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人绒毛膜促性腺激素(hCG)与人促黄体素(hLH),促卵泡素(hFSH)及促甲状腺素(hTSH)的结构近似,因而很难制备出与后三种糖蛋白激素无交叉反应的McAb。我们采用不同的免疫方法以及用不同分子量PEG作为融合剂,将hCG免疫的BALB/C小鼠脾细胞与SP2/O小鼠骨髓瘤细胞融合。在3次成功的融合中(融合率高于95％),从360个培养孔筛选出抗体阳性孔65个。经连续2～3次克隆化及液氮冻存后的多次再克隆化和传代,获得了12个持续分泌抗hCG McAb的杂交瘤株,并制备了抗体含量高达20～60mg/ml的大量小鼠腹水。
Human chorionic gonadotropin (hCG) is structurally similar to human hLH, hFSH and hTSH, making it difficult to prepare McAbs that do not cross-react with the last three glycoproteins . Using different immunization methods and immunization of hCG-immunized BALB / c mouse splenocytes with SP2 / O mouse myeloma cells using different molecular weight PEG as a fusogenic agent, In 3 successful fusions (fusion rate> 95%), 65 positive wells were screened from 360 culture wells. After two or three consecutive clones and several clonal re-cloning and passage after cryopreservation, twelve hybridomas secreting anti-hCG McAb were obtained and a large amount of antibody with the highest antibody content of 20-60 mg / ml Ascites in mice.