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杂交瘤技术的成功主要决定于能否获得抗原特异性淋巴细胞。本文作者提出了一种用于小分子量抗原的免疫方案,可使细胞融合后提高特异性杂交瘤的产生率。作者用生物合成的人胰岛素或结合于卵清蛋白的胰岛素免疫Balb/c雌性小鼠,初免用10μg免疫原,加弗氏完全佐剂作腹腔注射,每4周加强1次,共加强两次。将免疫小鼠分为8组,1～4组于融合前4天注射10μg半抗原,5～8组于融合前4天每天注射不同剂量的半抗原。然后将免疫小鼠脾脏制成单细胞悬液,与NS1骨髓瘤细胞按5∶1的比例融合,融合细胞培养于含次黄嘌
The success of hybridoma technology depends primarily on the availability of antigen-specific lymphocytes. The authors propose an immunization protocol for small molecular weight antigens to increase the rate of specific hybridomas after cell fusion. The authors used biosynthesis of human insulin or ovalbumin-conjugated insulin immunized Balb / c female mice, initial immunization with 10μg of immunogen, complete Freund’s adjuvant for intraperitoneal injection, once every 4 weeks to strengthen a total of two Times. The immunized mice were divided into 8 groups, 1 ~ 4 groups were injected with 10μg hapten 4 days before fusion, and 5 ~ 8 groups were injected with different doses of haptens 4 days before the fusion. Then, the spleens of the immunized mice were made into a single cell suspension and fused with NS1 myeloma cells at a ratio of 5: 1. The fused cells were cultured in a medium containing hypoxanthine