التفاصيل البيبلوغرافية
| العنوان: |
Assay for anti-INGAP antibodies |
| Document Number: |
20070184504 |
| تاريخ النشر: |
August 9, 2007 |
| Appl. No: |
11/656357 |
| Application Filed: |
January 22, 2007 |
| مستخلص: |
A solid phase assay is used for detecting antibodies to INGAP104-118 peptide, a 15-amino acid peptide that is the biologically active portion of islet neogenesis associated protein (INGAP). The isotype of the antibodies to INGAP104-118 peptide can be determined. A kit can also be used in the detection of anti-INGAP104-118 antibodies. Endogenous autoantibodies or antibody production during therapeutic treatment of a mammal with INGAP104-118 can be monitored. |
| Inventors: |
Vinik, Aaron I. (Norfolk, VA, US); Taylor-Fishwick, David A. (Norfolk, VA, US) |
| Claim: |
1. A method for detecting antibodies to INGAP104-118 peptide in a test sample and determining isotype of said antibodies, comprising: contacting a test sample from a human with INGAP104-118 peptide bound to a solid support, wherein the contacting is under conditions sufficient for binding an anti-INGAP104-118 antibody to the INGAP104-118 peptide; contacting the solid support with an isotype-specific antibody which specifically binds antibody molecules of one isotype of the human; and determining the isotype-specific antibody bound to the solid support, wherein detection antibody bound to the solid support indicates that the test sample contains antibodies to INGAP104-118 peptide and that the antibodies to INGAP104-118 peptide are of the one isotype; wherein the test sample is selected from the group consisting of blood, serum and plasma. |
| Claim: |
2. The method of claim 1 wherein the isotype-specific antibody comprises a detectable label. |
| Claim: |
3. The method of claim 2 wherein the detectable label is an enzyme. |
| Claim: |
4. The method of claim 3 wherein the enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase, and beta-galactosidase. |
| Claim: |
5. The method of claim 2 wherein the detectable label is selected from the group consisting of fluorescent molecules, chemiluminescent molecules, radioactive molecules, and dye molecules. |
| Claim: |
6. A kit for detecting an anti-INGAP104-118 antibody in a test sample of a mammal comprising: an INGAP104-118 peptide; and a detection antibody. |
| Claim: |
7. The kit of claim 6 wherein the test sample is selected from the group consisting of blood, serum and plasma. |
| Claim: |
8. The kit of claim 6 wherein the detection antibody specifically binds antibody molecules of all isotypes of a mammal. |
| Claim: |
9. The kit of claim 6 wherein the detection antibody is an isotype-specific antibody which specifically binds antibody molecules of one isotype of a mammal. |
| Claim: |
10. The kit of claim 6 wherein the detection antibody is detectably labeled. |
| Claim: |
11. The kit of claim 6 wherein the INGAP104-118 peptide is bound to a solid support. |
| Claim: |
12. The kit of claim 6 further comprising an anti-INGAP104-118 antibody. |
| Claim: |
13. The kit of claim 6 further comprising instructions for detecting antibodies to INGAP104-118 peptide using the peptide and the antibody. |
| Claim: |
14. The kit of claim 6 further comprising a standard curve for use in calculating amount of anti-INGAP104-118 antibody in a test sample. |
| Claim: |
15. The kit of claim 6 further comprising a buffer for washing unbound reagents. |
| Claim: |
16. The kit of claim 10 wherein the detectable label is selected from the group consisting of fluorescent molecules, chemiluminescent molecules, radioactive molecules, and dye molecules. |
| Claim: |
17. A method for detecting neutralizing antibodies in a patient undergoing treatment with an Islet Neogenesis Associated Protein, comprising: collecting a test sample from a patient being administered a therapeutic protein selected from the group comprising sequences 104-118 of SEQ ID NO 2, sequences 1-175 of SEQ ID No 2 or fragments thereof, contacting the test sample with peptide corresponding to the therapeutic protein bound to a solid support under conditions sufficient for binding an antibody to the therapeutics protein, contacting any test sample bound to the therapeutic protein on the solid support with a detection antibody which specifically binds to antibody molecules of all isotypes of the mammal; washing the solid substrate to remove any unbound detection antibody molecules, and determining the presence of detection antibody bound to the solid support, wherein the presence of any detection antibody bound to the solid support indicates that the test sample contains neutralizing antibodies to the therapeutic protein. |
| Claim: |
18. The method of claim 17 wherein amount of the detection antibody bound is compared to a standard curve generated using known amounts of an antibody to the therapeutic peptide. |
| Claim: |
19. The method of claim 17 wherein multiple test samples are used which are serial dilutions of the test sample of the mammal wherein the serial dilutions show relative measure of titre in a test sample or between test samples from different sources. |
| Claim: |
20. The method of claim 19 further comprising: determining a highest serial dilution at which a signal is generated, said highest serial dilution being the test sample used to evaluate titer of anti-INGAP104-118 peptide antibodies in the mammal. |
| Claim: |
21. The method of claim 20 wherein the highest serial dilution of a first mammalian test sample is compared to a highest serial dilution of a second mammalian test sample to provide a comparative measure of titer in the first and second mammalian test samples. |
| Claim: |
22. The method of claim 17 wherein the detection antibody comprises a detectable label. |
| Claim: |
23. The method of claim 22 wherein the detectable label is an enzyme. |
| Claim: |
24. The method of claim 23 wherein the enzyme is selected from the group consisting of alkaline phosphatase, horseradish peroxidase and beta-galactosidase. |
| Claim: |
25. The method of claim 25 wherein the detectable label is selected from the group consisting of fluorescent molecules, chemiluminescent molecules, radioactive molecules, and dye molecules. |
| Current U.S. Class: |
435007/920 |
| Current International Class: |
01 |
| رقم الانضمام: |
edspap.20070184504 |
| قاعدة البيانات: |
USPTO Patent Applications |
