小鼠体内抗牛血清白蛋白B细胞克隆对McAb筛选的影响

目的　探讨小鼠体内存在的抗牛血清白蛋白 B细胞克隆对抗原特异性 Mc Ab筛选的影响。方法　在Mc Ab筛选过程中 ,采用间接 EL ISA法 ,分别以人重组高效复合干扰素 (r SIFN- co)和 BSA为抗原包被酶标板 ,对杂交瘤细胞培养上清进行“双筛”,建立 r SIFN- co/ BSA双阳性杂交瘤细胞克隆和 r SIFN- co单阳性杂交瘤细胞克隆 ,并且对所建立的各株杂交瘤细胞培养上清和正常小鼠血清 ,用 BSA进行吸附抑制实验。结果　建立了 1株r SIFN- co/ BSA双阳性杂交瘤细胞株 ,3株 r SIFN- co单阳性杂交瘤细胞株 ,双阳性杂交瘤细胞株培养上清经 BSA预先吸附后 ,与 r SIFN- co的反应性被显著抑制 (P<0 .0 5 )。而且正常小鼠血清与 r SIFN- co呈阳性反应 ,经 BSA吸附后 ,与 r SIFN- co的反应也被显著抑制。结论　在杂交瘤细胞筛选中 ,存在对特异性抗原和用于封闭酶标板的BSA显示双阳性的杂交瘤细胞克隆 ,提示小鼠体内可能天然存在抗 BSA的 B细胞克隆 ,它们在 Mc Ab筛选过程中会引起假阳性反应 ,因而在杂交瘤细胞筛选过程中应该用特异性抗原和 BSA进行“双筛”。

The Effect of Anti-Bovine Serum Albumin B Cell Clone in Mice on Preparation of McAb

Objective To investigate the effect of anti bovine serum B cell clone in mice on preparation of McAb. Methods Supernatant of hybridoma cell was double screened with rSIFN co and BSA by indirect enzyme linked immunosorbent assay. After BSA absorption, the inhibition effects on culture supernatant of double positive hybridoma cell clone and rSIFN co single positive hybridoma cell clones and on normal mouse serum were investigated separately. Results One double positive cell clone and three rSIFN co single positive clones were established. The reaction of rSIFN co with the supernatant of the double positive clone was inhibited obviously on condition that the supernatant was absorbed with BSA in advance. Conclusion In the screening of hybridoma cell, there is rSIFN co and BSA double positive hybridoma cell clone, which suggests the possible existence of innate anti BSA B cell clone in mice; they can cause false positive reaction in the preparation of McAb. Thus, double screening with specific antigen and BSA is beneficial.