Swack J A, Karawya E, Albert W, Fedorko J, Minna J D, Wilson S H
Anal Biochem. 1985 May 15;147(1):10-21. doi: 10.1016/0003-2697(85)90003-x.
A panel of 12 hybridoma cell lines secreting monoclonal antibodies against alpha-polymerase were prepared by fusion of mouse myeloma cells and spleen cells of a rat immunized with homogeneous calf thymus alpha-polymerase. Hybridomas were selected and cloned on the basis of immunobinding to pure alpha-polymerase in solid phase radioimmunoassay. Antibodies secreted by these cells eventually were purified in milligram quantities from ascites fluids. These antibodies, all of the rat immunoglobulin M class, cross-reacted with alpha-polymerases from calf and monkey cells as revealed by immunobinding in radioimmunoassay and by immunoprecipitation of DNA polymerase activity. The antibodies were not capable of neutralizing the enzyme activity. With the methods described these antibodies may be used to immunoprecipitate alpha-polymerase from crude extracts of mammalian cells and to measure levels of the enzyme protein.
通过将小鼠骨髓瘤细胞与用纯小牛胸腺α-聚合酶免疫的大鼠脾细胞融合，制备了一组12种分泌抗α-聚合酶单克隆抗体的杂交瘤细胞系。基于在固相放射免疫测定中与纯α-聚合酶的免疫结合，选择并克隆杂交瘤。最终从腹水液中以毫克量纯化这些细胞分泌的抗体。这些抗体均为大鼠免疫球蛋白M类，通过放射免疫测定中的免疫结合和DNA聚合酶活性的免疫沉淀显示，它们与来自小牛和猴细胞的α-聚合酶发生交叉反应。这些抗体不能中和酶活性。用所述方法，这些抗体可用于从哺乳动物细胞粗提物中免疫沉淀α-聚合酶并测定酶蛋白水平。
