Monoclonal antibodies against native and denatured forms of estrogen-induced breast cancer protein (BCEI/pS2) obtained by expression in Escherichia coli

Several vectors were used to express the complementary DNA for breast cancer estrogen-induced BCEI (also called pS sub(2)) in Escherichia coli). The best results were obtained by using the pUR 290 expression vector after deletion of the sequence encoding the signal peptide of the protein. In these c...

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Bibliographic Details
Published in:Cancer research (Chicago, Ill.) Ill.), 1990-04, Vol.50 (8), p.2390-2396
Main Authors: PRUD'HOMME, J.-F, JOLIVET, A, PICHON, M.-F, SAVOURET, J.-F, MILGROM, E
Format: Article
Language:English
Subjects:
Antibodies, immunoglobulins
Biological and medical sciences
Fundamental and applied biological sciences. Psychology
Fundamental immunology
Molecular immunology
Monoclonal antibodies
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Summary:Several vectors were used to express the complementary DNA for breast cancer estrogen-induced BCEI (also called pS sub(2)) in Escherichia coli). The best results were obtained by using the pUR 290 expression vector after deletion of the sequence encoding the signal peptide of the protein. In these conditions, beta -galactosidase-BCEI/pS sub(2) fusion protein accounted for similar to 20% of total proteins in bacterial extracts. It was purified by chromatography on DEAE-Trisacryl or by gel electrophoresis and electroelution. Polyclonal antibodies were obtained by immunization of rabbits and goats, and monoclonal antibodies were raised in mice. The mBCEI sub(11) antibody was used in immunoaffinity chromatography to purify the peptide in a single step from culture media of estrogen-treated MCF-7 cells.
ISSN:0008-5472
1538-7445