Metal binding properties of a monoclonal antibody directed toward metal-chelate complexes
Metal binding properties of a monoclonal antibody directed toward metal-chelate complexes
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DOI：
10.1074/jbc.271.44.27677
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发表时间：
1996-11-01
影响因子：
4.8
通讯作者：
Blake, RC
中科院分区：
文献类型：
--
作者：
Blake, DA;Chakrabarti, P;Blake, RC
通过向BALB/c小鼠注射与载体蛋白共价偶联的金属螯合复合物，制备了识别镉-EDTA复合物的单克隆抗体。通过使用KinExA（TM）免疫测定仪器测量平衡结合常数，抗体与镉-和汞-EDTA络合物结合，平衡解离常数分别为21和26 nm。测试的所有其他金属EDTA络合物，包括Mn（II），In（III），Ni（II），Zn（II）、Ga（III）、Mg（II）和Al（III）的结合亲和力比镉-EDTA络合物的结合亲和力低20- 40，000倍。除汞和镁外，二价金属螯合物的结合与金属离子的大小密切相关，通过聚合酶链反应推导出重链和轻链可变区的氨基酸序列-相应基因的扩增区域并随后用于构建抗原结合区域的分子模型，2A 81 G5模型中镉结合的关键残基似乎是重链中的组氨酸96。
A monoclonal antibody that recognizes cadmium-EDTA complexes has been produced by the injection of BALB/c mice with a metal-chelate complex covalently coupled to a carrier protein, The ability of purified antibody to recognize 16 different metal-EDTA complexes was assessed by measuring equilibrium binding constants using a KinExA(TM) immunoassay instrument, The antibody bound to cadmium- and mercury-EDTA complexes with equilibrium dissociation constants of 21 and 26 nm, respectively, All other metal EDTA complexes tested, including those of Mn(II), In(III), Ni(II), Zn(II), Ga(III), Mg(II), and AI(III) bound with affinities from 20- to 40,000-fold less than that determined for the cadmium-EDTA complex, With the exception of mercury and magnesium, the binding of divalent metal-chelate complexes was well-correlated with the size of the metal ion, The amino acid sequences of the heavy and light chain variable regions were deduced from polymerase chain reaction-amplified regions of the corresponding genes and subsequently used to construct molecular models of the antigen binding region, The key residue for cadmium binding in the model for 2A81G5 appeared to be histidine 96 in the heavy chain.