摘要
将人肺腺癌细胞ＳＰＣ、Ａ５４９、Ａ４２７序贯免疫ＢＡＬＢ／Ｃ小鼠，取脾细胞与小鼠骨髓瘤细胞ＳＰ２／０融合。获得一株稳定分泌抗肺腺鳞癌单克隆抗体的杂交瘤细胞株──Ｓ５Ａ１０─２，制备了小鼠腹水抗体并进行了纯化，腹水效价为１．０２×１０７，亚型测定为ＩｇＧ１．免疫组化分析ｓ５Ａ１０－２单抗与肺腺、鳞癌和肺鳞癌有较好的选择反应性，与正常组织无交叉反应，ＷｅｓｔｅｒｎＢｌｏｔ测定Ｓ５Ａｌ０─２在ＳＰＣ细胞膜上的抗原分子量为４０Ｋｄ、５０Ｋｄ和８０Ｋｄ，在Ａ５４９，细胞膜上的抗原分子量为４３Ｋｄ，而与Ａ４２７细胞膜蛋白无明显反应。该抗体的亲和常数为８．４３×１０１０Ｍ－１，Ｓ５Ａ１０－２杂交瘤细胞株，经液氮冻存后复苏，其分泌抗体能力不受影响。
McAb S5A10-2 were prepared by fusing SP2/0 cells with spleen cells of BALB/c mice immunized with the human lung adenocarinoma cells SPC、A549 and A427.The titers of McAb in the ascites reached 1.02 ×107. The immunoglobulin class/subclass of McAb S5A10-2 was IgG1. Immunohostochemical study showed that S5A10-2 reacted with the majority of plumonary carcinoma tissues and not with normal tissues. Western blot analysis indicated that S5A10-2 recognized both SPC and A549 , but not A549, but not A427. The molecular weight of SPC cell membrane antigens were 40KD、50KD and 80KD and that A549 was 43KD. The affinity constant was 8.4× 1010M-1. After repeated frozen storage and revival, the S5A10-2 hybridoma cell line can steadily produce high titers of McAb.
出处
《肿瘤》
CAS
CSCD
北大核心
1995年第1期9-12,共4页
Tumor
关键词
肺肿瘤
腺癌
鳞癌
杂交瘤
抗体
单克隆抗体
研制
Lung adenocarcinoma, Lung squamous carcinoma, Hybridoma, Monoclonalantibody.
