Ghetie V, Ghetie M A, Uhr J W, Vitetta E S
Department of Microbiology, University of Texas Southwestern Medical Center, Dallas 75235.
J Immunol Methods. 1988 Sep 13;112(2):267-77. doi: 10.1016/0022-1759(88)90367-5.
In this report, we describe a method for the preparation of large amounts (grams) of immunotoxins (ITs) consisting of Fab' fragments of murine IgG1 monoclonal antibodies conjugated to chemically deglycosylated ricin A chain (dgA). The preparation of Fab' and dgA chain and the purification of the Fab'-dgA IT were accomplished by gel filtrations and affinity chromatography utilizing six Pharmacia Bioprocess columns (Sephadex G-25M, Sephacryl S-200HR and Blue Sepharose CL-4B) integrated into a semi-automatic chromatography system controlled by a Pharmacia C3-process controller. The final Fab'-dgA ITs were highly purified, potent, sterile and low in endotoxin concentration.
在本报告中，我们描述了一种制备大量（克级）免疫毒素（ITs）的方法，该免疫毒素由与化学去糖基化蓖麻毒素A链（dgA）偶联的鼠IgG1单克隆抗体的Fab'片段组成。Fab'和dgA链的制备以及Fab'-dgA IT的纯化通过凝胶过滤和亲和色谱法完成，使用了六个整合到由Pharmacia C3过程控制器控制的半自动色谱系统中的Pharmacia生物过程柱（Sephadex G-25M、Sephacryl S-200HR和Blue Sepharose CL-4B）。最终的Fab'-dgA ITs高度纯化、高效、无菌且内毒素浓度低。
