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Last Updated: March 28, 2024

Claims for Patent: 5,817,468


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Summary for Patent: 5,817,468
Title: Process for the identification or immunoassay of polypeptide antigens
Abstract:Process for the identification or assay of a particular polypeptide antigen X in which an immunological determination of the presence of said polypeptide antigen X is carried out in a sample, characterized in that the sample is treated in vitro using at least one chemical reagent reacting on an epsilon -NH.sub.2 function of a lysine residue or on a terminal alpha -NH.sub.2 function of a peptide in order to carry out the linking of said reagent to said polypeptide antigen X and in this way to obtain a modified polypeptide antigen X, (polypeptide antigen Xm), the immunological determination of the presence of said polypeptide antigen X in said sample is then carried out using at least one antibody produced by immunization against a polypeptide antigen Y identical to the polypeptide antigen Xm or to an antigenic fragment of this antigen Xm.
Inventor(s): Jean; Frederic (Marseilles, FR), Kertesz; Gilles (Marseilles, FR), Bourcier; Beatrice (Marseilles, FR)
Assignee: Immunotech (Marseilles, FR)
Application Number:08/773,498
Patent Claims:1. Process for the identification or assay of a particular polypeptide antigen X in which an immunological determination is carried out for the presence of said polypeptide antigen X in a sample, characterized in that the sample is treated in vitro using at least one modifying reagent

on an epsilon -NH.sub.2 function of a lysine residue or

on an alpha -NH.sub.2 function of a peptide,

in order to carry out the linking of said modifying agent to said polypeptide antigen X as a chemical modifying group and in this way to obtain a modified polypeptide antigen X, (polypeptide antigen Xm), wherein the chemical modifying group has a molecular weight less than 400 daltons and the immunological determination of the presence of said polypeptide antigen X in said sample is then carried out using at least one antibody specific for said polypeptide antigen Xm and non-specific for the modification itself, produced by immunization against a polypeptide antigen Y identical to the polypeptide antigen Xm or to an antigenic fragment of this antigen Xm.

2. Process according to claim 1 characterized in that a polypeptide antigen Y identical to the complete polypeptide antigen Xm is used to carry out the immunizations.

3. Process according to claim 1 characterized in that a polypeptide antigen Y corresponding to a fragment of the polypeptide antigen Xm is used to carry out the immunization and in this way to obtain antibodies directed against a particular region of the antigen Xm.

4. Process according to claim 1 characterized in that a single reagent for modification is used.

5. Process according to one of claim 1 characterized in that several reagents for modification are used.

6. Process according to claim 5 characterized in that the reagent or reagents are selected from: succinic anhydride, acetic anhydride, derivatives of the esters of N-hydroxy succinimide.

7. Process according to claim 1 characterized in that the sample is a biological fluid such as blood, serum, urine, sweat or tears.

8. Process according to claim 7 characterized in that the polypeptide antigen X is human corticotropic hormone (ACTH).

9. Process according to claim 7 characterized in that the polypeptide antigen is human parathyroid hormone (PTH).

10. Process according to claim 9 characterized in that the assay method is an immunoassay by competition or

a radioimmunometric assay using a radioactive, luminescent, fluorescent or enzymatic tracer, or

an immunoassay by particle agglutination,

said method using one or more monoclonal or polyclonal antibodies.

11. Process according to claim 6 characterized in that the sample is a tissue sample and the polypeptide antigen X is detected by an immunohistochemical method.

12. Process according to claim 6 characterized in that the sample is a cellular suspension and where the polypeptide antigen X is detected by a flow cytometry method.

13. A hybridoma producing antiacylated ACTH with succinic anhydride monoclonal antibodies, selected from the group consisting of hybridomas deposited at the Collection Nationale de Culture de Microorganismes on 12th Dec.,1995 under the accession numbers I-1648, I-1649 and I-1650.

14. An anti-acylated ACTH monoclonal antibody produced by a hybridoma as defined in claim 13.

15. Process according to claim 1 characterized in that the polypeptide antigen X is human corticotropic hormone (ACTH).

16. Process according to claim 1 characterized in that the polypeptide antigen is human parathyoid hormone (PTH).

17. Process according to claim 1 characterized in that the assay method is an immunoassay by competition or

a radioimmunometric assay using a radioactive, luminescent, fluorescent or enzymatic tracer, or

an immunoassay by particle agglutination,

said method using one or more monoclonal or polyclonal antibodies.

18. Process according to claim 1 characterized in that the sample is a tissue sample and the polypeptide antigen X is detected by an immunohistochemical method.

19. Process according to claim 1 characterized in that the sample is a cellular suspension and where the polypeptide antigen X is detected by a flow cytometry method.

20. A process for the identification or assay of a particular polypeptide antigen X in which an immunological determination is carried out for the presence of said polypeptide antigen X in a sample, characterized in that the sample is treated in vitro using at least one modifying reagent

on an epsilon -NH.sub.2 function of a lysine residue or

on an alpha -NH2 function of a peptide,

in order to carry out the linking of said modifying agent to said polypeptide antigen X as a chemical modifying group and in this way to obtain a modified polypeptide antigen X, designated polypeptide Xm, wherein the chemical modifying group has a molecular weight less than 400 daltons and the immunological determination of the presence of said polypeptide antigen X in said sample is then carried out using at least one antibody specific for said polypeptide antigen Xm and non-specific for the modification itself, produced by immunization against a polypeptide antigen Y coupled to a carrier protein via a coupling reagent, wherein said polypeptide antigen Y is identical to the polypeptide antigen Xm or to an antigenic fragment of polypeptide antigen Xm.

21. Process according to claim 20, wherein the assay is selected from the group consisting of an immunoassay by competition, a radioimmunometric assay using a radioactive, luminescent, fluorescent or enzymatic tracer, and an immunoassay by particle agglutination, said assay using one or more monoclonal or polyclonal antibodies.

22. Process according to claim 20 characterized in that a single reagent for modification is used.

23. Process according to claim 20 characterized in that several reagents for modification are used.

24. Process according to claim 23 characterized in that the reagent or reagents are selected from: succinic anhydride, acetic anhydride, derivatives of the esters of N-hydroxy succinimide.

25. Process according to claim 20 characterized in that the sample is a biological fluid such as blood, serum, urine, sweat or tears.

26. Process according to claim 20 characterized in that the polypeptide antigen X is human corticotropic hormone (ACTH).

27. Process according to claim 20 characterized in that the polypeptide antigen is human parathyoid hormone (PTH).

28. Process according to claim 20 characterized in that the sample is a tissue sample and the polypeptide antigen X is detected by an immunohistochemical method.

29. Process according to claim 20 characterized in that the sample is a cellular suspension and where the polypeptide antigen X is detected by a flow cytometry method.

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