Temponi M, Pupa S, Ferrone S
Department of Microbiology and Immunology, New York Medical College, Valhalla 10595.
J Immunol Methods. 1990 Feb 9;126(2):223-9. doi: 10.1016/0022-1759(90)90154-n.
A method is described to purify immunoreactive monoclonal antibodies from radiolabeled monoclonal antibody preparations. The method is based on incubation of radiolabeled monoclonal antibodies with insolubilized anti-idiotypic monoclonal antibodies to idiotopes within the antigen-combining site of monoclonal antibodies to be purified and elution of bound monoclonal antibodies with a low pH buffer. The immunoreactive fraction of the purified monoclonal antibodies was at least 82%; the yield was at least 73%. The purification procedure did not cause any detectable change in the affinity constant of the eluted monoclonal antibodies. The method is simple and rapid; the requirement for anti-idiotypic monoclonal antibodies to idiotopes within the antigen-combining site of the antibodies to be purified is not likely to represent a major limitation in the broad application of the present method, since the hybridoma technology has greatly facilitated the development of anti-idiotypic monoclonal antibodies.
本文描述了一种从放射性标记的单克隆抗体制剂中纯化免疫反应性单克隆抗体的方法。该方法基于将放射性标记的单克隆抗体与针对待纯化单克隆抗体抗原结合位点内独特型表位的不溶性抗独特型单克隆抗体一起孵育，并用低pH缓冲液洗脱结合的单克隆抗体。纯化后的单克隆抗体的免疫反应性部分至少为82%；产率至少为73%。纯化过程未导致洗脱的单克隆抗体的亲和常数发生任何可检测到的变化。该方法简单快速；对于针对待纯化抗体抗原结合位点内独特型表位的抗独特型单克隆抗体的需求，不太可能成为本方法广泛应用的主要限制因素，因为杂交瘤技术极大地促进了抗独特型单克隆抗体的开发。
