US2002155545A1PendingUtilityA1
Chemokines expressed in pancreas
Est. expiryFeb 17, 2015(expired)· nominal 20-yr term from priority
A61P 43/00A61K 38/00C07K 16/24C12Q 1/68G01N 33/6893A61P 29/00C07K 14/523C07K 1/22C12Q 2600/118G01N 2800/067G01N 33/6863
54
PatentIndex Score
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Claims
Abstract
The present invention provides nucleotide and amino acid sequences that identify and encode novel expressed chemokines (PANEC-1 and PANEC-2) from human pancreas cells. The present invention also provides for antisense molecules to the nucleotide sequences which encode PANEC-1 and PANEC-2, expression vectors for the production of purified PANEC-1 and PANEC-2, antibodies capable of binding specifically to PANEC-1 and PANEC-2, hybridization probes or oligonucleotides for the detection of PANEC-1- or PANEC-2- encoding nucleotide sequences, genetically engineered host cells for the expression of PANEC-1 and PANEC-2, diagnostic tests for chemokine activation based on PANEC-1- and PANEC-2- encoding nucleic acid molecules and antibodies capable of binding specifically to the protein.
Claims
exact text as granted — not AI-modifiedWhat is claimed is:
1 . An isolated polypeptide selected from the group consisting of:
a) a polypeptide comprising an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4, b) a polypeptide comprising a naturally occurring polypeptide comprising an amino acid sequence at least 90% identical to an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4, c) a biologically active fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4, and d) an immunogenic fragment of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
2 . An isolated polypeptide of claim 1 , having a sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
3 . An isolated polynucleotide encoding a polypeptide of claim 1 .
4 . An isolated polynucleotide encoding a polypeptide of claim 2 .
5 . An isolated polynucleotide of claim 4 , having a sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:3.
6 . A recombinant polynucleotide comprising a promoter sequence operably linked to a polynucleotide of claim 3 .
7 . A cell transformed with a recombinant polynucleotide of claim 6 .
8 . A transgenic organism comprising a recombinant polynucleotide of claim 6 .
# SEQUENCE LISTING
(1) GENERAL INFORMATION:
(iii) NUMBER OF SEQUENCES: 11
(2) INFORMATION FOR SEQ ID NO: 1:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 289 base
#pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human Panc
#reas
(B) CLONE: 223187
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#1:
ATGAAGGTCT CCGCAGCACT TCTGTGGCTG CTGCTCATAG CAGCTGCCTT CA
#GCCCCCAG 60
GGGCTCACTG GGCCAGCTTC TGTCCCAACC ACCTGCTGCT TTAACCTGGC CA
#ATAGGAAG 120
ATACCCCTTC AGCGACTAGA GAGCTACAGG AGAATCACCA GTGGCAAATG TC
#CCCAGAAA 180
GCTGTGATCT TCAAGACCAA ACTGGCCAAG GATATCTGTG CCGACCCCAA GA
#AGAAGTGG 240
GTGCAGGATT CCATGAAGTA TCTGGACCAA AAATCTCCAA CTCCAAAGC
# 289
(2) INFORMATION FOR SEQ ID NO: 2:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 97 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human Panc
#reas
(B) CLONE: 223187
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#2:
Met Lys Val Ser Ala Ala Leu Leu Trp Leu Le
#u Leu Ile Ala Ala Ala
1 5
# 10
# 15
Phe Ser Pro Gln Gly Leu Thr Gly Pro Ala Se
#r Val Pro Thr Thr Cys
20
# 25
# 30
Cys Phe Asn Leu Ala Asn Arg Lys Ile Pro Le
#u Gln Arg Leu Glu Ser
35
# 40
# 45
Tyr Arg Arg Ile Thr Ser Gly Lys Cys Pro Gl
#n Lys Ala Val Ile Phe
50
# 55
# 60
Lys Thr Lys Leu Ala Lys Asp Ile Cys Ala As
#p Pro Lys Lys Lys Trp
65
#70
#75
#80
Val Gln Asp Ser Met Lys Tyr Leu Asp Gln Ly
#s Ser Pro Thr Pro Lys
85
# 90
# 95
Pro
(2) INFORMATION FOR SEQ ID NO: 3:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 402 base
#pairs
(B) TYPE: nucleic acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: cDNA
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human Panc
#reas
(B) CLONE: 226152
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#3:
ATGGCTCAGT CACTGGCTCT GAGCCTCCTT ATCCTGGTTC TGGCCTTTGG CA
#TCCCCAGG 60
ACCCAAGGCA GTGATGGAGG GGCTCAGGAC TGTTGCCTCA AGTACAGCCA AA
#GGAAGATT 120
CCCGCCAAGG TTGTCCGCAG CTACCGGAAG CAGGAACCAA GCTTAGGCTG CT
#CCATCCCA 180
GCTATCCTGT TCTTGCCCCG CAAGCGCTCT CAGGCAGAGC TATGTGCAGA CC
#CAAAGGAG 240
CTCTGGGTGC AGCAGCTGAT GCAGCATCTG GACAAGACAC CATCCCCACA GA
#AACCAGCC 300
CAGGGCTGCA GGAAGGACAG GGGGGCCTCC AAGACTGGCA AGAAAGGAAA GG
#GCTCCAAA 360
GGCTGCAAGA GGACTGAGCG GTCACAGACC CCTAAAGGGC CA
#
# 402
(2) INFORMATION FOR SEQ ID NO: 4:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 134 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: Human Panc
#reas
(B) CLONE: 226152
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#4:
Met Ala Gln Ser Leu Ala Leu Ser Leu Leu Il
#e Leu Val Leu Ala Phe
1 5
# 10
# 15
Gly Ile Pro Arg Thr Gln Gly Ser Asp Gly Gl
#y Ala Gln Asp Cys Cys
20
# 25
# 30
Leu Lys Tyr Ser Gln Arg Lys Ile Pro Ala Ly
#s Val Val Arg Ser Tyr
35
# 40
# 45
Arg Lys Gln Glu Pro Ser Leu Gly Cys Ser Il
#e Pro Ala Ile Leu Phe
50
# 55
# 60
Leu Pro Arg Lys Arg Ser Gln Ala Glu Leu Cy
#s Ala Asp Pro Lys Glu
65
#70
#75
#80
Leu Trp Val Gln Gln Leu Met Gln His Leu As
#p Lys Thr Pro Ser Pro
85
# 90
# 95
Gln Lys Pro Ala Gln Gly Cys Arg Lys Asp Ar
#g Gly Ala Ser Lys Thr
100
# 105
# 110
Gly Lys Lys Gly Lys Gly Ser Lys Gly Cys Ly
#s Arg Thr Glu Arg Ser
115
# 120
# 125
Gln Thr Pro Lys Gly Pro
130
(2) INFORMATION FOR SEQ ID NO: 5:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 97 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#5:
Met Lys Val Ser Ala Ala Leu Leu Ala Leu Le
#u Leu Ile Ala Ala Ala
1 5
# 10
# 15
Phe Cys Pro Gln Gly Leu Ala Gln Pro Asp Gl
#y Val Asp Thr Pro Thr
20
# 25
# 30
Thr Cys Cys Phe Asn Tyr Ile Asn Arg Lys Il
#e Pro Arg Gln Arg Leu
35
# 40
# 45
Glu Ser Tyr Arg Arg Ile Thr Ser Ser Lys Cy
#s Ser Lys Pro Ala Val
50
# 55
# 60
Ile Phe Lys Thr Lys Arg Ala Lys Gln Val Cy
#s Ala Asp Pro Lys Glu
65
#70
#75
#80
Lys Trp Val Gln Asp Ser Met Lys His Leu As
#p Lys Gln Thr Pro Lys
85
# 90
# 95
Pro
(2) INFORMATION FOR SEQ ID NO: 6:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 92 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: MIP-1a
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#6:
Met Gln Val Ser Thr Ala Ala Leu Ala Val Le
#u Leu Cys Thr Met Ala
1 5
# 10
# 15
Leu Cys Asn Gln Phe Ser Ala Ser Leu Ala Al
#a Asp Thr Pro Thr Ala
20
# 25
# 30
Cys Cys Phe Ser Tyr Thr Ser Arg Gln Ile Pr
#o Gln Asn Phe Ile Ala
35
# 40
# 45
Asp Tyr Phe Glu Thr Ser Ser Gln Cys Ser Ly
#s Pro Gly Val Ile Phe
50
# 55
# 60
Leu Thr Lys Arg Ser Arg Gln Val Cys Ala As
#p Pro Ser Glu Glu Trp
65
#70
#75
#80
Val Gln Lys Tyr Val Ser Asp Leu Glu Leu Se
#r Ala
85
# 90
(2) INFORMATION FOR SEQ ID NO: 7:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 92 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: MIP-1b
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#7:
Met Lys Leu Cys Val Thr Val Leu Ser Leu Le
#u Met Leu Val Ala Ala
1 5
# 10
# 15
Phe Cys Ser Pro Ala Leu Ser Ala Pro Met Gl
#y Ser Asp Pro Pro Thr
20
# 25
# 30
Ala Cys Cys Phe Ser Tyr Thr Ala Arg Lys Le
#u Pro Arg Asn Phe Val
35
# 40
# 45
Val Asp Tyr Tyr Glu Thr Ser Ser Leu Cys Se
#r Gln Pro Ala Val Val
50
# 55
# 60
Phe Gln Thr Lys Arg Ser Lys Gln Val Cys Al
#a Asp Pro Ser Glu Ser
65
#70
#75
#80
Trp Val Gln Glu Tyr Val Tyr Asp Leu Glu Le
#u Asn
85
# 90
(2) INFORMATION FOR SEQ ID NO: 8:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 91 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: RANTES
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#8:
Met Lys Val Ser Ala Ala Arg Leu Ala Val Il
#e Leu Ile Ala Thr Ala
1 5
# 10
# 15
Leu Cys Ala Pro Ala Ser Ala Ser Pro Tyr Se
#r Ser Asp Thr Thr Pro
20
# 25
# 30
Cys Cys Phe Ala Tyr Ile Ala Arg Pro Leu Pr
#o Arg Ala His Ile Lys
35
# 40
# 45
Glu Tyr Phe Tyr Thr Ser Gly Lys Cys Ser As
#n Pro Ala Val Val Phe
50
# 55
# 60
Val Thr Arg Lys Asn Arg Gln Val Cys Ala As
#n Pro Glu Lys Lys Trp
65
#70
#75
#80
Val Arg Glu Tyr Ile Asn Ser Leu Glu Met Se
#r
85
# 90
(2) INFORMATION FOR SEQ ID NO: 9:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 99 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: MCP-1
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#9:
Met Lys Val Ser Ala Ala Leu Leu Cys Leu Le
#u Leu Ile Ala Ala Thr
1 5
# 10
# 15
Phe Ile Pro Gln Gly Leu Ala Gln Pro Asp Al
#a Ile Asn Ala Pro Val
20
# 25
# 30
Thr Cys Cys Tyr Asn Phe Thr Asn Arg Lys Il
#e Ser Val Gln Arg Leu
35
# 40
# 45
Ala Ser Tyr Arg Arg Ile Thr Ser Ser Lys Cy
#s Pro Lys Glu Ala Val
50
# 55
# 60
Ile Phe Lys Thr Ile Val Ala Lys Glu Ile Cy
#s Ala Asp Pro Lys Gln
65
#70
#75
#80
Lys Trp Val Gln Asp Ser Met Asp His Leu As
#p Lys Gln Thr Gln Thr
85
# 90
# 95
Pro Lys Thr
(2) INFORMATION FOR SEQ ID NO: 10:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 77 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: MCP-2
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#10:
Ala Gln Pro Asp Ser Val Ser Ile Pro Ile Th
#r Cys Cys Phe Asn Val
1 5
# 10
# 15
Ile Asn Arg Lys Ile Pro Ile Gln Arg Leu Gl
#u Ser Tyr Thr Arg Ile
20
# 25
# 30
Thr Asn Ile Gln Cys Pro Lys Glu Ala Val Il
#e Phe Lys Thr Lys Arg
35
# 40
# 45
Gly Lys Glu Val Cys Ala Asp Pro Lys Glu Ar
#g Trp Val Arg Asp Ser
50
# 55
# 60
Met Lys His Leu Asp Gln Ile Phe Gln Asn Le
#u Lys Pro
65
#70
#75
(2) INFORMATION FOR SEQ ID NO: 11:
(i) SEQUENCE CHARACTERISTICS:
(A) LENGTH: 109 amino
#acids
(B) TYPE: amino acid
(C) STRANDEDNESS: single
(D) TOPOLOGY: linear
(ii) MOLECULE TYPE: peptide
(vi) ORIGINAL SOURCE:
(vii) IMMEDIATE SOURCE:
(A) LIBRARY: GenBank
(B) CLONE: MCP-3
(xi) SEQUENCE DESCRIPTION: SEQ ID NO:
#11:
Met Trp Lys Pro Met Pro Ser Pro Ser Asn Me
#t Lys Ala Ser Ala Ala
1 5
# 10
# 15
Leu Leu Cys Leu Leu Leu Thr Ala Ala Ala Ph
#e Ser Pro Gln Gly Leu
20
# 25
# 30
Ala Gln Pro Val Gly Ile Asn Thr Ser Thr Th
#r Cys Cys Tyr Arg Phe
35
# 40
# 45
Ile Asn Lys Lys Ile Pro Lys Gln Arg Leu Gl
#u Ser Tyr Arg Arg Thr
50
# 55
# 60
Thr Ser Ser His Cys Pro Arg Glu Ala Val Il
#e Phe Lys Thr Lys Leu
65
#70
#75
#80
Asp Lys Glu Ile Cys Ala Asp Pro Thr Gln Ly
#s Trp Val Gln Asp Phe
85
# 90
# 95
Met Lys His Leu Asp Lys Lys Thr Gln Thr Pr
#o Lys Leu
100
# 105
9 . A method for producing a polypeptide of claim 1 , the method comprising:
a) culturing a cell under conditions suitable for expression of the polypeptide, wherein said cell is transformed with a recombinant polynucleotide, and said recombinant polynucleotide comprises a promoter sequence operably linked to a polynucleotide encoding the polypeptide of claim 1 , and b) recovering the polypeptide so expressed.
10 . A method of claim 9 , wherein the polypeptide comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
11 . An isolated antibody which specifically binds to a polypeptide of claim 1 .
12 . An isolated polynucleotide comprising a sequence selected from the group consisting of:
a) a polynucleotide comprising a polynucleotide sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:3, b) a naturally occurring polynucleotide comprising a polynucleotide sequence at least 90% identical to a polynucleotide sequence selected from the group consisting of SEQ ID NO:1 and SEQ ID NO:3, c) a polynucleotide having a sequence complementary to a polynucleotide of a), d) a polynucleotide having a sequence complementary to a polynucleotide of b) and e) an RNA equivalent of a)-d).
13 . An isolated polynucleotide comprising at least 60 contiguous nucleotides of a polynucleotide of claim 12 .
14 . A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 12 , the method comprising:
a) hybridizing the sample with a probe comprising at least 20 contiguous nucleotides comprising a sequence complementary to said target polynucleotide in the sample, and which probe specifically hybridizes to said target polynucleotide, under conditions whereby a hybridization complex is formed between said probe and said target polynucleotide or fragments thereof, and b) detecting the presence or absence of said hybridization complex, and, optionally, if present, the amount thereof.
15 . A method of claim 14 , wherein the probe comprises at least 60 contiguous nucleotides.
16 . A method for detecting a target polynucleotide in a sample, said target polynucleotide having a sequence of a polynucleotide of claim 12 , the method comprising:
a) amplifying said target polynucleotide or fragment thereof using polymerase chain reaction amplification, and b) detecting the presence or absence of said amplified target polynucleotide or fragment thereof, and, optionally, if present, the amount thereof.
17 . A composition comprising a polypeptide of claim 1 and a pharmaceutically acceptable excipient.
18 . A composition of claim 17 , wherein the polypeptide has an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
19 . A method for treating a disease or condition associated with decreased expression of functional PANEC, comprising administering to a patient in need of such treatment the composition of claim 17 .
20 . A method for screening a compound for effectiveness as an agonist of a polypeptide of claim 1 , the method comprising:
a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting agonist activity in the sample.
21 . A composition comprising an agonist compound identified by a method of claim 20 and a pharmaceutically acceptable excipient.
22 . A method for treating a disease or condition associated with decreased expression of functional PANEC, comprising administering to a patient in need of such treatment a composition of claim 21 .
23 . A method for screening a compound for effectiveness as an antagonist of a polypeptide of claim 1 , the method comprising:
a) exposing a sample comprising a polypeptide of claim 1 to a compound, and b) detecting antagonist activity in the sample.
24 . A composition comprising an antagonist compound identified by a method of claim 23 and a pharmaceutically acceptable excipient.
25 . A method for treating a disease or condition associated with overexpression of functional PANEC, comprising administering to a patient in need of such treatment a composition of claim 24 .
26 . A method of screening for a compound that specifically binds to the polypeptide of claim 1 , the method comprising:
a) combining the polypeptide of claim 1 with at least one test compound under suitable conditions, and b) detecting binding of the polypeptide of claim 1 to the test compound, thereby identifying a compound that specifically binds to the polypeptide of claim 1 .
27 . A method of screening for a compound that modulates the activity of the polypeptide of claim 1 , said method comprising:
a) combining the polypeptide of claim 1 with at least one test compound under conditions permissive for the activity of the polypeptide of claim 1 , b) assessing the activity of the polypeptide of claim 1 in the presence of the test compound, and c) comparing the activity of the polypeptide of claim 1 in the presence of the test compound with the activity of the polypeptide of claim 1 in the absence of the test compound, wherein a change in the activity of the polypeptide of claim 1 in the presence of the test compound is indicative of a compound that modulates the activity of the polypeptide of claim 1 .
28 . A method for screening a compound for effectiveness in altering expression of a target polynucleotide, wherein said target polynucleotide comprises a polynucleotide sequence of claim 5 , the method comprising:
a) exposing a sample comprising the target polynucleotide to a compound, under conditions suitable for the expression of the target polynucleotide, b) detecting altered expression of the target polynucleotide, and c) comparing the expression of the target polynucleotide in the presence of varying amounts of the compound and in the absence of the compound.
29 . A method for assessing toxicity of a test compound, the method comprising:
a) treating a biological sample containing nucleic acids with the test compound, b) hybridizing the nucleic acids of the treated biological sample with a probe comprising at least 20 contiguous nucleotides of a polynucleotide of claim 12 under conditions whereby a specific hybridization complex is formed between said probe and a target polynucleotide in the biological sample, said target polynucleotide comprising a polynucleotide sequence of a polynucleotide of claim 12 or fragment thereof, c) quantifying the amount of hybridization complex, and d) comparing the amount of hybridization complex in the treated biological sample with the amount of hybridization complex in an untreated biological sample, wherein a difference in the amount of hybridization complex in the treated biological sample is indicative of toxicity of the test compound.
30 . A diagnostic test for a condition or disease associated with the expression of PANEC in a biological sample, the method comprising:
a) combining the biological sample with an antibody of claim 11 , under conditions suitable for the antibody to bind the polypeptide and form an antibody:polypeptide complex, and b) detecting the complex, wherein the presence of the complex correlates with the presence of the polypeptide in the biological sample.
31 . The antibody of claim 11 , wherein the antibody is:
a) a chimeric antibody, b) a single chain antibody, c) a Fab fragment, d) a F(ab') 2 fragment, or e) a humanized antibody.
32 . A composition comprising an antibody of claim 11 and an acceptable excipient.
33 . A method of diagnosing a condition or disease associated with the expression of PANEC in a subject, comprising administering to said subject an effective amount of the composition of claim 32 .
34 . A composition of claim 32 , wherein the antibody is labeled.
35 . A method of diagnosing a condition or disease associated with the expression of PANEC in a subject, comprising administering to said subject an effective amount of the composition of claim 34 .
36 . A method of preparing a polyclonal antibody with the specificity of the antibody of claim 11 , the method comprising:
a) immunizing an animal with a polypeptide having an amino acid sequence of selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibodies from said animal, and c) screening the isolated antibodies with the polypeptide, thereby identifying a polyclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
37 . An antibody produced by a method of claim 36 .
38 . A composition comprising the antibody of claim 37 and a suitable carrier.
39 . A method of making a monoclonal antibody with the specificity of the antibody of claim 11 , the method comprising:
a) immunizing an animal with a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4, or an immunogenic fragment thereof, under conditions to elicit an antibody response, b) isolating antibody producing cells from the animal, c) fusing the antibody producing cells with immortalized cells to form monoclonal antibody-producing hybridoma cells, d) culturing the hybridoma cells, and e) isolating from the culture monoclonal antibody which binds specifically to a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
40 . A monoclonal antibody produced by a method of claim 39 .
41 . A composition comprising the monoclonal antibody of claim 40 and a suitable carrier.
42 . The antibody of claim 11 , wherein the antibody is produced by screening a Fab expression library.
43 . The antibody of claim 11 , wherein the antibody is produced by screening a recombinant immunoglobulin library.
44 . A method of detecting a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4 in a sample, the method comprising:
a) incubating the antibody of claim 11 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) detecting specific binding, wherein specific binding indicates the presence of a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4 in the sample.
45 . A method of purifying a polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4 from a sample, the method comprising:
a) incubating the antibody of claim 11 with a sample under conditions to allow specific binding of the antibody and the polypeptide, and b) separating the antibody from the sample and obtaining the purified polypeptide having an amino acid sequence selected from the group consisting of SEQ ID NO:2 and SEQ ID NO:4.
46 . A microarray wherein at least one element of the microarray is a polynucleotide of claim 13 .
47 . A method of generating an expression profile of a sample which contains polynucleotides,, the method comprising:
a) labeling the polynucleotides of the sample, b) contacting the elements of the microarray of claim 46 with the labeled polynucleotides of the sample under conditions suitable for the formation of a hybridization complex, and c) quantifying the expression of the polynucleotides in the sample.
48 . An array comprising different nucleotide molecules affixed in distinct physical locations on a solid substrate, wherein at least one of said nucleotide molecules comprises a first oligonucleotide or polynucleotide sequence specifically hybridizable with at least 30 contiguous nucleotides of a target polynucleotide, and wherein said target polynucleotide is a polynucleotide of claim 12 .
49 . An array of claim 48 , wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 30 contiguous nucleotides of said target polynucleotide.
50 . An array of claim 48 , wherein said first oligonucleotide or polynucleotide sequence is completely complementary to at least 60 contiguous nucleotides of said target polynucleotide.
51 . An array of claim 48 , wherein said first oligonucleotide or polynucleotide sequence is completely complementary to said target plynucleotide.
52 . An array of claim 48 , which is a microarray.
53 . An array of claim 48 , further comprising said target polynucleotide hybridized to a nucleotide molecule comprising said first oligonucleotide or polynucleotide sequence.
54 . An array of claim 48 , wherein a linker joins at least one of said nucleotide molecules to said solid substrate.
55 . An array of claim 48 , wherein each distinct physical location on the substrate contains multiple nucleotide molecules, and the multiple nucleotide molecules at any single distinct physical location have the same sequence, and each distinct physical location on the substrate contains nucleolide molecules having a sequence which differs from the sequence of nucleotide molecules at another distinct physical location on the substrate.
56 . A polypeptide of claim 1 , comprising the amino acid sequence of SEQ ID NO:2.
57 . A polypeptide of claim 1 , comprising the amino acid sequence of SEQ ID NO:4.
58 . A polynucleotide of claim 12 , comprising the polynucleotide sequence of SEQ ID NO:1.
59 . A polynucleotide of claim 12 , comprising the polynucleotide sequence of SEQ ID NO:3.Join the waitlist — get patent alerts
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