Isolation and characterization of the human adenocarcinoma-associated glycoprotein gp40.
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.
Sportsman
J R
JR
Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, Indiana 46285.
Taber
L D
LD
Slisz
M C
MC
Apelgren
L D
LD
Bumol
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TF
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm / Q000032:analysis / Q000302:isolation & purification*; D015153:Blotting, Western; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D007124:Immunoenzyme Techniques; D008562:Membrane Glycoproteins / Q000032:analysis / Q000302:isolation & purification*; D012685:Sepharose; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D008562:Membrane Glycoproteins; C035267:glycoprotein 40 antigen, human; D012685:Sepharose