Sportsman J R, Taber L D, Slisz M C, Apelgren L D, Bumol T F
Lilly Research Laboratories, A Division of Eli Lilly and Company, Indianapolis, Indiana 46285.
Biotechnol Appl Biochem. 1988 Dec;10(6):536-44.
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.
人腺癌相关抗原gp40是一种细胞表面糖蛋白，可被鼠单克隆抗体KS1/4识别。利用KS1/4-琼脂糖亲和基质从人肺腺癌细胞系P3-UCLA的组织培养细胞或裸鼠异种移植瘤的去污剂裂解物中纯化gp40。这一单步免疫亲和层析步骤得到了纯度约为95%的抗原制剂，并用免疫化学和酶学技术对其进行了进一步表征。结果表明，gp40分子同时具有复杂型和高甘露糖型寡糖，约占表观分子量的16%。该抗原制剂适用于气相N端氨基酸测序，并测定了N端的前16个残基。尽管存在相当大的分子异质性，但gp40显示出单一的N端序列。
