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Antigenic modulation and internalization of monoclonal antibody to human colonic carcinoma cells detected by enzyme‐linked immunosorbent assay
Author(s) -
Yokota T.,
Otsuji E.,
Noguchi A.,
Yamaguchi T.,
Sawai K.,
Takahashi T.
Publication year - 1989
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.2910440626
Subject(s) - monoclonal antibody , biotinylation , internalization , microbiology and biotechnology , antigen , antibody , biology , cell culture , epitope , staining , biotin , chemistry , cell , biochemistry , immunology , genetics
We studied antigenic modulation and internalization of monoclonal antibody (MAb) A7 using the enzyme‐linked immunosorbent assay (ELISA) and biotin‐labelled antibody‐staining techniques. Incubation of the colonic SW1116 cell line with an excess of MAb A7 induced modulation of the cell‐surface antigen. When the line was assayed by ELISA, a change in cellular reactivity with MAb A7 was seen after 1 hr. After 24 hr, the cellular reactivity showed a 52% decrease in absorbance. Modulation was inhibited by 0.1 % sodium azide and acetone fixation, suggesting that this is an energy‐dependent phenomenon. The internalization of biotinylated MAb A7 was examined. Internalized biotinylated MAb A7 was detected in cells fixed before labelling with avidin‐biotin peroxidase complex. It was observed that the amount of MAb A7 which remained associated with the cell surface had decreased since A7 was internalized.