Ghetie V, Swindell E, Uhr J W, Vitetta E S
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235-9048.
J Immunol Methods. 1993 Nov 5;166(1):117-22. doi: 10.1016/0022-1759(93)90335-5.
A method for the preparation of immunotoxins (ITs) containing one and two molecules of deglycosylated ricin A chain (dgA) bound to each molecule of anti-human CD22 mouse IgG1 monoclonal antibody is described. The cross-linking of antibody and dgA molecules was accomplished with N-succinimidyl-oxycarbonyl-alpha-methyl-(2-pyridyldithio) toluene. The two ITs were purified by successive chromatographies on Blue-Sepharose, Sephacryl S-200, Blue-Sepharose with NaCl gradient and Sephacryl S-300. The IT with two dgAs was seven-fold more cytotoxic to CD22+ Daudi cells than that containing one molecule of dgA.
描述了一种制备免疫毒素（ITs）的方法，该免疫毒素包含与每分子抗人CD22小鼠IgG1单克隆抗体结合的一分子和两分子去糖基化蓖麻毒素A链（dgA）。抗体和dgA分子的交联是用N-琥珀酰亚胺基-氧羰基-α-甲基-（2-吡啶二硫基）甲苯完成的。通过在Blue-Sepharose、Sephacryl S-200、含NaCl梯度的Blue-Sepharose和Sephacryl S-300上连续进行色谱法对这两种ITs进行纯化。含有两分子dgA的IT对CD22+ Daudi细胞的细胞毒性比含有一分子dgA的IT高7倍。
