Process and reagent for the determination of the luteinizing hormone and monoclonal antibodies suitable thereof
First Claim
1. Hybridoma cell line which produces monoclonal antibodies which specifically bind to luteinising hormone and cross react with TSH, HCG and FSH to an extent of less than 3% as determined using an enzyme linked immunosorbent assay, said cell line prepared by(i) intraperitoneally immunizing a host animal with human luteinizing hormone;
- (ii) removing the spleen of said host animal;
(iii) fusing cells of said spleen with myeloma cells;
(iv) separating fused cells from non-fused cells by culturing in selection medium;
(v) screening supernatant of fused cells for production of antibodies by mixing said supernatant with human luteinizing hormone to bind antibody in said supernatant to human luteinizing hormone and with labelled antibody against antibodies which bind to human luteinizing hormone to identify said human luteinizing hormone binding antibodies, and(vi) repeating step (v) with a glycoprotein hormone selected from the group consisting of TSH, HCG and FSH and with a labelled antibody, to determine human luteinizing hormone specific antibodies which cross react to an extend less than 3% with said glycoprotein hormone.
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Abstract
The present invention provides an immunological process for the determination of the luteinizing hormone (LH), wherein at least one monoclonal antibody is used which is directed against LH and cross-reacts with other glycoprotein hormones to an extent of less than 3%.
The present invention also provides a reagent for the determination of the luteinizing hormone, wherein it contains at least one monoclonal antibody which is directed against LH and cross-reacts with other glycoprotein hormones to an extent of less than 3%.
Furthermore, the present invention provides a monoclonal antibody against the luteinizing hormone and a process and a hybridoma cell line for producing it.
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Citations
33 Claims
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1. Hybridoma cell line which produces monoclonal antibodies which specifically bind to luteinising hormone and cross react with TSH, HCG and FSH to an extent of less than 3% as determined using an enzyme linked immunosorbent assay, said cell line prepared by
(i) intraperitoneally immunizing a host animal with human luteinizing hormone; -
(ii) removing the spleen of said host animal; (iii) fusing cells of said spleen with myeloma cells; (iv) separating fused cells from non-fused cells by culturing in selection medium; (v) screening supernatant of fused cells for production of antibodies by mixing said supernatant with human luteinizing hormone to bind antibody in said supernatant to human luteinizing hormone and with labelled antibody against antibodies which bind to human luteinizing hormone to identify said human luteinizing hormone binding antibodies, and (vi) repeating step (v) with a glycoprotein hormone selected from the group consisting of TSH, HCG and FSH and with a labelled antibody, to determine human luteinizing hormone specific antibodies which cross react to an extend less than 3% with said glycoprotein hormone. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33)
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Specification