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由于大多数抗原呈可溶状态,根据抗原抗体复合物原理,可将单克隆抗体应用于免疫检测。多数情况下必须在实验系统中提供固相,一般是采用抗原或抗体包被在玻璃珠、试管壁等因相上,或加入PEG将抗原抗体复合物沉淀等方法,操作较繁琐。根据分泌抗体的细胞膜上嵌有大量具免疫活性的抗体,本文提出一种将抗体直接固于分泌抗体的杂交瘤细胞上的新方法。具体作法是将活跃分泌单抗的杂交瘤细胞悬浮于磷酸缓冲液中,加入戊二醛,终浓度为2.5%(V/V)。置4℃固定30分钟,然后加入甘氨酸,
Since most antigens are soluble, monoclonal antibodies can be used in immunoassays based on the principle of antigen-antibody complexes. In most cases, the solid phase must be provided in the experimental system. Generally, the antigen or antibody is coated on the glass beads, the wall of the test tube or the like, or the method of precipitating the antigen-antibody complex by adding PEG is more complicated. According to the antibody-secreting cell membrane embedded with a large number of immunocompetent antibodies, this paper presents a new method of directly immobilizing antibodies on antibody-secreting hybridoma cells. Specifically, monoclonal antibody-secreting hybridoma cells were suspended in phosphate buffer and glutaraldehyde was added at a final concentration of 2.5% (V / V). Set 4 ℃ fixed 30 minutes, then add glycine,