Anderson L J, Coombs R A, Tsou C, Hierholzer J C
J Clin Microbiol. 1984 Jun;19(6):934-6. doi: 10.1128/jcm.19.6.934-936.1984.
To characterize the specificity of monoclonal antibodies against respiratory syncytial virus, we developed a technique which combined the biotin-avidin system, immunoprecipitation, and transblot electrophoresis. The viral proteins were first biotinylated and then immunoprecipitated with a monoclonal antibody or respiratory syncytial virus immune serum, separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis, and electrophoretically transferred to nitrocellulose paper. The protein bands were located on the paper with avidin-peroxidase plus a precipitable substrate. This procedure gave reproducible results qualitatively similar to those obtained by radioimmunoprecipitation but without the cost, risk of radiation exposure, or disposal problems of radioisotopes. This procedure should have widespread applications.
为了鉴定抗呼吸道合胞病毒单克隆抗体的特异性，我们开发了一种结合生物素-抗生物素蛋白系统、免疫沉淀和转印电泳的技术。首先将病毒蛋白生物素化，然后用单克隆抗体或呼吸道合胞病毒免疫血清进行免疫沉淀，通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳分离，再电泳转移至硝酸纤维素纸上。用抗生物素蛋白-过氧化物酶加可沉淀底物在纸上定位蛋白条带。该方法产生的定性结果与放射免疫沉淀法获得的结果相似且具有可重复性，但没有放射免疫沉淀法的成本、辐射暴露风险或放射性同位素的处置问题。该方法应具有广泛的应用。
