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Generation of monoclonal antibodies reactive with nuclear proteins of human primary breast tumors
Author(s) -
Dinh BaoLinh,
Tremblay Monique,
Paradis Diane,
Roy PaulEmile,
Tetu Bernard
Publication year - 1988
Publication title -
international journal of cancer
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.475
H-Index - 234
eISSN - 1097-0215
pISSN - 0020-7136
DOI - 10.1002/ijc.2910410207
Subject(s) - monoclonal antibody , microbiology and biotechnology , immunoperoxidase , biology , antibody , nuclear protein , staining , pathology , breast carcinoma , gel electrophoresis , breast cancer , cancer , immunology , biochemistry , medicine , genetics , gene , transcription factor
Nuclear proteins were extracted from purified nuclei of human primary breast tumors (BrT) and bladder tumors and of human normal breast, kidney and lymphocytes by enzymatic treatment. SDS‐Polyacrylamide gel electrophoresis of nuclear proteins from breast tumors showed different bands in the molecular weight zones from 25 to 220 kDa which were absent or present only as traces in normal breast tissue. Murine monoclonal antibodies (MAbs) have been produced using nuclear extracts of human primary breast tumors as immunogens. Approximately 2,000 hybridomas were generated from 5 hybridizations. According to their reactivity to BrT nuclear extracts and mammary carcinoma cell line MCF‐7, seven hybridomas were selected and cloned. They were further characterized with histological immunoperoxidase assays of formaldehyde‐fixed BrT paraffin tissue sections. MAb 6A3 particularly gave strong nuclear staining with all BrT specimens while MAb ID8 showed both nuclear and cytoplasmic staining with only some of them. Specimens from mammoplasty did not react with these MAbs. Immunoblotting of BrT nuclear extracts as developed with MAbs 6A3 and ID8 revealed major protein bands with molecular weight of 120 and 130 kDa. The potential use of these MAb‐defined BrT‐related nuclear proteins as markers for human breast cancer was suggested.