Cheng W C, Gupta R K, Morton D L
Immunol Invest. 1985 Aug;14(4):367-78. doi: 10.3109/08820138509022672.
Immune complexes (IC) from the serum of a melanoma patient were partially purified with DEAE Affigel Blue. The IC were radioiodinated and then immobilized with activated Sepharose 4B. Immunologically bound components in the IC were dissociated with 3M MgCl2 followed by gel filtration chromatography. A distinct antigen component of about 700,000 daltons was obtained by refractionation of a region of the chromatogram containing antigenic activity by Sephacryl S-300 column chromatography. Though the antigenic component behaved as a single entity in the gel filtration, it consisted of at least five polypeptide chains that were revealed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis under reducing conditions. However, 92,000 and 41,000 dalton subunits were predominant. This procedure may be useful for isolating tumor antigens from circulating IC of cancer patients.
用DEAE交联琼脂糖蓝对一名黑色素瘤患者血清中的免疫复合物(IC)进行部分纯化。对IC进行放射性碘化,然后用活化的琼脂糖凝胶4B固定。IC中免疫结合的成分先用3M氯化镁解离,然后进行凝胶过滤色谱分析。通过用Sephacryl S - 300柱色谱对色谱图中含有抗原活性的区域进行重分离,获得了一种约700,000道尔顿的独特抗原成分。尽管该抗原成分在凝胶过滤中表现为单一实体,但在还原条件下通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳显示它至少由五条多肽链组成。然而,92,000道尔顿和41,000道尔顿的亚基占主导。该方法可能有助于从癌症患者的循环IC中分离肿瘤抗原。