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UC729-6细胞与淋巴结细胞一次性融合产生了几株人-人杂交瘤,对其所分泌和合成大分子类型和性质进行了鉴定。VLN3G2杂交瘤株生长4天后,分泌的IgG比细胞浆高4倍。而IgM的分布与IgG正相反。与VLN3G2不同,VLN5C7在同一时间内,胞浆中所含IgG与IgM的量比分泌型的量高出几倍。这二株杂交瘤所分泌的IgG和JgM,只有IgG对靶A431细胞表面抗原有免疫反应性。还有一株为VLN1H12,所分泌的IgM对靶A431细胞有免疫活性,未测到有IgG的分泌。反复冻融杂交瘤细胞制备的胞浆蛋白,NP-40的细胞膜提取获得膜蛋白,用ELISA检测各杂交瘤上清的分泌蛋白,以了解细胞和细胞外观察到的免疫蛋白的变异性。用于融合的亲本UC-729-6细胞系只产生微量无免疫活性的IgM。对这些杂交瘤上清的分子筛柱层折提示有完整IgG和IgM分子,缺乏游离重链或同时含有μ和γ量链的杂交抗体。用抗硫-蛋氨酸内标记VLN3G2杂交瘤细胞说明不仅有非免疫球蛋白存在,而且在培养上清有小分子量A蛋白结合多肽存在。从VLN3G2培养液,胞浆及细胞膜分离到的硫-蛋氨酸整合IgG和IgM抗体,也表明可与菌体A蛋白结合。因此本文结论就是,人-人杂交瘤分泌的MCAbs量的变异不是由于这些分子的合成减少所引起的。
One-time fusion of UC729-6 cells with lymph node cells resulted in several human-human hybridomas that identified the type and nature of their secreted and synthesized macromolecules. After 4 days of growth of the VLN3G2 hybridoma strain, secreted IgG was 4-fold more cytosolic. The distribution of IgM and IgG is the opposite. Unlike VLN3G2, VLN5C7 has several times more secreted amounts of IgG and IgM than cytoplasm at the same time. The IgG and JgM secreted by these two hybridomas were only IgG immunoreactive with the target A431 cell surface antigen. There is also a strain of VLN1H12, secreted by the IgM target A431 cells have immune activity, did not measure the secretion of IgG. The cytoplasmic proteins prepared from hybridoma cells were repeatedly frozen and thawed, and the membrane proteins were extracted from the cell membrane of NP-40. The secreted protein of each hybridoma supernatant was detected by ELISA to understand the variability of the cellular and extracellular immune proteins. The parent UC-729-6 cell line used for fusion produced only minimal amounts of non-immunocompetent IgM. Molecular sieve column stacking on these hybridoma supernatants suggests intact IgG and IgM molecules, lacking free heavy chains or hybrid antibodies containing both the [mu] and [gamma] chains. The anti-S-methionine VLN3G2 hybridoma cells showed that not only the non-immunoglobulin, but also the presence of small molecular weight protein A binding polypeptide in the culture supernatant. Sulfur-methionine-integrated IgG and IgM antibodies isolated from the VLN3G2 broth, cytoplasm and cell membranes also showed binding to the Mycoprotein A protein. Therefore, the conclusion of this paper is that the variation in the amount of MCAbs secreted by human-human hybridomas is not due to the reduced synthesis of these molecules.