Wylie D E, Lu D, Carlson L D, Carlson R, Babacan K F, Schuster S M, Wagner F W
School of Biological Sciences, University of Nebraska, Lincoln 68588.
Proc Natl Acad Sci U S A. 1992 May 1;89(9):4104-8. doi: 10.1073/pnas.89.9.4104.
Monoclonal antibodies (mAbs) that react with soluble mercuric ions have been produced by injection of BALB/c mice with a hapten-carrier complex designed to maximize exposure of the metal to the immune system. Three hybridomas producing antibodies that reacted with bovine serum albumin (BSA)-glutathione-HgCl, but not with BSA-glutathione, were isolated from the spleen of a mouse given multiple injections with glutathione-HgCl conjugated to keyhole limpet hemocyanin. Stable subclones were established from two of these antibodies, designated mAb 4A10 and mAb 1F10. The binding of both antibodies to immobilized BSA-glutathione-HgCl was inhibited by soluble HgCl2, and dissociation constants for mercuric chloride binding were 2.3 and 3.7 nM for mAbs 4A10 and 1F10, respectively. Both antibodies bound mercuric acetate with similar affinities, demonstrating that the antibodies were capable of binding to mercuric ions in the presence of a different counterion than the one used in the immunogen. Reactions were not observed with other metal cations by either antibody. These data demonstrate the successful induction of antibodies that react very specifically with mercuric ions in solution regardless of the presence of a carrier.
通过向BALB/c小鼠注射一种旨在使金属最大程度暴露于免疫系统的半抗原-载体复合物,已产生了与可溶性汞离子发生反应的单克隆抗体(mAb)。从多次注射与钥孔戚血蓝蛋白偶联的谷胱甘肽-HgCl₂的小鼠脾脏中,分离出了三种产生与牛血清白蛋白(BSA)-谷胱甘肽-HgCl₂反应但不与BSA-谷胱甘肽反应的抗体的杂交瘤。从其中两种抗体建立了稳定的亚克隆,分别命名为单克隆抗体4A10和单克隆抗体1F10。可溶性HgCl₂抑制了两种抗体与固定化的BSA-谷胱甘肽-HgCl₂的结合,单克隆抗体4A10和1F10与氯化汞结合的解离常数分别为2.3 nM和3.7 nM。两种抗体以相似的亲和力结合醋酸汞,表明这些抗体在存在与免疫原中所用不同抗衡离子的情况下能够结合汞离子。两种抗体均未观察到与其他金属阳离子发生反应。这些数据证明成功诱导出了无论是否存在载体都能与溶液中的汞离子非常特异性反应的抗体。