Cullen S E, Schwartz B D
J Immunol. 1976 Jul;117(1):136-42.
Mouse alloantigens, including the histocompatibility (H-2) and immune response linked (Ia) antigen molecules, can be readily isolated by a new precipitation technique utilizing protein A-bearing, fixed Staphylococci. The antigens are prepared by radiolabeling mouse spleen cells with 3H-leucine, solubilizing with the non-ionic detergent Non-Idet P-40, and purifying by affinity chromatography with lentil lectin coupled to Sapharose. The antigen preparations are mixed with appropriate alloantisera, and immune complexes formed in the mixture are then bound by the protein A sites on the Staphylococci. The organisms, Staphylococcus aureus, Cowan I strain (ATCC 12598), are heat inactivated and fixed, and are a stable, uniform IgG-binding reagent, especially when stored frozen. The antigen-antibody complexes are easily eluted from the organisms for electrophoretic analysis. The precipitation mediated by the organisms is more efficient, rapid, and artifact-free than the traditional "sandwich" precipitation technique involving anti-globulin reagents.
小鼠同种异体抗原，包括组织相容性（H-2）和免疫反应相关（Ia）抗原分子，可通过一种利用带有蛋白A的固定葡萄球菌的新沉淀技术轻松分离出来。抗原的制备方法是先用3H-亮氨酸对小鼠脾细胞进行放射性标记，然后用非离子去污剂Non-Idet P-40使其溶解，并通过与偶联到琼脂糖上的扁豆凝集素进行亲和层析来纯化。将抗原制剂与适当的同种异体抗血清混合，混合物中形成的免疫复合物随后被葡萄球菌上的蛋白A位点结合。金黄色葡萄球菌考恩I菌株（ATCC 12598）经过热灭活和固定处理，是一种稳定、均匀的IgG结合试剂，尤其是在冷冻保存时。抗原-抗体复合物很容易从这些微生物中洗脱出来用于电泳分析。与涉及抗球蛋白试剂的传统“夹心”沉淀技术相比，由这些微生物介导的沉淀更高效、快速且无假象。
