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Isolation and characterization of the human adenocarcinoma‐associated glycoprotein gp40
Author(s) -
Sportsman JR,
Taber LD,
Slisz MC,
Apelgren LD,
Bumol TF
Publication year - 1988
Publication title -
biotechnology and applied biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.468
H-Index - 70
eISSN - 1470-8744
pISSN - 0885-4513
DOI - 10.1111/j.1470-8744.1988.tb00041.x
Subject(s) - monoclonal antibody , antigen , microbiology and biotechnology , biology , glycoprotein , biochemistry , cell culture , sepharose , antibody , enzyme , chemistry , immunology , genetics
The human adenocarcinoma‐associated antigen gp40 is a cell surface glycoprotein recognized by murine monoclonal antibody KS1/4. A KS1/4‐Sepharose affinity matrix was utilized to purify gp40 from detergent lysates of either tissue culture cells or nude mouse xenograft tumors of the human lung adenocarcinoma cell line P3‐UCLA. This single immunoaffinity chromatography step yielded an antigen preparation of approximately 95% purity which was further characterized by immunochemical and enzymatic techniques. The gp40 molecule was shown to have both complex and high‐mannose oligosaccharides comprising some 16% of the apparent molecular weight. The antigen preparation was suitable for gas‐phase N‐terminal amino acid sequencing and the first 16 residues of the N‐terminus were determined. Despite considerable molecular heterogeneity, gp40 shows a single N‐terminal sequence.