Direct Enantiomer Separation in Immunoaffinity Systems

描述(申请人提供)：生物体在几乎所有的相互作用水平上区分手性化合物的对映体，例如药物，并对它们做出不同的反应。虽然一种对映体可能表现出预期的活性，但另一种可能会引起严重的药理学和毒理学副作用，或充当拮抗剂。然而，在500种最畅销的药物中，超过一半的药物和目前正在开发的药物中约三分之二是手性的。自1988年以来，FDA不仅要求定量测定手性药物的立体异构体组成，还要求进行立体化学特异性鉴定试验，以及单个对映体的药理学和药代动力学表征。因此，发展精细的分析技术和选择器来精确测定生物活性手性分子及其纯化是非常有意义的。在色谱或电泳系统中，对映体的直接拆分是基于它们与手性选择剂的不同相互作用。尽管目前市面上有大约200种手性选择剂，但这些选择剂通常不是为特定的分离问题量身定做的，识别合适的选择剂仍然是一项乏味的反复试验。人们很早就知道抗体可以区分手性化合物的不同对映体。由于抗体几乎可以针对任何感兴趣的化合物而产生，我们对利用抗体的立体选择性和特异性来生产定制的手性固定相的想法很感兴趣。我们建议开发和研究将选择性与易用性、分析时间短和长期稳定性相结合的免疫亲和系统，因此适合于常规的对映体分离。立体选择性抗体或其片段将固定在传统的和高流通型固体支持材料上，并用于高压和电动分离技术，如高效液相色谱、微型LC、CE、CEC、OTEC和OTLC。开发在温和、等规度条件下分离对映体有利于蛋白质稳定性和活性的方法特别令人感兴趣。将优化层析参数，以调整分离条件以适应抗体和分析物之间的相互作用强度。

DESCRIPTION (provided by applicant): Living organisms discriminate between the enantiomers of chiral compounds, e.g., drugs, at virtually all levels of interaction, and respond differently to them. While one enantiomer may exhibit a desired activity, the other may cause severe pharmacologic and toxicologic side effects, or act as an antagonist. However, more than half of the 500 top selling drugs and about two-thirds of the drugs presently in development are chiral. Since 1988 the FDA demands not only the quantitative determination of the stereo isomeric composition of chiral drugs, but also stereochemically specific identity tests, as well as pharmacologic and pharmacokinetic characterization of the individual enantiomers. The development of refined analytical techniques and selectors for the precise determination of bioactive chiral molecules and their purification is therefore of great interest. Direct resolution of enantiomers in chromatographic or electrophoretic systems is based on their differential interactions with a chiral selector. Although approximately 200 chiral selectors are currently commercially available, these selectors are generally not tailor-made for a specific separation problem, and the identification of a suitable selector is still a tedious trial-and-error exercise. It has long been known that antibodies can differentiate between the enantiomers of chiral compounds. Since antibodies can furthermore be raised against virtually any compound of interest, we are intrigued by the idea of exploiting the stereoselectivity and specificity of antibodies for the production of tailor-made chiral stationary phases. We propose to develop and study immunoaffinity systems that combine selectivity with ease of use, short analysis times, and long-term stability, and are therefore suitable for routine enantiomer separation. Stereoselective antibodies, or fragments thereof, will be immobilized on conventional and high flow through type solid support materials and employed in pressure- and electrodriven separation techniques such as HPLC, micro-LC, CE, CEC, OTEC, and OTLC. The development of methods that allow enantiomer separation under mild, isocratic conditions favorable for protein stability and activity is of special interest. Chromatographic parameters will be optimized in order to adjust separation conditions to the strength of interaction between antibody and analyte.