Method for selecting hybridomas producing antibodies specific to

Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...

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4351722, 43524027, 435948, 436548, 436813, 530808, 935 90, 935104, C07K 1514, G01N 33577

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048373060

ABSTRACT:
The invention provides a method for selecting hybridomas which produce antibodies specific to domains of a cell surface antigen which is usually not accessible at the surface of intact cells. The method employs the screening of the hybridoma clones obtained for the production of antibodies specific against the cell surface antigen by use of immunoblotting analysis, namely by screening the clones against antigen immobilized on a solid substrate such as nitrocellulose. The invention also includes those hybridomas and monoclonal antibodies when produced according to the method. The method provides monoclonal antibodies to P-glycoprotein surface antigen correlated with multidrug resistance. The antibodies are used to obtain a cDNA probe which in turn was used to select a cDNA clone encoding for a portion of the P-glycoprotein including the C-terminal end. The C-terminal portion of the P-glycoprotein comprising about 239 amino acids and localized to the cytoplasmic side of the plasma membrane contains the epitopes for binding of the monoclonal antibodies.

REFERENCES:
F. B. Armstrong, Biochemistry, 2nd Ed., Oxford University Press, New York, 1983, pp. 218-219.
C. A. Molinaro et al., in R. M. Nakamura et al (Eds.), Immunologic Analysis, Masson Publishing, U.S.A., New York, 1982, pp. 129-143.
C. J. O'Hara et al., Immunology Letters, 5, 15-18, 1982.
C. J. O'Hara et al., Journ. Clin. Immunol., 4, 403-441, 1984.
J. R. Riordan et al., Journ. Biol. Chem., 254, 12701-12705, 1979.
C. J. O'Hara; Characterization of Monoclonal Antibodies Demonstrating Specificity for Drug-Resistant Tumour Cells; PhD. Thesis Sep., 1983; pp. 5.1-5.5

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