Direct Enantiomer Separation in Immunoaffinity Systems

免疫亲和系统中的直接对映体分离

• 批准号: 6556266
• 负责人: OLIVER HOFSTETTER
• 金额: $ 14.29万
• 依托单位: NORTHERN ILLINOIS UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-02-01 至 2006-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6556266
• 关键词: SDS polyacrylamide gel electrophoresis; aminoacid; capillary electrophoresis; chemical stability; high performance liquid chromatography; immobilized enzymes; immunoaffinity chromatography; stereochemistry; stereoisomer; technology /technique development

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.

描述（由申请人提供）：生物体区分手性化合物的对映体，例如，药物，在几乎所有的相互作用水平，并作出不同的反应。虽然一种对映异构体可能表现出所需的活性，但另一种可能导致严重的药理学和毒理学副作用，或作为拮抗剂。然而，500种最畅销的药物中有一半以上和目前正在开发的药物中约三分之二是手性药物。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，而且要求进行立体化学特异性鉴定试验，以及对单个对映体的药理学和药代动力学表征。因此，开发用于精确测定生物活性手性分子及其纯化的精细分析技术和选择剂具有很大的意义。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差异相互作用。虽然目前约有200种手性选择剂可商购获得，但这些选择剂通常不是针对特定分离问题定制的，并且合适选择剂的鉴定仍然是繁琐的试错练习。早已知道抗体可以区分手性化合物的对映体。由于抗体可以进一步针对几乎任何感兴趣的化合物，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免疫亲和系统，结合联合收割机的选择性，易于使用，分析时间短，长期稳定性，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常规和高流通型固体支持材料上，并用于压力驱动和电驱动分离技术，如HPLC、微LC、CE、CEC、OTEC和OTLC。开发的方法，允许对映体分离在温和，等度条件下有利于蛋白质的稳定性和活性是特别感兴趣的。将优化色谱参数，以根据抗体和分析物之间的相互作用强度调整分离条件。

项目成果

Stereoselective antibodies to free alpha-hydroxy acids.

针对游离 α-羟基酸的立体选择性抗体。

• DOI: 10.1002/jmr.741
• 发表时间: 2005
• 期刊: Journal of molecular recognition : JMR.
• 作者: Zeleke,TigabuK;Zeleke,JessicaM;Hofstetter,Heike;Hofstetter,Oliver
• 通讯作者: Hofstetter,Oliver

Effect of the mobile phase on antibody-based enantiomer separations of amino acids in high-performance liquid chromatography.

流动相对高效液相色谱中基于抗体的氨基酸对映体分离的影响。

• 发表时间: 2004
• 期刊: Journal of chromatography. A
• 作者: Hofstetter,Oliver;Lindstrom,Heather;Hofstetter,Heike
• 通讯作者: Hofstetter,Heike

New developments in the production and use of stereoselective antibodies.

• DOI: 10.1002/chir.20099
• 发表时间: 2005
• 期刊: Chirality
• 影响因子: 2
• 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

Enantiomer separation of alpha-hydroxy acids in high-performance immunoaffinity chromatography.

高效免疫亲和色谱法中 α-羟基酸的对映体分离。

• DOI: 10.1016/j.jpba.2007.10.004
• 发表时间: 2008
• 期刊: Journal of pharmaceutical and biomedical analysis
• 影响因子: 3.4
• 作者: Franco,ElliottJ;Hofstetter,Heike;Hofstetter,Oliver
• 通讯作者: Hofstetter,Oliver

Enantiomer separation of amino acids in immunoaffinity micro LC-MS.

在免疫亲和微型 LC-MS 中分离氨基酸的对映体。

• DOI: 10.1002/chir.20286
• 发表时间: 2006
• 期刊: Chirality
• 影响因子: 2
• 作者: Zeleke,JessicaM;Smith,GregoryB;Hofstetter,Heike;Hofstetter,Oliver
• 通讯作者: Hofstetter,Oliver