure 38 shows the amino acid sequence (SEQ ID NO:38) derived from the coding sequence of SEQ
  • Figure 39 shows a nucleotide sequence (SEQ ID NO:39) of a native sequence PRO5000 cDNA, wherein SEQ ID NO:39 is a clone designated herein as "DNA59774-2665".
  • Figure 40 shows the amino acid sequence (SEQ ID NO:40) derived from the coding sequence of SEQ ID NO:39 shown in Figure 39.
  • Figure 41 shows a nucleotide sequence (SEQ ID NO:41) of a native sequence PRO 1881 cDNA, wherein SEQ ID NO:41 is a clone designated herein as "DNA60281-2518".
  • Figure 42 shows the amino acid sequence (SEQ ID NO: 42) derived from the coding sequence of SEQ ID NO:41 shown in Figure 41.
  • Figure 43 shows a nucleotide sequence (SEQ ID NO: 43) of a native sequence PR04314 cDNA, wherein
  • SEQ ID NO:43 is a clone designated herein as "DNA60736-2559".
  • Figure 44 shows the amino acid sequence (SEQ ID NO:44) derived from the coding sequence of SEQ ID NO: 43 shown in Figure 43.
  • Figure 45 shows a nucleotide sequence (SEQ ID NO:45) of a native sequence PR04987 cDNA, wherein SEQ ID NO:45 is a clone designated herein as "DNA61875-2653".
  • Figure 46 shows the amino acid sequence (SEQ ID NO: 46) derived from the coding sequence of SEQ ID NO: 45 shown in Figure 45.
  • Figure 47 shows a nucleotide sequence (SEQ ID NO: 47) of a native sequence PR04313 cDNA, wherein SEQ ID NO:47 is a clone designated herein as "DNA62312-2558".
  • Figure 48 shows the amino acid sequence (SEQ ID NO: 48) derived from the coding sequence of SEQ ID NO: 47 shown in Figure 47.
  • Figure 49 shows a nucleotide sequence (SEQ ID NO: 49) of a native sequence PR04799 cDNA, wherein SEQ ID NO:49 is a clone designated herein as "DNA62849-1604" .
  • Figure 50 shows the amino acid sequence (SEQ ID NO:50) derived from the coding sequence of SEQ ID NO:49 shown in Figure 49.
  • Figure 51 shows a nucleotide sequence (SEQ ID NO : 51 ) of a native sequence PR04995 cDN A , wherein SEQ ID NO:51 is a clone designated herein as "DNA66307-2661 ".
  • Figure 52 shows the amino acid sequence (SEQ ID NO:52) derived from the coding sequence of SEQ
  • Figure 53 shows a nucleotide sequence (SEQ ID NO:53) of a native sequence PR01341 cDNA, wherein SEQ ID NO:53 is a clone designated herein as "DNA66677-2535".
  • Figure 54 shows the amino acid sequence (SEQ ID NO:54) derived from the coding sequence of SEQ ID NO:53 shown in Figure 53.
  • Figure 55 shows a nucleotide sequence (SEQ ID NO:55) of a native sequence PR01777 cDNA, wherein SEQ ID NO:55 is a clone designated herein as "DNA71235-1706".
  • Figure 56 shows the amino acid sequence (SEQ ID NO:56) derived from the coding sequence of SEQ ID NO:55 shown in Figure 55.
  • Figure 57 shows a nucleotide sequence (SEQ ID NO:57) of a native sequence PRO3580 cDNA, wherein
  • SEQ ID NO:57 is a clone designated herein as "DNA71289-2547".
  • Figure 58 shows the amino acid sequence (SEQ ID NO:58) derived from the coding sequence of SEQ ID NO:57 shown in Figure 57.
  • Figure 59 shows a nucleotide sequence (SEQ ID NO:59) of a native sequence PR01779 cDNA, wherein SEQ ID N0:59 is a clone designated herein as "DNA73775-1707" .
  • Figure 60 shows the amino acid sequence (SEQ ID NO: 60) derived from the coding sequence of SEQ ID NO:59 shown in Figure 59.
  • Figure 61 shows a nucleotide sequence (SEQ ID NO: 61) of a native sequence PRO 1754 cDNA, wherein SEQ ID NO:61 is a clone designated herein as "DNA76385-1692".
  • Figure 62 shows the amino acid sequence (SEQ ID NO: 62) derived from the coding sequence of SEQ
  • Figure 63 shows a nucleotide sequence (SEQ ID NO: 63) of a native sequence PROl 906 cDNA, wherein SEQ ID NO:63 is a clone designated herein as "DNA76395-2527".
  • Figure 64 shows the amino acid sequence (SEQ ID NO: 64) derived from the coding sequence of SEQ ID NO: 63 shown in Figure 63.
  • Figure 65 shows a nucleotide sequence (SEQ ID NO:65) of a native sequence PRO1870 cDNA, wherein SEQ ID NO:65 is a clone designated herein as "DNA77622-2516".
  • Figure 66 shows the amino acid sequence (SEQ ID NO: 66) derived from the coding sequence of SEQ ID NO: 65 shown in Figure 65.
  • Figure 67 shows a nucleotide sequence (SEQ ID NO: 67) of a native sequence PR04329 cDNA, wherein SEQ ID NO:67 is a clone designated herein as "DNA77629-2573".
  • Figure 68 shows the amino acid sequence (SEQ ID NO:68) derived from the coding sequence of SEQ ID NO:67 shown in Figure 67.
  • Figure 69 shows a nucleotide sequence (SEQ ID NO: 69) of a native sequence PR04979 cDNA, wherein SEQ ID NO:69 is a clone designated herein as "DNA77645-2648".
  • Figure 70 shows the amino acid sequence (SEQ ID NO: 70) derived from the coding sequence of SEQ ID NO: 69 shown in Figure 69.
  • Figure 71 shows a nucleotide sequence (SEQ ID NO : 71 ) of a native sequence PRO 1885 cDN A , wherein
  • SEQ ID NO:71 is a clone designated herein as "DNA79302-2521".
  • Figure 72 shows the amino acid sequence (SEQ ID NO: 72) derived from the coding sequence of SEQ ID NO:71 shown in Figure 71.
  • Figure 73 shows a nucleotide sequence (SEQ ID NO:73) of a native sequence PR01882 cDNA, wherein SEQ ID NO:73 is a clone designated herein as "DNA79865-2519".
  • Figure 74 shows the amino acid sequence (SEQ ID NO: 74) derived from the coding sequence of SEQ ID NO:73 shown in Figure 73.
  • Figure 75 shows a nucleotide sequence (SEQ ID NO:75) of a native sequence PR04989 cDNA, wherein SEQ ID NO:75 is a clone designated herein as "DNA80135-2655".
  • Figure 76 shows the amino acid sequence (SEQ ID NO: 76) derived from the coding sequence of SEQ
  • Figure 77 shows a nucleotide sequence (SEQ ID NO : 77) of a native sequence PR04323 cDN A , wherein SEQ ID NO:77 is a clone designated herein as "DNA80794-2568".
  • Figure 78 shows the amino acid sequence (SEQ ID NO: 78) derived from the coding sequence of SEQ ID NO:77 shown in Figure 77.
  • Figure 79 shows a nucleotide sequence (SEQ ID NO: 79) of a native sequence PRO 1886 cDNA, wherein SEQ ID NO:79 is a clone designated herein as "DNA80796-2523".
  • Figure 80 shows the amino acid sequence (SEQ ID NO: 80) derived from the coding sequence of SEQ ID NO:79 shown in Figure 79.
  • Figure 81 shows a nucleotide sequence (SEQ ID NO:81) of a native sequence PR04395 cDNA, wherein
  • SEQ ID NO:81 is a clone designated herein as "DNA80840-2605 " .
  • Figure 82 shows the amino acid sequence (SEQ ID NO: 82) derived from the coding sequence of SEQ ID NO:81 shown in Figure 81.
  • Figure 83 shows a nucleotide sequence (SEQ ID NO:83) of a native sequence PR01782 cDNA, wherein SEQ ID NO:83 is a clone designated herein as "DNA80899-2501 " .
  • Figure 84 shows the amino acid sequence (SEQ ID NO: 84) derived from the coding sequence of SEQ ID NO:83 shown in Figure 83.
  • Figure 85 shows a nucleotide sequence (SEQ ID NO:85) of a native sequence PR04338 cDNA, wherein SEQ ID NO:85 is a clone designated herein as "DNA81228-2580".
  • Figure 86 shows the amino acid sequence (SEQ ID NO: 86) derived from the coding sequence of SEQ ID NO:85 shown in Figure 85.
  • Figure 87 shows a nucleotide sequence (SEQ ID NO:87) of a native sequence PR04341 cDNA, wherein SEQ ID NO:87 is a clone designated herein as "DNA81761-2583 " .
  • Figure 88 shows the amino acid sequence (SEQ ID NO: 88) derived from the coding sequence of SEQ ID NO:87 shown in Figure 87.
  • Figure 89 shows a nucleotide sequence (SEQ ID NO:89) of a native sequence PRO5990 cDNA, wherein SEQ ID NO:89 is a clone designated herein as "DNA96042-2682".
  • Figure 90 shows the amino acid sequence (SEQ ID NO: 90) derived from the coding sequence of SEQ
  • Figure 91 shows a nucleotide sequence (SEQ ID NO:91) of a native sequence PR03438 cDNA, wherein SEQ ID NO:91 is a clone designated herein as "DNA82364-2538".
  • Figure 92 shows the amino acid sequence (SEQ ID NO: 92) derived from the coding sequence of SEQ ID NO:91 shown in Figure 91.
  • Figure 93 shows a nucleotide sequence (SEQ ID NO:93) of a native sequence PR04321 cDNA, wherein SEQ ID NO:93 is a clone designated herein as "DNA82424-2566".
  • Figure 94 shows the amino acid sequence (SEQ ID NO: 94) derived from the coding sequence of SEQ ID NO: 93 shown in Figure 93.
  • Figure 95 shows a nucleotide sequence (SEQ ID NO:95) of a native sequence PRO4304 cDNA, wherein
  • SEQ ID NO:95 is a clone designated herein as "DNA82430-2557”.
  • Figure 96 shows the amino acid sequence (SEQ ID NO: 96) derived from the coding sequence of SEQ ID NO: 95 shown in Figure 95.
  • Figure 97 shows a nucleotide sequence (SEQ ID NO : 97) of a native sequence PRO 1801 cDN A , wherein SEQ ID NO:97 is a clone designated herein as "DNA83500-2506" .
  • Figure 98 shows the amino acid sequence (SEQ ID NO: 98) derived from the coding sequence of SEQ ID NO: 97 shown in Figure 97.
  • Figure 99 shows a nucleotide sequence (SEQ ID NO: 99) of a native sequence PRO4403 cDNA, wherein SEQ ID NO:99 is a clone designated herein as "DNA83509-2612".
  • Figure 100 shows the amino acid sequence (SEQ ID NO: 100) derived from the coding sequence of SEQ
  • Figure 101 shows a nucleotide sequence (SEQ ID NO: 101) of a native sequence PR04324 cDNA, wherein SEQ ID NO: 101 is a clone designated herein as "DNA83560-2569".
  • Figure 102 shows the amino acid sequence (SEQ ID NO: 102) derived from the coding sequence of SEQ ID NO: 101 shown in Figure 101.
  • Figure 103 shows a nucleotide sequence (SEQ ID NO: 103) of a native sequence PRO4303 cDNA, wherein SEQ ID NO: 103 is a clone designated herein as "DNA84139-2555".
  • Figure 104 shows the amino acid sequence (SEQ ID NO: 104) derived from the coding sequence of SEQ ID NO: 103 shown in Figure 103.
  • Figure 105 shows a nucleotide sequence (SEQ ID NO: 105) of a native sequence PRO4305 cDNA, wherein SEQ ID NO: 105 is a clone designated herein as "DNA84141-2556" .
  • Figure 106 shows the amino acid sequence (SEQ ID NO: 106) derived from the coding sequence of SEQ ID NO: 105 shown in Figure 105.
  • Figure 107 shows a nucleotide sequence (SEQ ID NO: 107) of a native sequence PRO4404 cDNA, wherein SEQ ID NO: 107 is a clone designated herein as "DNA84142-2613".
  • Figure 108 shows the amino acid sequence (SEQ ID NO: 108) derived from the coding sequence of SEQ ID NO: 107 shown in Figure 107.
  • Figure 109 shows a nucleotide sequence (SEQ ID NO: 109) of a native sequence PRO 1884 cDNA, wherein SEQ ID NO: 109 is a clone designated herein as "DNA84318-2520".
  • Figure 110 shows the amino acid sequence (SEQ ID NO: 110) derived from the coding sequence of SEQ ID NO: 109 shown in Figure 109.
  • Figure 111 shows a nucleotide sequence (SEQ ID NO: 111) of a native sequence PR04349 cDNA, wherein SEQ ID NO: 111 is a clone designated herein as "DNA84909-2590" .
  • Figure 112 shows the amino acid sequence (SEQ ID NO: 112) derived from the coding sequence of SEQ ID NO: 111 shown in Figure 111.
  • Figure 113 shows a nucleotide sequence (SEQ ID NO: 113) of a native sequence PRO4401 cDNA, wherein SEQ ID NO: 113 is a clone designated herein as "DNA84912-2610".
  • Figure 114 shows the amino acid sequence (SEQ ID NO : 114) derived from the coding sequence of SEQ
  • Figure 115 shows a nucleotide sequence (SEQ ID NO: 115) of a native sequence PR01867 cDNA, wherein SEQ ID NO: 115 is a clone designated herein as "DNA84925-2514".
  • Figure 116 shows the amino acid sequence (SEQ ID NO: 116) derived from the coding sequence of SEQ ID NO: 115 shown in Figure 115.
  • Figure 117 shows a nucleotide sequence (SEQ ID NO: 117) of a native sequence PR04319 cDNA, wherein SEQ ID NO: l 17 is a clone designated herein as "DNA84928-2564".
  • Figure 118 shows the amino acid sequence (SEQ ID NO: 118) derived from the coding sequence of SEQ ID NO: 117 shown in Figure 117.
  • Figure 119 shows a nucleotide sequence (SEQ ID NO: 119) of a native sequence PR04991 cDNA, wherein SEQ ID NO: 119 is a clone designated herein as "DNA84932-2657”.
  • Figure 120 shows the amino acid sequence (SEQ ID NO: 120) derived from the coding sequence of SEQ ID NO: 119 shown in Figure 119.
  • Figure 121 shows a nucleotide sequence (SEQ ID NO: 121) of a native sequence PR04398 cDNA, wherein SEQ ID NO: 121 is a clone designated herein as "DNA86592-2607" .
  • Figure 122 shows the amino acid sequence (SEQ ID NO: 122) derived from the coding sequence of SEQ ID NO: 121 shown in Figure 121.
  • Figure 123 shows a nucleotide sequence (SEQ ID NO: 123) of a native sequence PR04346 cDNA, wherein SEQ ID NO: 123 is a clone designated herein as "DNA86594-2587".
  • Figure 124 shows the amino acid sequence (SEQ ID NO: 124) derived from the coding sequence of SEQ ID NO: 123 shown in Figure 123.
  • Figure 125 shows a nucleotide sequence (SEQ ID NO: 125) of a native sequence PRO4350 cDNA, wherein SEQ ID NO: 125 is a clone designated herein as "DNA86647-2591 " .
  • Figure 126 shows the amino acid sequence (SEQ ID NO: 126) derived from the coding sequence of SEQ ID NO: 125 shown in Figure 125.
  • Figure 127 shows a nucleotide sequence (SEQ ID NO: 127) of a native sequence PR04318 cDNA, wherein SEQ ID NO:127 is a clone designated herein as "DNA87185-2563".
  • Figure 128 shows the amino acid sequence (SEQ ID NO : 128) derived from the coding sequence of SEQ
  • Figure 129 shows a nucleotide sequence (SEQ ID NO: 129) of a native sequence PRO4340 cDNA, wherein SEQ ID NO: 129 is a clone designated herein as "DNA87656-2582".
  • Figure 130 shows the amino acid sequence (SEQ ID NO: 130) derived from the coding sequence of SEQ ID NO: 129 shown in Figure 129.
  • Figure 131 shows a nucleotide sequence (SEQ ID NO: 131) of a native sequence PRO4400 cDNA, wherein SEQ ID NO:131 is a clone designated herein as "DNA87974-2609".
  • Figure 132 shows the amino acid sequence (SEQ ID NO: 132) derived from the coding sequence of SEQ ID NO: 131 shown in Figure 131.
  • Figure 133 shows a nucleotide sequence (SEQ ID NO: 133) of a native sequence PRO4320 cDNA, wherein SEQ ID NO: 133 is a clone designated herein as "DNA88001-2565".
  • Figure 134 shows the amino acid sequence (SEQ ID NO: 134) derived from the coding sequence of SEQ ID NO: 133 shown in Figure 133.
  • Figure 135 shows a nucleotide sequence (SEQ ID NO: 135) of a native sequence PRO4409 cDNA, wherein SEQ ID NO: 135 is a clone designated herein as "DNA88004-2575" .
  • Figure 136 shows the amino acid sequence (SEQ ID NO : 136) derived from the coding sequence of SEQ ID NO: 135 shown in Figure 135.
  • Figure 137 shows a nucleotide sequence (SEQ ID NO: 137) of a native sequence PR04399 cDNA, wherein SEQ ID NO: 137 is a clone designated herein as "DNA89220-2608".
  • Figure 138 shows the amino acid sequence (SEQ ID NO: 138) derived from the coding sequence of SEQ
  • Figure 139 shows a nucleotide sequence (SEQ ID NO: 139) of a native sequence PR04418 cDNA, wherein SEQ ID NO: 139 is a clone designated herein as "DNA89947-2618".
  • Figure 140 shows the amino acid sequence (SEQ ID NO: 140) derived from the coding sequence of SEQ ID NO: 139 shown in Figure 139.
  • Figure 141 shows a nucleotide sequence (SEQ ID NO: 141) of a native sequence PRO4330 cDNA, wherein SEQ ID NO: 141 is a clone designated herein as "DNA90842-2574".
  • Figure 142 shows the amino acid sequence (SEQ ID NO: 142) derived from the coding sequence of SEQ ID NO: 141 shown in Figure 141.
  • Figure 143 shows a nucleotide sequence (SEQ ID NO: 143) of a native sequence PR04339 cDNA, wherein SEQ ID NO: 143 is a clone designated herein as "DNA91775-2581 " .
  • Figure 144 shows the amino acid sequence (SEQ ID NO: 144) derived from the coding sequence of SEQ ID NO: 143 shown in Figure 143.
  • Figure 145 shows a nucleotide sequence (SEQ ID NO: 145) of a native sequence PR04326 cDNA, wherein SEQ ID NO: 145 is a clone designated herein as "DNA91779-2571 ".
  • Figure 146 shows the amino acid sequence (SEQ ID NO: 146) derived from the coding sequence of SEQ ID NO: 145 shown in Figure 145.
  • Figure 147 shows a nucleotide sequence (SEQ ID NO: 147) of a native sequence PRO6014 cDNA, wherein SEQ ID NO: 147 is a clone designated herein as "DNA92217-2697".
  • Figure 148 shows the amino acid sequence (SEQ ID NO: 148) derived from the coding sequence of SEQ ID NO: 147 shown in Figure 147.
  • Figure 149 shows a nucleotide sequence (SEQ ID NO: 149) of a native sequence PR03446 cDNA, wherein SEQ ID NO: 149 is a clone designated herein as "DNA92219-2541 " .
  • Figure 150 shows the amino acid sequence (SEQ ID NO: 150) derived from the coding sequence of SEQ ID NO: 149 shown in Figure 149.
  • Figure 151 shows a nucleotide sequence (SEQ ID NO:151) of a native sequence PR04322 cDNA, wherein SEQ ID NO: 151 is a clone designated herein as "DNA92223-2567" .
  • Figure 152 shows the amino acid sequence (SEQ ID NO: 152) derived from the coding sequence of SEQ
  • Figure 153 shows a nucleotide sequence (SEQ ID NO: 153) of a native sequence PR04381 cDNA, wherein SEQ ID NO: 153 is a clone designated herein as "DNA92225-2603" .
  • Figure 154 shows the amino acid sequence (SEQ ID NO : 154) derived from the coding sequence of SEQ ID NO: 153 shown in Figure 153.
  • Figure 155 shows a nucleotide sequence (SEQ ID NO: 155) of a native sequence PR04348 cDNA, wherein SEQ ID NO: 155 is a clone designated herein as "DNA92232-2589".
  • Figure 156 shows the amino acid sequence (SEQ ID NO: 156) derived from the coding sequence of SEQ ID NO: 155 shown in Figure 155.
  • Figure 157 shows a nucleotide sequence (SEQ ID NO: 157) of a native sequence PR04371 cDNA, wherein SEQ ID NO: 157 is a clone designated herein as "DNA92233-2599".
  • Figure 158 shows the amino acid sequence (SEQ ID NO: 158) derived from the coding sequence of SEQ ID NO: 157 shown in Figure 157.
  • Figure 159 shows a nucleotide sequence (SEQ ID NO: 159) of a native sequence PR03742 cDNA, wherein SEQ ID NO: 159 is a clone designated herein as "DNA92243-2549" .
  • Figure 160 shows the amino acid sequence (SEQ ID NO: 160) derived from the coding sequence of SEQ ID NO: 159 shown in Figure 159.
  • Figure 161 shows a nucleotide sequence (SEQ ID NO: 161) of a native sequence PR05773 cDNA, wherein SEQ ID NO: 161 is a clone designated herein as "DNA92253-2671".
  • Figure 162 shows the amino acid sequence (SEQ ID NO: 162) derived from the coding sequence of SEQ ID NO: 161 shown in Figure 161.
  • Figure 163 shows a nucleotide sequence (SEQ ID NO: 163) of a native sequence PR05774 cDNA, wherein SEQ ID NO: 163 is a clone designated herein as "DNA92254-2672".
  • Figure 164 shows the amino acid sequence (SEQ ID NO: 164) derived from the coding sequence of SEQ ID NO: 163 shown in Figure 163.
  • Figure 165 shows a nucleotide sequence (SEQ ID NO: 165) of a native sequence PR04343 cDNA, wherein SEQ ID NO: 165 is a clone designated herein as "DNA92255-2584".
  • Figure 166 shows the amino acid sequence (SEQ ID NO : 166) derived from the coding sequence of SEQ
  • Figure 167 shows a nucleotide sequence (SEQ ID NO: 167) of a native sequence PR04325 cDNA, wherein SEQ ID NO: 167 is a clone designated herein as "DNA92269-2570".
  • Figure 168 shows the amino acid sequence (SEQ ID NO: 168) derived from the coding sequence of SEQ ID NO : 167 shown in Figure 167.
  • Figure 169 shows a nucleotide sequence (SEQ ID NO: 169) of a native sequence PR04347 cDNA, wherein SEQ ID NO: 169 is a clone designated herein as "DNA92288-2588".
  • Figure 170 shows the amino acid sequence (SEQ ID NO: 170) derived from the coding sequence of SEQ ID NO: 169 shown in Figure 169.
  • Figure 171 shows a nucleotide sequence (SEQ ID NO: 171) of a native sequence PR03743 cDNA, wherein SEQ ID NO:171 is a clone designated herein as "DNA92290-2550".
  • Figure 172 shows the amino acid sequence (SEQ ID NO: 172) derived from the coding sequence of SEQ ID NO: 171 shown in Figure 171.
  • Figure 173 shows a nucleotide sequence (SEQ ID NO: 173) of a native sequence PR04426 cDNA, wherein SEQ ID NO: 173 is a clone designated herein as "DNA93012-2622".
  • Figure 174 shows the amino acid sequence (SEQ ID NO : 174) derived from the coding sequence of SEQ ID NO: 173 shown in Figure 173.
  • Figure 175 shows a nucleotide sequence (SEQ ID NO: 175) of a native sequence PRO4500 cDNA, wherein SEQ ID NO: 175 is a clone designated herein as "DNA93020-2642".
  • Figure 176 shows the amino acid sequence (SEQ ID NO : 176) derived from the coding sequence of SEQ
  • Figure 177 shows a nucleotide sequence (SEQ ID NO: 177) of a native sequence PR04389 cDNA, wherein SEQ ID NO: 177 is a clone designated herein as "DNA94830-2604".
  • Figure 178 shows the amino acid sequence (SEQ ID NO: 178) derived from the coding sequence of SEQ ID NO : 177 shown in Figure 177.
  • Figure 179 shows a nucleotide sequence (SEQ ID NO: 179) of a native sequence PR04337 cDNA, wherein SEQ ID NO: 179 is a clone designated herein as "DNA94833-2579".
  • Figure 180 shows the amino acid sequence (SEQ ID NO : 180) derived from the coding sequence of SEQ ID NO: 179 shown in Figure 179.
  • Figure 181 shows a nucleotide sequence (SEQ ID NO: 181) of a native sequence PR04992 cDNA, wherein SEQ ID NO: 181 is a clone designated herein as "DNA94838-2658" .
  • Figure 182 shows the amino acid sequence (SEQ ID NO: 182) derived from the coding sequence of SEQ ID NO: 181 shown in Figure 181.
  • Figure 183 shows a nucleotide sequence (SEQ ID NO: 183) of a native sequence PR05996 cDNA, wherein SEQ ID NO: 183 is a clone designated herein as "DNA94844-2686".
  • Figure 184 shows the amino acid sequence (SEQ ID NO: 184) derived from the coding sequence of SEQ ID NO: 183 shown in Figure 183.
  • Figure 185 shows a nucleotide sequence (SEQ ID NO: 185) of a native sequence PR04345 cDNA, wherein SEQ ID NO: 185 is a clone designated herein as "DNA94854-2586" .
  • Figure 186 shows the amino acid sequence (SEQ ID NO : 186) derived from the coding sequence of SEQ ID NO: 185 shown in Figure 185.
  • Figure 187 shows a nucleotide sequence (SEQ ID NO: 187) of a native sequence PR04978 cDNA, wherein SEQ ID NO: 187 is a clone designated herein as "DNA95930" .
  • Figure 188 shows the amino acid sequence (SEQ ID NO: 188) derived from the coding sequence of SEQ ID NO: 187 shown in Figure 187.
  • Figure 189 shows a nucleotide sequence (SEQ ID NO: 189) of a native sequence PRO5780 cDNA, wherein SEQ ID NO: 189 is a clone designated herein as "DNA96868-2677".
  • Figure 190 shows the amino acid sequence (SEQ ID NO : 190) derived from the coding sequence of SEQ
  • Figure 191 shows a nucleotide sequence (SEQ ID NO: 191) of a native sequence PR05992 cDNA, wherein SEQ ID NO: 191 is a clone designated herein as "DNA96871-2683".
  • Figure 192 shows the amino acid sequence (SEQ ID NO: 192) derived from the coding sequence of SEQ ID NO: 191 shown in Figure 191.
  • Figure 193 shows a nucleotide sequence (SEQ ID NO: 193) of a native sequence PR04428 cDNA, wherein SEQ ID NO: 193 is a clone designated herein as "DNA96880-2624".
  • Figure 194 shows the amino acid sequence (SEQ ID NO : 194) derived from the coding sequence of SEQ ID NO: 193 shown in Figure 193.
  • Figure 195 shows a nucleotide sequence (SEQ ID NO: 195) of a native sequence PR04994 cDNA, wherein SEQ ID NO: 195 is a clone designated herein as "DNA96986-2660" .
  • Figure 196 shows the amino acid sequence (SEQ ID NO : 196) derived from the coding sequence of SEQ ID NO: 195 shown in Figure 195.
  • Figure 197 shows a nucleotide sequence (SEQ ID NO: 197) of a native sequence PR05995 cDNA, wherein SEQ ID NO: 197 is a clone designated herein as "DNA96988-2685 " .
  • Figure 198 shows the amino acid sequence (SEQ ID NO : 198) derived from the coding sequence of SEQ ID NO: 197 shown in Figure 197.
  • Figure 199 shows a nucleotide sequence (SEQ ID NO: 199) of a native sequence PRO6094 cDNA, wherein SEQ ID NO: 199 is a clone designated herein as "DNA96995-2709".
  • Figure 200 shows the amino acid sequence (SEQ ID NO: 200) derived from the coding sequence of SEQ ID NO: 199 shown in Figure 199.
  • Figure 201 shows a nucleotide sequence (SEQ ID NO:201) of a native sequence PR04317 cDNA, wherein SEQ ID NO:201 is a clone designated herein as "DNA97004-2562".
  • Figure 202 shows the amino acid sequence (SEQ ID NO: 202) derived from the coding sequence of SEQ ID NO: 201 shown in Figure 201.
  • Figure 203 shows a nucleotide sequence (SEQ ID NO:203) of a native sequence PR05997 cDNA, wherein SEQ ID NO:203 is a clone designated herein as "DNA97005-2687".
  • Figure 204 shows the amino acid sequence (SEQ ID NO:204) derived from the coding sequence of SEQ
  • Figure 205 shows a nucleotide sequence (SEQ ID NO:205) of a native sequence PRO5005 cDNA, wherein SEQ ID NO:205 is a clone designated herein as "DNA97009-2668".
  • Figure 206 shows the amino acid sequence (SEQ ID NO: 206) derived from the coding sequence of SEQ ID NO:205 shown in Figure 205.
  • Figure 207 shows a nucleotide sequence (SEQ ID NO:207) of a native sequence PRO5004 cDNA, wherein SEQ ID NO:207 is a clone designated herein as "DNA97013-2667".
  • Figure 208 shows the amino acid sequence (SEQ ID NO: 208) derived from the coding sequence of SEQ ID NO: 207 shown in Figure 207.
  • Figure 209 shows a nucleotide sequence (SEQ ID NO: 209) of a native sequence PRO6001 cDNA, wherein SEQ ID NO:209 is a clone designated herein as "DNA98380-2690".
  • Figure 210 shows the amino acid sequence (SEQ ID NO : 210) derived from the coding sequence of SEQ ID NO:209 shown in Figure 209.
  • Figure 211 shows a nucleotide sequence (SEQ ID NO:211) of a native sequence PRO6013 cDNA, wherein SEQ ID NO:211 is a clone designated herein as "DNA98561-2696".
  • Figure 212 shows the amino acid sequence (SEQ ID NO:212) derived from the coding sequence of SEQ ID NO:211 shown in Figure 211.
  • Figure 213 shows a nucleotide sequence (SEQ ID NO:213) of a native sequence PRO4502 cDNA, wherein SEQ ID NO:213 is a clone designated herein as "DNA98575-2644".
  • Figure 214 shows the amino acid sequence (SEQ ID NO : 214) derived from the coding sequence of SEQ
  • Figure 215 shows a nucleotide sequence (SEQ ID NO:215) of a native sequence PRO6007 cDNA, wherein SEQ ID NO:215 is a clone designated herein as "DNA98593-2694".
  • Figure 216 shows the amino acid sequence (SEQ ID NO:216) derived from the coding sequence of SEQ ID NO:215 shown in Figure 215.
  • Figure 217 shows a nucleotide sequence (SEQ ID NO:217) of a native sequence PRO6028 cDNA, wherein SEQ ID NO:217 is a clone designated herein as "DNA98600-2703".
  • Figure 218 shows the amino acid sequence (SEQ ID NO:218) derived from the coding sequence of SEQ ID NO:217 shown in Figure 217.
  • Figure 219 shows a nucleotide sequence (SEQ ID NO:219) of a native sequence PRO 100 cDNA, wherein SEQ ID NO:219 is a clone designated herein as "DNA99333".
  • Figure 220 shows the amino acid sequence (SEQ ID NO: 220) derived from the coding sequence of SEQ ID NO:219 shown in Figure 219.
  • Figure 221 shows a nucleotide sequence (SEQ ID NO: 221) of a native sequence PR04327 cDNA, wherein SEQ ID NO:221 is a clone designated herein as "DNA99391-2572".
  • Figure 222 shows the amino acid sequence (SEQ ID NO: 222) derived from the coding sequence of SEQ ID NO: 221 shown in Figure 221.
  • Figure 223 shows a nucleotide sequence (SEQ ID NO:223) of a native sequence PR04315 cDNA, wherein SEQ ID NO:223 is a clone designated herein as "DNA99393-2560" .
  • Figure 224 shows the amino acid sequence (SEQ ID NO: 224) derived from the coding sequence of SEQ ID NO: 223 shown in Figure 223.
  • Figure 225 shows a nucleotide sequence (SEQ ID NO:225) of a native sequence PR05993 cDNA, wherein SEQ ID NO:225 is a clone designated herein as "DNA 100276-2684" .
  • Figure 226 shows the amino acid sequence (SEQ ID NO: 226) derived from the coding sequence of SEQ ID NO:225 shown in Figure 225.
  • Figure 227 shows a nucleotide sequence (SEQ ID NO: 227) of a native sequence PRO4503 cDNA, wherein SEQ ID NO:227 is a clone designated herein as "DNA100312-2645".
  • Figure 228 shows the amino acid sequence (SEQ ID NO: 228) derived from the coding sequence of SEQ
  • Figure 229 shows a nucleotide sequence (SEQ ID NO: 229) of a native sequence PR04976 cDNA, wherein SEQ ID NO:229 is a clone designated herein as "DNA 100902-2646" .
  • Figure 230 shows the amino acid sequence (SEQ ID NO:230) derived from the coding sequence of SEQ ID NO:229 shown in Figure 229.
  • Figure 231 shows a nucleotide sequence (SEQ ID NO:231) of a native sequence PR05798 cDNA, wherein SEQ ID NO:231 is a clone designated herein as "DNA 102899-2679".
  • Figure 232 shows the amino acid sequence (SEQ ID NO:232) derived from the coding sequence of SEQ ID NO:231 shown in Figure 231.
  • Figure 233 shows a nucleotide sequence (SEQ ID NO:233) of a native sequence PR06242 cDNA, wherein SEQ ID NO:233 is a clone designated herein as "DNA104875-2720".
  • Figure 234 shows the amino acid sequence (SEQ ID NO:234) derived from the coding sequence of SEQ ID NO:233 shown in Figure 233.
  • Figure 235 shows a nucleotide sequence (SEQ ID NO:235) of a native sequence PRO6095 cDNA, wherein SEQ ID NO:235 is a clone designated herein as "DNA105680-2710" .
  • Figure 236 shows the amino acid sequence (SEQ ID NO:236) derived from the coding sequence of SEQ ID NO:235 shown in Figure 235.
  • Figure 237 shows a nucleotide sequence (SEQ ID NO:237) of a native sequence PRO6093 cDNA, wherein SEQ ID NO:237 is a clone designated herein as "DNA105779-2708".
  • Figure 238 shows the amino acid sequence (SEQ ID NO:238) derived from the coding sequence of SEQ ID NO: 237 shown in Figure 237.
  • Figure 239 shows a nucleotide sequence (SEQ ID NO:239) of a native sequence PRO6012 cDNA, wherein SEQ ID NO:239 is a clone designated herein as "DNA 105794-2695".
  • Figure 240 shows the amino acid sequence (SEQ ID NO: 240) derived from the coding sequence of SEQ ID NO:239 shown in Figure 239.
  • Figure 241 shows a nucleotide sequence (SEQ ID NO:241) of a native sequence PRO6027 cDNA, wherein SEQ ID NO:241 is a clone designated herein as "DNA105838-2702".
  • Figure 242 shows the amino acid sequence (SEQ ID NO : 242) derived from the coding sequence of SEQ
  • Figure 243 shows a nucleotide sequence (SEQ ID NO:243) of a native sequence PR06181 cDNA, wherein SEQ ID NO:243 is a clone designated herein as "DNA107698-2715".
  • Figure 244 shows the amino acid sequence (SEQ ID NO:244) derived from the coding sequence of SEQ ID NO:243 shown in Figure 243.
  • Figure 245 shows a nucleotide sequence (SEQ ID NO:245) of a native sequence PRO6097 cDNA, wherein SEQ ID NO:245 is a clone designated herein as "DNA107701-2711".
  • Figure 246 shows the amino acid sequence (SEQ ID NO:246) derived from the coding sequence of SEQ ID NO: 245 shown in Figure 245.
  • Figure 247 shows a nucleotide sequence (SEQ ID NO: 247) of a native sequence PRO6090 cDNA, wherein SEQ ID NO:247 is a clone designated herein as "DNA107781-2707".
  • Figure 248 shows the amino acid sequence (SEQ ID NO:248) derived from the coding sequence of SEQ ID NO: 247 shown in Figure 247.
  • Figure 249 shows a nucleotide sequence (SEQ ID NO: 249) of a native sequence PR07171 cDNA, wherein SEQ ID NO:249 is a clone designated herein as "DNA 108670-2744" .
  • Figure 250 shows the amino acid sequence (SEQ ID NO:250) derived from the coding sequence of SEQ ID NO:249 shown in Figure 249.
  • Figure 251 shows a nucleotide sequence (SEQ ID NO:251) of a native sequence PR06258 cDNA, wherein SEQ ID NO:251 is a clone designated herein as "DNA 108688-2725".
  • Figure 252 shows the amino acid sequence (SEQ ID NO:252) derived from the coding sequence of SEQ
  • Figure 253 shows a nucleotide sequence (SEQ ID NO:253) of a native sequence PRO9820 cDNA, wherein SEQ ID NO:253 is a clone designated herein as "DNA108769-2765".
  • Figure 254 shows the amino acid sequence (SEQ ID NO:254) derived from the coding sequence of SEQ ID NO:253 shown in Figure 253.
  • Figure 255 shows a nucleotide sequence (SEQ ID NO:255) of a native sequence PR06243 cDNA, wherein SEQ ID NO:255 is a clone designated herein as "DNA108935-2721 ".
  • Figure 256 shows the amino acid sequence (SEQ ID NO:256) derived from the coding sequence of SEQ ID NO:255 shown in Figure 255.
  • Figure 257 shows a nucleotide sequence (SEQ ID NO:257) of a native sequence PR06182 cDNA, wherein SEQ ID NO:257 is a clone designated herein as "DNA 110700-2716".
  • Figure 258 shows the amino acid sequence (SEQ ID NO: 258) derived from the coding sequence of SEQ ID NO:257 shown in Figure 257.
  • Figure 259 shows a nucleotide sequence (SEQ ID NO:259) of a native sequence PRO6079 cDNA, wherein SEQ ID NO:259 is a clone designated herein as "DNA111750-2706".
  • Figure 260 shows the amino acid sequence (SEQ ID NO: 260) derived from the coding sequence of SEQ ID NO:259 shown in Figure 259.
  • Figure 261 shows a nucleotide sequence (SEQ ID NO: 261) of a native sequence PR07434 cDNA, wherein SEQ ID NO:261 is a clone designated herein as "DNA123430-2755".
  • Figure 262 shows the amino acid sequence (SEQ ID NO: 262) derived from the coding sequence of SEQ ID NO:261 shown in Figure 261.
  • Figure 263 shows a nucleotide sequence (SEQ ID NO:263) of a native sequence PR09865 cDNA, wherein SEQ ID NO: 263 is a clone designated herein as "DNA 125154-2785".
  • Figure 264 shows the amino acid sequence (SEQ ID NO: 264) derived from the coding sequence of SEQ ID NO:263 shown in Figure 263.
  • Figure 265 shows a nucleotide sequence (SEQ ID NO:265) of a native sequence PR09828 cDNA, wherein SEQ ID NO:265 is a clone designated herein as "DNA142238-2768" .
  • Figure 266 shows the amino acid sequence (SEQ ID NO: 266) derived from the coding sequence of SEQ
  • Figure 267 shows a nucleotide sequence (SEQ ID NO: 267) of a native sequence PRO 196 cDNA, wherein SEQ ID NO:267 is a clone designated herein as "DNA22779-1130" .
  • Figure 268 shows the amino acid sequence (SEQ ID NO: 268) derived from the coding sequence of SEQ ID NO: 267 shown in Figure 267.
  • Figure 269 shows a nucleotide sequence (SEQ ID NO: 269) of a native sequence PRO 197 cDNA, wherein SEQ ID NO:269 is a clone designated herein as "DNA22780-1078".
  • Figure 270 shows the amino acid sequence (SEQ ID NO:270) derived from the coding sequence of SEQ ID NO:269 shown in Figure 269.
  • Figure 271 shows a nucleotide sequence (SEQ ID NO:271) of a native sequence PR0195 cDNA, wherein SEQ ID NO:271 is a clone designated herein as "DNA26847-1395" .
  • Figure 272 shows the amino acid sequence (SEQ ID NO : 272) derived from the coding sequence of SEQ ID NO:271 shown in Figure 271.
  • Figure 273 shows a nucleotide sequence (SEQ ID NO:273) of a native sequence PR0187 cDNA, wherein SEQ ID NO:273 is a clone designated herein as "DNA27864-1155 " .
  • Figure 274 shows the amino acid sequence (SEQ ID NO: 274) derived from the coding sequence of SEQ ID NO:273 shown in Figure 273.
  • Figure 275 shows a nucleotide sequence (SEQ ID NO:275) of a native sequence PR0182 cDNA, wherein SEQ ID NO:275 is a clone designated herein as "DNA27865-1091 ".
  • Figure 276 shows the amino acid sequence (SEQ ID NO: 276) derived from the coding sequence of SEQ ID NO: 275 shown in Figure 275.
  • Figure 277 shows a nucleotide sequence (SEQ ID NO: 277) of a native sequence PRO 188 cDNA, wherein SEQ ID NO:277 is a clone designated herein as "DNA28497-1130" .
  • Figure 278 shows the amino acid sequence (SEQ ID NO:278) derived from the coding sequence of SEQ ID NO: 277 shown in Figure 277.
  • Figure 279 shows a nucleotide sequence (SEQ ID NO: 279) of a native sequence PRO 183 cDNA, wherein SEQ ID NO:279 is a clone designated herein as "DNA28498".
  • Figure 280 shows the amino acid sequence (SEQ ID NO: 280) derived from the coding sequence of SEQ
  • Figure 281 shows a nucleotide sequence (SEQ ID NO:281) of a native sequence PR0184 cDNA, wherein SEQ ID NO:281 is a clone designated herein as "DNA28500”.
  • Figure 282 shows the amino acid sequence (SEQ ID NO: 282) derived from the coding sequence of SEQ ID NO:281 shown in Figure 281.
  • Figure 283 shows a nucleotide sequence (SEQ ID NO:283) of a native sequence PR0185 cDNA, wherein SEQ ID NO:283 is a clone designated herein as "DNA28503".
  • Figure 284 shows the amino acid sequence (SEQ ID NO:284) derived from the coding sequence of SEQ ID NO:283 shown in Figure 283.
  • Figure 285 shows a nucleotide sequence (SEQ ID NO:285) of a native sequence PRO200 cDNA, wherein SEQ ID NO:285 is a clone designated herein as "DNA29101-1122".
  • Figure 286 shows the amino acid sequence (SEQ ID NO:286) derived from the coding sequence of SEQ ID NO:285 shown in Figure 285.
  • Figure 287 shows a nucleotide sequence (SEQ ID NO: 287) of a native sequence PRO202 cDNA, wherein SEQ ID NO:287 is a clone designated herein as "DNA30869" .
  • Figure 288 shows the amino acid sequence (SEQ ID NO: 288) derived from the coding sequence of SEQ ID NO:287 shown in Figure 287.
  • Figure 289 shows a nucleotide sequence (SEQ ID NO: 289) of a native sequence PR0214 cDNA, wherein SEQ ID NO:289 is a clone designated herein as "DNA32286-1191".
  • Figure 290 shows the amino acid sequence (SEQ ID NO: 290) derived from the coding sequence of SEQ
  • Figure 291 shows a nucleotide sequence (SEQ ID NO:291) of a native sequence PR0215 cDNA, wherein SEQ ID NO:291 is a clone designated herein as "DNA32288-1132".
  • Figure 292 shows the amino acid sequence (SEQ ID NO: 292) derived from the coding sequence of SEQ ID NO:291 shown in Figure 291.
  • Figure 293 shows a nucleotide sequence (SEQ ID NO:293) of a native sequence PR0219 cDNA, wherein SEQ ID NO:293 is a clone designated herein as "DNA32290-1164".
  • Figure 294 shows the amino acid sequence (SEQ ID NO: 294) derived from the coding sequence of SEQ ID NO:293 shown in Figure 293.
  • Figure 295 shows a nucleotide sequence (SEQ ID NO:295) of a native sequence PR0211 cDNA, wherein SEQ ID NO:295 is a clone designated herein as "DNA32292-1131 " .
  • Figure 296 shows the amino acid sequence (SEQ ID NO:296) derived from the coding sequence of SEQ ID NO:295 shown in Figure 295.
  • Figure 297 shows a nucleotide sequence (SEQ ID NO: 297) of a native sequence PRO220 cDNA, wherein SEQ ID NO:297 is a clone designated herein as "DNA32298-1132".
  • Figure 298 shows the amino acid sequence (SEQ ID NO: 298) derived from the coding sequence of SEQ ID NO: 297 shown in Figure 297.
  • Figure 299 shows a nucleotide sequence (SEQ ID NO:299) of a native sequence PR0366 cDNA, wherein SEQ ID NO:299 is a clone designated herein as "DNA33085-1110" .
  • Figure 300 shows the amino acid sequence (SEQ ID NO: 300) derived from the coding sequence of SEQ ID NO:299 shown in Figure 299.
  • Figure 301 shows a nucleotide sequence (SEQ ID NO:301) of a native sequence PR0216 cDNA, wherein SEQ ID NO:301 is a clone designated herein as "DNA33087-1158" .
  • Figure 302 shows the amino acid sequence (SEQ ID NO:302) derived from the coding sequence of SEQ ID NO:301 shown in Figure 301.
  • Figure 303 shows a nucleotide sequence (SEQ ID NO:303) of a native sequence PR0221 cDNA, wherein SEQ ID NO:303 is a clone designated herein as "DNA33089-1132" .
  • Figure 304 shows the amino acid sequence (SEQ ID NO:304) derived from the coding sequence of SEQ
  • Figure 305 shows a nucleotide sequence (SEQ ID NO:305) of a native sequence PR0228 cDNA, wherein SEQ ID NO:305 is a clone designated herein as "DNA33092-1202".
  • Figure 306 shows the amino acid sequence (SEQ ID NO:306) derived from the coding sequence of SEQ ID NO:305 shown in Figure 305.
  • Figure 307 shows a nucleotide sequence (SEQ ID NO:307) of a native sequence PR0217 cDNA, wherein SEQ ID NO:307 is a clone designated herein as "DNA33094-1131 " .
  • Figure 308 shows the amino acid sequence (SEQ ID NO:308) derived from the coding sequence of SEQ ID NO: 307 shown in Figure 307.
  • Figure 309 shows a nucleotide sequence (SEQ ID NO:309) of a native sequence PR0222 cDNA, wherein SEQ ID NO:309 is a clone designated herein as "DNA33107-1135" .
  • Figure 310 shows the amino acid sequence (SEQ ID NO : 310) derived from the coding sequence of SEQ ID NO:309 shown in Figure 309.
  • Figure 311 shows a nucleotide sequence (SEQ ID NO:311) of a native sequence PR0224 cDNA, wherein SEQ ID NO:311 is a clone designated herein as "DNA33221-1133".
  • Figure 312 shows the amino acid sequence (SEQ ID NO : 312) derived from the coding sequence of SEQ ID NO:311 shown in Figure 311.
  • Figure 313 shows a nucleotide sequence (SEQ ID NO:313) of a native sequence PRO230 cDNA, wherein SEQ ID NO:313 is a clone designated herein as "DNA33223-1136".
  • Figure 314 shows the amino acid sequence (SEQ ID NO:314) derived from the coding sequence of SEQ ID NO:313 shown in Figure 313.
  • Figure 315 shows a nucleotide sequence (SEQ ID NO:315) of a native sequence PR0198 cDNA, wherein SEQ ID NO:315 is a clone designated herein as "DNA33457-1078".
  • Figure 316 shows the amino acid sequence (SEQ ID NO:316) derived from the coding sequence of SEQ ID NO:315 shown in Figure 315.
  • Figure 317 shows a nucleotide sequence (SEQ ID NO:317) of a native sequence PR0226 cDNA, wherein SEQ ID NO:317 is a clone designated herein as "DNA33460-1166".
  • Figure 318 shows the amino acid sequence (SEQ ID NO : 318) derived from the coding sequence of SEQ
  • Figure 319 shows a nucleotide sequence (SEQ ID NO:319) of a native sequence PR0261 cDNA, wherein SEQ ID NO:319 is a clone designated herein as "DNA33473-1176".
  • Figure 320 shows the amino acid sequence (SEQ ID NO:320) derived from the coding sequence of SEQ ID NO:319 shown in Figure 319.
  • Figure 321 shows a nucleotide sequence (SEQ ID NO:321) of a native sequence PR0242 cDNA, wherein SEQ ID NO:321 is a clone designated herein as "DNA33785-1143".
  • Figure 322 shows the amino acid sequence (SEQ ID NO:322) derived from the coding sequence of SEQ ID NO:321 shown in Figure 321.
  • Figure 323 shows a nucleotide sequence (SEQ ID NO:323) of a native sequence PR0227 cDNA, wherein SEQ ID NO:323 is a clone designated herein as "DNA33786-1132".
  • Figure 324 shows the amino acid sequence (SEQ ID NO:324) derived from the coding sequence of SEQ ID NO:323 shown in Figure 323.
  • Figure 325 shows a nucleotide sequence (SEQ ID NO:325) of a native sequence PR0237 cDNA, wherein SEQ ID NO:325 is a clone designated herein as "DNA34353-1428" .
  • Figure 326 shows the amino acid sequence (SEQ ID NO:326) derived from the coding, sequence of SEQ ID NO:325 shown in Figure 325.
  • Figure 327 shows a nucleotide sequence (SEQ ID NO:327) of a native sequence PR0241 cDNA, wherein SEQ ID NO:327 is a clone designated herein as "DNA34392-1170".
  • Figure 328 shows the amino acid sequence (SEQ ID NO:328) derived from the coding sequence of SEQ
  • Figure 329 shows a nucleotide sequence (SEQ ID NO:329) of a native sequence PR0231 cDNA, wherein SEQ ID NO:329 is a clone designated herein as "DNA34434-1139".
  • Figure 330 shows the amino acid sequence (SEQ ID NO:330) derived from the coding sequence of SEQ ID NO:329 shown in Figure 329.
  • Figure 331 shows a nucleotide sequence (SEQ ID NO:331) of a native sequence PR0235 cDNA, wherein SEQ ID NO:331 is a clone designated herein as "DNA35558-1167".
  • Figure 332 shows the amino acid sequence (SEQ ID NO:332) derived from the coding sequence of SEQ ID NO:331 shown in Figure 331.
  • Figure 333 shows a nucleotide sequence (SEQ ID NO:333) of a native sequence PR0323 cDNA, wherein SEQ ID NO:333 is a clone designated herein as "DNA35595-1228" .
  • Figure 334 shows the amino acid sequence (SEQ ID NO: 334) derived from the coding sequence of SEQ ID NO:333 shown in Figure 333.
  • Figure 335 shows a nucleotide sequence (SEQ ID NO:335) of a native sequence PR0245 cDNA, wherein SEQ ID NO:335 is a clone designated herein as "DNA35638-1216" .
  • Figure 336 shows the amino acid sequence (SEQ ID NO:336) derived from the coding sequence of SEQ ID NO:335 shown in Figure 335.
  • Figure 337 shows a nucleotide sequence (SEQ ID NO:337) of a native sequence PR0246 cDNA, wherein SEQ ID NO:337 is a clone designated herein as "DNA35639-1172".
  • Figure 338 shows the amino acid sequence (SEQ ID NO:338) derived from the coding sequence of SEQ ID NO:337 shown in Figure 337.
  • Figure 339 shows a nucleotide sequence (SEQ ID NO:339) of a native sequence PR0288 cDNA, wherein SEQ ID NO:339 is a clone designated herein as "DNA35663-1129" .
  • Figure 340 shows the amino acid sequence (SEQ ID NO:340) derived from the coding sequence of SEQ ID NO:339 shown in Figure 339.
  • Figure 341 shows a nucleotide sequence (SEQ ID NO:341) of a native sequence PR0248 cDNA, wherein SEQ ID NO:341 is a clone designated herein as "DNA35674-1142".
  • Figure 342 shows the amino acid sequence (SEQ ID NO:342) derived from the coding sequence of SEQ
  • Figure 343 shows a nucleotide sequence (SEQ ID NO:343) of a native sequence PR0257 cDNA, wherein SEQ ID NO:343 is a clone designated herein as "DNA35841-1173" .
  • Figure 344 shows the amino acid sequence (SEQ ID NO:344) derived from the coding sequence of SEQ ID NO:343 shown in Figure 343.
  • Figure 345 shows a nucleotide sequence (SEQ ID NO:345) of a native sequence PR0172 cDNA, wherein SEQ ID NO:345 is a clone designated herein as "DNA35916-1161 ".
  • Figure 346 shows the amino acid sequence (SEQ ID NO: 346) derived from the coding sequence of SEQ ID NO:345 shown in Figure 345.
  • Figure 347 shows a nucleotide sequence (SEQ ID NO:347) of a native sequence PR0258 cDNA, wherein SEQ ID NO:347 is a clone designated herein as "DNA35918-1174" .
  • Figure 348 shows the amino acid sequence (SEQ ID NO:348) derived from the coding sequence of SEQ ID NO:347 shown in Figure 347.
  • Figure 349 shows a nucleotide sequence (SEQ ID NO:349) of a native sequence PR0265 cDNA, wherein SEQ ID NO:349 is a clone designated herein as "DNA36350-1158" .
  • Figure 350 shows the amino acid sequence (SEQ ID NO:350) derived from the coding sequence of SEQ ID NO:349 shown in Figure 349.
  • Figure 351 shows a nucleotide sequence (SEQ ID NO:351) of a native sequence PR0326 cDNA, wherein SEQ ID NO:351 is a clone designated herein as "DNA37140-1234" .
  • Figure 352 shows the amino acid sequence (SEQ ID NO:352) derived from the coding sequence of SEQ ID NO:351 shown in Figure 351.
  • Figure 353 shows a nucleotide sequence (SEQ ID NO:353) of a native sequence PR0266 cDNA, wherein SEQ ID NO:353 is a clone designated herein as "DNA37150-1178" .
  • Figure 354 shows the amino acid sequence (SEQ ID NO:354) derived from the coding sequence of SEQ ID NO:353 shown in Figure 353.
  • Figure 355 shows a nucleotide sequence (SEQ ID NO:355) of a native sequence PR0269 cDNA, wherein SEQ ID NO:355 is a clone designated herein as "DN A38260- 1180" .
  • Figure 356 shows the amino acid sequence (SEQ ID NO:356) derived from the coding sequence of SEQ
  • Figure 357 shows a nucleotide sequence (SEQ ID NO:357) of a native sequence PR0285 cDNA, wherein SEQ ID NO:357 is a clone designated herein as "DNA40021-1154" .
  • Figure 358 shows the amino acid sequence (SEQ ID NO:358) derived from the coding sequence of SEQ ID NO:357 shown in Figure 357.
  • Figure 359 shows a nucleotide sequence (SEQ ID NO:359) of a native sequence PR0328 cDNA, wherein SEQ ID NO:359 is a clone designated herein as "DNA40587-1231 ".
  • Figure 360 shows the amino acid sequence (SEQ ID NO:360) derived from the coding sequence of SEQ ID NO:359 shown in Figure 359.
  • Figure 361 shows a nucleotide sequence (SEQ ID NO:361) of a native sequence PR0344 cDNA, wherein SEQ ID NO:361 is a clone designated herein as "DNA40592-1242".
  • Figure 362 shows the amino acid sequence (SEQ ID NO:362) derived from the coding sequence of SEQ ID NO:361 shown in Figure 361.
  • Figure 363 shows a nucleotide sequence (SEQ ID NO:363) of a native sequence PR0272 cDNA, wherein SEQ ID NO:363 is a clone designated herein as "DNA40620-1183".
  • Figure 364 shows the amino acid sequence (SEQ ID NO: 364) derived from the coding sequence of SEQ ID NO:363 shown in Figure 363.
  • Figure 365 shows a nucleotide sequence (SEQ ID NO:365) of a native sequence PRO301 cDNA, wherein SEQ ID NO:365 is a clone designated herein as "DNA40628-1216" .
  • Figure 366 shows the amino acid sequence (SEQ ID NO:366) derived from the coding sequence of SEQ
  • Figure 367 shows a nucleotide sequence (SEQ ID NO:367) of a native sequence PR0331 cDNA, wherein SEQ ID NO:367 is a clone designated herein as "DNA40981-1234" .
  • Figure 368 shows the amino acid sequence (SEQ ID NO:368) derived from the coding sequence of SEQ ID NO:367 shown in Figure 367.
  • Figure 369 shows a nucleotide sequence (SEQ ID NO:369) of a native sequence PR0332 cDNA, wherein SEQ ID NO:369 is a clone designated herein as "DNA40982-1235".
  • Figure 370 shows the amino acid sequence (SEQ ID NO:370) derived from the coding sequence of SEQ ID NO:369 shown in Figure 369.
  • Figure 371 shows a nucleotide sequence (SEQ ID NO:371) of a native sequence PR0353 cDNA, wherein SEQ ID NO:371 is a clone designated herein as "DNA41234- 1242" .
  • Figure 372 shows the amino acid sequence (SEQ ID NO: 372) derived from the coding sequence of SEQ ID NO:371 shown in Figure 371.
  • Figure 373 shows a nucleotide sequence (SEQ ID NO:373) of a native sequence PRO310 cDNA, wherein SEQ ID NO:373 is a clone designated herein as "DNA43046-1225" .
  • Figure 374 shows the amino acid sequence (SEQ ID NO: 374) derived from the coding sequence of SEQ ID NO: 373 shown in Figure 373.
  • Figure 375 shows a nucleotide sequence (SEQ ID NO:375) of a native sequence PR0337 cDNA, wherein SEQ ID NO:375 is a clone designated herein as "DNA43316-1237".
  • Figure 376 shows the amino acid sequence (SEQ ID NO:376) derived from the coding sequence of SEQ ID NO: 375 shown in Figure 375.
  • Figure 377 shows a nucleotide sequence (SEQ ID NO:377) of a native sequence PR0346 cDNA, wherein SEQ ID NO:377 is a clone designated herein as "DNA44167- 1243".
  • Figure 378 shows the amino acid sequence (SEQ ID NO: 378) derived from the coding sequence of SEQ ID NO: 377 shown in Figure 377.
  • Figure 379 shows a nucleotide sequence (SEQ ID NO:379) of a native sequence PRO350 cDNA, wherein SEQ ID NO:379 is a clone designated herein as "DNA44175-1314".
  • Figure 380 shows the amino acid sequence (SEQ ID NO:380) derived from the coding sequence of SEQ
  • Figure 381 shows a nucleotide sequence (SEQ ID NO:381) of a native sequence PR0526 cDNA, wherein SEQ ID NO:381 is a clone designated herein as "DNA44184-1319" .
  • Figure 382 shows the amino acid sequence (SEQ ID NO:382) derived from the coding sequence of SEQ ID NO:381 shown in Figure 381.
  • Figure 383 shows a nucleotide sequence (SEQ ID NO:383) of a native sequence PR0381 cDNA, wherein SEQ ID NO:383 is a clone designated herein as "DNA44194-1317".
  • Figure 384 shows the amino acid sequence (SEQ ID NO: 384) derived from the coding sequence of SEQ ID NO:383 shown in Figure 383.
  • Figure 385 shows a nucleotide sequence (SEQ ID NO:385) of a native sequence PR0846 cDNA, wherein SEQ ID NO:385 is a clone designated herein as "DNA44196-1353".
  • Figure 386 shows the amino acid sequence (SEQ ID NO:386) derived from the coding sequence of SEQ ID NO:385 shown in Figure 385.
  • Figure 387 shows a nucleotide sequence (SEQ ID NO:387) of a native sequence PR0363 cDNA, wherein SEQ ID NO:387 is a clone designated herein as "DNA45419-1252" .
  • Figure 388 shows the amino acid sequence (SEQ ID NO:388) derived from the coding sequence of SEQ ID NO:387 shown in Figure 387.
  • Figure 389 shows a nucleotide sequence (SEQ ID NO:389) of a native sequence PR0365 cDNA, wherein SEQ ID NO:389 is a clone designated herein as "DNA46777-1253".
  • Figure 390 shows the amino acid sequence (SEQ ID NO:390) derived from the coding sequence of SEQ ID NO:389 shown in Figure 389.
  • Figure 391 shows a nucleotide sequence (SEQ ID NO:391) of a native sequence PRO1310 cDNA, wherein SEQ ID NO:391 is a clone designated herein as "DNA47394-1572".
  • Figure 392 shows the amino acid sequence (SEQ ID NO:392) derived from the coding sequence of SEQ ID NO:391 shown in Figure 391.
  • Figure 393 shows a nucleotide sequence (SEQ ID NO:393) of a native sequence PR0731 cDNA, wherein SEQ ID NO:393 is a clone designated herein as "DNA48331-1329".
  • Figure 394 shows the amino acid sequence (SEQ ID NO:394) derived from the coding sequence of SEQ
  • Figure 395 shows a nucleotide sequence (SEQ ID NO:395) of a native sequence PR0322 cDNA, wherein SEQ ID NO:395 is a clone designated herein as "DNA48336-1309".
  • Figure 396 shows the amino acid sequence (SEQ ID NO:396) derived from the coding sequence of SEQ ID NO: 395 shown in Figure 395.
  • Figure 397 shows a nucleotide sequence (SEQ ID NO:397) of a native sequence PR0536 cDNA, wherein SEQ ID NO:397 is a clone designated herein as "DNA49142-1430".
  • Figure 398 shows the amino acid sequence (SEQ ID NO:398) derived from the coding sequence of SEQ ID NO:397 shown in Figure 397.
  • Figure 399 shows a nucleotide sequence (SEQ ID NO:399) of a native sequence PR0719 cDNA, wherein SEQ ID NO:399 is a clone designated herein as "DNA49646-1327".
  • Figure 400 shows the amino acid sequence (SEQ ID NO: 400) derived from the coding sequence of SEQ ID NO:399 shown in Figure 399.
  • Figure 401 shows a nucleotide sequence (SEQ ID NO: 401) of a native sequence PR0619 cDNA, wherein SEQ ID NO:401 is a clone designated herein as "DNA49821-1562".
  • Figure 402 shows the amino acid sequence (SEQ ID NO: 402) derived from the coding sequence of SEQ ID NO:401 shown in Figure 401.
  • Figure 403 shows a nucleotide sequence (SEQ ID NO:403) of a native sequence PR0771 cDNA, wherein SEQ ID NO:403 is a clone designated herein as "DNA49829-1346".
  • Figure 404 shows the amino acid sequence (SEQ ID NO:404) derived from the coding sequence of SEQ
  • Figure 405 shows a nucleotide sequence (SEQ ID NO:405) of a native sequence PRO1083 cDNA, wherein SEQ ID NO:405 is a clone designated herein as "DNA50921-1458".
  • Figure 406 shows the amino acid sequence (SEQ ID NO: 406) derived from the coding sequence of SEQ ID NO:405 shown in Figure 405.
  • Figure 407 shows a nucleotide sequence (SEQ ID NO: 407) of a native sequence PR0862 cDNA, wherein SEQ ID NO:407 is a clone designated herein as "DNA52187-1354".
  • Figure 408 shows the amino acid sequence (SEQ ID NO: 408) derived from the coding sequence of SEQ ID NO:407 shown in Figure 407.
  • Figure 409 shows a nucleotide sequence (SEQ ID NO:409) of a native sequence PR0733 cDNA, wherein SEQ ID NO:409 is a clone designated herein as "DNA52196-1348" .
  • Figure 410 shows the amino acid sequence (SEQ ID NO:410) derived from the coding sequence of SEQ ID NO:409 shown in Figure 409.
  • Figure 411 shows a nucleotide sequence (SEQ ID NO:411) of a native sequence PROl 188 cDNA, wherein SEQ ID NO:411 is a clone designated herein as "DNA52598-1518" .
  • Figure 412 shows the amino acid sequence (SEQ ID NO:412) derived from the coding sequence of SEQ ID NO:411 shown in Figure 411.
  • Figure 413 shows a nucleotide sequence (SEQ ID NO:413) of a native sequence PRO770 cDNA, wherein SEQ ID NO:413 is a clone designated herein as "DNA54228-1366" .
  • Figure 414 shows the amino acid sequence (SEQ ID NO:414) derived from the coding sequence of SEQ ID NO:413 shown in Figure 413.
  • Figure 415 shows a nucleotide sequence (SEQ ID NO:415) of a native sequence PRO 1080 cDNA, wherein SEQ ID NO:415 is a clone designated herein as "DNA56047-1456" .
  • Figure 416 shows the amino acid sequence (SEQ ID NO:416) derived from the coding sequence of SEQ ID NO:415 shown in Figure 415.
  • Figure 417 shows a nucleotide sequence (SEQ ID NO:417) of a native sequence PRO1017 cDNA, wherein SEQ ID NO:417 is a clone designated herein as "DNA56112-1379".
  • Figure 418 shows the amino acid sequence (SEQ ID NO : 418) derived from the coding sequence of SEQ
  • Figure 419 shows a nucleotide sequence (SEQ ID NO:419) of a native sequence PRO1016 cDNA, wherein SEQ ID NO:419 is a clone designated herein as "DNA56113-1378".
  • Figure 420 shows the amino acid sequence (SEQ ID NO: 420) derived from the coding sequence of SEQ ID NO:419 shown in Figure 419.
  • Figure 421 shows a nucleotide sequence (SEQ ID NO: 421) of a native sequence PR0792 cDNA, wherein SEQ ID NO:421 is a clone designated herein as "DNA56352-1358".
  • Figure 422 shows the amino acid sequence (SEQ ID NO: 422) derived from the coding sequence of SEQ ID NO:421 shown in Figure 421.
  • Figure 423 shows a nucleotide sequence (SEQ ID NO:423) of a native sequence PR0938 cDNA, wherein SEQ ID NO:423 is a clone designated herein as "DNA56433-1406".
  • Figure 424 shows the amino acid sequence (SEQ ID NO: 424) derived from the coding sequence of SEQ ID NO:423 shown in Figure 423.
  • Figure 425 shows a nucleotide sequence (SEQ ID NO: 425) of a native sequence PRO 1012 cDNA, wherein SEQ ID NO:425 is a clone designated herein as "DNA56439-1376".
  • Figure 426 shows the amino acid sequence (SEQ ID NO:426) derived from the coding sequence of SEQ ID NO:425 shown in Figure 425.
  • Figure 427 shows a nucleotide sequence (SEQ ID NO:427) of a native sequence PRO1008 cDNA, wherein SEQ ID NO:427 is a clone designated herein as "DNA57530-1375".
  • Figure 428 shows the amino acid sequence (SEQ ID NO: 428) derived from the coding sequence of SEQ ID NO: 427 shown in Figure 427.
  • Figure 429 shows a nucleotide sequence (SEQ ID NO: 429) of a native sequence PRO 1075 cDNA, wherein SEQ ID NO:429 is a clone designated herein as "DNA57689-1385 " .
  • Figure 430 shows the amino acid sequence (SEQ ID NO:430) derived from the coding sequence of SEQ ID NO:429 shown in Figure 429.
  • Figure 431 shows a nucleotide sequence (SEQ ID NO: 431) of a native sequence PRO 1007 cDNA, wherein SEQ ID NO:431 is a clone designated herein as "DNA57690-1374".
  • Figure 432 shows the amino acid sequence (SEQ ID NO:432) derived from the coding sequence of SEQ
  • Figure 433 shows a nucleotide sequence (SEQ ID NO:433) of a native sequence PRO1056 cDNA, wherein SEQ ID NO:433 is a clone designated herein as "DNA57693-1424".
  • Figure 434 shows the amino acid sequence (SEQ ID NO: 434) derived from the coding sequence of SEQ ID NO:433 shown in Figure 433.
  • Figure 435 shows a nucleotide sequence (SEQ ID NO:435) of a native sequence PR0791 cDNA, wherein SEQ ID NO:435 is a clone designated herein as "DNA57838-1337".
  • Figure 436 shows the amino acid sequence (SEQ ID NO:436) derived from the coding sequence of SEQ ID NO:435 shown in Figure 435.
  • Figure 437 shows a nucleotide sequence (SEQ ID NO:437) of a native sequence PROl 111 cDNA, wherein SEQ ID NO:437 is a clone designated herein as "DNA58721-1475".
  • Figure 438 shows the amino acid sequence (SEQ ID NO:438) derived from the coding sequence of SEQ ID NO: 437 shown in Figure 437.
  • Figure 439 shows a nucleotide sequence (SEQ ID NO:439) of a native sequence PR0812 cDNA, wherein SEQ ID NO:439 is a clone designated herein as "DNA59205-1421 " .
  • Figure 440 shows the amino acid sequence (SEQ ID NO:440) derived from the coding sequence of SEQ ID NO:439 shown in Figure 439.
  • Figure 441 shows a nucleotide sequence (SEQ ID NO:441) of a native sequence PRO1066 cDNA, wherein SEQ ID NO:441 is a clone designated herein as "DNA59215-1425”.
  • Figure 442 shows the amino acid sequence (SEQ ID NO:442) derived from the coding sequence of SEQ
  • Figure 443 shows a nucleotide sequence (SEQ ID NO:443) of a native sequence PROl 185 cDNA, wherein SEQ ID NO:443 is a clone designated herein as "DNA59220-1514".
  • Figure 444 shows the amino acid sequence (SEQ ID NO: 444) derived from the coding sequence of SEQ ID NO:443 shown in Figure 443.
  • Figure 445 shows a nucleotide sequence (SEQ ID NO:445) of a native sequence PRO1031 cDNA, wherein SEQ ID NO:445 is a clone designated herein as "DNA59294-1381".
  • Figure 446 shows the amino acid sequence (SEQ ID NO:446) derived from the coding sequence of SEQ ID NO: 445 shown in Figure 445.
  • Figure 447 shows a nucleotide sequence (SEQ ID NO:447) of a native sequence PRO1360 cDNA, wherein SEQ ID NO:447 is a clone designated herein as "DNA59488-1603" .
  • Figure 448 shows the amino acid sequence (SEQ ID NO:448) derived from the coding sequence of SEQ ID NO: 447 shown in Figure 447.
  • Figure 449 shows a nucleotide sequence (SEQ ID NO:449) of a native sequence PRO1309 cDNA, wherein SEQ ID NO:449 is a clone designated herein as "DNA59588-1571 ".
  • Figure 450 shows the amino acid sequence (SEQ ID NO: 450) derived from the coding sequence of SEQ ID NO: 449 shown in Figure 449.
  • Figure 451 shows a nucleotide sequence (SEQ ID NO:451) of a native sequence PROl 107 cDNA, wherein SEQ ID NO:451 is a clone designated herein as "DNA59606-1471 ".
  • Figure 452 shows the amino acid sequence (SEQ ID NO:452) derived from the coding sequence of SEQ ID NO: 451 shown in Figure 451.
  • Figure 453 shows a nucleotide sequence (SEQ ID NO:453) of a native sequence PR0836 cDNA, wherein SEQ ID NO:453 is a clone designated herein as "DNA59620-1463".
  • Figure 454 shows the amino acid sequence (SEQ ID NO:454) derived from the coding sequence of SEQ ID NO:453 shown in Figure 453.
  • Figure 455 shows a nucleotide sequence (SEQ ID NO:455) of a native sequence PROl 132 cDNA, wherein SEQ ID NO:455 is a clone designated herein as "DNA59767-1489".
  • Figure 456 shows the amino acid sequence (SEQ ID NO:456) derived from the coding sequence of SEQ
  • Figure 457 shows a nucleotide sequence (SEQ ID NO:457) of a native sequence PROl 131 cDNA, wherein SEQ ID NO:457 is a clone designated herein as "DNA59777-1480".
  • Figure 458 shows the amino acid sequence (SEQ ID NO:458) derived from the coding sequence of SEQ ID NO:457 shown in Figure 457.
  • Figure 459 shows a nucleotide sequence (SEQ ID NO:459) of a native sequence PROl 130 cDNA, wherein SEQ ID NO:459 is a clone designated herein as "DNA59814-1486" .
  • Figure 460 shows the amino acid sequence (SEQ ID NO: 460) derived from the coding sequence of SEQ ID NO:459 shown in Figure 459.
  • Figure 461 shows a nucleotide sequence (SEQ ID NO:461) of a native sequence PR0844 cDNA, wherein SEQ ID NO:461 is a clone designated herein as "DNA59839-1461 ".
  • Figure 462 shows the amino acid sequence (SEQ ID NO: 462) derived from the coding sequence of SEQ ID NO:461 shown in Figure 461.
  • Figure 463 shows a nucleotide sequence (SEQ ID NO:463) of a native sequence PROl 154 cDNA, wherein SEQ ID NO:463 is a clone designated herein as "DNA59846-1503".
  • Figure 464 shows the amino acid sequence (SEQ ID NO: 464) derived from the coding sequence of SEQ ID NO: 463 shown in Figure 463.
  • Figure 465 shows a nucleotide sequence (SEQ ID NO:465) of a native sequence PROl 181 cDNA, wherein SEQ ID NO:465 is a clone designated herein as "DNA59847-1511 ".
  • Figure 466 shows the amino acid sequence (SEQ ID NO:466) derived from the coding sequence of SEQ ID NO: 465 shown in Figure 465.
  • Figure 467 shows a nucleotide sequence (SEQ ID NO: 467) of a native sequence PROl 126 cDNA, wherein SEQ ID NO:467 is a clone designated herein as "DNA60615-1483".
  • Figure 468 shows the amino acid sequence (SEQ ID NO:468) derived from the coding sequence of SEQ ID NO: 467 shown in Figure 467.
  • Figure 469 shows a nucleotide sequence (SEQ ID NO:469) of a native sequence PROl 186 cDNA, wherein SEQ ID NO:469 is a clone designated herein as "DNA60621-1516".
  • Figure 470 shows the amino acid sequence (SEQ ID NO : 470) derived from the coding sequence of SEQ
  • Figure 471 shows a nucleotide sequence (SEQ ID NO:471) of a native sequence PROl 198 cDNA, wherein SEQ ID NO:471 is a clone designated herein as "DNA60622-1525".
  • Figure 472 shows the amino acid sequence (SEQ ID NO:472) derived from the coding sequence of SEQ ID NO:471 shown in Figure 471.
  • Figure 473 shows a nucleotide sequence (SEQ ID NO:473) of a native sequence PROl 159 cDNA, wherein SEQ ID NO:473 is a clone designated herein as "DNA60627-1508".
  • Figure 474 shows the amino acid sequence (SEQ ID NO: 474) derived from the coding sequence of SEQ ID NO: 473 shown in Figure 473.
  • Figure 475 shows a nucleotide sequence (SEQ ID NO:475) of a native sequence PR01265 cDNA, wherein SEQ ID NO:475 is a clone designated herein as "DNA60764-1533".
  • Figure 476 shows the amino acid sequence (SEQ ID NO: 476) derived from the coding sequence of SEQ ID NO: 475 shown in Figure 475.
  • Figure 477 shows a nucleotide sequence (SEQ ID NO:477) of a native sequence PRO1250 cDNA, wherein SEQ ID NO:477 is a clone designated herein as "DNA60775-1532".
  • Figure 478 shows the amino acid sequence (SEQ ID NO:478) derived from the coding sequence of SEQ ID NO: 477 shown in Figure 477.
  • Figure 479 shows a nucleotide sequence (SEQ ID NO: 479) of a native sequence PRO 1475 cDNA, wherein SEQ ID NO:479 is a clone designated herein as "DNA61185-1646".
  • Figure 480 shows the amino acid sequence (SEQ ID NO:480) derived from the coding sequence of SEQ
  • Figure 481 shows a nucleotide sequence (SEQ ID NO: 481) of a native sequence PR01312 cDNA, wherein SEQ ID NO:481 is a clone designated herein as "DNA61873-1574" .
  • Figure 482 shows the amino acid sequence (SEQ ID NO: 482) derived from the coding sequence of SEQ ID NO:481 shown in Figure 481.
  • Figure 483 shows a nucleotide sequence (SEQ ID NO:483) of a native sequence PRO1308 cDNA, wherein SEQ ID NO:483 is a clone designated herein as "DNA62306-1570" .
  • Figure 484 shows the amino acid sequence (SEQ ID NO:484) derived from the coding sequence of SEQ ID NO:483 shown in Figure 483.
  • Figure 485 shows a nucleotide sequence (SEQ ID NO:485) of a native sequence PR01326 cDNA, wherein SEQ ID NO:485 is a clone designated herein as "DNA62808-1582" .
  • Figure 486 shows the amino acid sequence (SEQ ID NO:486) derived from the coding sequence of SEQ ID NO:485 shown in Figure 485.
  • Figure 487 shows a nucleotide sequence (SEQ ID NO:487) of a native sequence PROl 192 cDNA, wherein SEQ ID NO:487 is a clone designated herein as "DNA62814-1521 " .
  • Figure 488 shows the amino acid sequence (SEQ ID NO: 488) derived from the coding sequence of SEQ ID NO:487 shown in Figure 487.
  • Figure 489 shows a nucleotide sequence (SEQ ID NO: 489) of a native sequence PRO 1246 cDNA, wherein SEQ ID NO:489 is a clone designated herein as "DNA64885-1529" .
  • Figure 490 shows the amino acid sequence (SEQ ID NO: 490) derived from the coding sequence of SEQ ID NO:489 shown in Figure 489.
  • Figure 491 shows a nucleotide sequence (SEQ ID NO:491) of a native sequence PR01356 cDNA, wherein SEQ ID NO:491 is a clone designated herein as "DNA64886-1601" .
  • Figure 492 shows the amino acid sequence (SEQ ID NO: 492) derived from the coding sequence of SEQ ID NO: 491 shown in Figure 491.
  • Figure 493 shows a nucleotide sequence (SEQ ID NO:493) of a native sequence PR01275 cDNA, wherein SEQ ID NO:493 is a clone designated herein as "DNA64888-1542" .
  • Figure 494 shows the amino acid sequence (SEQ ID NO : 494) derived from the coding sequence of SEQ
  • Figure 495 shows a nucleotide sequence (SEQ ID NO:495) of a native sequence PR01274 cDNA, wherein SEQ ID NO:495 is a clone designated herein as "DNA64889-1541 " .
  • Figure 496 shows the amino acid sequence (SEQ ID NO: 496) derived from the coding sequence of SEQ ID NO:495 shown in Figure 495.
  • Figure 497 shows a nucleotide sequence (SEQ ID NO:497) of a native sequence PR01358 cDNA, wherein SEQ ID NO:497 is a clone designated herein as "DNA64890-1612".
  • Figure 498 shows the amino acid sequence (SEQ ID NO: 498) derived from the coding sequence of SEQ ID NO:497 shown in Figure 497.
  • Figure 499 shows a nucleotide sequence (SEQ ID NO:499) of a native sequence PR01286 cDNA, wherein SEQ ID NO:499 is a clone designated herein as "DNA64903-1553".
  • Figure 500 shows the amino acid sequence (SEQ ID NO: 500) derived from the coding sequence of SEQ ID NO:499 shown in Figure 499.
  • Figure 501 shows a nucleotide sequence (SEQ ID NO:501) of a native sequence PR01294 cDNA, wherein SEQ ID NO:501 is a clone designated herein as "DNA64905-1558".
  • Figure 502 shows the amino acid sequence (SEQ ID NO:502) derived from the coding sequence of SEQ ID NO:501 shown in Figure 501.
  • Figure 503 shows a nucleotide sequence (SEQ ID NO:503) of a native sequence PR01273 cDNA, wherein SEQ ID NO:503 is a clone designated herein as "DNA65402-1540".
  • Figure 504 shows the amino acid sequence (SEQ ID NO:504) derived from the coding sequence of SEQ ID NO:503 shown in Figure 503.
  • Figure 505 shows a nucleotide sequence (SEQ ID NO:505) of a native sequence PR01279 cDNA, wherein SEQ ID NO:505 is a clone designated herein as "DNA65405-1547".
  • Figure 506 shows the amino acid sequence (SEQ ID NO:506) derived from the coding sequence of SEQ ID NO:505 shown in Figure 505.
  • Figure 507 shows a nucleotide sequence (SEQ ID NO: 507) of a native sequence PROl 195 cDNA, wherein SEQ ID NO:507 is a clone designated herein as "DNA65412-1523" .
  • Figure 508 shows the amino acid sequence (SEQ ID NO : 508) derived from the coding sequence of SEQ
  • Figure 509 shows a nucleotide sequence (SEQ ID NO:509) of a native sequence PR01271 cDNA, wherein SEQ ID NO:509 is a clone designated herein as "DNA66309-1538" .
  • Figure 510 shows the amino acid sequence (SEQ ID NO: 510) derived from the coding sequence of SEQ ID NO:509 shown in Figure 509.
  • Figure 511 shows a nucleotide sequence (SEQ ID NO:511) of a native sequence PR01338 cDNA, wherein SEQ ID NO:511 is a clone designated herein as "DNA66667-1596".
  • Figure 512 shows the amino acid sequence (SEQ ID NO:512) derived from the coding sequence of SEQ ID NO:511 shown in Figure 511.
  • Figure 513 shows a nucleotide sequence (SEQ ID NO:513) of a native sequence PR01343 cDNA, wherein SEQ ID NO:513 is a clone designated herein as "DNA66675-1587".
  • Figure 514 shows the amino acid sequence (SEQ ID NO: 514) derived from the coding sequence of SEQ ID NO:513 shown in Figure 513.
  • Figure 515 shows a nucleotide sequence (SEQ ID NO:515) of a native sequence PR01434 cDNA, wherein SEQ ID NO:515 is a clone designated herein as "DNA68818-2536" .
  • Figure 516 shows the amino acid sequence (SEQ ID NO: 516) derived from the coding sequence of SEQ ID NO:515 shown in Figure 515.
  • Figure 517 shows a nucleotide sequence (SEQ ID NO:517) of a native sequence PR01418 cDNA, wherein SEQ ID NO:517 is a clone designated herein as "DNA68864-1629".
  • Figure 518 shows the amino acid sequence (SEQ ID NO : 518) derived from the coding sequence of SEQ
  • Figure 519 shows a nucleotide sequence (SEQ ID NO:519) of a native sequence PR01387 cDNA, wherein SEQ ID NO:519 is a clone designated herein as "DNA68872-1620".
  • Figure 520 shows the amino acid sequence (SEQ ID NO:520) derived from the coding sequence of SEQ ID NO:519 shown in Figure 519.
  • Figure 521 shows a nucleotide sequence (SEQ ID NO:521) of a native sequence PR01384 cDNA, wherein SEQ ID NO:521 is a clone designated herein as "DNA71159-1617".
  • Figure 522 shows the amino acid sequence (SEQ ID NO:522) derived from the coding sequence of SEQ ID NO: 521 shown in Figure 521.
  • Figure 523 shows a nucleotide sequence (SEQ ID NO:523) of a native sequence PR01565 cDNA, wherein SEQ ID NO:523 is a clone designated herein as "DNA73727-1673" .
  • Figure 524 shows the amino acid sequence (SEQ ID NO:524) derived from the coding sequence of SEQ ID NO:523 shown in Figure 523.
  • Figure 525 shows a nucleotide sequence (SEQ ID NO:525) of a native sequence PR01474 cDNA, wherein SEQ ID NO:525 is a clone designated herein as "DNA73739-1645".
  • Figure 526 shows the amino acid sequence (SEQ ID NO: 526) derived from the coding sequence of SEQ ID NO: 525 shown in Figure 525.
  • Figure 527 shows a nucleotide sequence (SEQ ID NO:527) of a native sequence PR01917 cDNA, wherein SEQ ID NO:527 is a clone designated herein as "DNA76400-2528".
  • Figure 528 shows the amino acid sequence (SEQ ID NO:528) derived from the coding sequence of SEQ ID NO:527 shown in Figure 527.
  • Figure 529 shows a nucleotide sequence (SEQ ID NO:529) of a native sequence PR01787 cDNA, wherein SEQ ID NO:529 is a clone designated herein as "DNA76510-2504".
  • Figure 530 shows the amino acid sequence (SEQ ID NO:530) derived from the coding sequence of SEQ ID NO:529 shown in Figure 529.
  • Figure 531 shows a nucleotide sequence (SEQ ID NO:531) of a native sequence PR01556 cDNA, wherein SEQ ID NO:531 is a clone designated herein as "DNA76529-1666".
  • Figure 532 shows the amino acid sequence (SEQ ID NO:532) derived from the coding sequence of SEQ
  • Figure 533 shows a nucleotide sequence (SEQ ID NO:533) of a native sequence PR01561 cDNA, wherein SEQ ID NO:533 is a clone designated herein as "DNA76538-1670".
  • Figure 534 shows the amino acid sequence (SEQ ID NO: 534) derived from the coding sequence of SEQ ID NO: 533 shown in Figure 533.
  • Figure 535 shows a nucleotide sequence (SEQ ID NO:535) of a native sequence PR01693 cDNA, wherein SEQ ID NO:535 is a clone designated herein as "DNA77301-1708".
  • Figure 536 shows the amino acid sequence (SEQ ID NO:536) derived from the coding sequence of SEQ ID NO:535 shown in Figure 535.
  • Figure 537 shows a nucleotide sequence (SEQ ID NO:537) of a native sequence PR01868 cDNA, wherein SEQ ID NO:537 is a clone designated herein as "DNA77624-2515".
  • Figure 538 shows the amino acid sequence (SEQ ID NO: 538) derived from the coding sequence of SEQ ID NO:537 shown in Figure 537.
  • Figure 539 shows a nucleotide sequence (SEQ ID NO:539) of a native sequence PRO1890 cDNA, wherein SEQ ID NO:539 is a clone designated herein as "DNA79230-2525".
  • Figure 540 shows the amino acid sequence (SEQ ID NO: 540) derived from the coding sequence of SEQ ID NO:539 shown in Figure 539.
  • Figure 541 shows a nucleotide sequence (SEQ ID NO:541) of a native sequence PR01887 cDNA, wherein SEQ ID NO:541 is a clone designated herein as "DNA79862-2522".
  • Figure 542 shows the amino acid sequence (SEQ ID NO:542) derived from the coding sequence of SEQ ID NO:541 shown in Figure 541.
  • Figure 543 shows a nucleotide sequence (SEQ ID NO:543) of a native sequence PR04353 cDNA, wherein SEQ ID NO:543 is a clone designated herein as "DNA80145-2594".
  • Figure 544 shows the amino acid sequence (SEQ ID NO:544) derived from the coding sequence of SEQ ID NO: 543 shown in Figure 543.
  • Figure 545 shows a nucleotide sequence (SEQ ID NO:545) of a native sequence PROl 801 cDNA, wherein SEQ ID NO:545 is a clone designated herein as "DNA83500-2506".
  • Figure 546 shows the amino acid sequence (SEQ ID NO : 546) derived from the coding sequence of SEQ
  • Figure 547 shows a nucleotide sequence (SEQ ID NO:547) of a native sequence PR04357 cDNA, wherein SEQ ID NO:547 is a clone designated herein as "DNA84917-2597" .
  • Figure 548 shows the amino acid sequence (SEQ ID NO:548) derived from the coding sequence of SEQ ID NO: 547 shown in Figure 547.
  • Figure 549 shows a nucleotide sequence (SEQ ID NO:549) of a native sequence PRO4302 cDNA, wherein SEQ ID NO:549 is a clone designated herein as "DNA92218-2554".
  • Figure 550 shows the amino acid sequence (SEQ ID NO:550) derived from the coding sequence of SEQ ID NO:549 shown in Figure 549.
  • PRO polypeptide and "PRO” as used herein and when immediately followed by a numerical designation refer to various polypeptides, wherein the complete designation (i.e. , PRO/number) refers to specific polypeptide sequences as described herein.
  • PRO/number polypeptide and
  • PRO/number wherein the term “number” is provided as an actual numerical designation as used herein encompass native sequence polypeptides and polypeptide variants (which are further defined herein).
  • the PRO polypeptides described herein may be isolated from a variety of sources, such as from human tissue types or from another source, or prepared by recombinant or synthetic methods.
  • the term "PRO polypeptide” refers to each individual PRO/number polypeptide disclosed herein. All disclosures in this specification which refer to the "PRO polypeptide” refer to each of the polypeptides individually as well as jointly. For example, descriptions of the preparation of, purification of, derivation of, formation of antibodies to or against, administration of, compositions containing, treatment of a disease with, etc. , pertain to each polypeptide of the invention individually.
  • the term “PRO polypeptide” also includes variants of the PRO/number polypeptides disclosed herein.
  • a “native sequence PRO polypeptide” comprises a polypeptide having the same amino acid sequence as the corresponding PRO polypeptide derived from nature. Such native sequence PRO polypeptides can be isolated from nature or can be produced by recombinant or synthetic means.
  • the term "native sequence PRO polypeptide” specifically encompasses naturally-occurring truncated or secreted forms of the specific PRO polypeptide (e.g. , an extracellular domain sequence), naturally-occurring variant forms (e.g., alternatively spliced forms) and naturally-occurring allelic variants of the polypeptide.
  • the native sequence PRO polypeptides disclosed herein are mature or full-length native sequence polypeptides comprising the full-length amino acids sequences shown in the accompanying figures. Start and stop codons are shown in bold font and underlined in the figures. However, while the PRO polypeptide disclosed in the accompanying figures are shown to begin with methionine residues designated herein as amino acid position 1 in the figures, it is conceivable and possible that other methionine residues located either upstream or downstream from the amino acid position 1 in the figures may be employed as the starting amino acid residue for the PRO polypeptides.
  • the PRO polypeptide "extracellular domain” or “ECD” refers to a form of the PRO polypeptide which is essentially free of the transmembrane and cytoplasmic domains. Ordinarily, a PRO polypeptide ECD will have less than 1 % of such transmembrane and/or cytoplasmic domains and preferably, will have less than 0.5% of such domains. It will be understood that any transmembrane domains identified for the PRO polypeptides of the present invention are identified pursuant to criteria routinely employed in the art for identifying that type of hydrophobic domain. The exact boundaries of a transmembrane domain may vary but most likely by no more than about 5 amino acids at either end of the domain as initially identified herein.
  • an extracellular domain of a PRO polypeptide may contain from about 5 or fewer amino acids on either side of the transmembrane domain/extracellular domain boundary as identified in the Examples or specification and such polypeptides, with or without the associated signal peptide, and nucleic acid encoding them, are comtemplated by the present invention.
  • cleavage of a signal sequence from a secreted polypeptide is not entirely uniform, resulting in more than one secreted species.
  • These mature polypeptides, where the signal peptide is cleaved within no more than about 5 amino acids on either side of the C-terminal boundary of the signal peptide as identified herein, and the polynucleotides encoding them, are contemplated by the present invention.
  • PRO polypeptide variant means an active PRO polypeptide as defined above or below having at least about 80% amino acid sequence identity with a full-length native sequence PRO polypeptide sequence as disclosed herein, a PRO polypeptide sequence lacking the signal peptide as disclosed herein, an extracellular domain of a PRO polypeptide, with or without the signal peptide, as disclosed herein or any other fragment of a full-length PRO polypeptide sequence as disclosed herein.
  • Such PRO polypeptide variants include, for instance, PRO polypeptides wherein one or more amino acid residues are added, or deleted, at the N- or C- terminus of the full-length native amino acid sequence.
  • a PRO polypeptide variant will have at least about 80% amino acid sequence identity, alternatively at least about 81 % amino acid sequence identity, alternatively at least about 82% amino acid sequence identity, alternatively at least about 83% amino acid sequence identity, alternatively at least about 84% amino acid sequence identity, alternatively at least about 85 % amino acid sequence identity, alternatively at least about 86% amino acid sequence identity, alternatively at least about 87% amino acid sequence identity, alternatively at least about 88% amino acid sequence identity, alternatively at least about 89% amino acid sequence identity, alternatively at least about 90% amino acid sequence identity, alternatively at least about 91 % amino acid sequence identity, alternatively at least about 92% amino acid sequence identity, alternatively at least about 93 % amino acid sequence identity, alternatively at least about 94% amino acid sequence identity, alternatively at least about 95 % amino acid sequence identity, alternatively at least about 96% amino acid sequence identity, alternatively at least about 97% amino acid sequence identity, alternatively at least about 98% amino acid sequence identity and alternatively at least about 99% amino acid sequence identity,
  • PRO variant polypeptides are at least about 10 amino acids in length, alternatively at least about 20 amino acids in length, alternatively at least about 30 amino acids in length, alternatively at least about 40 amino acids in length, alternatively at least about 50 amino acids in length, alternatively at least about 60 amino acids in length, alternatively at least about 70 amino acids in length, alternatively at least about 80 amino acids in length, alternatively at least about 90 amino acids in length, alternatively at least about 100 amino acids in length, alternatively at least about 150 amino acids in length, alternatively at least about 200 amino acids in length, alternatively at least about 300 amino acids in length, or more.
  • Percent (%) amino acid sequence identity with respect to the PRO polypeptide sequences identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical with the amino acid residues in the specific PRO polypeptide sequence, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity, and not considering any conservative substitutions as part of the sequence identity. Alignment for purposes of determining percent amino acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software. Those skilled in the art can determine appropriate parameters for measuring alignment, including any algorithms needed to achieve maximal alignment over the full length of the sequences being compared.
  • % amino acid sequence identity values are generated using the sequence comparison computer program ALIGN-2, wherein the complete source code for the ALIGN-2 program is provided in Table 1 below.
  • the ALIGN-2 sequence comparison computer program was authored by Genentech, Inc. and the source code shown in Table 1 below has been filed with user documentation in the U.S. Copyright Office, Washington D.C., 20559, where it is registered under U.S. Copyright Registration No. TXU510087.
  • the ALIGN-2 program is publicly available through Genentech, Inc., South San Francisco, California or may be compiled from the source code provided in Table 1 below.
  • the ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.
  • % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B is calculated as follows:
  • Tables 2 and 3 demonstrate how to calculate the % amino acid sequence identity of the amino acid sequence designated "Comparison Protein” to the amino acid sequence designated "PRO” , wherein “PRO” represents the amino acid sequence of a hypothetical PRO polypeptide of interest, “Comparison Protein” represents the amino acid sequence of a polypeptide against which the "PRO” polypeptide of interest is being compared, and "X, " Y” and “Z” each represent different hypothetical amino acid residues.
  • a % amino acid sequence identity value is determined by dividing (a) the number of matching identical amino acid residues between the amino acid sequence of the PRO polypeptide of interest having a sequence derived from the native PRO polypeptide and the comparison amino acid sequence of interest (i.e. , the sequence against which the PRO polypeptide of interest is being compared which may be a PRO variant polypeptide) as determined by WU-BLAST-2 by (b) the total number of amino acid residues of the PRO polypeptide of interest.
  • amino acid sequence A is the comparison amino acid sequence of interest and the amino acid sequence B is the amino acid sequence of the PRO polypeptide of interest.
  • Percent amino acid sequence identity may also be determined using the sequence comparison program NCBI-BLAST2 (Altschul et al. , Nucleic Acids Res. 25:3389-3402 (1997)).
  • NCBI-BLAST2 sequence comparison program may be downloaded from http://www.ncbi.nlm.nih.gov or otherwise obtained from the National Institute of Health, Bethesda, MD.
  • % amino acid sequence identity of a given amino acid sequence A to, with, or against a given amino acid sequence B is calculated as follows:
  • PRO variant polynucleotide or "PRO variant nucleic acid sequence” means a nucleic acid molecule which encodes an active PRO polypeptide as defined below and which has at least about 80% nucleic acid sequence identity with a nucleotide acid sequence encoding a full-length native sequence PRO polypeptide sequence as disclosed herein, a full-length native sequence PRO polypeptide sequence lacking the signal peptide as disclosed herein, an extracellular domain of a PRO polypeptide, with or without the signal peptide, as disclosed herein or any other fragment of a full-length PRO polypeptide sequence as disclosed herein.
  • a PRO variant polynucleotide will have at least about 80% nucleic acid sequence identity, alternatively at least about 81 % nucleic acid sequence identity, alternatively at least about 82% nucleic acid sequence identity, alternatively at least about 83 % nucleic acid sequence identity, alternatively at least about 84 % nucleic acid sequence identity, alternatively at least about 85% nucleic acid sequence identity, alternatively at least about 86% nucleic acid sequence identity, alternatively at least about 87% nucleic acid sequence identity, alternatively at least about 88% nucleic acid sequence identity, alternatively at least about 89% nucleic acid sequence identity, alternatively at least about 90% nucleic acid sequence identity, alternatively at least about 91 % nucleic acid sequence identity, alternatively at least about 92% nucleic acid sequence identity, alternatively at least about 93 % nucleic acid sequence identity, alternatively at least about 94% nucleic acid sequence identity, alternatively at least about 95% nucleic acid sequence identity, alternatively at least about 96%
  • PRO variant polynucleotides are at least about 30 nucleotides in length, alternatively at least about 60 nucleotides in length, alternatively at least about 90 nucleotides in length, alternatively at least about 120 nucleotides in length, alternatively at least about 150 nucleotides in length, alternatively at least about 180 nucleotides in length, alternatively at least about 210 nucleotides in length, alternatively at least about 240 nucleotides in length, alternatively at least about 270 nucleotides in length, alternatively at least about 300 nucleotides in length, alternatively at least about 450 nucleotides in length, alternatively at least about 600 nucleotides in length, alternatively at least about 900 nucleotides in length, or more.
  • Percent (%) nucleic acid sequence identity with respect to PRO-encoding nucleic acid sequences identified herein is defined as the percentage of nucleotides in a candidate sequence that are identical with the nucleotides in the PRO nucleic acid sequence of interest, after aligning the sequences and introducing gaps, if necessary, to achieve the maximum percent sequence identity. Alignment for purposes of determining percent nucleic acid sequence identity can be achieved in various ways that are within the skill in the art, for instance, using publicly available computer software such as BLAST, BLAST-2, ALIGN or Megalign (DNASTAR) software.
  • % nucleic acid sequence identity values are generated using the sequence comparison computer program ALIGN-2, wherein the complete source code for the ALIGN-2 program is provided in Table 1 below.
  • the ALIGN-2 sequence comparison computer program was authored by Genentech, Inc. and the source code shown in Table 1 below has been filed with user documentation in the U.S. Copyright Office, Washington D.C. , 20559, where it is registered under U.S. Copyright Registration No. TXU510087.
  • the ALIGN-2 program is publicly available through Genentech, Inc., South San Francisco, California or may be compiled from the source code provided in Table 1 below.
  • the ALIGN-2 program should be compiled for use on a UNIX operating system, preferably digital UNIX V4.0D. All sequence comparison parameters are set by the ALIGN-2 program and do not vary.
  • the % nucleic acid sequence identity of a given nucleic acid sequence C to, with, or against a given nucleic acid sequence D is calculated as follows:
  • Tables 4 and 5 demonstrate how to calculate the % nucleic acid sequence identity of the nucleic acid sequence designated "Comparison DNA” to the nucleic acid sequence designated "PRO-DNA”, wherein "PRO-DNA” represents a hypothetical PRO-encoding nucleic acid sequence of interest, “Comparison DNA” represents the nucleotide sequence of a nucleic acid molecule against which the "PRO-DNA” nucleic acid molecule of interest is being compared, and "N", “L” and “V” each represent different hypothetical nucleotides.
  • a % nucleic acid sequence identity value is determined by dividing (a) the number of matching identical nucleotides between the nucleic acid sequence of the PRO polypeptide-encoding nucleic acid molecule of interest having a sequence derived from the native sequence PRO polypeptide-encoding nucleic acid and the comparison nucleic acid molecule of interest (i.e. , the sequence against which the PRO polypeptide-encoding nucleic acid molecule of interest is being compared which may be a variant PRO polynucleotide) as determined by WU-BLAST-2 by (b) the total number of nucleotides of the PRO polypeptide-encoding nucleic acid molecule of interest.
  • nucleic acid sequence A is the comparison nucleic acid molecule of interest and the nucleic acid sequence B is the nucleic acid sequence of the PRO polypeptide- encoding nucleic acid molecule of interest.
  • Percent nucleic acid sequence identity may also be determined using the sequence comparison program
  • NCBI-BLAST2 (Altschul et al., Nucleic Acids Res. 25:3389-3402 (1997)).
  • the NCBI-BLAST2 sequence comparison program may be downloaded from http://www.ncbi.nlm.nih.gov or otherwise obtained from the National Institute of Health, Bethesda, MD.
  • % nucleic acid sequence identity of a given nucleic acid sequence C to, with, or against a given nucleic acid sequence D is calculated as follows:
  • the % nucleic acid sequence identity of C to D will not equal the % nucleic acid sequence identity of D to C.
  • PRO variant polynucleotides are nucleic acid molecules that encode an active PRO polypeptide and which are capable of hybridizing, preferably under stringent hybridization and wash conditions, to nucleotide sequences encoding a full-length PRO polypeptide as disclosed herein.
  • PRO variant polypeptides may be those that are encoded by a PRO variant polynucleotide.
  • Isolated, " when used to describe the various polypeptides disclosed herein, means polypeptide that has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials that would typically interfere with diagnostic or therapeutic uses for the polypeptide, and may include enzymes, hormones, and other proteinaceous or non-proteinaceous solutes.
  • the polypeptide will be purified (1) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (2) to homogeneity by SDS-PAGE under non-reducing or reducing conditions using Coomassie blue or, preferably, silver stain.
  • Isolated polypeptide includes polypeptide in situ within recombinant cells, since at least one component of the PRO polypeptide natural environment will not be present. Ordinarily, however, isolated polypeptide will be prepared by at least one purification step.
  • An "isolated" PRO polypeptide-encoding nucleic acid or other polypeptide-encoding nucleic acid is a nucleic acid molecule that is identified and separated from at least one contaminant nucleic acid molecule with which it is ordinarily associated in the natural source of the polypeptide-encoding nucleic acid.
  • An isolated polypeptide-encoding nucleic acid molecule is other than in the form or setting in which it is found in nature. Isolated polypeptide-encoding nucleic acid molecules therefore are distinguished from the specific polypeptide- encoding nucleic acid molecule as it exists in natural cells.
  • an isolated polypeptide-encoding nucleic acid molecule includes polypeptide-encoding nucleic acid molecules contained in cells that ordinarily express the polypeptide where, for example, the nucleic acid molecule is in a chromosomal location different from that of natural cells.
  • control sequences refers to DNA sequences necessary for the expression of an operably linked coding sequence in a particular host organism.
  • the control sequences that are suitable for prokaryotes include a promoter, optionally an operator sequence, and a ribosome binding site.
  • Eukaryotic cells are known to utilize promoters, polyadenylation signals, and enhancers.
  • Nucleic acid is "operably linked" when it is placed into a functional relationship with another nucleic acid sequence.
  • DNA for a presequence or secretory leader is operably linked to DNA for a polypeptide if it is expressed as a preprotein that participates in the secretion of the polypeptide;
  • a promoter or enhancer is operably linked to a coding sequence if it affects the transcription of the sequence; or
  • a ribosome binding site is operably linked to a coding sequence if it is positioned so as to facilitate translation.
  • "operably linked” means that the DNA sequences being linked are contiguous, and, in the case of a secretory leader, contiguous and in reading phase.
  • antibody is used in the broadest sense and specifically covers, for example, single anti-PRO monoclonal antibodies (including agonist, antagonist, and neutralizing antibodies), anti-PRO antibody compositions with polyepitopic specificity, single chain anti-PRO antibodies, and fragments of anti-PRO antibodies (see below).
  • monoclonal antibody refers to an antibody obtained from a population of substantially homogeneous antibodies, i.e. , the individual antibodies comprising the population are identical except for possible naturally-occurring mutations that may be present in minor amounts.
  • “Stringency” of hybridization reactions is readily determinable by one of ordinary skill in the art, and generally is an empirical calculation dependent upon probe length, washing temperature, and salt concentration. In general, longer probes require higher temperatures for proper annealing, while shorter probes need lower temperatures. Hybridization generally depends on the ability of denatured DNA to reanneal when complementary strands are present in an environment below their melting temperature. The higher the degree of desired homology between the probe and hybridizable sequence, the higher the relative temperature which can be used. As a result, it follows that higher relative temperatures would tend to make the reaction conditions more stringent, while lower temperatures less so. For additional details and explanation of stringency of hybridization reactions, see Ausubel et al., Current Protocols in Molecular Biology. Wiley Interscience Publishers, (1995).
  • “Stringent conditions” or “high stringency conditions”, as defined herein, may be identified by those that: (1) employ low ionic strength and high temperature for washing, for example 0.015 M sodium chloride/0.0015 M sodium citrate/0.1 % sodium dodecyl sulfate at 50°C; (2) employ during hybridization a denaturing agent, such as formamide, for example, 50% (v/v) formamide with 0.1 % bovine serum albumin/0.1 % Ficoll/0.1 % polyvinylpyrrolidone/50mM sodium phosphate buffer at pH 6.5 with 750 mM sodium chloride, 75 mM sodium citrate at 42°C; or (3) employ 50% formamide, 5 x SSC (0.75 M NaCI, 0.075 M sodium citrate), 50 mM sodium phosphate (pH 6.8), 0.1 % sodium pyrophosphate, 5 x Denhardt's solution, sonicated salmon sperm DNA (50 ⁇ g/ml), 0.1% SDS, and 10%
  • Modely stringent conditions may be identified as described by Sambrook et al., Molecular Cloning: A Laboratory Manual. New York: Cold Spring Harbor Press, 1989, and include the use of washing solution and hybridization conditions (e.g., temperature, ionic strength and %SDS) less stringent that those described above.
  • washing solution and hybridization conditions e.g., temperature, ionic strength and %SDS
  • An example of moderately stringent conditions is overnight incubation at 37°C in a solution comprising: 20% formamide, 5 x SSC (150 mM NaCI, 15 mM trisodium citrate), 50 mM sodium phosphate (pH
  • epitope tagged when used herein refers to a chimeric polypeptide comprising a PRO polypeptide fused to a "tag polypeptide" .
  • the tag polypeptide has enough residues to provide an epitope against which an antibody can be made, yet is short enough such that it does not interfere with activity of the polypeptide to which it is fused.
  • the tag polypeptide preferably also is fairly unique so that the antibody does not substantially cross-react with other epitopes.
  • Suitable tag polypeptides generally have at least six amino acid residues and usually between about 8 and 50 amino acid residues (preferably, between about 10 and 20 amino acid residues).
  • immunoadhesin designates antibody-like molecules which combine the binding specificity of a heterologous protein (an “adhesin”) with the effector functions of immunoglobulin constant domains.
  • the immunoadhesins comprise a fusion of an amino acid sequence with the desired binding specificity which is other than the antigen recognition and binding site of an antibody (i.e., is “heterologous"), and an immunoglobulin constant domain sequence.
  • the adhesin part of an immunoadhesin molecule typically is a contiguous amino acid sequence comprising at least the binding site of a receptor or a ligand.
  • the immunoglobulin constant domain sequence in the immunoadhesin may be obtained from any immunoglobulin, such as IgG-1, IgG-2, IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE, IgD or IgM.
  • immunoglobulin such as IgG-1, IgG-2, IgG-3, or IgG-4 subtypes, IgA (including IgA-1 and IgA-2), IgE, IgD or IgM.
  • Active or “activity” for the purposes herein refers to form(s) of a PRO polypeptide which retain a biological and/or an immunological activity of native or naturally-occurring PRO, wherein "biological” activity refers to a biological function (either inhibitory or stimulatory) caused by a native or naturally-occurring PRO other than the ability to induce the production of an antibody against an antigenic epitope possessed by a native or naturally-occurring PRO and an "immunological” activity refers to the ability to induce the production of an antibody against an antigenic epitope possessed by a native or naturally-occurring PRO.
  • antagonist is used in the broadest sense, and includes any molecule that partially or fully blocks, inhibits, or neutralizes a biological activity of a native PRO polypeptide disclosed herein.
  • agonist is used in the broadest sense and includes any molecule that mimics a biological activity of a native PRO polypeptide disclosed herein.
  • Suitable agonist or antagonist molecules specifically include agonist or antagonist antibodies or antibody fragments, fragments or amino acid sequence variants of native PRO polypeptides, peptides, antisense oligonucleotides, small organic molecules, etc.
  • Methods for identifying agonists or antagonists of a PRO polypeptide may comprise contacting a PRO polypeptide with a candidate agonist or antagonist molecule and measuring a detectable change in one or more biological activities normally associated with the PRO polypeptide.
  • Treatment refers to both therapeutic treatment and prophylactic or preventative measures, wherein the object is to prevent or slow down (lessen) the targeted pathologic condition or disorder.
  • Those in need of treatment include those already with the disorder as well as those prone to have the disorder or those in whom the disorder is to be prevented.
  • “Chronic” administration refers to administration of the agent(s) in a continuous mode as opposed to an acute mode, so as to maintain the initial therapeutic effect (activity) for an extended period of time.
  • “Intermittent” administration is treatment that is not consecutively done without interruption, but rather is cyclic in nature.
  • “Mammal” for purposes of treatment refers to any animal classified as a mammal, including humans, domestic and farm animals, and zoo, sports, or pet animals, such as dogs, cats, cattle, horses, sheep, pigs, goats, rabbits, etc. Preferably, the mammal is human.
  • Administration "in combination with" one or more further therapeutic agents includes simultaneous (concurrent) and consecutive administration in any order.
  • Carriers as used herein include pharmaceutically acceptable carriers, excipients, or stabilizers which are nontoxic to the cell or mammal being exposed thereto at the dosages and concentrations employed. Often the physiologically acceptable carrier is an aqueous pH buffered solution.
  • physiologically acceptable carriers include buffers such as phosphate, citrate, and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptide; proteins, such as serum albumin, gelatin, or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone; amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides, and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt- forming counterions such as sodium; and/or nonionic surfactants such as TWEENTM, polyethylene glycol (PEG), and PLURONICSTM.
  • buffers such as phosphate, citrate, and other organic acids
  • antioxidants including ascorbic acid
  • proteins such as serum albumin
  • Antibody fragments comprise a portion of an intact antibody, preferably the antigen binding or variable region of the intact antibody.
  • antibody fragments include Fab, Fab', F(ab') 2 , and Fv fragments; diabodies; linear antibodies (Zapata et al., Protein Eng. 8(10): 1057-1062 [1995]); single-chain antibody molecules; and multispecific antibodies formed from antibody fragments.
  • Papain digestion of antibodies produces two identical antigen-binding fragments, called “Fab” fragments, each with a single antigen-binding site, and a residual “Fc” fragment, a designation reflecting the ability to crystallize readily.
  • Pepsin treatment yields an F(ab') 2 fragment that has two antigen-combining sites and is still capable of cross-linking antigen.
  • Fv is the minimum antibody fragment which contains a complete antigen-recognition and -binding site. This region consists of a dimer of one heavy- and one light-chain variable domain in tight, non-covalent association. It is in this configuration that the three CDRs of each variable domain interact to define an antigen- binding site on the surface of the V H -V L dimer.
  • the six CDRs confer antigen-binding specificity to the antibody.
  • a single variable domain or half of an Fv comprising only three CDRs specific for an antigen
  • the Fab fragment also contains the constant domain of the light chain and the first constant domain (CHI) of the heavy chain.
  • Fab fragments differ from Fab' fragments by the addition of a few residues at the carboxy terminus of the heavy chain CHI domain including one or more cysteines from the antibody hinge region .
  • Fab ' -SH is the designation herein for Fab ' in which the cysteine residue(s) of the constant domains bear a free thiol group.
  • F(ab') 2 antibody fragments originally were produced as pairs of Fab' fragments which have hinge cysteines between them. Other chemical couplings of antibody fragments are also known.
  • the "light chains" of antibodies (immunoglobulins) from any vertebrate species can be assigned to one of two clearly distinct types, called kappa and lambda, based on the amino acid sequences of their constant domains.
  • immunoglobulins can be assigned to different classes. There are five major classes of immunoglobulins: IgA, IgD, IgE, IgG, and IgM , and several of these may be further divided into subclasses (isotypes) , e.g., IgGl , IgG2, IgG3 , IgG4 , IgA , and IgA2.
  • Single-chain Fv or “sFv” antibody fragments comprise the V H and V L domains of antibody, wherein these domains are present in a single polypeptide chain.
  • the Fv polypeptide further comprises a polypeptide linker between the V H and V L domains which enables the sFv to form the desired structure for antigen binding .
  • diabodies refers to small antibody fragments with two antigen-binding sites, which fragments comprise a heavy-chain variable domain (V H ) connected to a light-chain variable domain (VJ in the same polypeptide chain (V H -V L ).
  • V H heavy-chain variable domain
  • VJ light-chain variable domain
  • linker that is too short to allow pairing between the two domains on the same chain, the domains are forced to pair with the complementary domains of another chain and create two antigen-binding sites.
  • Diabodies are described more fully in, for example, EP 404,097; WO 93/11161 ; and Hollinger et al., Proc. Natl. Acad. Sci. USA. 90:6444-6448 (1993).
  • an “isolated” antibody is one which has been identified and separated and/or recovered from a component of its natural environment. Contaminant components of its natural environment are materials which would interfere with diagnostic or therapeutic uses for the antibody, and may include enzymes, hormones, and other proteinaceous or nonproteinaceous solutes.
  • the antibody will be purified (1) to greater than 95% by weight of antibody as determined by the Lowry method, and most preferably more than 99% by weight, (2) to a degree sufficient to obtain at least 15 residues of N-terminal or internal amino acid sequence by use of a spinning cup sequenator, or (3) to homogeneity by SDS-PAGE under reducing or nonreducing conditions using Coomassie blue or, preferably, silver stain.
  • Isolated antibody includes the antibody in situ within recombinant cells since at least one component of the antibody's natural environment will not be present. Ordinarily, however, isolated antibody will be prepared by at least one purification step.
  • An antibody that "specifically binds to” or is “specific for” a particular polypeptide or an epitope on a particular polypeptide is ope that binds to that particular polypeptide or epitope on a particular polypeptide without substantially binding to any other polypeptide or polypeptide epitope.
  • label when used herein refers to a detectable compound or composition which is conjugated directly or indirectly to the antibody so as to generate a "labeled” antibody.
  • the label may be detectable by itself (e.g. radioisotope labels or fluorescent labels) or, in the case of an enzymatic label, may catalyze chemical alteration of a substrate compound or composition which is detectable.
  • solid phase is meant a non-aqueous matrix to which the antibody of the present invention can adhere. Examples of solid phases encompassed herein include those formed partially or entirely of glass (e.g. , controlled pore glass), polysaccharides (e.g., agarose), polyacrylamides, polystyrene, polyvinyl alcohol and silicones.
  • the solid phase can comprise the well of an assay plate; in others it is a purification column (e.g., an affinity chromatography column).
  • This term also includes a discontinuous solid phase of discrete particles, such as those described in U.S. Patent No. 4,275,149.
  • a “liposome” is a small vesicle composed of various types of lipids, phospholipids and/or surfactant which is useful for delivery of a drug (such as a PRO polypeptide or antibody thereto) to a mammal.
  • a drug such as a PRO polypeptide or antibody thereto
  • the components of the liposome are commonly arranged in a bilayer formation, similar to the lipid arrangement of biological membranes.
  • a "small molecule” is defined herein to have a molecular weight below about 500 Daltons.
  • an “effective amount” of a polypeptide disclosed herein or an agonist or antagonist thereof is an amount sufficient to carry out a specifically stated purpose.
  • An “effective amount” may be determined empirically and in a routine manner, in relation to the stated purpose.
  • filel and file2 are two dna or two protein sequences.
  • Max file length is 65535 (limited by unsigned short x in the jmp struct)
  • a sequence with 1/3 or more of its elements ACGTU is assumed to be DNA
  • the program may create a tmp file in /tmp to hold info about traceback.
  • dumpblockO * putline() put out a line (name, [num], seq, [num]): dumpblockO
  • static nm matches in core — for checking */ static lmax; /* lengths of stripped file names */ static UP]; /* jmp index for a path */ static nc[2]; /* number at start of current line */ static ni[2]; /* current elem number — for gapping */ static siz[2]; static char *ps[2]; /* ptr to current element */ static char *po[2]; /* ptr to next output char slot */ static char out[2][P LINE]; /* output line */ static char starfP LINE]; /* set by stars() *//
  • *ps[i] toupper(*ps[i]); po[i] + +; ps[i] + + ;
  • # include "nw.h” ⁇ include ⁇ sys/file.h> char *jname " /tmp/homgXXXXX " ; /* tmp file for jmps */
  • *py+ + toupper(*px); if (index("ATGCU",*(py-l))) natgc + + ; ⁇
  • the present invention provides newly identified and isolated nucleotide sequences encoding polypeptides referred to in the present application as PRO polypeptides.
  • cDNAs encoding various PRO polypeptides have been identified and isolated, as disclosed in further detail in the Examples below. It is noted that proteins produced in separate expression rounds may be given different PRO numbers but the UNQ number is unique for any given DNA and the encoded protein, and will not be changed.
  • PRO/number the protein encoded by the full length native nucleic acid molecules disclosed herein as well as all further native homologues and variants included in the foregoing definition of PRO, will be referred to as "PRO/number" , regardless of their origin or mode of preparation.
  • PRO variants can be prepared.
  • PRO variants can be prepared by introducing appropriate nucleotide changes into the PRO DNA, and/or by synthesis of the desired PRO polypeptide.
  • amino acid changes may alter post-translational processes of the PRO, such as changing the number or position of glycosylation sites or altering the membrane anchoring characteristics.
  • Variations in the native full-length sequence PRO or in various domains of the PRO described herein can be made, for example, using any of the techniques and guidelines for conservative and non-conservative mutations set forth, for instance, in U.S. Patent No. 5,364,934.
  • Variations may be a substitution, deletion or insertion of one or more codons encoding the PRO that results in a change in the amino acid sequence of the PRO as compared with the native sequence PRO.
  • the variation is by substitution of at least one amino acid with any other amino acid in one or more of the domains of the PRO.
  • Guidance in determining which amino acid residue may be inserted, substituted or deleted without adversely affecting the desired activity may be found by comparing the sequence of the PRO with that of homologous known protein molecules and minimizing the number of amino acid sequence changes made in regions of high homology.
  • Amino acid substitutions can be the result of replacing one amino acid with another amino acid having similar structural and/or chemical properties, such as the replacement of a leucine with a serine, i.e., conservative amino acid replacements.
  • Insertions or deletions may optionally be in the range of about 1 to 5 amino acids. The variation allowed may be determined by systematically making insertions, deletions or substitutions of amino acids in the sequence and testing the resulting variants for activity exhibited by the full-length or mature native sequence.
  • PRO polypeptide fragments are provided herein. Such fragments may be truncated at the N-terminus or C-terminus, or may lack internal residues, for example, when compared with a full length native protein. Certain fragments lack amino acid residues that are not essential for a desired biological activity of the PRO polypeptide.
  • PRO fragments may be prepared by any of a number of conventional techniques. Desired peptide fragments may be chemically synthesized. An alternative approach involves generating PRO fragments by enzymatic digestion, e.g., by treating the protein with an enzyme known to cleave proteins at sites defined by particular amino acid residues, or by digesting the DNA with suitable restriction enzymes and isolating the desired fragment. Yet another suitable technique involves isolating and amplifying a DNA fragment encoding a desired polypeptide fragment, by polymerase chain reaction (PCR). Oligonucleotides that define the desired termini of the DNA fragment are employed at the 5' and 3' primers in the PCR. Preferably, PRO polypeptide fragments share at least one biological and/or immunological activity with the native PRO polypeptide disclosed herein.
  • PCR polymerase chain reaction
  • conservative substitutions of interest are shown in Table 6 under the heading of preferred substitutions. If such substitutions result in a change in biological activity, then more substantial changes, denominated exemplary substitutions in Table 6, or as further described below in reference to amino acid classes, are introduced and the products screened.
  • Val (V) ile; leu; met; phe; ala; norleucine leu Substantial modifications in function or immunological identity of the PRO polypeptide are accomplished by selecting substitutions that differ significantly in their effect on maintaining (a) the structure of the polypeptide backbone in the area of the substitution, for example, as a sheet or helical conformation, (b) the charge or hydrophobicity of the molecule at the target site, or (c) the bulk of the side chain.
  • Naturally occurring residues are divided into groups based on common side-chain properties: (1) hydrophobic: norleucine, met, ala, val, leu, ile;
  • Non-conservative substitutions will entail exchanging a member of one of these classes for another class. Such substituted residues also may be introduced into the conservative substitution sites or, more preferably, into the remaining (non-conserved) sites.
  • oligonucleotide-mediated (site- directed) mutagenesis alanine scanning, and PCR mutagenesis.
  • Site-directed mutagenesis [Carter et al. , Nucl. Acids Res .. 13:4331 (1986); Zoller et al., Nucl. Acids Res .. 10:6487 (1987)]
  • cassette mutagenesis [Wells et al.. Gene. 34:315 (1985)]
  • restriction selection mutagenesis [Wells et al., Philos. Trans. R. Soc. London Ser A.
  • Scanning amino acid analysis can also be employed to identify one or more amino acids along a contiguous sequence.
  • preferred scanning amino acids are relatively small, neutral amino acids.
  • Such amino acids include alanine, glycine, serine, and cysteine.
  • Alanine is typically a preferred scanning amino acid among this group because it eliminates the side-chain beyond the beta-carbon and is less likely to alter the main- chain conformation of the variant [Cunningham and Wells, Science. 244: 1081-1085 (1989)]. Alanine is also typically preferred because it is the most common amino acid.
  • Covalent modifications of PRO are included within the scope of this invention.
  • One type of covalent modification includes reacting targeted amino acid residues of a PRO polypeptide with an organic derivatizing agent that is capable of reacting with selected side chains or the N- or C- terminal residues of the PRO.
  • Derivatization with bifunctional agents is useful, for instance, for crosslinking PRO to a water-insoluble support matrix or surface for use in the method for purifying anti-PRO antibodies, and vice-versa.
  • crosslinking agents include, e.g., l,l-bis(diazoacetyl)-2-phenylethane, glutaraldehyde, N-hydroxysuccinimide esters, for example, esters with 4-azidosalicylic acid, homobifunctional imidoesters, including disuccinimidyl esters such as 3,3'-dithiobis(succinimidylpropionate), bifunctional maleimides such as bis-N-maleimido-1,8- octane and agents such as methyl-3-[(p-azidophenyl)dithio]propioimidate.
  • Another type of covalent modification of the PRO polypeptide included within the scope of this invention comprises altering the native glycosylation pattern of the polypeptide.
  • "Altering the native glycosylation pattern” is intended for purposes herein to mean deleting one or more carbohydrate moieties found in native sequence PRO (either by removing the underlying glycosylation site or by deleting the glycosylation by chemical and/or enzymatic means), and/or adding one or more glycosylation sites that are not present in the native sequence PRO.
  • the phrase includes qualitative changes in the glycosylation of the native proteins, involving a change in the nature and proportions of the various carbohydrate moieties present.
  • Addition of glycosylation sites to the PRO polypeptide may be accomplished by altering the amino acid sequence.
  • the alteration may be made, for example, by the addition of, or substitution by, one or more serine or threonine residues to the native sequence PRO (for O-linked glycosylation sites).
  • the PRO amino acid sequence may optionally be altered through changes at the DNA level, particularly by mutating the DNA encoding the PRO polypeptide at preselected bases such that codons are generated that will translate into the desired amino acids.
  • Another means of increasing the number of carbohydrate moieties on the PRO polypeptide is by chemical or enzymatic coupling of glycosides to the polypeptide. Such methods are described in the art, e.g., in WO 87/05330 published 11 September 1987, and in Aplin and Wriston, CRC Crit. Rev. Biochem.. pp. 259- 306 (1981).
  • Removal of carbohydrate moieties present on the PRO polypeptide may be accomplished chemically or enzymatically or by mutational substitution of codons encoding for amino acid residues that serve as targets for glycosylation.
  • Chemical deglycosylation techniques are known in the art and described, for instance, by Haki uddin, et al., Arch. Biochem. Biophvs.. 259:52 (1987) and by Edge et al., Anal. Biochem.. 118: 131 (1981).
  • Enzymatic cleavage of carbohydrate moieties on polypeptides can be achieved by the use of a variety of endo- and exo-glycosidases as described by Thotakura et al., Meth. Enzymol.. 138:350 (1987).
  • PRO polypeptide to one of a variety of nonproteinaceous polymers, e.g., polyethylene glycol (PEG), polypropylene glycol, or polyoxyalkylenes, in the manner set forth in U.S. Patent Nos. 4,640,835; 4,496,689; 4,301 , 144; 4,670,417; 4,791 , 192 or 4, 179,337.
  • PEG polyethylene glycol
  • polypropylene glycol polypropylene glycol
  • polyoxyalkylenes polyoxyalkylenes
  • the PRO of the present invention may also be modified in a way to form a chimeric molecule comprising PRO fused to another, heterologous polypeptide or amino acid sequence.
  • such a chimeric molecule comprises a fusion of the PRO with a tag polypeptide which provides an epitope to which an anti-tag antibody can selectively bind.
  • the epitope tag is generally placed at the amino- or carboxyl- terminus of the PRO. The presence of such epitope-tagged forms of the PRO can be detected using an antibody against the tag polypeptide. Also, provision of the epitope tag enables the PRO to be readily purified by affinity purification using an anti-tag antibody or another type of affinity matrix that binds to the epitope tag.
  • tag polypeptides and their respective antibodies are well known in the art.
  • poly-histidine poly-his
  • poly-histidine-glycine poly-his-glycine tags
  • flu HA tag polypeptide and its antibody 12CA5 [Field et al., Mol. Cell. Biol.. 8:2159-2165 (1988)]
  • c-myc tag and the 8F9, 3C7, 6E10, G4, B7 and 9E10 antibodies thereto [Evan et al.. Molecular and Cellular Biology. 5:3610-3616 (1985)]
  • Herpes Simplex virus glycoprotein D (gD) tag and its antibody [Paborsky et al. , Protein Engineering. 3(6):547- 553 (1990)].
  • tag polypeptides include the Flag-peptide [Hopp et al. , BioTechnology. 6: 1204-1210 (1988)]; the KT3 epitope peptide [Martin et al., Science. 255: 192-194 (1992)]; an ⁇ -tubulin epitope peptide [Skinner et al., J. Biol. Chem.. 266: 15163-15166 (1991)]; and the T7 gene 10 protein peptide tag [Lutz- Freyermuth et al., Proc. Natl. Acad. Sci. USA. 87:6393-6397 (1990)].
  • the chimeric molecule may comprise a fusion of the PRO with an immunoglobulin or a particular region of an immunoglobulin.
  • an immunoglobulin also referred to as an "immunoadhesin”
  • a fusion could be to the Fc region of an IgG molecule.
  • the Ig fusions preferably include the substitution of a soluble (transmembrane domain deleted or inactivated) form of a PRO polypeptide in place of at least one variable region within an Ig molecule.
  • the immunoglobulin fusion includes the hinge, CH2 and CH3, or the hinge, CHI, CH2 and CH3 regions of an IgG 1 molecule.
  • PRO sequence or portions thereof, may be produced by direct peptide synthesis using solid-phase techniques [see, e.g., Stewart et al.. Solid-Phase Peptide Synthesis. W.H. Freeman Co., San Francisco, CA (1969); Merrifield, J. Am. Chem. Soc. 85:2149-2154 (1963)]. In vitro protein synthesis may be performed using manual techniques or by automation.
  • Automated synthesis may be accomplished, for instance, using an Applied Biosystems Peptide Synthesizer (Foster City, CA) using manufacturer's instructions.
  • Various portions of the PRO may be chemically synthesized separately and combined using chemical or enzymatic methods to produce the full-length PRO.
  • DNA encoding PRO may be obtained from a cDNA library prepared from tissue believed to possess the PRO mRNA and to express it at a detectable level. Accordingly, human PRO DNA can be conveniently obtained from a cDNA library prepared from human tissue, such as described in the Examples.
  • the PRO- encoding gene may also be obtained from a genomic library or by known synthetic procedures (e.g. , automated nucleic acid synthesis). Libraries can be screened with probes (such as antibodies to the PRO or oligonucleotides of at least about 20-80 bases) designed to identify the gene of interest or the protein encoded by it. Screening the cDNA or genomic library with the selected probe may be conducted using standard procedures, such as described in
  • the Examples below describe techniques for screening a cDNA library.
  • the oligonucleotide sequences selected as probes should be of sufficient length and sufficiently unambiguous that false positives are minimized.
  • the oligonucleotide is preferably labeled such that it can be detected upon hybridization to DNA in the library being screened.
  • Methods of labeling are well known in the art, and include the use of radiolabels like 32 P-labeled ATP, biotinylation or enzyme labeling.
  • Hybridization conditions including moderate stringency and high stringency, are provided in Sambrook et al., supra.
  • Sequences identified in such library screening methods can be compared and aligned to other known sequences deposited and available in public databases such as GenBank or other private sequence databases. Sequence identity (at either the amino acid or nucleotide level) within defined regions of the molecule or across the full-length sequence can be determined using methods known in the art and as described herein.
  • Nucleic acid having protein coding sequence may be obtained by screening selected cDNA or genomic libraries using the deduced amino acid sequence disclosed herein for the first time, and, if necessary, using conventional primer extension procedures as described in Sambrook et al., supra, to detect precursors and processing intermediates of mRNA that may not have been reverse-transcribed into cDNA.
  • Host cells are transfected or transformed with expression or cloning vectors described herein for PRO production and cultured in conventional nutrient media modified as appropriate for inducing promoters, selecting transformants, or amplifying the genes encoding the desired sequences.
  • the culture conditions such as media, temperature, pH and the like, can be selected by the skilled artisan without undue experimentation. In general, principles, protocols, and practical techniques for maximizing the productivity of cell cultures can be found in Mammalian Cell Biotechnology: a Practical Approach. M. Butler, ed. (IRL Press, 1991) and Sambrook et al., supra.
  • Methods of eukaryotic cell transfection and prokaryotic cell transformation are known to the ordinarily skilled artisan, for example, CaCl 2 , CaP0 4 , liposome-mediated and electroporation.
  • transformation is performed using standard techniques appropriate to such cells.
  • the calcium treatment employing calcium chloride, as described in Sambrook et al., supra, or electroporation is generally used for prokaryotes.
  • Infection with Agrobacterium tumefaciens is used for transformation of certain plant cells, as described by Shaw et al., Gene. 23:315 (1983) and WO 89/05859 published 29 June 1989.
  • polybrene polyornithine
  • Suitable host cells for cloning or expressing the DNA in the vectors herein include prokaryote, yeast, or higher eukaryote cells.
  • Suitable prokaryotes include but are not limited to eubacteria, such as Gram-negative or Gram-positive organisms, for example, Enterobacteriaceae such as E. coli.
  • E. coli strains are publicly available, such as E.
  • coli K12 strain MM294 ATCC 31,446
  • E. coli X1776 ATCC 31,537)
  • E. coli strain W3110 ATCC 27,325)
  • K5 772 ATCC 53,635
  • Other suitable prokaryotic host cells include Enterobacteriaceae such as Escherichia, e.g., E. coli, Enterobacter, Erwinia, Klebsiella, Proteus, Salmonella, e.g., Salmonella typhimurium, Serratia, e.g., Serratia marcescans, and Shigella, as well as Bacilli such as B. subtilis and B. licheniformis (e.g., B.
  • Strain W3110 is one particularly preferred host or parent host because it is a common host strain for recombinant DNA product fermentations. Preferably, the host cell secretes minimal amounts of proteolytic enzymes.
  • strain W3110 may be modified to effect a genetic mutation in the genes encoding proteins endogenous to the host, with examples of such hosts including E . coli W3110 strain 1 A2, which has the complete genotype tonA ; E.
  • E. coli W3110 strain 9E4 which has the complete genotype tonA ptr3
  • E. coli W3110 strain 27C7 (ATCC 55,244), which has the complete genotype tonAptr3 phoA El 5 (argF-lac)169 degP ompTkari
  • E. coli W3110 strain 37D6 which has the complete genotype tonA ptr3 phoA El 5 (argF-lac)169 degP ompT rbs7 ilvG kan r
  • E. coli W3110 strain 40B4 which is strain 37D6 with a non-kanamycin resistant degP deletion mutation; and an__.
  • coli strain having mutant periplasmic protease disclosed in U.S. Patent No.4,946,783 issued 7 August 1990.
  • in vitro methods of cloning e.g., PCR or other nucleic acid polymerase reactions, are suitable.
  • eukaryotic microbes such as filamentous fungi or yeast are suitable cloning or expression hosts for PRO-encoding vectors. Saccharomyces cerevisiae is a commonly used lower eukaryotic host microorganism. Others include Schizosaccharomyces pombe (Beach and Nurse, Nature. 290: 140 [1981]; EP 139,383 published 2 May 1985); Kluyveromyces hosts (U.S.
  • Patent No. 4,943,529 Fleer et al., Bio/Technology. 9:968-975 (1991)
  • K. lactis MW98-8C, CBS683, CBS4574; Louvencourt et al., J. Bacteriol.. 154(2):737-742 [1983]
  • K. fragilis ATCC 12,424)
  • K. bulgaricus ATCC 16,045)
  • K. wickeramii ATCC 24,178
  • K. waltii ATCC 56,500
  • K. drosophilarum ATCC 36,906; Van den Berg et al., Bio/Technology. 8: 135 (1990)
  • K. thermotolerans and K.
  • Schwanniomyces such as Schwanniomyces occidentalis (EP 394,538 published 31 October 1990); and filamentous fungi such as, e.g., Neurospora, Penicillium, Tolypocladium WO 91/00357 published lOJanuary 1991), and Aspergillus hosts such as A. nidulans (Ballance et al., Biochem. Biophvs. Res. Commun.. 112:284-289 [1983]; Tilburn et al., Gene. 26:205-221 [19831: Yeltonet al.. Proc. Natl. Acad. Sci. USA.
  • Methylotropic yeasts are suitable herein and include, but are not limited to, yeast capable of growth on methanol selected from the genera consisting of Hansenula, Candida, Kloeckera, Pichia, Saccharomyces, Torulopsis, and Rhodotorula. A list of specific species that are exemplary of this class of yeasts may be found in C. Anthony, The Biochemistry of Methylotrophs. 269 (1982). Suitable host cells for the expression of glycosylated PRO are derived from multicellular organisms.
  • invertebrate cells include insect cells such as Drosophila S2 and Spodoptera Sf9, as well as plant cells.
  • useful mammalian host cell lines include Chinese hamster ovary (CHO) and COS cells. More specific examples include monkey kidney CV1 line transformed by SV40 (COS-7, ATCC CRL 1651); human embryonic kidney line (293 or 293 cells subcloned for growth in suspension culture, Graham et al., J. Gen Virol .. 36:59 (1977)); Chinese hamster ovary cellsADHFR (CHO, Urlaub and Chasin, Proc. Natl. Acad. Sci. USA. 77:4216 (1980)); mouse sertoli cells (TM4, Mather, Biol. Reprod..
  • the nucleic acid (e.g., cDNA or genomic DNA) encoding PRO may be inserted into a replicable vector for cloning (amplification of the DNA) or for expression.
  • a replicable vector for cloning (amplification of the DNA) or for expression.
  • the vector may, for example, be in the form of a plasmid, cosmid, viral particle, or phage.
  • the appropriate nucleic acid sequence may be inserted into the vector by a variety of procedures. In general, DNA is inserted into an appropriate restriction endonuclease site(s) using techniques known in the art.
  • Vector components generally include, but are not limited to, one or more of a signal sequence, an origin of replication, one or more marker genes, an enhancer element, a promoter, and a transcription termination sequence.
  • the PRO may be produced recombinantly not only directly, but also as a fusion polypeptide with a heterologous polypeptide, which may be a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • a heterologous polypeptide which may be a signal sequence or other polypeptide having a specific cleavage site at the N-terminus of the mature protein or polypeptide.
  • the signal sequence may be a component of the vector, or it may be a part of the PRO-encoding DNA that is inserted into the vector.
  • the signal sequence may be a prokaryotic signal sequence selected, for example, from the group of the alkaline phosphatase, penicillinase, lpp, or heat-stable enterotoxin II leaders.
  • the signal sequence may be, e.g., the yeast invertase leader, alpha factor leader (including Saccharomyces and Kluyveromyces ⁇ -factor leaders, the latter described in U.S. Patent No. 5,010, 182), or acid phosphatase leader, the C. albicans glucoamylase leader (EP 362,179 published 4 April 1990), or the signal described in WO 90/13646 published 15 November 1990.
  • mammalian signal sequences may be used to direct secretion of the protein, such as signal sequences from secreted polypeptides of the same or related species, as well as viral secretory leaders.
  • Both expression and cloning vectors contain a nucleic acid sequence that enables the vector to replicate in one or more selected host cells. Such sequences are well known for a variety of bacteria, yeast, and viruses.
  • the origin of replication from the plasmid pBR322 is suitable for most Gram-negative bacteria, the 2 ⁇ plasmid origin is suitable for yeast, and various viral origins (SV40, polyoma, adenovirus, VSV or BPV) are useful for cloning vectors in mammalian cells.
  • Expression and cloning vectors will typically contain a selection gene, also termed a selectable marker.
  • Typical selection genes encode proteins that (a) confer resistance to antibiotics or other toxins, e.g., ampicillin, neomycin, methotrexate, or tetracycline, (b) complement auxotrophic deficiencies, or (c) supply critical nutrients not available from complex media, e.g., the gene encoding D-alanine racemase for Bacilli.
  • suitable selectable markers for mammalian cells are those that enable the identification of cells competent to take up the PRO-encoding nucleic acid, such as DHFR or thymidine kinase.
  • An appropriate host cell when wild-type DHFR is employed is the CHO cell line deficient in DHFR activity, prepared and propagated as described by Urlaub et al.. Proc. Natl. Acad. Sci. USA. 77:4216 (1980).
  • a suitable selection gene for use in yeast is the trpl gene present in the yeast plasmid YRp7 [Stinchcomb et al., Nature. 282:39 (1979); Kingsman et al., Gene. 7: 141 (1979); Tschemper et al., Gene.
  • the trpl gene provides a selection marker for a mutant strain of yeast lacking the ability to grow in tryptophan, for example, ATCC No. 44076 or PEP4-1 [Jones, Genetics. 85: 12 (1977)].
  • Expression and cloning vectors usually contain a promoter operably linked to the PRO-encoding nucleic acid sequence to direct mRNA synthesis. Promoters recognized by a variety of potential host cells are well known. Promoters suitable for use with prokaryotic hosts include the ⁇ -lactamase and lactose promoter systems [Chang et al., Nature. 275:615 (1978); Goeddel et al., Nature. 281:544 (1979)], alkaline phosphatase, a tryptophan (trp) promoter system [Goeddel, Nucleic Acids Res.. 8:4057 (1980); EP 36,776], and hybrid promoters such as the tac promoter [deBoer et al.
  • Promoters for use in bacterial systems also will contain a Shine-Dalgarno (S.D.) sequence operably linked to the DNA encoding PRO.
  • suitable promoting sequences for use with yeast hosts include the promoters for 3- phosphoglycerate kinase [Hitzeman et al., J. Biol. Chem.. 255:2073 (1980)] or other glycolytic enzymes [Hess et al. , J. Adv. Enzyme Reg..
  • yeast promoters which are inducible promoters having the additional advantage of transcription controlled by growth conditions, are the promoter regions for alcohol dehydrogenase 2, isocytochrome C, acid phosphatase, degradative enzymes associated with nitrogen metabolism, metallothionein, glyceraldehyde-3- phosphate dehydrogenase, and enzymes responsible for maltose and galactose utilization. Suitable vectors and promoters for use in yeast expression are further described in EP 73,657.
  • PRO transcription from vectors in mammalian host cells is controlled, for example, by promoters obtained from the genomes of viruses such as polyoma virus, fowlpox virus (UK 2,211,504 published 5 July 1989), adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, hepatitis-B virus and Simian Virus 40 (SV40), from heterologous mammalian promoters, e.g., the actin promoter or an immunoglobulin promoter, and from heat-shock promoters, provided such promoters are compatible with the host cell systems.
  • viruses such as polyoma virus, fowlpox virus (UK 2,211,504 published 5 July 1989), adenovirus (such as Adenovirus 2), bovine papilloma virus, avian sarcoma virus, cytomegalovirus, a retrovirus, he
  • Enhancers are cis-acting elements of DNA, usually about from 10 to 300 bp, that act on a promoter to increase its transcription.
  • Many enhancer sequences are now known from mammalian genes (globin, elastase, albumin, ⁇ -fetoprotein, and insulin).
  • an enhancer from a eukaryotic cell virus. Examples include the SV40 enhancer on the late side of the replication origin (bp 100-270), the cytomegalovirus early promoter enhancer, the polyoma enhancer on the late side of the replication origin, and adenovirus enhancers.
  • the enhancer may be spliced into the vector at a position 5' or 3' to the PRO coding sequence, but is preferably located at a site 5' from the promoter.
  • Expression vectors used in eukaryotic host cells will also contain sequences necessary for the termination of transcription and for stabilizing the mRNA. Such sequences are commonly available from the 5' and, occasionally 3 ' , untranslated regions of eukaryotic or viral DNAs or cDNAs. These regions contain nucleotide segments transcribed as polyadenylated fragments in the untranslated portion of the mRNA encoding PRO.
  • Gene amplification and/or expression may be measured in a sample directly, for example, by conventional Southern blotting, Northern blotting to quantitate the transcription of mRNA [Thomas, Proc. Natl. Acad. Sci. USA. 77:5201-5205 (1980)], dot blotting (DNA analysis), or in situ hybridization, using an appropriately labeled probe, based on the sequences provided herein.
  • antibodies may be employed that can recognize specific duplexes, including DNA duplexes, RNA duplexes, and DNA-RNA hybrid duplexes or DNA-protein duplexes.
  • the antibodies in turn may be labeled and the assay may be carried out where the duplex is bound to a surface, so that upon the formation of duplex on the surface, the presence of antibody bound to the duplex can be detected.
  • Gene expression alternatively, may be measured by immunological methods, such as immunohistochemical staining of cells or tissue sections and assay of cell culture or body fluids, to quantitate directly the expression of gene product.
  • Antibodies useful for immunohistochemical staining and/or assay of sample fluids may be either monoclonal or polyclonal, and may be prepared in any mammal.
  • the antibodies may be prepared against a native sequence PRO polypeptide or against a synthetic peptide based on the DNA sequences provided herein or against exogenous sequence fused to PRO DNA and encoding a specific antibody epitope. 5. Purification of Polypeptide
  • PRO may be recovered from culture medium or from host cell lysates. If membrane-bound, it can be released from the membrane using a suitable detergent solution (e.g. Triton-X 100) or by enzymatic cleavage. Cells employed in expression of PRO can be disrupted by various physical or chemical means, such as freeze-thaw cycling, sonication, mechanical disruption, or cell lysing agents. It may be desired to purify PRO from recombinant cell proteins or polypeptides.
  • a suitable detergent solution e.g. Triton-X 100
  • Cells employed in expression of PRO can be disrupted by various physical or chemical means, such as freeze-thaw cycling, sonication, mechanical disruption, or cell lysing agents. It may be desired to purify PRO from recombinant cell proteins or polypeptides.
  • the following procedures are exemplary of suitable purification procedures: by fractionation on an ion-exchange column; ethanol precipitation; reverse phase HPLC; chromatography on silica or on a cation-exchange resin such as DEAE; chromatofocusing; SDS-PAGE; ammonium sulfate precipitation; gel filtration using, for example, Sephadex G-75; protein A Sepharose columns to remove contaminants such as IgG; and metal chelating columns to bind epitope-tagged forms of the PRO.
  • Various methods of protein purification may be employed and such methods are known in the art and described for example in Deutscher, Methods in Enzvmologv. 182 (1990); Scopes, Protein Purification: Principles and Practice. Springer- Verlag, New York (1982).
  • the purification step(s) selected will depend, for example, on the nature of the production process used and the particular PRO produced.
  • PRO Nucleotide sequences (or their complement) encoding PRO have various applications in the art of molecular biology, including uses as hybridization probes, in chromosome and gene mapping and in the generation of anti-sense RNA and DNA.
  • PRO nucleic acid will also be useful for the preparation of PRO polypeptides by the recombinant techniques described herein.
  • the full-length native sequence PRO gene, or portions thereof, may be used as hybridization probes for a cDNA library to isolate the full-length PRO cDNA or to isolate still other cDNAs (for instance, those encoding naturally-occurring variants of PRO or PRO from other species) which have a desired sequence identity to the native PRO sequence disclosed herein.
  • the length of the probes will be about 20 to about 50 bases.
  • the hybridization probes may be derived from at least partially novel regions of the full length native nucleotide sequence wherein those regions may be determined without undue experimentation or from genomic sequences including promoters, enhancer elements and introns of native sequence PRO.
  • a screening method will comprise isolating the coding region of the PRO gene using the known DNA sequence to synthesize a selected probe of about 40 bases.
  • Hybridization probes may be labeled by a variety of labels, including radionucleotides such as 32 P or 35 S, or enzymatic labels such as alkaline phosphatase coupled to the probe via avidin/biotin coupling systems. Labeled probes having a sequence complementary to that of the PRO gene of the present invention can be used to screen libraries of human cDNA, genomic DNA or mRNA to determine which members of such libraries the probe hybridizes to. Hybridization techniques are described in further detail in the Examples below.
  • any EST sequences disclosed in the present application may similarly be employed as probes, using the methods disclosed herein.
  • Other useful fragments of the PRO nucleic acids include antisense or sense oligonucleotides comprising a singe-stranded nucleic acid sequence (either RNA or DNA) capable of binding to target PRO mRNA (sense) or PRO DNA (antisense) sequences.
  • Antisense or sense oligonucleotides, according to the present invention comprise a fragment of the coding region of PRO DNA. Such a fragment generally comprises at least about 14 nucleotides, preferably from about 14 to 30 nucleotides.
  • binding of antisense or sense oligonucleotides to target nucleic acid sequences results in the formation of duplexes that block transcription or translation of the target sequence by one of several means, including enhanced degradation of the duplexes, premature termination of transcription or translation, or by other means.
  • the antisense oligonucleotides thus may be used to block expression of PRO proteins.
  • Antisense or sense oligonucleotides further comprise oligonucleotides having modified sugar-phosphodiester backbones (or other sugar linkages, such as those described in WO 91/06629) and wherein such sugar linkages are resistant to endogenous nucleases.
  • Such oligonucleotides with resistant sugar linkages are stable in vivo (i.e., capable of resisting enzymatic degradation) but retain sequence specificity to be able to bind to target nucleotide sequences.
  • Other examples of sense or antisense oligonucleotides include those oligonucleotides which are covalently linked to organic moieties, such as those described in WO 90/10048, and other moieties that increases affinity of the oligonucleotide for a target nucleic acid sequence, such as poly-(L-lysine).
  • intercalating agents such as ellipticine, and alkylating agents or metal complexes may be attached to sense or antisense oligonucleotides to modify binding specificities of the antisense or sense oligonucleotide for the target nucleotide sequence.
  • Antisense or sense oligonucleotides may be introduced into a cell containing the target nucleic acid sequence by any gene transfer method, including, for example, CaP0 4 -mediated DNA transfection, electroporation, or by using gene transfer vectors such as Epstein-Barr virus.
  • an antisense or sense oligonucleotide is inserted into a suitable retro viral vector.
  • a cell containing the target nucleic acid sequence is contacted with the recombinant retroviral vector, either in vivo or ex vivo.
  • Suitable retroviral vectors include, but are not limited to, those derived from the murine retrovirus M-MuLV, N2 (a retrovirus derived from M-MuLV), or the double copy vectors designated DCT5A, DCT5B and DCT5C (see WO 90/13641).
  • Sense or antisense oligonucleotides also may be introduced into a cell containing the target nucleotide sequence by formation of a conjugate with a ligand binding molecule, as described in WO 91/04753.
  • Suitable ligand binding molecules include, but are not limited to, cell surface receptors, growth factors, other cytokines, or other ligands that bind to cell surface receptors.
  • conjugation of the ligand binding molecule does not substantially interfere with the ability of the ligand binding molecule to bind to its corresponding molecule or receptor, or block entry of the sense or antisense oligonucleotide or its conjugated version into the cell.
  • a sense or an antisense oligonucleotide may be introduced into a cell containing the target nucleic acid sequence by formation of an oligonucleotide-lipid complex, as described in WO 90/10448.
  • the sense or antisense oligonucleotide-lipid complex is preferably dissociated within the cell by an endogenous lipase.
  • Antisense or sense RNA or DNA molecules are generally at least about 5 bases in length, about 10 bases in length, about 15 bases in length, about 20 bases in length, about 25 bases in length, about 30 bases in length, about 35 bases in length, about 40 bases in length, about 45 bases in length, about 50 bases in length, about 55 bases in length, about 60 bases in length, about 65 bases in length, about 70 bases in length, about 75 bases in length, about 80 bases in length, about 85 bases in length, about 90 bases in length, about 95 bases in length, about 100 bases in length, or more.
  • the probes may also be employed in PCR techniques to generate a pool of sequences for identification of closely related PRO coding sequences.
  • Nucleotide sequences encoding a PRO can also be used to construct hybridization probes for mapping the gene which encodes that PRO and for the genetic analysis of individuals with genetic disorders.
  • the nucleotide sequences provided herein may be mapped to a chromosome and specific regions of a chromosome using known techniques, such as in situ hybridization, linkage analysis against known chromosomal markers, and hybridization screening with libraries.
  • the PRO can be used in assays to identify the other proteins or molecules involved in the binding interaction. By such methods, inhibitors of the receptor/ligand binding interaction can be identified. Proteins involved in such binding interactions can also be used to screen for peptide or small molecule inhibitors or agonists of the binding interaction. Also, the receptor PRO can be used to isolate correlative ligand(s). Screening assays can be designed to find lead compounds that mimic the biological activity of a native PRO or a receptor for PRO. Such screening assays will include assays amenable to high-throughput screening of chemical libraries, making them particularly suitable for identifying small molecule drug candidates.
  • Small molecules contemplated include synthetic organic or inorganic compounds.
  • the assays can be performed in a variety of formats, including protein-protein binding assays, biochemical screening assays, immunoassays and cell based assays, which are well characterized in the art.
  • Nucleic acids which encode PRO or its modified forms can also be used to generate either transgenic animals or "knock out" animals which, in turn, are useful in the development and screening of therapeutically useful reagents.
  • a transgenic animal e.g., a mouse or rat
  • a transgenic animal is an animal having cells that contain a transgene, which transgene was introduced into the animal or an ancestor of the animal at a prenatal, e.g., an embryonic stage.
  • a transgene is a DNA which is integrated into the genome of a cell from which a transgenic animal develops.
  • cDNA encoding PRO can be used to clone genomic DNA encoding PRO in accordance with established techniques and the genomic sequences used to generate transgenic animals that contain cells which express DNA encoding PRO.
  • Methods for generating transgenic animals, particularly animals such as mice or rats, have become conventional in the art and are described, for example, in U.S. Patent
  • Transgenic animals that include a copy of a transgene encoding PRO introduced into the germ line of the animal at an embryonic stage can be used to examine the effect of increased expression of DNA encoding PRO. Such animals can be used as tester animals for reagents thought to confer protection from, for example, pathological conditions associated with its overexpression. In accordance with this facet of the invention, an animal is treated with the reagent and a reduced incidence of the pathological condition, compared to untreated animals bearing the transgene, would indicate a potential therapeutic intervention for the pathological condition.
  • non-human homologues of PRO can be used to construct a PRO "knock out" animal which has a defective or altered gene encoding PRO as a result of homologous recombination between the endogenous gene encoding PRO and altered genomic DNA encoding PRO introduced into an embryonic stem cell of the animal.
  • cDNA encoding PRO can be used to clone genomic DNA encoding PRO in accordance with established techniques. A portion of the genomic DNA encoding PRO can be deleted or replaced with another gene, such as a gene encoding a selectable marker which can be used to monitor integration.
  • flanking DNA typically, several kilobases of unaltered flanking DNA (both at the 5' and 3' ends) are included in the vector [see e.g., Thomas and Capecchi, Cell. 51 :503 (1987) for a description of homologous recombination vectors] .
  • the vector is introduced into an embryonic stem cell line (e.g. , by electroporation) and cells in which the introduced DNA has homologously recombined with the endogenous DNA are selected [see e.g., Li et al., Cell. 69:915 (1992)].
  • the selected cells are then injected into a blastocyst of an animal (e.g., a mouse or rat) to form aggregation chimeras [see e.g., Bradley, in Teratocarcinomas and Embryonic Stem Cells: A Practical Approach, E. J. Robertson, ed. (IRL, Oxford, 1987), pp. 113-152].
  • a chimeric embryo can then be implanted into a suitable pseudopregnant female foster animal and the embryo brought to term to create a "knock out" animal.
  • Progeny harboring the homologously recombined DNA in their germ cells can be identified by standard techniques and used to breed animals in which all cells of the animal contain the homologously recombined DNA. Knockout animals can be characterized for instance, for their ability to defend against certain pathological conditions and for their development of pathological conditions due to absence of the PRO polypeptide.
  • Nucleic acid encoding the PRO polypeptides may also be used in gene therapy.
  • genes are introduced into cells in order to achieve in vivo synthesis of a therapeutically effective genetic product, for example for replacement of a defective gene.
  • Gene therapy includes both conventional gene therapy where a lasting effect is achieved by a single treatment, and the administration of gene therapeutic agents, which involves the one time or repeated administration of a therapeutically effective DNA or mRNA.
  • Antisense RNAs and DNAs can be used as therapeutic agents for blocking the expression of certain genes in vivo. It has already been shown that short antisense oligonucleotides can be imported into cells where they act as inhibitors, despite their low intracellular concentrations caused by their restricted uptake by the cell membrane.
  • oligonucleotides can be modified to enhance their uptake, e.g. by substituting their negatively charged phosphodiester groups by uncharged groups.
  • nucleic acids there are a variety of techniques available for introducing nucleic acids into viable cells.
  • the techniques vary depending upon whether the nucleic acid is transferred into cultured cells in vitro, or in vivo in the cells of the intended host.
  • Techniques suitable for the transfer of nucleic acid into mammalian cells in vitro include the use of liposomes, electroporation, microinjection, cell fusion, DEAE-dextran, the calcium phosphate precipitation method, etc.
  • the currently preferred in vivo gene transfer techniques include transfection with viral (typically retroviral) vectors and viral coat protein-liposome mediated transfection (Dzau et al., Trends in Biotechnology 11, 205-210 [1993]).
  • the nucleic acid source with an agent that targets the target cells, such as an antibody specific for a cell surface membrane protein or the target cell, a ligand for a receptor on the target cell, etc.
  • an agent that targets the target cells such as an antibody specific for a cell surface membrane protein or the target cell, a ligand for a receptor on the target cell, etc.
  • proteins which bind to a cell surface membrane protein associated with endocytosis may be used for targeting and/or to facilitate uptake, e.g. capsid proteins or fragments thereof tropic for a particular cell type, antibodies for proteins which undergo internalization in cycling, proteins that target intracellular localization and enhance intracellular half-life.
  • the technique of receptor-mediated endocytosis is described, for example, by Wu et al., J. Biol. Chem.
  • PRO polypeptides described herein may also be employed as molecular weight markers for protein electrophoresis purposes and the isolated nucleic acid sequences may be used for recombinantly expressing those markers.
  • nucleic acid molecules encoding the PRO polypeptides or fragments thereof described herein are useful for chromosome identification.
  • there exists an ongoing need to identify new chromosome markers since relatively few chromosome marking reagents, based upon actual sequence data are presently available.
  • Each PRO nucleic acid molecule of the present invention can be used as a chromosome marker.
  • PRO polypeptides and nucleic acid molecules of the present invention may also be used diagnostically for tissue typing, wherein the PRO polypeptides of the present invention may be differentially expressed in one tissue as compared to another, preferably in a diseased tissue as compared to a normal tissue of the same tissue type.
  • PRO nucleic acid molecules will find use for generating probes for PCR, Northern analysis, Southern analysis and Western analysis.
  • PRO polypeptides described herein may also be employed as therapeutic agents.
  • the PRO polypeptides of the present invention can be formulated according to known methods to prepare pharmaceutically useful compositions, whereby the PRO product hereof is combined in admixture with a pharmaceutically acceptable carrier vehicle.
  • Therapeutic formulations are prepared for storage by mixing the active ingredient having the desired degree of purity with optional physiologically acceptable carriers, excipients or stabilizers (Remington's Pharmaceutical Sciences 16th edition, Osol, A. Ed. (1980)), in the form of lyophilized formulations or aqueous solutions.
  • Acceptable carriers, excipients or stabilizers are nontoxic to recipients at the dosages and concentrations employed, and include buffers such as phosphate, citrate and other organic acids; antioxidants including ascorbic acid; low molecular weight (less than about 10 residues) polypeptides; proteins, such as serum albumin, gelatin or immunoglobulins; hydrophilic polymers such as polyvinylpyrrolidone, amino acids such as glycine, glutamine, asparagine, arginine or lysine; monosaccharides, disaccharides and other carbohydrates including glucose, mannose, or dextrins; chelating agents such as EDTA; sugar alcohols such as mannitol or sorbitol; salt-forming counterions such as sodium; and/or nonionic surfactants such as TWEENTM, PLURONICSTM or PEG.
  • buffers such as phosphate, citrate and other organic acids
  • antioxidants including ascorbic acid
  • compositions to be used for in vivo administration must be sterile. This is readily accomplished by filtration through sterile filtration membranes, prior to or following lyophilization and reconstitution.
  • Therapeutic compositions herein generally are placed into a container having a sterile access port, for example, an intravenous solution bag or vial having a stopper pierceable by a hypodermic injection needle.
  • the route of administration is in accord with known methods, e.g. injection or infusion by intravenous, intraperitoneal, intracerebral, intramuscular, intraocular, intraarterial or intralesional routes, topical administration, or by sustained release systems.
  • Dosages and desired drug concentrations of pharmaceutical compositions of the present invention may vary depending on the particular use envisioned.
  • the determination of the appropriate dosage or route of administration is well within the skill of an ordinary physician. Animal experiments provide reliable guidance for the determination of effective doses for human therapy.
  • Interspecies scaling of effective doses can be performed following the principles laid down by Mordenti, J. and Chappell, W. "The use of interspecies scaling in toxicokinetics" In Toxicokinetics and New Drug Development, Yacobi et al., Eds., Pergamon Press, New York 1989, pp. 42-96.
  • normal dosage amounts may vary from about 10 ng/kg to up to 100 mg/kg of mammal body weight or more per day, preferably about 1 ⁇ g/kg/day to 10 mg/kg/day, depending upon the route of administration.
  • Guidance as to particular dosages and methods of delivery is provided in the literature; see, for example, U.S. Pat. Nos. 4,657,760; 5,206,344; or 5,225,212. It is anticipated that different formulations will be effective for different treatment compounds and different disorders, that administration targeting one organ or tissue, for example, may necessitate delivery in a manner different from that to another organ or tissue.
  • microencapsulation of the PRO polypeptide is contemplated.
  • Microencapsulation of recombinant proteins for sustained release has been successfully performed with human growth hormone (rhGH), interferon- (rhlFN- ), interleukin-2, and MN rgpl20. Johnson et al., Nat. Med.. 2:795-799 (1996); Yasuda, Biomed. Ther.. 27: 1221-1223 (1993); Hora et al. , Bio/Technology.
  • the sustained-release formulations of these proteins were developed using poly-lactic-coglycolic acid (PLGA) polymer due to its biocompatibility and wide range of biodegradable properties.
  • PLGA poly-lactic-coglycolic acid
  • the degradation products of PLGA, lactic and glycolic acids, can be cleared quickly within the human body.
  • the degradability of this polymer can be adjusted from months to years depending on its molecular weight and composition. Lewis, "Controlled release of bioactive agents from lactide/glycolide polymer," in: M. Chasin and R. Langer (Eds.), Biodegradable Polymers as Drug Delivery Systems (Marcel Dekker: New York, 1990), pp. 1-41.
  • This invention encompasses methods of screening compounds to identify those that mimic the PRO polypeptide (agonists) or prevent the effect of the PRO polypeptide (antagonists).
  • Screening assays for antagonist drug candidates are designed to identify compounds that bind or complex with the PRO polypeptides encoded by the genes identified herein, or otherwise interfere with the interaction of the encoded polypeptides with other cellular proteins.
  • Such screening assays will include assays amenable to high-throughput screening of chemical libraries, making them particularly suitable for identifying small molecule drug candidates.
  • the assays can be performed in a variety of formats, including protein-protein binding assays, biochemical screening assays, immunoassays, and cell-based assays, which are well characterized in the art. All assays for antagonists are common in that they call for contacting the drug candidate with a PRO polypeptide encoded by a nucleic acid identified herein under conditions and for a time sufficient to allow these two components to interact.
  • the interaction is binding and the complex formed can be isolated or detected in the reaction mixture.
  • the PRO polypeptide encoded by the gene identified herein or the drug candidate is immobilized on a solid phase, e.g., on a microtiter plate, by covalent or non-covalent attachments.
  • Non-covalent attachment generally is accomplished by coating the solid surface with a solution of the PRO polypeptide and drying.
  • an immobilized antibody e.g. , a monoclonal antibody, specific for the PRO polypeptide to be immobilized can be used to anchor it to a solid surface.
  • the assay is performed by adding the non-immobilized component, which may be labeled by a detectable label, to the immobilized component, e.g., the coated surface containing the anchored component.
  • the non-reacted components are removed, e.g., by washing, and complexes anchored on the solid surface are detected.
  • the detection of label immobilized on the surface indicates that complexing occurred.
  • complexing can be detected, for example, by using a labeled antibody specifically binding the immobilized complex.
  • the candidate compound interacts with but does not bind to a particular PRO polypeptide encoded by a gene identified herein, its interaction with that polypeptide can be assayed by methods well known for detecting protein-protein interactions.
  • assays include traditional approaches, such as, e.g., cross-linking, co- immunoprecipitation, and co-purification through gradients or chromatographic columns.
  • protein- protein interactions can be monitored by using a yeast-based genetic system described by Fields and co-workers (Fields and Song, Nature (London * ) . 340:245-246 (1989); Chien et al., Proc. Natl. Acad. Sci. USA.
  • yeast GAL4 consist of two physically discrete modular domains, one acting as the DNA-binding domain, the other one functioning as the transcription-activation domain.
  • the yeast expression system described in the foregoing publications (generally referred to as the "two-hybrid system") takes advantage of this property, and employs two hybrid proteins, one in which the target protein is fused to the DNA-binding domain of GAL4, and another, in which candidate activating proteins are fused to the activation domain.
  • GALl-t ⁇ cZ reporter gene under control of a GAL4-activated promoter depends on reconstitution of GAL4 activity via protein-protein interaction. Colonies containing interacting polypeptides are detected with a chromogenic substrate for ⁇ -galactosidase.
  • a complete kit (MATCHMAKERTM) for identifying protein-protein interactions between two specific proteins using the two- hybrid technique is commercially available from Clontech. This system can also be extended to map protein domains involved in specific protein interactions as well as to pinpoint amino acid residues that are crucial for these interactions.
  • a reaction mixture is prepared containing the product of the gene and the intra- or extracellular component under conditions and for a time allowing for the interaction and binding of the two products.
  • a candidate compound to inhibit binding, the reaction is run in the absence and in the presence of the test compound.
  • a placebo may be added to a third reaction mixture, to serve as positive control.
  • the binding (complex formation) between the test compound and the intra- or extracellular component present in the mixture is monitored as described hereinabove. The formation of a complex in the control reaction(s) but not in the reaction mixture containing the test compound indicates that the test compound interferes with the interaction of the test compound and its reaction partner.
  • the PRO polypeptide may be added to a cell along with the compound to be screened for a particular activity and the ability of the compound to inhibit the activity of interest in the presence of the PRO polypeptide indicates that the compound is an antagonist to the PRO polypeptide.
  • antagonists may be detected by combining the PRO polypeptide and a potential antagonist with membrane-bound PRO polypeptide receptors or recombinant receptors under appropriate conditions for a competitive inhibition assay.
  • the PRO polypeptide can be labeled, such as by radioactivity, such that the number of PRO polypeptide molecules bound to the receptor can be used to determine the effectiveness of the potential antagonist.
  • the gene encoding the receptor can be identified by numerous methods known to those of skill in the art, for example, ligand panning and FACS sorting. Coligan et al., Current Protocols in Immun.. 1(2): Chapter 5 (1991).
  • expression cloning is employed wherein polyadenylated RNA is prepared from a cell responsive to the PRO polypeptide and a cDNA library created from this RNA is divided into pools and used to transfect COS cells or other cells that are not responsive to the PRO polypeptide. Transfected cells that are grown on glass slides are exposed to labeled PRO polypeptide.
  • the PRO polypeptide can be labeled by a variety of means including iodination or inclusion of a recognition site for a site-specific protein kinase. Following fixation and incubation, the slides are subjected to autoradiographic analysis. Positive pools are identified and sub-pools are prepared and re-transfected using an interactive sub-pooling and re-screening process, eventually yielding a single clone that encodes the putative receptor.
  • labeled PRO polypeptide can be photoaffinity- linked with cell membrane or extract preparations that express the receptor molecule. Cross-linked material is resolved by PAGE and exposed to X-ray film. The labeled complex containing the receptor can be excised, resolved into peptide fragments, and subjected to protein micro-sequencing. The amino acid sequence obtained from micro- sequencing would be used to design a set of degenerate oligonucleotide probes to screen a cDNA library to identify the gene encoding the putative receptor. In another assay for antagonists, mammalian cells or a membrane preparation expressing the receptor would be incubated with labeled PRO polypeptide in the presence of the candidate compound.
  • potential antagonists include an oligonucleotide that binds to the fusions of immunoglobulin with PRO polypeptide, and, in particular, antibodies including, without limitation, poly- and monoclonal antibodies and antibody fragments, single-chain antibodies, anti-idiotypic antibodies, and chimeric or humanized versions of such antibodies or fragments, as well as human antibodies and antibody fragments.
  • a potential antagonist may be a closely related protein, for example, a mutated form of the PRO polypeptide that recognizes the receptor but imparts no effect, thereby competitively inhibiting the action of the PRO polypeptide.
  • Another potential PRO polypeptide antagonist is an antisense RNA or DNA construct prepared using antisense technology, where, e.g., an antisense RNA or DNA molecule acts to block directly the translation of mRNA by hybridizing to targeted mRNA and preventing protein translation.
  • Antisense technology can be used to control gene expression through triple-helix formation or antisense DNA or RNA, both of which methods are based on binding of a polynucleotide to DNA or RNA.
  • the 5' coding portion of the polynucleotide sequence, which encodes the mature PRO polypeptides herein, is used to design an antisense RNA oligonucleotide of from about 10 to 40 base pairs in length.
  • a DNA oligonucleotide is designed to be complementary to a region of the gene involved in transcription (triple helix - see Lee et al., Nucl. Acids Res.. 3:173 (1979); Cooney et al., Science. 241 : 456 (1988); Dervan et al., Science. 251 : 1360 (1991)), thereby preventing transcription and the production of the PRO polypeptide.
  • the antisense RNA oligonucleotide hybridizes to the mRNA in vivo and blocks translation of the mRNA molecule into the PRO polypeptide (antisense - Okano, Neurochem..
  • oligodeoxynucleotides as Antisense Inhibitors of Gene Expression (CRC Press: Boca Raton, FL, 1988).
  • the oligonucleotides described above can also be delivered to cells such that the antisense RNA or DNA may be expressed in vivo to inhibit production of the PRO polypeptide.
  • antisense DNA oligodeoxyribonucleotides derived from the translation-initiation site, e.g., between about -10 and + 10 positions of the target gene nucleotide sequence, are preferred.
  • Potential antagonists include small molecules that bind to the active site, the receptor binding site, or growth factor or other relevant binding site of the PRO polypeptide, thereby blocking the normal biological activity of the PRO polypeptide.
  • small molecules include, but are not limited to, small peptides or peptide-like molecules, preferably soluble peptides, and synthetic non-peptidyl organic or inorganic compounds.
  • Ribozymes are enzymatic RNA molecules capable of catalyzing the specific cleavage of RNA. Ribozymes act by sequence-specific hybridization to the complementary target RNA, followed by endonucleolytic cleavage. Specific ribozyme cleavage sites within a potential RNA target can be identified by known techniques. For further details see, e.g. , Rossi, Current Biology. 4:469-471 (1994), and PCT publication No. WO 97/33551 (published September 18, 1997).
  • Nucleic acid molecules in triple-helix formation used to inhibit transcription should be single-stranded and composed of deoxynucleotides.
  • the base composition of these oligonucleotides is designed such that it promotes triple-helix formation via Hoogsteen base-pairing rules, which generally require sizeable stretches of purines or pyrimidines on one strand of a duplex.
  • These small molecules can be identified by any one or more of the screening assays discussed hereinabove and/or by any other screening techniques well known for those skilled in the art.
  • Diagnostic and therapeutic uses of the herein disclosed molecules may also be based upon the positive functional assay hits disclosed and described below.
  • the present invention further provides anti-PRO antibodies.
  • Exemplary antibodies include polyclonal, monoclonal, humanized, bispecific, and heteroconjugate antibodies.
  • the anti-PRO antibodies may comprise polyclonal antibodies. Methods of preparing polyclonal antibodies are known to the skilled artisan. Polyclonal antibodies can be raised in a mammal, for example, by one or more injections of an immunizing agent and, if desired, an adjuvant. Typically, the immunizing agent and/or adjuvant will be injected in the mammal by multiple subcutaneous or intraperitoneal injections.
  • the immunizing agent may include the PRO polypeptide or a fusion protein thereof. It may be useful to conjugate the immunizing agent to a protein known to be immunogenic in the mammal being immunized.
  • immunogenic proteins include but are not limited to keyhole limpet hemocyanin, serum albumin, bovine thyroglobulin, and soybean trypsin inhibitor.
  • adjuvants which may be employed include Freund's complete adjuvant and MPL-TDM adjuvant (monophosphoryl Lipid A, synthetic trehalose dicorynomycolate).
  • the immunization protocol may be selected by one skilled in the art without undue experimentation.
  • the anti-PRO antibodies may, alternatively, be monoclonal antibodies.
  • Monoclonal antibodies may be prepared using hybridoma methods, such as those described by Kohler and Milstein, Nature. 256:495 (1975).
  • a hybridoma method a mouse, hamster, or other appropriate host animal, is typically immunized with an immunizing agent to elicit lymphocytes that produce or are capable of producing antibodies that will specifically bind to the immunizing agent.
  • the lymphocytes may be immunized in vitro.
  • the immunizing agent will typically include the PRO polypeptide or a fusion protein thereof.
  • PBLs peripheral blood lymphocytes
  • spleen cells or lymph node cells are used if non-human mammalian sources are desired.
  • the lymphocytes are then fused with an immortalized cell line using a suitable fusing agent, such as polyethylene glycol, to form a hybridoma cell [Goding, Monoclonal Antibodies: Principles and Practice. Academic Press, (1986) pp. 59-103].
  • Immortalized cell lines are usually transformed mammalian cells, particularly myeloma cells of rodent, bovine and human origin.
  • rat or mouse myeloma cell lines are employed.
  • the hybridoma cells may be cultured in a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells.
  • a suitable culture medium that preferably contains one or more substances that inhibit the growth or survival of the unfused, immortalized cells.
  • the culture medium for the hybridomas typically will include hypoxanthine, aminopterin, and thymidine (“HAT medium”), which substances prevent the growth of HGPRT-deficient cells.
  • Preferred immortalized cell lines are those that fuse efficiently, support stable high level expression of antibody by the selected antibody-producing cells, and are sensitive to a medium such as HAT medium. More preferred immortalized cell lines are murine myeloma lines, which can be obtained, for instance, from the Salk Institute Cell Distribution Center, San Diego, California and the American Type Culture Collection, Manassas, Virginia. Human myeloma and mouse-human heteromyeloma cell lines also have been described for the production of human monoclonal antibodies [Kozbor, J. Immunol.. 133:3001 (1984); Brodeur et al. , Monoclonal Antibody Production Techniques and Applications, Marcel Dekker, Inc., New York, (1987) pp. 51-63].
  • the culture medium in which the hybridoma cells are cultured can then be assayed for the presence of monoclonal antibodies directed against PRO.
  • the binding specificity of monoclonal antibodies produced by the hybridoma cells is determined by immunoprecipitation or by an in vitro binding assay, such as radioimmunoassay (RIA) or enzyme-linked immunoabsorbent assay (ELISA).
  • RIA radioimmunoassay
  • ELISA enzyme-linked immunoabsorbent assay
  • the binding affinity of the monoclonal antibody can, for example, be determined by the Scatchard analysis of Munson and Pollard, Anal. Biochem.. 107:220 (1980).
  • the clones may be subcloned by limiting dilution procedures and grown by standard methods [Goding, supra] .
  • Suitable culture media for this purpose include, for example, Dulbecco's Modified Eagle's Medium and RPMI-1640 medium.
  • the hybridoma cells may be grown in vivo as ascites in a mammal.
  • the monoclonal antibodies secreted by the subclones may be isolated or purified from the culture medium or ascites fluid by conventional immunoglobulin purification procedures such as, for example, protein A-Sepharose, hydroxylapatite chromatography, gel electrophoresis, dialysis, or affinity chromatography.
  • the monoclonal antibodies may also be made by recombinant DNA methods, such as those described in U.S. Patent No. 4,816,567.
  • DNA encoding the monoclonal antibodies of the invention can be readily isolated and sequenced using conventional procedures (e.g., by using oligonucleotide probes that are capable of binding specifically to genes encoding the heavy and light chains of murine antibodies).
  • the hybridoma cells of the invention serve as a preferred source of such DNA.
  • the DNA may be placed into expression vectors, which are then transfected into host cells such as simian COS cells, Chinese hamster ovary (CHO) cells, or myeloma cells that do not otherwise produce immunoglobulin protein, to obtain the synthesis of monoclonal antibodies in the recombinant host cells.
  • the DNA also may be modified, for example, by substituting the coding sequence for human heavy and light chain constant domains in place of the homologous murine sequences [U.S. Patent No. 4,816,567; Morrison et al., supra] or by covalently joining to the immunoglobulin coding sequence all or part of the coding sequence for a non-immunoglobulin polypeptide.
  • Such a non-immunoglobulin polypeptide can be substituted for the constant domains of an antibody of the invention, or can be substituted for the variable domains of one antigen-combining site of an antibody of the invention to create a chimeric bivalent antibody.
  • the antibodies may be monovalent antibodies. Methods for preparing monovalent antibodies are well known in the art. For example, one method involves recombinant expression of immunoglobulin light chain and modified heavy chain. The heavy chain is truncated generally at any point in the Fc region so as to prevent heavy chain crosslinking. Alternatively, the relevant cysteine residues are substituted with another amino acid residue or are deleted so as to prevent crosslinking.
  • In vitro methods are also suitable for preparing monovalent antibodies. Digestion of antibodies to produce fragments thereof, particularly, Fab fragments, can be accomplished using routine techniques known in the art.
  • the anti-PRO antibodies of the invention may further comprise humanized antibodies or human antibodies.
  • Humanized forms of non-human (e.g., murine) antibodies are chimeric immunoglobulins, immunoglobulin chains or fragments thereof (such as Fv, Fab, Fab', F(ab') 2 or other antigen-binding subsequences of antibodies) which contain minimal sequence derived from non-human immunoglobulin.
  • Humanized antibodies include human immunoglobulins (recipient antibody) in which residues from a complementary determining region (CDR) of the recipient are replaced by residues from a CDR of a non-human species (donor antibody) such as mouse, rat or rabbit having the desired specificity, affinity and capacity.
  • CDR complementary determining region
  • donor antibody non-human species
  • Fv framework residues of the human immunoglobulin are replaced by corresponding non-human residues.
  • Humanized antibodies may also comprise residues which are found neither in the recipient antibody nor in the imported CDR or framework sequences.
  • the humanized antibody will comprise substantially all of at least one, and typically two, variable domains, in which all or substantially all of the CDR regions correspond to those of a non-human immunoglobulin and all or substantially all of the FR regions are those of a human immunoglobulin consensus sequence.
  • the humanized antibody optimally also will comprise at least a portion of an immunoglobulin constant region (Fc), typically that of a human immunoglobulin [Jones et al., Nature. 321:522-525 (1986); Riechmann et al. , Nature. 332:323-329 (1988); and Presta, Curr. Op.
  • a humanized antibody has one or more amino acid residues introduced into it from a source which is non-human. These non- human amino acid residues are often referred to as "import" residues, which are typically taken from an "import” variable domain.
  • Humanization can be essentially performed following the method of Winter and co-workers [Jones et al., Nature. 321:522-525 (1986); Riechmann et al., Nature, 332:323-327 (1988); Verhoeyen et al., Science. 239: 1534-1536 (1988)], by substituting rodent CDRs or CDR sequences for the corresponding sequences of a human antibody.

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EP06000581A EP1666494A1 (en) 1999-12-01 2000-12-01 Secreted and transmembrane polypeptides and nucleic acids encoding the same
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EP06000585A EP1661996A1 (en) 1999-12-01 2000-12-01 Secreted and transmembrane polypeptides and nucleic acids encoding the same
EP06000583A EP1686134A3 (en) 1999-12-01 2000-12-01 Secreted and transmembrane polypeptides and nucleic acids encoding the same
EP06000588A EP1690873A3 (en) 1999-12-01 2000-12-01 Composition and methods for the diagnosis of tumours
EP06000587A EP1690872A3 (en) 1999-12-01 2000-12-01 Composition and methods for the diagnosis of tumours
EP06000584A EP1669371A3 (en) 1999-12-01 2000-12-01 Composition and methods for the diagnosis of tumours
EP05025102A EP1672070A3 (en) 1999-12-01 2000-12-01 Secreted and transmembrane polypeptides and nucleic acids encoding the same
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WOPCT/US99/30911 1999-02-20
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PCT/US1999/028301 WO2000032776A2 (en) 1998-12-01 1999-12-01 Secreted amd transmembrane polypeptides and nucleic acids encoding the same
WOPCT/US99/28301 1999-12-01
PCT/US1999/028634 WO2000036102A2 (en) 1998-12-16 1999-12-01 Secreted and transmembrane polypeptides and nucleic acids encoding the same
PCT/US1999/028565 WO2000037638A2 (en) 1998-12-22 1999-12-02 Methods and compositions for inhibiting neoplastic cell growth
WOPCT/US99/28564 1999-12-02
PCT/US1999/028551 WO2000053750A1 (en) 1999-03-08 1999-12-02 Compositions and methods for the treatment of tumors
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PCT/US1999/028564 WO2000055319A1 (en) 1999-03-12 1999-12-02 Methods and compositions for inhibiting neoplastic cell growth
US17026299P 1999-12-09 1999-12-09
US170262P 1999-12-09
PCT/US1999/030095 WO2000037640A2 (en) 1998-12-22 1999-12-16 Compositions and methods for the treatment of tumor
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PCT/US1999/030911 WO2000075316A1 (en) 1999-06-02 1999-12-20 Methods and compositions for inhibiting neoplastic cell growth
PCT/US1999/030999 WO2001005836A1 (en) 1999-07-20 1999-12-20 Polypeptidic compositions and methods for the treatment of tumors
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PCT/US2000/004914 WO2000075327A1 (en) 1999-06-02 2000-02-24 Methods and compositions for inhibiting neoplastic cell growth
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US18720200P 2000-03-03 2000-03-03
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PCT/US2000/006319 WO2000053760A2 (en) 1999-03-12 2000-03-10 Method of preventing the death of retinal neurons and treating ocular diseases
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PCT/US2000/013705 WO2000073445A2 (en) 1999-06-02 2000-05-17 Interleukin-1-receptor associated kinase-3 (irak3)
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PCT/US2000/020710 WO2001009327A2 (en) 1999-07-28 2000-07-28 Method of preventing the injury or death of retinal cells and treating ocular diseases
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Families Citing this family (129)

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US6812327B1 (en) 1996-10-25 2004-11-02 Human Genome Sciences, Inc. Neutrokine-alpha polypeptides
US8212004B2 (en) 1999-03-02 2012-07-03 Human Genome Sciences, Inc. Neutrokine-alpha fusion proteins
US7053190B2 (en) 1997-03-07 2006-05-30 Human Genome Sciences, Inc. Secreted protein HRGDF73
WO2001066740A2 (en) * 2000-03-03 2001-09-13 Genentech, Inc. Compositions and methods for the treatment of immune related diseases
US20020137890A1 (en) 1997-03-31 2002-09-26 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
US7057017B2 (en) 1997-04-16 2006-06-06 Millennium Pharmaceuticals, Inc. Human dickkopf-related protein and nucleic acid molecules and uses therefor
US6506569B1 (en) 1997-05-30 2003-01-14 Human Genome Sciences, Inc. Antibodies to human tumor necrosis factor receptor TR10
US8007798B2 (en) 1997-11-21 2011-08-30 Genentech, Inc. Treatment of complement-associated disorders
US7419663B2 (en) 1998-03-20 2008-09-02 Genentech, Inc. Treatment of complement-associated disorders
US7192589B2 (en) 1998-09-16 2007-03-20 Genentech, Inc. Treatment of inflammatory disorders with STIgMA immunoadhesins
US8088386B2 (en) 1998-03-20 2012-01-03 Genentech, Inc. Treatment of complement-associated disorders
US7446181B2 (en) 1998-01-15 2008-11-04 Millennium Pharmaceuticals, Inc. Antibodies that bind human Dickkopf-1 proteins
JP2002518010A (ja) 1998-06-16 2002-06-25 ヒューマン ジノーム サイエンシーズ， インコーポレイテッド ９４個のヒト分泌タンパク質
US20070014787A1 (en) 1998-07-15 2007-01-18 Human Genome Sciences, Inc. 71 human secreted proteins
US20020172678A1 (en) * 2000-06-23 2002-11-21 Napoleone Ferrara EG-VEGF nucleic acids and polypeptides and methods of use
US7264801B2 (en) 1998-08-11 2007-09-04 Genentech, Inc. EG-VEGF nucleic acids and polypeptides and method of use
CA2340884A1 (en) 1998-08-25 2000-03-02 Human Genome Sciences, Inc. 49 human secreted proteins
US6344541B1 (en) 1998-09-25 2002-02-05 Amgen Inc. DKR polypeptides
US7014996B1 (en) 1998-10-02 2006-03-21 Diadexus, Inc. Method of diagnosing, monitoring, staging, imaging and treating gynecologic cancers
US20030095967A1 (en) 1999-01-25 2003-05-22 Mackay Fabienne BAFF, inhibitors thereof and their use in the modulation of B-cell response and treatment of autoimmune disorders
US6630142B2 (en) 1999-05-03 2003-10-07 Zymogenetics, Inc. Method of treating fibroproliferative disorders
US6468543B1 (en) 1999-05-03 2002-10-22 Zymogenetics, Inc. Methods for promoting growth of bone using ZVEGF4
US7034132B2 (en) 2001-06-04 2006-04-25 Anderson David W Therapeutic polypeptides, nucleic acids encoding same, and methods of use
US6951738B2 (en) 1999-07-16 2005-10-04 Human Genome Sciences, Inc. Human tumor necrosis factor receptors TR13 and TR14
ES2355428T3 (es) 1999-08-12 2011-03-25 Agensys, Inc. Antígeno transmembranal de lectina tipo c expresado en cáncer de próstata humano y usos del mismo.
AU6063900A (en) * 1999-09-10 2001-04-10 Millennium Pharmaceuticals, Inc. Novel genes encoding proteins having prognostic, diagnostic, preventive, therapeutic, and other uses
ATE463510T1 (de) 1999-09-29 2010-04-15 Teijin Ltd Polypeptide und für diese kodierende gene
CA2386858C (en) 1999-10-28 2011-12-20 Agensys, Inc. 36p6d5: secreted tumor antigen
GB0001898D0 (en) * 2000-01-27 2000-03-22 Smithkline Beecham Plc Novel compounds
WO2001066766A2 (en) * 2000-03-06 2001-09-13 Darwin Molecular Corporation A matrix metalloproteinase (mmp-25) expressed in skin cell
JP2002335964A (ja) * 2000-04-27 2002-11-26 Takeda Chem Ind Ltd 新規タンパク質およびその用途
WO2001083775A2 (en) * 2000-05-04 2001-11-08 Incyte Genomics, Inc. Proteases
EP1280909A2 (en) * 2000-05-12 2003-02-05 Lexicon Genetics Incorporated Human lipocalin homologs and polynucleotides encoding the same
US7220840B2 (en) 2000-06-16 2007-05-22 Human Genome Sciences, Inc. Antibodies that immunospecifically bind to B lymphocyte stimulator protein
MXPA02012434A (es) 2000-06-16 2004-09-06 Cambridge Antibody Tech Anticuerpos que se unen inmunoespecificamente a estimulador de linfocitos ii.
EP1299538A2 (en) * 2000-06-20 2003-04-09 Incyte Genomics, Inc. Secreted proteins
EP1303536B1 (en) 2000-07-19 2010-03-17 Advanced Research And Technology Institute, Inc. Novel fibroblast growth factor (fgf23) and methods for use
US6984522B2 (en) 2000-08-03 2006-01-10 Regents Of The University Of Michigan Isolation and use of solid tumor stem cells
ES2344679T3 (es) 2000-08-11 2010-09-03 Kyowa Hakko Kirin Co., Ltd. Polipeptidos que controlan el metabolismo del acido fosforico, el metabolismo del calcio, el metabolismo de la calcificacion y de la vitamina d y moleculas de adn que los codifican.
WO2002016411A2 (en) 2000-08-18 2002-02-28 Human Genome Sciences, Inc. Binding polypeptides and methods based thereon
AU2001292601A1 (en) * 2000-09-08 2002-03-22 Millennium Pharmaceuticals, Inc. Human matrix metalloproteinase
US20020119139A1 (en) * 2000-10-11 2002-08-29 Michel Lazdunski Cloning and recombinant expression of mammalian group XII secreted phospholipase A2
US6531297B2 (en) * 2000-10-20 2003-03-11 Applera Corporation Isolated human drug-metabolizing proteins, nucleic acid molecules encoding human drug-metabolizing proteins, and uses thereof
US6949371B2 (en) 2000-10-20 2005-09-27 Applera Corporation Isolated human drug-metabolizing proteins, nucleic acid molecules encoding human drug-metabolizing proteins, and uses thereof
WO2002034920A2 (en) * 2000-10-24 2002-05-02 Bayer Aktiengesellschaft Regulation of human ceruloplasmin-like protein
WO2002036625A2 (en) 2000-11-03 2002-05-10 The Regents Of The University Of California Prokineticin polypeptides, related compositions and methods
WO2002057450A2 (en) * 2000-11-29 2002-07-25 Curagen Corporation Proteins and nucleic acids encoding same
US7109000B2 (en) 2001-03-08 2006-09-19 Curagen Corporation Proteins and nucleic acids encoding same
EP1409655A4 (en) * 2001-03-12 2005-04-06 Incyte Genomics Inc SUPERFAMILY PROTEINS OF IMMUNOGLOBULINS
WO2002083921A2 (en) 2001-04-10 2002-10-24 Agensys, Inc. Nuleic acids and corresponding proteins useful in the detection and treatment of various cancers
US20030013151A1 (en) * 2001-05-17 2003-01-16 Pe Corporation (Ny) Isolated human secreted proteins, nucleic acid molecules encoding human secreted proteins, and uses thereof
US20050272120A1 (en) 2001-06-20 2005-12-08 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2003000113A2 (en) * 2001-06-20 2003-01-03 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
WO2003002737A1 (fr) * 2001-06-27 2003-01-09 Riken Nouvelle proteine inhibitrice de la topoisomerase humaine 2$g(a) et utilisation associee
WO2003006673A2 (en) * 2001-07-11 2003-01-23 Texas Biotechnology Corporation A nucleic acid encoding a human junctional adhesion protein (jam3)
WO2003018805A1 (fr) * 2001-08-23 2003-03-06 Japan Science And Technology Agency Regulateurs de la proliferation de cellules souches hematopoietiques et polynucleotides codant ces regulateurs
US20030219823A1 (en) * 2001-09-07 2003-11-27 Alsobrook John P. Therapeutic polypeptides, nucleic acids encoding same, and methods of use
AU2002340118B2 (en) * 2001-10-04 2007-09-13 Immunex Corporation UL16 Binding protein 4
US7888478B2 (en) 2002-09-11 2011-02-15 Genentech, Inc. Compositions and methods for the treatment of tumor of hematopoietic origin
US7858330B2 (en) 2001-10-19 2010-12-28 Genentech, Inc. Compositions and methods for the treatment of tumor of hematopoietic origin
ATE384783T1 (de) 2001-10-19 2008-02-15 Zymogenetics Inc Dimerisierter wachstumsfaktor sowie materialien und verfahren zu seiner herstellung
US20050123925A1 (en) 2002-11-15 2005-06-09 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
EP1463525B1 (en) * 2001-12-19 2008-09-17 Genentech, Inc. Compositions and methods for the diagnosis and treatment of tumor
CN100519584C (zh) 2001-12-28 2009-07-29 麒麟医药株式会社 抗成纤维细胞生长因子23的抗体
AU2003210996A1 (en) 2002-02-11 2003-09-04 Zymogenetics, Inc. Materials and methods for preparing dimeric growth factors
EP3115470B1 (en) 2002-03-13 2018-07-18 Genomic Health, Inc. Gene expression profiling in biopsied tumor tissues
US7507808B2 (en) 2002-12-12 2009-03-24 Isis Pharmaceuticals, Inc. Modulation of endothelial lipase expression
CN101613406A (zh) * 2002-06-06 2009-12-30 肿瘤疗法科学股份有限公司 与人结肠癌相关的基因和多肽
US20040224378A1 (en) * 2002-06-07 2004-11-11 Athersys, Inc. Methods for using ADAMTS-12, an integrin and metalloprotease with thrombspondin motifs
WO2004022087A1 (ja) * 2002-09-04 2004-03-18 Takeda Pharmaceutical Company Limited 疾患関連遺伝子の用途
DE10242146A1 (de) * 2002-09-04 2004-03-18 Eberhard-Karls-Universität Tübingen Universitätsklinikum Antikörper zur Identifizierung und/oder Isolierung von hämatopoetischen Stammzellen
JP2006512903A (ja) * 2002-09-11 2006-04-20 ジェネンテック・インコーポレーテッド 免疫関連疾患の治療のための新規組成物と方法
AU2003270621A1 (en) * 2002-09-13 2004-04-30 Richard D. Cummings CORE 1 Beta3-GALACTOSYLTRANSFERASE SPECIFIC MOLECULAR CHAPERONES, NUCLEIC ACIDS, AND METHODS OF USE THEREOF
CA2499843A1 (en) * 2002-09-25 2004-04-08 Genentech, Inc. Novel compositions and methods for the treatment of psoriasis
US7662917B2 (en) 2002-09-30 2010-02-16 Ludwig Institute For Cancer Research Human sarcoma-associated antigens
DE60325628D1 (de) 2002-09-30 2009-02-12 Oncotherapy Science Inc Gene und polypeptide in verbindung mit menschlichen pankreaskarzinomen
US7560537B2 (en) 2002-09-30 2009-07-14 Ludwig Institute For Cancer Research Human sarcoma-associated antigens
JP2004135605A (ja) * 2002-10-18 2004-05-13 Nissui Pharm Co Ltd 脂肪細胞分化関連遺伝子およびタンパク質
AU2003295598B2 (en) 2002-11-15 2009-12-24 Genomic Health, Inc. Gene expression profiling of EGFR positive cancer
DE10254601A1 (de) 2002-11-22 2004-06-03 Ganymed Pharmaceuticals Ag Differentiell in Tumoren exprimierte Genprodukte und deren Verwendung
US20040231909A1 (en) 2003-01-15 2004-11-25 Tai-Yang Luh Motorized vehicle having forward and backward differential structure
WO2004074518A1 (en) 2003-02-20 2004-09-02 Genomic Health, Inc. Use of intronic rna to measure gene expression
NZ541480A (en) 2003-03-12 2008-03-28 Genentech Inc Use of Bv8 and/or EG-VEGF to promote hematopoiesis
WO2004085666A2 (en) * 2003-03-20 2004-10-07 Incyte Corporation Secreted proteins
HUE050365T2 (hu) 2003-06-24 2022-09-28 Genomic Health Inc Rákkiújulás valószínûségének elõrejelzése
ES2905579T3 (es) 2003-07-10 2022-04-11 Genomic Health Inc Algoritmo del perfil de expresión y prueba para el pronóstico de la recaída del cáncer de mama
JP2005168338A (ja) * 2003-12-09 2005-06-30 Japan Science & Technology Agency 新規タンパク質カルフォグリン、これをコードする遺伝子、薬剤、及び標的物質
US20050196782A1 (en) 2003-12-23 2005-09-08 Kiefer Michael C. Universal amplification of fragmented RNA
WO2005068497A1 (en) * 2004-01-06 2005-07-28 The United States Of America As Represented By The Secretary, Department Of Health And Human Services Iis: a novel ifn-gamma-inducible protein, member of the secretoglobin family, that modulates cellular invasion and migration
US20050186577A1 (en) 2004-02-20 2005-08-25 Yixin Wang Breast cancer prognostics
DK2163650T3 (en) 2004-04-09 2015-11-02 Genomic Health Inc Genekspressionsmarkører for prediction of response to chemotherapy
DE102004024617A1 (de) 2004-05-18 2005-12-29 Ganymed Pharmaceuticals Ag Differentiell in Tumoren exprimierte Genprodukte und deren Verwendung
WO2005118641A1 (en) * 2004-06-04 2005-12-15 Applied Research Systems Ars Holding N.V. Splice variant of unc5h2
EP2474317A1 (en) 2004-06-24 2012-07-11 Biogen Idec MA Inc. Treatment of conditions involving demyelination
US7587279B2 (en) 2004-07-06 2009-09-08 Genomic Health Method for quantitative PCR data analysis system (QDAS)
EP2336177A1 (en) 2004-08-04 2011-06-22 Amgen, Inc Antibodies to DKK-1
DK1815014T3 (da) 2004-11-05 2012-06-18 Genomic Health Inc Molekylære indikatorer for brystcancerprognose og forudsigelse af behandlingsrespons
CA2585571C (en) 2004-11-05 2020-01-21 Genomic Health, Inc. Predicting response to chemotherapy using gene expression markers
WO2006130135A1 (en) * 2005-05-31 2006-12-07 Genentech, Inc. Compositions and methods for the treatment of tumor of hematopoietic origin
JP2009502123A (ja) 2005-07-08 2009-01-29 バイオジェン・アイデック・エムエイ・インコーポレイテッド Ｓｐ３５抗体およびその使用
US9168286B2 (en) 2005-10-13 2015-10-27 Human Genome Sciences, Inc. Methods and compositions for use in treatment of patients with autoantibody positive disease
KR20150055628A (ko) 2005-11-04 2015-05-21 제넨테크, 인크. 안질환 치료를 위한 보체 경로 억제제의 용도
EP1790664A1 (en) 2005-11-24 2007-05-30 Ganymed Pharmaceuticals AG Monoclonal antibodies against claudin-18 for treatment of cancer
US8211649B2 (en) 2006-03-31 2012-07-03 Human Genome Sciences, Inc. Methods of diagnosing and prognosing hodgkin's lymphoma
HUE028379T2 (en) 2006-09-29 2016-12-28 Oncomed Pharm Inc Preparations and procedures for the diagnosis and treatment of cancer
DK2097455T3 (en) 2006-11-02 2014-12-15 Genentech Inc HUMANIZED ANTI-FACTOR D ANTIBODIES
US8128926B2 (en) 2007-01-09 2012-03-06 Biogen Idec Ma Inc. Sp35 antibodies and uses thereof
US7883705B2 (en) 2007-02-14 2011-02-08 Kyowa Hakko Kirin Co., Ltd. Anti FGF23 antibody and a pharmaceutical composition comprising the same
AR066660A1 (es) 2007-05-23 2009-09-02 Genentech Inc Prevencion y tratamiento de condiciones del ojo asociadas con su complemento
US9143569B2 (en) 2008-02-21 2015-09-22 Dexcom, Inc. Systems and methods for processing, transmitting and displaying sensor data
CR20170001A (es) 2008-04-28 2017-08-10 Genentech Inc Anticuerpos anti factor d humanizados
TWI508973B (zh) 2008-05-06 2015-11-21 建南德克公司 親和力成熟之ＣＲＩｇ變體
JP2011527572A (ja) 2008-07-09 2011-11-04 バイオジェン・アイデック・エムエイ・インコーポレイテッド Ｌｉｎｇｏ抗体または断片を含む組成物
TR201818814T4 (tr) 2009-10-16 2019-01-21 Oncomed Pharm Inc Dll4 antagonist antikorlarının ve anti-hipertansif ajanların terapötik kombinasyonu ve kullanımı.
US8551479B2 (en) 2010-09-10 2013-10-08 Oncomed Pharmaceuticals, Inc. Methods for treating melanoma
JP5891561B2 (ja) * 2011-06-03 2016-03-23 学校法人自治医科大学 ミトコンドリア膜タンパク質群およびそれらをコードする遺伝子群
ES2707580T3 (es) 2011-09-23 2019-04-04 Oncomed Pharm Inc Agentes de unión a VEGF/DLL4 y usos de los mismos
WO2013167153A1 (en) 2012-05-09 2013-11-14 Ganymed Pharmaceuticals Ag Antibodies useful in cancer diagnosis
CA2873623C (en) 2012-05-14 2021-11-09 Biogen Idec Ma Inc. Lingo-2 antagonists for treatment of conditions involving motor neurons
WO2014071018A1 (en) 2012-10-31 2014-05-08 Oncomed Pharmaceuticals, Inc. Methods and monitoring of treatment with a dll4 antagonist
US10093978B2 (en) 2013-08-12 2018-10-09 Genentech, Inc. Compositions for detecting complement factor H (CFH) and complement factor I (CFI) polymorphisms
MA39934A (fr) 2014-05-01 2017-03-08 Hoffmann La Roche Variants d'anticorps anti-facteur d et leurs utilisations
WO2016070051A2 (en) 2014-10-31 2016-05-06 Oncomed Pharmaceuticals, Inc. Combination therapy for treatment of disease
AU2016205197B2 (en) 2015-01-08 2021-10-21 Biogen Ma Inc. LINGO-1 antagonists and uses for treatment of demyelinating disorders
JP6967003B2 (ja) 2015-09-23 2021-11-17 メレオ バイオファーマ ５ インコーポレイテッド がんの処置のための方法および組成物
WO2017075173A2 (en) 2015-10-30 2017-05-04 Genentech, Inc. Anti-factor d antibodies and conjugates
WO2017075252A1 (en) 2015-10-30 2017-05-04 Genentech, Inc. Anti-factor d antibody variant conjugates and uses thereof
SG10202111399YA (en) 2015-12-22 2021-11-29 Immatics Biotechnologies Gmbh Peptides and combination of peptides for use in immunotherapy against breast cancer and other cancers
GB201522667D0 (en) 2015-12-22 2016-02-03 Immatics Biotechnologies Gmbh Novel peptides and combination of peptides for use in immunotherapy against breast cancer and other cancers

Family Cites Families (116)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US3773919A (en) 1969-10-23 1973-11-20 Du Pont Polylactide-drug mixtures
US4179337A (en) 1973-07-20 1979-12-18 Davis Frank F Non-immunogenic polypeptides
FR2413974A1 (fr) 1978-01-06 1979-08-03 David Bernard Sechoir pour feuilles imprimees par serigraphie
US4275149A (en) 1978-11-24 1981-06-23 Syva Company Macromolecular environment control in specific receptor assays
US4657760A (en) 1979-03-20 1987-04-14 Ortho Pharmaceutical Corporation Methods and compositions using monoclonal antibody to human T cells
JPS6023084B2 (ja) 1979-07-11 1985-06-05 味の素株式会社 代用血液
US4399216A (en) 1980-02-25 1983-08-16 The Trustees Of Columbia University Processes for inserting DNA into eucaryotic cells and for producing proteinaceous materials
ZA811368B (en) 1980-03-24 1982-04-28 Genentech Inc Bacterial polypedtide expression employing tryptophan promoter-operator
US4485045A (en) 1981-07-06 1984-11-27 Research Corporation Synthetic phosphatidyl cholines useful in forming liposomes
NZ201705A (en) 1981-08-31 1986-03-14 Genentech Inc Recombinant dna method for production of hepatitis b surface antigen in yeast
US4640835A (en) 1981-10-30 1987-02-03 Nippon Chemiphar Company, Ltd. Plasminogen activator derivatives
US4943529A (en) 1982-05-19 1990-07-24 Gist-Brocades Nv Kluyveromyces as a host strain
US4870009A (en) 1982-11-22 1989-09-26 The Salk Institute For Biological Studies Method of obtaining gene product through the generation of transgenic animals
AU2353384A (en) 1983-01-19 1984-07-26 Genentech Inc. Amplification in eukaryotic host cells
US4713339A (en) 1983-01-19 1987-12-15 Genentech, Inc. Polycistronic expression vector construction
NZ207394A (en) 1983-03-08 1987-03-06 Commw Serum Lab Commission Detecting or determining sequence of amino acids
US4816567A (en) 1983-04-08 1989-03-28 Genentech, Inc. Recombinant immunoglobin preparations
DD266710A3 (de) 1983-06-06 1989-04-12 Ve Forschungszentrum Biotechnologie Verfahren zur biotechnischen Herstellung van alkalischer Phosphatase
US4544545A (en) 1983-06-20 1985-10-01 Trustees University Of Massachusetts Liposomes containing modified cholesterol for organ targeting
AU3145184A (en) 1983-08-16 1985-02-21 Zymogenetics Inc. High expression of foreign genes in schizosaccharomyces pombe
US4496689A (en) 1983-12-27 1985-01-29 Miles Laboratories, Inc. Covalently attached complex of alpha-1-proteinase inhibitor with a water soluble polymer
US4736866A (en) 1984-06-22 1988-04-12 President And Fellows Of Harvard College Transgenic non-human mammals
US4879231A (en) 1984-10-30 1989-11-07 Phillips Petroleum Company Transformation of yeasts of the genus pichia
DE3675588D1 (de) 1985-06-19 1990-12-20 Ajinomoto Kk Haemoglobin, das an ein poly(alkenylenoxid) gebunden ist.
US5206344A (en) 1985-06-26 1993-04-27 Cetus Oncology Corporation Interleukin-2 muteins and polymer conjugation thereof
US4676980A (en) 1985-09-23 1987-06-30 The United States Of America As Represented By The Secretary Of The Department Of Health And Human Services Target specific cross-linked heteroantibodies
WO1987005330A1 (en) 1986-03-07 1987-09-11 Michel Louis Eugene Bergh Method for enhancing glycoprotein stability
GB8610600D0 (en) 1986-04-30 1986-06-04 Novo Industri As Transformation of trichoderma
US4791192A (en) 1986-06-26 1988-12-13 Takeda Chemical Industries, Ltd. Chemically modified protein with polyethyleneglycol
US5773249A (en) * 1986-11-04 1998-06-30 Protein Polymer Technologies, Inc. High molecular weight collagen-like protein polymers
US4946783A (en) 1987-01-30 1990-08-07 President And Fellows Of Harvard College Periplasmic protease mutants of Escherichia coli
US5010182A (en) 1987-07-28 1991-04-23 Chiron Corporation DNA constructs containing a Kluyveromyces alpha factor leader sequence for directing secretion of heterologous polypeptides
IL87737A (en) 1987-09-11 1993-08-18 Genentech Inc Method for culturing polypeptide factor dependent vertebrate recombinant cells
GB8724885D0 (en) 1987-10-23 1987-11-25 Binns M M Fowlpox virus promotors
AU633248B2 (en) 1987-12-21 1993-01-28 University Of Toledo, The Agrobacterium mediated transformation of germinating plant seeds
AU4005289A (en) 1988-08-25 1990-03-01 Smithkline Beecham Corporation Recombinant saccharomyces
GB8823869D0 (en) 1988-10-12 1988-11-16 Medical Res Council Production of antibodies
US5225538A (en) 1989-02-23 1993-07-06 Genentech, Inc. Lymphocyte homing receptor/immunoglobulin fusion proteins
US5009772A (en) 1989-02-27 1991-04-23 Kerr-Mcgee Corporation Solvent extraction process
EP0462145B1 (en) 1989-03-07 1994-04-27 Genentech, Inc. Covalent conjugates of lipid and oligonucleotide
FR2646437B1 (fr) 1989-04-28 1991-08-30 Transgene Sa Nouvelles sequences d'adn, leur application en tant que sequence codant pour un peptide signal pour la secretion de proteines matures par des levures recombinantes, cassettes d'expression, levures transformees et procede de preparation de proteines correspondant
DE394538T1 (de) 1989-04-28 1991-04-11 Rhein Biotech Ges. Fuer Biotechnologische Prozesse Und Produkte Mbh, 4000 Duesseldorf Hefezellen der schwanniomyces-gattung.
CA2055435A1 (en) 1989-05-10 1990-11-11 Eli Gilboa Stably transformed eucaryotic cells comprising a foreign transcribable dna under the control of a pol iii promoter
EP0402226A1 (en) 1989-06-06 1990-12-12 Institut National De La Recherche Agronomique Transformation vectors for yeast yarrowia
DE3920358A1 (de) 1989-06-22 1991-01-17 Behringwerke Ag Bispezifische und oligospezifische, mono- und oligovalente antikoerperkonstrukte, ihre herstellung und verwendung
DK0479909T3 (da) 1989-06-29 1997-04-07 Medarex Inc Bispecifikke reagenser til AIDS-behandling
FR2649120B1 (fr) 1989-06-30 1994-01-28 Cayla Nouvelle souche et ses mutants de champignons filamenteux, procede de production de proteines recombinantes a l'aide de ladite souche et souches et proteines obtenues selon ce procede
WO1991004753A1 (en) 1989-10-02 1991-04-18 Cetus Corporation Conjugates of antisense oligonucleotides and therapeutic uses thereof
US5013556A (en) 1989-10-20 1991-05-07 Liposome Technology, Inc. Liposomes with enhanced circulation time
US5225212A (en) 1989-10-20 1993-07-06 Liposome Technology, Inc. Microreservoir liposome composition and method
JPH05504553A (ja) 1989-10-24 1993-07-15 ギリアド　サイエンシズ，インコーポレイテッド 新規の結合を有するオリゴヌクレオチドアナログ
ES2246502T3 (es) 1990-08-29 2006-02-16 Genpharm International, Inc. Animales no humanos transgenicos capaces de producir anticuerpos heterologos.
US5625126A (en) 1990-08-29 1997-04-29 Genpharm International, Inc. Transgenic non-human animals for producing heterologous antibodies
US5661016A (en) 1990-08-29 1997-08-26 Genpharm International Inc. Transgenic non-human animals capable of producing heterologous antibodies of various isotypes
US5633425A (en) 1990-08-29 1997-05-27 Genpharm International, Inc. Transgenic non-human animals capable of producing heterologous antibodies
US5545806A (en) 1990-08-29 1996-08-13 Genpharm International, Inc. Ransgenic non-human animals for producing heterologous antibodies
US5122469A (en) 1990-10-03 1992-06-16 Genentech, Inc. Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins
US5206161A (en) 1991-02-01 1993-04-27 Genentech, Inc. Human plasma carboxypeptidase B
JPH06507398A (ja) 1991-05-14 1994-08-25 リプリジェン　コーポレーション Ｈｉｖ感染治療のための異種複合抗体
WO1993008829A1 (en) 1991-11-04 1993-05-13 The Regents Of The University Of California Compositions that mediate killing of hiv-infected cells
DK0617706T3 (da) 1991-11-25 2001-11-12 Enzon Inc Multivalente antigenbindende proteiner
EP0669836B1 (en) 1992-11-13 1996-07-03 Idec Pharmaceuticals Corporation Therapeutic application of chimeric and radiolabeled antibodies to human b lymphocyte restricted differentiation antigen for treatment of b cell lymphoma
DE69329295T2 (de) 1992-12-02 2001-03-15 Alkermes Controlled Therapeutics, Inc. Wachstumhormon enthaltende mikrosphaeren mit kontrollierter freisetzung
US5536637A (en) 1993-04-07 1996-07-16 Genetics Institute, Inc. Method of screening for cDNA encoding novel secreted mammalian proteins in yeast
PT779806E (pt) 1994-09-09 2001-02-28 Takeda Chemical Industries Ltd Preparacao de libertacao sustentada contendo um sal metalico de um peptido
US5731168A (en) 1995-03-01 1998-03-24 Genentech, Inc. Method for making heteromultimeric polypeptides
JPH11506740A (ja) 1995-06-07 1999-06-15 アルカームズ コントロールド セラピューティックス，インコーポレイテッド ヒト成長ホルモンの徐放性組成物
ZA965368B (en) 1995-07-14 1997-01-14 Novo Nordisk As A pharmaceutical formulation
US5707829A (en) * 1995-08-11 1998-01-13 Genetics Institute, Inc. DNA sequences and secreted proteins encoded thereby
CN1211279A (zh) * 1996-01-10 1999-03-17 科里克萨有限公司 用于治疗和诊断乳腺癌的组合物和方法
US6458939B1 (en) 1996-03-15 2002-10-01 Millennium Pharmaceuticals, Inc. Compositions and methods for the diagnosis, prevention, and treatment of neoplastic cell growth and proliferation
US5824504A (en) * 1996-09-26 1998-10-20 Elshourbagy; Nabil A. Human 7-transmembrane receptor and DNA
US5871965A (en) * 1996-10-25 1999-02-16 Incyte Pharmaceuticals, Inc. Guanylate binding proteins
AU4885297A (en) * 1996-11-13 1998-06-03 Protegene Inc. Human proteins having transmembrane domains and DNAs encoding these prot eins
JPH10295388A (ja) * 1997-02-28 1998-11-10 Japan Tobacco Inc 哺乳動物由来生理活性タンパク
WO1998056804A1 (en) * 1997-06-13 1998-12-17 Human Genome Sciences, Inc. 86 human secreted proteins
EP1394252A3 (en) * 1997-03-07 2004-03-10 Human Genome Sciences, Inc. 70 human secreted proteins
WO1998039446A2 (en) * 1997-03-07 1998-09-11 Human Genome Sciences, Inc. 70 human secreted proteins
ZA982968B (en) * 1997-04-09 1998-10-27 Corixa Corp Compositions and methods for the treatment and diagnosis of breast cancer
AU6956798A (en) * 1997-04-10 1998-10-30 Genetics Institute Inc. Secreted expressed sequence tags (sests)
ATE361933T1 (de) * 1997-08-01 2007-06-15 Serono Genetics Inst Sa 5' ests für sekretierte proteine aus gehirngeweben
JP2002537756A (ja) * 1997-08-21 2002-11-05 ジェネティックス・インスチチュート・インコーポレーテッド 分泌蛋白
WO2002024888A2 (en) * 2000-09-01 2002-03-28 Genentech, Inc. Secreted and transmembrane polypeptides and nucleic acids encoding the same
KR100502596B1 (ko) * 1997-09-17 2005-07-22 제넨테크, 인크. 분비 폴리펩티드와 막횡단 폴리펩티드 및 이들을 코딩하는핵산
AU9312198A (en) * 1997-09-17 1999-04-05 Genentech Inc. Genes amplified in tumours, antibodies against the proteins encoded thereby, andtheir use in diagnosis and treatment of cancer
KR100386046B1 (ko) * 1997-12-03 2003-06-02 제넨테크, 인크. 폴리펩티드 및 이를 암호화하는 핵산
ID27813A (id) * 1998-01-28 2001-04-26 Corixa Corp Senyawa-senyawa untuk terapi dan diagnosa kanker paru-paru dan metoda untuk penggunaannya
JP2002537757A (ja) * 1998-08-24 2002-11-12 アルファジーン・インコーポレイテッド 分泌タンパク質及びそれらをコードするポリヌクレオチド
WO2000021991A1 (en) * 1998-10-15 2000-04-20 Genetics Institute, Inc. SECRETED EXPRESSED SEQUENCE TAGS (sESTs)
WO2001053455A2 (en) * 1999-12-23 2001-07-26 Hyseq, Inc. Novel nucleic acids and polypeptides
AU2390800A (en) * 1998-12-23 2000-07-12 Genetics Institute Inc. Secreted proteins
AU3395900A (en) * 1999-03-12 2000-10-04 Human Genome Sciences, Inc. Human lung cancer associated gene sequences and polypeptides
CA2383592A1 (en) * 1999-03-31 2000-10-05 Curagen Corporation 2384891 acids including open reading frames encoding polypeptides; orfx
EP1173566A2 (en) * 1999-04-23 2002-01-23 Incyte Genomics, Inc. Human membrane-associated proteins
AU5047800A (en) * 1999-05-28 2000-12-18 Ludwig Institute For Cancer Research Breast, gastric and prostate cancer associated antigens and uses therefor
JP2003518920A (ja) * 1999-07-02 2003-06-17 カイロン コーポレイション 新規なヒト遺伝子および遺伝子発現産物
EP1130094A3 (en) * 1999-07-08 2001-11-21 Helix Research Institute Primers for synthesizing full length cDNA clones and their use
EP1074617A3 (en) * 1999-07-29 2004-04-21 Research Association for Biotechnology Primers for synthesising full-length cDNA and their use
US6780627B1 (en) * 2000-01-31 2004-08-24 Millennium Pharmaceuticals, Inc. 22438, 23553, 25278, and 26212 novel human sulfatases
AU7721500A (en) * 1999-09-29 2001-04-30 Human Genome Sciences, Inc. Colon and colon cancer associated polynucleotides and polypeptides
AU7999300A (en) * 1999-10-07 2001-05-10 Zymogenetics Inc. Novel human phosphodiesterase zcytor13
WO2001053312A1 (en) * 1999-12-23 2001-07-26 Hyseq, Inc. Novel nucleic acids and polypeptides
CA2398003A1 (en) * 2000-01-28 2001-08-02 Incyte Genomics, Inc. Phosphodiesterases
WO2001054472A2 (en) * 2000-01-31 2001-08-02 Human Genome Sciences, Inc. Nucleic acids, proteins, and antibodies
WO2001055326A2 (en) * 2000-01-31 2001-08-02 Human Genome Sciences, Inc. Nucleic acids, proteins, and antibodies
CA2392757A1 (en) * 2000-01-31 2001-08-02 Human Genome Sciences Inc. Nucleic acids, proteins, and antibodies
CA2395676A1 (en) * 2000-01-31 2001-08-02 Human Genome Sciences, Inc. Nucleic acids, proteins, and antibodies
US20030149238A1 (en) * 2000-02-14 2003-08-07 Pankaj Agarwal Novel compounds
US6436703B1 (en) * 2000-03-31 2002-08-20 Hyseq, Inc. Nucleic acids and polypeptides
WO2001074836A1 (en) * 2000-03-31 2001-10-11 Hyseq, Inc. Novel bone marrow nucleic acids and polypeptides
JP2004528003A (ja) * 2000-06-30 2004-09-16 インサイト・ゲノミックス・インコーポレイテッド 細胞外マトリクスおよび細胞接着分子
EP1343886A2 (en) * 2000-07-11 2003-09-17 Corixa Corporation Compositions and methods for the therapy and diagnosis of lung cancer
EP1346040A2 (en) * 2000-09-11 2003-09-24 Nuvelo, Inc. Novel nucleic acids and polypeptides
US20040044184A1 (en) * 2000-10-27 2004-03-04 Baughn Mariah R Cytoskeleton-associated proteins
DE10055285A1 (de) * 2000-11-08 2002-06-06 Deutsches Krebsforsch Neue Marker für die Diagnose und Therapie von Tumoren
EP1661986A1 (en) * 2000-11-17 2006-05-31 Nuvelo, Inc. Nucleic acids and polypeptides enocoded thereby that are member of the kallikrein gene familyNovel nucleic acids and polypeptides

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See references of WO0140466A3 *

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