Generation of human monoclonal antibodies to cancer-associated antigens using limited numbers of patient lymphocytes.

Abstract:

:A limiting dilution method for the efficient transformation by Epstein-Barr virus (EBV) of human B lymphocytes has been applied to the production of human monoclonal antibodies to ovarian cancer-associated antigens. Limited numbers (e.g., 2 X 10(5)) of EBV-infected B lymphocytes from ovarian cancer patient spleen, lymph node, tumor, ascites and blood were successfully transformed using this method. An immunofiltration assay system was employed to identify EBV transformants secreting IgM antibody which reacted selectively with ovarian cancer patient ascites tumor cells, but not with a mixture of normal cell types. A miniature Western blot assay was utilized to screen for IgG reactivity to protein species in detergent extracts of ovarian cancer tumor cells. EBV-transformed cells selected after screening were then fused with heteromyeloma fusion partner SHM-D33 resulting in efficient recovery of hybridomas secreting MAb of the desired specificity. Human MAbs which selectively react with antigens associated with ovarian cancer tumor cells were obtained.

journal_name

J Immunol Methods

authors

Smith LH,Yin A,Bieber M,Teng NN

doi

10.1016/0022-1759(87)90274-2

subject

Has Abstract

pub_date

1987-12-24 00:00:00

pages

263-73

issue

2

eissn

0022-1759

issn

1872-7905

pii

0022-1759(87)90274-2

journal_volume

105

pub_type

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