Peng Dan-dan, Ning Yun-shan, Li Yan, Hong Yan-hua, Wang Yun-dan, Wu Fen-fang, Li Ming
Institute of Tropical Medicine, Southern Medical University, Guangzhou 510515, China.
Di Yi Jun Yi Da Xue Xue Bao. 2005 Jan;25(1):40-3.
To prepare monoclonal antibodies (mAbs) against multiple antigens by single cell fusion.
BALB/c mice were immunized with the multiple antigens, namely alpha fetoprotein (AFP), carcinoembryonic antigen (CEA), HBsAgiHBcAg and HBeAg, and hybridomas were employed using PEG as the fusing agent. The hybridoma cells were respectively screened with AFP, CEA, HBsAg, HBcAg and HBeAg by enzyme-linked immunosorbent assay and limited dilution. The mAbs were purified by protein G affinity chromatography, its subtype was identified, the affinity constants (K(a)) were determined and the specificity was analyzed by Western blotting.
Twenty hybridoma cell lines were obtained by single cell fusion, including 5 cell lines against AFP, 6 against CEA, 3 against HBsAg, 4 against HBcAg, and 2 against HBeAg. The subtypes of some hybridoma cell lines positive for the mAbs were identified as the immunoglobulin G1 (IgG1), with K(a) ranging from 1x10(9) M(-1) to x10(11) M(-1). Western blot analysis showed that all the mAbs strongly and specifically bound to their respective antigens.
The mAbs against multiple antigens have been obtained by single cell-fusion, which increases the production of mAbs and reduces the time of preparation.
通过单细胞融合制备针对多种抗原的单克隆抗体（mAb）。
用甲胎蛋白（AFP）、癌胚抗原（CEA）、乙肝表面抗原（HBsAg）/乙肝核心抗原（HBcAg）和乙肝e抗原等多种抗原免疫BALB/c小鼠，并以聚乙二醇（PEG）作为融合剂制备杂交瘤。通过酶联免疫吸附测定法和有限稀释法，分别用AFP、CEA、HBsAg、HBcAg和HBeAg对杂交瘤细胞进行筛选。用蛋白G亲和层析法纯化单克隆抗体，鉴定其亚型，测定亲和常数（K(a)），并用蛋白质印迹法分析其特异性。
通过单细胞融合获得了20株杂交瘤细胞系，其中5株针对AFP，6株针对CEA，3株针对HBsAg，4株针对HBcAg，2株针对HBeAg。鉴定出部分产生单克隆抗体的杂交瘤细胞系的亚型为免疫球蛋白G1（IgG1），K(a)范围为1×10⁹ M⁻¹至×10¹¹ M⁻¹。蛋白质印迹分析表明，所有单克隆抗体均能与其各自的抗原发生强烈且特异性的结合。
通过单细胞融合获得了针对多种抗原的单克隆抗体，提高了单克隆抗体的制备效率并缩短了制备时间。
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