Electrospray ionization-ion mobility spectrometry identified monoclonal antibodies that bind exclusively to either the monomeric or a dimeric form of prostate specific antigen.
Electrospray ionization-ion mobility spectrometry identified monoclonal antibodies that bind exclusively to either the monomeric or a dimeric form of prostate specific antigen.
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电喷雾电离离子迁移率识别单克隆抗体，这些抗体仅与前列腺特异性抗原的单体或二聚体形式结合。
DOI：
10.1021/ac301527v
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发表时间：
2012-08-07
影响因子：
7.4
通讯作者：
Blake, Diane A.
中科院分区：
文献类型：
--
作者：
Blake, Robert C., II;Blake, Diane A.
大分子离子迁移谱用于区分仅与单体前列腺特异性抗原结合的单克隆抗体（克隆M612165）和仅与仅占总蛋白6.8%的抗原二聚体形式结合的不同单克隆抗体（克隆M612166）。在存在过量抗原的情况下，M612165的迁移率光谱被代表抗体混合物（包括1或2当量蛋白抗原）的复合光谱替代。在类似情况下，M612166的迁移率谱被代表抗体混合物（包括2或4当量蛋白抗原）的复合谱所替代。当暴露于两种抗体中的任一种时，前列腺特异性抗原的迁移率谱显示在一种情况下单体抗原和在另一种情况下二聚体抗原伴随减少。虽然灵敏的动力学排阻试验证明两种抗体的抗原结合行为存在较大差异，但仅这些功能研究不足以揭示所观察到差异的可能结构来源。大分子离子迁移率的测量被证明是一个有用的和翔实的补充功能的研究，了解复杂的大分子相互作用。
Macroion mobility spectrometry was used to distinguish between a monoclonal antibody (clone M612165) that bound exclusively to monomeric prostate specific antigen and a different monoclonal antibody (clone M612166) that bound exclusively to a dimeric form of the antigen that only comprised 6.8% of the total protein. In the presence of excess antigen, the mobility spectrum of M612165 was replaced by a composite spectrum that represented a mixture of antibodies that included either one or two equivalents of the protein antigen. In similar circumstances, the mobility spectrum of M612166 was replaced by a composite spectrum that represented a mixture of antibodies that included either two or four equivalents of the protein antigen. When exposed to either of the two antibodies, the mobility spectrum of the prostate specific antigen showed a concomitant decrease in the monomeric antigen in one case and in the dimeric antigen in the other case. While sensitive kinetic exclusion assays demonstrated large differences in the antigen binding behavior of the two antibodies, these functional studies alone were insufficient to reveal the likely structural origins of the observed differences. Macroion mobility measurements were shown to be a useful and informative complement to functional studies in understanding complex macromolecular interactions.
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