New developments in the production and use of stereoselective antibodies.
New developments in the production and use of stereoselective antibodies.
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DOI：
10.1002/chir.20099
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发表时间：
2005
期刊：
影响因子：
2
通讯作者：
H. Hofstetter;James R Cary;Pierre P. Eleniste;Jay K Hertweck;H. Lindstrom;Daniel I Ranieri;Gregory B Smith;L. P. Undesser;Jessica M Zeleke;Tigabu K Zeleke;O. Hofstetter
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文献类型：
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作者：
H. Hofstetter;James R Cary;Pierre P. Eleniste;Jay K Hertweck;H. Lindstrom;Daniel I Ranieri;Gregory B Smith;L. P. Undesser;Jessica M Zeleke;Tigabu K Zeleke;O. Hofstetter
本文介绍了使用经典的免疫学和现代分子生物学技术的立体选择性抗体的生产。立体选择性抗体对α-羟基酸提出了在兔子和小鼠，并与以前生产的抗α-氨基酸抗体进行比较。发现两种类型的抗体都将联合收割机立体选择性与类别特异性结合。序列分析显示，具有相反立体选择性的抗体可以在亲和力成熟过程中从共同的祖先或独立地使用非同源结合位点形成。首次采用噬菌体展示技术获得了具有立体选择性的抗体片段。立体选择性抗体作为手性选择剂的多功能性通过将其应用于几种免疫传感器和手性色谱中来证明。一个简单的，基于膜的光学传感器允许检测对映体杂质在1/2,000水平（99.9%ee）。基于二氧化硅的抗体手性固定相可用于在标准尺寸柱中分离脂肪族氨基酸的对映体，而小型化柱允许与MS检测器连接。
This article describes the production of stereoselective antibodies using both classical immunological and modern molecular biological techniques. Stereoselective antibodies against alpha-hydroxy acids were raised in rabbits and mice and compared with previously produced anti-alpha-amino acid antibodies. It was found that both types of antibodies combine stereoselectivity with class-specificity. Sequence analyses revealed that antibodies with opposing stereoselectivities can be formed during the affinity maturation process from a common progenitor or independently using nonhomologous binding sites. For the first time, phage display was employed to obtain stereoselective antibody fragments. The versatility of stereoselective antibodies as chiral selectors was demonstrated by applying them in several immunosensors and in chiral chromatography. A simple, membrane-based optical sensor allowed detection of enantiomeric impurities at the 1/2,000 level (99.9% ee). Silica-based antibody chiral stationary phases could be used for enantiomer separation of aliphatic amino acids in standard-sized columns, while miniaturized columns allowed interfacing with an MS-detector.