Mazzaferro P K, Mayers G L
J Immunol Methods. 1981;46(3):327-36. doi: 10.1016/0022-1759(81)90318-5.
Two bifunctional reagents were shown to be useful in the coupling of immunogens to the surface of either nucleated or non-nucleated cells. The heterobifunctional reagent methyl- para -hydroxybenzimidate was used to couple aromatic amino-haptens to the surface of SRBC which resulted in a stable and sensitive target cell capable of detecting as little as 20 pg of purified anti-hapten antibody in the hemolytic spot assay. The multifunctional reagent 1, 3, 5-trichlorotriazine yielded similar results when coupling amino-haptens to the surface of SRBC for use in the hemolytic spot assay. This reagent was also used to couple protein to the surface of SRBC which were able to detect as low as 1 ng of purified anti-protein antibody in the hemolytic spot assay. The sensitized SRBC produced using either of these coupling reagents were shown to remain stable for several months giving reproducible results from one test to another. Lastly, 1, 3, 5-trichlorotriazine was used to couple the hapten 4-aminophthalate to the surface of nucleated cells with retention of greater than 90% cell viability with continued growth and cellular division in culture.
两种双功能试剂被证明可用于将免疫原偶联到有核细胞或无核细胞表面。异双功能试剂对羟基苯甲酸甲酯用于将芳香族氨基半抗原偶联到SRBC表面，这产生了一种稳定且灵敏的靶细胞，在溶血斑试验中能够检测低至20 pg的纯化抗半抗原抗体。多功能试剂1,3,5-三氯三嗪在将氨基半抗原偶联到SRBC表面用于溶血斑试验时也产生了类似结果。该试剂还用于将蛋白质偶联到SRBC表面，这些SRBC在溶血斑试验中能够检测低至1 ng的纯化抗蛋白质抗体。使用这两种偶联试剂中的任何一种产生的致敏SRBC被证明可在几个月内保持稳定，从一次试验到另一次试验都能得到可重复的结果。最后，1,3,5-三氯三嗪用于将半抗原4-氨基邻苯二甲酸偶联到有核细胞表面，细胞活力保留大于90%，在培养中持续生长并进行细胞分裂。
