What is claimed is:
1 A combination comprising a plurality of cDNAs wherein the cDNAs are SEQ ID NOs:1-25 and the complements of SEQ ID NOs:1-25.
2. An isolated cDNA comprising a polynucleotide having the nucleic acid sequence of SEQ ID No:8.
3. A method for detecting differential expression of one or more cDNAs in a sample containing nucleic acids, the method comprising:
a) hybridizing the combination of claim 1 with nucleic acids of the sample, thereby forming one or more hybridization complexes;
b) detecting the hybridization complexes; and
c) comparing the hybridization complexes with those of a standard, wherein differences between the standard and sample hybridization complexes indicate differential expression of cDNAs in the sample.
4. The method of claim 3, wherein the nucleic acids of the sample are amplified prior to hybridization.
5. The method of claim 3, wherein the sample is from a subject with a disorder associated with atherosclerosis.
6. The method of claim 3, wherein the sample is from aorta, arteries, arterioles, endothelial cells, plaque, or blood.
7. The method of claim 3 wherein the combination is immobilized on a substrate.
8. The method of claim 3 wherein differential expression is diagnostic of angina pectoris, coronary artery disease, myocardial infarction, hypertension, transient cerebral ischemia, mesenteric ischemia, peripheral vascular disease, renal artery stenosis, or stroke.
9. The method of claim 6 wherein the substrate is a microarray.
10. A method of using cDNAs to screen a sample to identify a ligand which specifically binds a cDNA, the method comprising:
a) combining the combination of claim 1 with the sample under conditions to allow specific binding; and
b) detecting specific binding between each cDNA and at least ligand, thereby identifying a ligand that specifically binds to each cDNA.
11. The method of claim 10 wherein the ligands are DNA molecules, proteins, RNA molecules, or transcription factors.
12. A vector containing the cDNA of claim 2.
13. A host cell containing the vector of claim 12.
14. A method for producing a protein, the method comprising the steps of:
a) culturing the host cell of claim 13 under conditions for expression of protein; and
b) recovering the protein from the host cell culture.
15. A protein comprising a polypeptide having the amino acid of SEQ ID NO:26.
16. A method for using a protein to screen a plurality of molecules or compounds to identify at least one ligand which specifically binds the protein, the method comprising:
a) combining the protein encoded by a cDNA of claim 1 with the plurality of molecules or compounds under conditions to allow specific binding; and
b) detecting specific binding between the protein and a molecule or compound, thereby identifying a ligand which specifically binds the protein.
17. The method of claim 16 wherein the plurality of molecules or compounds is selected from agonists, antagonists, antibodies, DNA molecules, small molecule drugs, immunoglobulins, inhibitors, mimetics, peptide nucleic acids, peptides, pharmaceutical agents, proteins, RNA molecules, and ribozymes.
18. A method of using a protein to produce an antibody, the method comprising:
a) immunizing an animal with the protein encoded by a cDNA of claim 1 under conditions to elicit an antibody response;
b) isolating animal antibodies; and
c) screening the isolated antibodies with the protein, thereby identifying an antibody which specifically binds the protein.
19. A antibody produced by the method of claim 18.
20. A method for using an antibody to detect gene expression in a sample, the method comprising:
a) combining the antibody of claim 19 with a sample under conditions which allow the formation of antibody:protein complexes; and
b) detecting complex formation, wherein complex formation indicates expression of the protein in the sample.
The claims below are in addition to those above.
All refrences to claim(s) which appear below refer to the numbering after this setence.
1. A liquid pharmaceutical formulation of an isolated IL-6 antibody, comprising at least one light chain variable region and one heavy chain variable region, said light chain variable region comprising:
a complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO:132 comprising the sequence S-X1-X2-X3-X4-V-X5-Y-M-Y, wherein X1 is A or G, X2 is S or R, X3 is H, I, S, or Y, X4 is S or Y, and X5 is S or F;
a CDRL2 amino acid sequence of SEQ ID NO:133 comprising the sequence D-X6-S-X7-L-X8-S, wherein X6 is F, L, M, or T, X7 is N or E, and X8 is A or T; and
a CDRL3 amino acid sequence of SEQ ID NO:134 comprising the sequence X9-X10-W-S-G-Y-P-Y-T, wherein X9 is M, C, or S, and X10 is Q or C; said heavy chain variable region comprising:
a complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO:135 comprising the sequence G-F-X11-X12-S-X13-F-A-X14-S, wherein X11 is T or Q, X12 is F, S, or T, X13 is S or P, and X14 is L or M;
a CDRH2 amino acid sequence of SEQ ID NO:136 comprising the sequence K-X15-S-X16-G-G-S-X17-X18-Y-X19-X20-D-T-X21-X22-X23, wherein X15 is A or I, X16 is S or P, X17 is Y or W, X18 is T, E, or Y, X19 is Y or F, X20 is P, S, D, or F, X21 is V or D, X22 is T or A, and X23 is G or P; and
a CDRH3 amino acid sequence of SEQ ID NO:137 comprising the sequence Q-L-W-G-X24-Y-A-L-D-X25, wherein X24 is S, Y, T, or N, and X25 is Y, T, F, or I.
2. The pharmaceutical formulation of claim 1, comprising acetate buffer, sorbitol, and polysorbate 20.
3. The pharmaceutical formulation of claim 2, wherein the pH is about 5.0.
4. The pharmaceutical formulation of claim 1, wherein the isolated IL-6 antibody comprises a light chain variable region amino acid sequence of SEQ ID NO:97 and a heavy chain variable region amino acid sequence of SEQ ID NO:99.
5. The pharmaceutical formulation of claim 4, comprising acetate buffer, sorbitol, and polysorbate 20.
6. The pharmaceutical formulation of claim 5, wherein the pH is about 5.0.
7. The pharmaceutical formulation of claim 1, wherein the isolated IL-6 antibody comprises a complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO:3, a CDRL2 amino acid sequence of SEQ ID NO:21, a CDRL3 amino acid sequence of SEQ ID NO:29, a complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO:39, a CDRH2 amino acid sequence of SEQ ID NO:59, and a CDRH3 amino acid sequence of SEQ ID NO:89.
8. The pharmaceutical formulation of claim 7, comprising acetate buffer, sorbitol, and polysorbate 20.
9. The pharmaceutical formulation of claim 8, wherein the pH is about 5.0.
10. A liquid pharmaceutical formulation, comprising an isolated IL-6 antibody having a complementarity determining region light chain 1 (CDRL1) amino acid sequence of SEQ ID NO:3, a CDRL2 amino acid sequence of SEQ ID NO:21, a CDRL3 amino acid sequence of SEQ ID NO:29, a complementarity determining region heavy chain 1 (CDRH1) amino acid sequence of SEQ ID NO:39, a CDRH2 amino acid sequence of SEQ ID NO:59, and a CDRH3 amino acid sequence of SEQ ID NO:89, acetate buffer, sorbitol, and polysorbate 20, wherein the pH is about 5.0.
11. A liquid pharmaceutical formulation, comprising an isolated IL-6 antibody having a light chain variable region amino acid sequence of SEQ ID NO:97 and a heavy chain variable region amino acid sequence of SEQ ID NO:99, acetate buffer, sorbitol, and polysorbate 20, wherein the pH is about 5.0.