Process for sex determination in man by use of monoclonal antibodies to the H-Y antigen

• US 4,680,258 A
• Filed: 08/09/1983
• Issued: 07/14/1987
• Est. Priority Date: 03/24/1981
• Status: Expired due to Term

• Current Assignee: Sloan-Kettering Institute For Cancer Research (Memorial Sloan-Kettering Cancer Center)
• Original Assignee: Sloan-Kettering Institute For Cancer Research (Memorial Sloan-Kettering Cancer Center)
• Inventors: Tada, Nobuhiko, Koo, Gloria C., Hammerling, Ulrich
• Primary Examiner(s): Kepplinger, Esther M.
• Application Number:
  US06/521,635
• Time in Patent Office: 1,435 Days
• Field of Search: 435/7, 435/172, 435/64, 435/4, 435/172.2, 435/948, 435/240, 424/85, 436/548, 436/510, 436/518, 436/540, 436/542, 436/545, 436/811, 436/828, 436/821, 260/112 R, 260/112 B, 935/103, 935/110, 530/387
• US Class Current: 435/7.21
• CPC Class Codes: C07K 16/28 against receptors, cell surface antigens or cell surface determinants
• CPC Class Codes: Y10S 435/948 using viruses or cell lines
• CPC Class Codes: Y10S 436/821 involving complement factors or complement systems
• CPC Class Codes: Y10S 436/828 Protein A

Abstract

Production of anti-H-Y hybridoma cell lines, and the use of the antibodies to determine the presence of H-Y antigen to indicate the sex of the proband inclusive of fetus, newborn and adult humans.

Claims

1. Hybridoma cell lines selected from the group consisting of ATCC HB 9070, ATCC HB 9071 and ATCC HB 9072 producing monoclonal antibodies 12/44, 12/49, and 11/6 which specifically bind to the H-Y antigen.

2. Process for determining gender of a fetus carried in a female, comprising obtaining a sample of blood or amniotic fluid from the female and measuring the H-Y antigen in said sample by contacting said sample with monoclonal anti-H-Y antibodies selected from the group consisting of 12/44, 12/49, and 6/11 produced by hybridoma cell line ATCC HB 9070, ATCC HB 9071 or ATCC HB 9072 and observing reaction between said sample and said monoclonal antibodies, presence of said reaction indicating that the gender of said fetus is male absence of said reaction indicating the gender of said fetus is female.

3. Process of claim 2 wherein the step of determining whether or not a reaction occurs comprises a direct radioimmuno assay using radioactive labeled monoclonal anti-H-Y antibodies, or an indirect assay, using unlabeled anti-H-Y antibody and radioactively labeled anti-immunoglobulin.

4. Process of claim 2 wherein the step of determining whether or not a reaction occurs comprises a Protein A rosette binding assay.

5. Process of claim 2 wherein said reaction is detected by enzyme-linked antibody binding assay (ELISA).

6. Process of claim 2 wherein said reaction is detected by an assay selected from the group consisting of direct and indirect immunofluorescence assays.

7. Process of claim 3 wherein the step of determining whether or not a reaction occurs comprises a complement-dependent cytotoxicity reaction.

8. Process of claim 2 wherein the step of determining whether or not a reaction occurs comprises a neutralization assay with cell bound H-Y antigen and further comprises measurement of whether neutralization of anti-H-Y antibody occurs.

9. Process of claim 3 wherein said sample comprises human amniotic cells whereby the presence of H-Y positive cells indicates a male fetus and the absence of H-Y positive cells a female fetus.

10. Process of claim 9 wherein the step of determining whether a reaction occurs comprises an immunofluorescence flow cytometry assay.

11. Process of claim 2 wherein said sample comprises human amniotic fluid whereby the presence of H-Y antigen signals a male fetus and the absence of H-Y a female fetus.

12. Process of claim 2 or 8 wherein said sample comprises serum of a pregnant woman, whereby the presence of H-Y antigen will indicate a male fetus.

13. Process of claim 2 wherein said sample comprises peripheral blood cells of a pregnant woman, whereby the presence of H-Y positive cells with indicate a male fetus.

14. Process of claim 13 wherein the step of determining whether a reaction occurs comprises an immunofluorescence flow cytometry assay.

15. Process of claim 2 wherein the step of determining whether a reaction occurs comprises an immunofluorescence flow cytometry assay.

16. Process of claim 2, wherein the step of determining whether a reaction occurs comprises an assay method selected from the group consisting of radioimmuno assay, enzyme-linked antibody binding assay (ELISA), direct or indirect immunofluorescence assay, and immunofluorescence flow cytometry assay.

17. Monoclonal antibodies which specifically bind to H-Y antigen selected from the group consisting of 12/44, 12/49, and 6/11 produced by hybridoma cell line ATCC HB 9070, ATCC HB 9071 or ATCC HB 9072.