Monoclonal antibodies 50B4 and 50E6 recognize two distinct epitopes of human p85 glycoprotein (CDw44). Both epitopes are destroyed by reduction of the purified gycoprotein as demonstrated by inhibition of cellular radioimmunoassay and Western blot analysis. Endoglycosidase F treated p85 glycoprotein, with an apparent molecular weight of 73,000 is still reactive with both monoclonal antibodies. Thus both epitopes are conformational determinats of the polypeptide chain. A rabbit antibody produced against purified native p85 glycoprotein also reacted only with the non-reduced form of p85. Repeated immunizations with SDS-dissociated and reduced p85 yielded a polyclonal antibody reactive by Western blot analysis with reduced and non-reduced forms of p85 glycoprotein. When a HOON leukemia cell line cDNA expression library was screened with this polyclonal antibody, two cDNA clones were isolated which reacted specifically with the antiserum and not with the control non-immune serum. Preliminary characterization of these clones indicates that they are p85-related.
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August 01 1988
Several epitopes of p85 glycoprotein (CDw44) are dependent on intact disulphide bonds. Isolation of cDNA clones requires a polyclonal antibody raised against the reduced protein
Ian Rogers;
Ian Rogers
1Department of Immunology, University of Toronto, Toronto M5G 1X8
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Giacomo D'Agostaro;
Giacomo D'Agostaro
2Department of Medical Genetics, University of Toronto, Toronto M5G 1X8
2Lab. Biophysics, CRE Enea, S.P. Anguillarese, 301 Casaccia, 00100 Roma
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Sonia Vera;
Sonia Vera
1Department of Immunology, University of Toronto, Toronto M5G 1X8
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Michelle Letarte
Michelle Letarte
1Department of Immunology, University of Toronto, Toronto M5G 1X8
1Division of Immunology, the Hospital for Sick Children, 555 University Avenue, Toronto M5G 1X8
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Publisher: Portland Press Ltd
Received:
April 22 1988
Online ISSN: 1573-4935
Print ISSN: 0144-8463
© 1988 Plenum Publishing Corporation
1988
Biosci Rep (1988) 8 (4): 359–368.
Article history
Received:
April 22 1988
Citation
Ian Rogers, Giacomo D'Agostaro, Sonia Vera, Michelle Letarte; Several epitopes of p85 glycoprotein (CDw44) are dependent on intact disulphide bonds. Isolation of cDNA clones requires a polyclonal antibody raised against the reduced protein. Biosci Rep 1 August 1988; 8 (4): 359–368. doi: https://doi.org/10.1007/BF01115227
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