Low Molecular Weight Displacers for Protein Purification

用于蛋白质纯化的低分子量置换剂

• 批准号: 6894594
• 负责人: STEVEN Michael CRAMER
• 金额: $ 51.61万
• 依托单位: RENSSELAER POLYTECHNIC INSTITUTE
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-01 至 2007-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6894594
• 关键词: bioengineering /biomedical engineering; chromatography; electrophoresis; enzyme linked immunosorbent assay; high performance liquid chromatography; high throughput technology; ion exchange chromatography; molecular sieving; molecular weight; oligonucleotides; physical separation; polymers; protein purification; technology /technique development

DESCRIPTION (provided by applicant): In the second grant we demonstrated that novel low molecular weight (MW) displacers can be successfully employed for high resolution/high-throughput protein purifications in ion exchange systems. We demonstrated that low molecular weight displacers can also be used for oligonucleotide purification and that the stationary phase has a significant effect on the efficacy of various low molecular weight displacers. Preliminary results indicate that high-throughput screening can be successfully employed to evaluate a wide variety of displacer chemistries. Further results from our laboratory indicate that this data can be employed in concert with novel quantitative structure efficacy relationship models to both predict and interpret displacer efficacy. Finally, preliminary results in our laboratory indicate that chemically selective displacers can be identified which can dramatically increase the selectivity of ion exchange systems. In the next phase of this project we will incorporate these new lines of research into our ongoing investigation into the rational design of displacers for the purification of biomolecules. We will incorporate the tools of high-throughput screening and synthesis, and quantitative structure activity relationship models into the research and we will address several critical issues related to the design of both high affinity and chemically selective low MW displacers for proteins and oligonucleotides in ion exchange systems. The next phase of the project will include high throughput screening, quantitative structure activity relationship models, displacer synthesis, chemoenzymatic synthesis of displacer libraries, the development of high affinity displacers for cation and anion exchange systems, development of selective displacers, and the application of these displacers for purification of biomolecules from complex mixtures. The synthetic work will include structural variants of low MW displacers identified in the second grant as well as variants of other promising candidates recently identified in our laboratory. The displacers developed in this research will be employed in a detailed investigation of the interplay between displacer chemistry and its ability to act as a high affinity or selective displacers for various classes of separation problems. This research will evaluate several hypotheses related to displacer design. The low MW displacers developed in this research will be employed for the purification of proteins and oligonucleotides from complex biological mixtures. Finally, these high affinity displacers will be employed to develop a generic displacement technique for purification of a range of proteins with possible applications to proteomics. The proposed research will provide significant insight into the design of efficient low MW displacers for protein and oligonucleotide purification and will enable more efficient purification of a wide range of biopharmaceuticals. Furthermore, this work will facilitate the use of this high resolution/high throughput technique by medical researchers at the bench scale for the rapid purification of novel biomolecules from dilute complex mixtures.

描述（由申请人提供）：在第二项资助中，我们证明了新型低分子量（MW）置换剂可以成功地用于离子交换系统中的高分辨率/高通量蛋白质纯化。我们证明低分子量置换剂也可用于寡核苷酸纯化，并且固定相对各种低分子量置换剂的功效具有显着影响。初步结果表明，高通量筛选可成功用于评估各种置换剂化学成分。我们实验室的进一步结果表明，这些数据可以与新颖的定量结构功效关系模型结合使用，以预测和解释置换剂功效。最后，我们实验室的初步结果表明可以识别化学选择性置换剂，这可以显着提高离子交换系统的选择性。在该项目的下一阶段，我们将把这些新的研究领域纳入我们正在进行的对用于纯化生物分子的置换器的合理设计的调查中。我们将把高通量筛选和合成工具以及定量结构活性关系模型纳入研究中，我们将解决与离子交换中蛋白质和寡核苷酸的高亲和力和化学选择性低分子量置换剂设计相关的几个关键问题系统。项目下一阶段将包括高通量筛选、定量结构活性关系模型、置换剂合成、置换剂库的化学酶合成、用于阳离子和阴离子交换系统的高亲和力置换剂的开发、选择性置换剂的开发以及这些置换剂的应用。用于从复杂混合物中纯化生物分子的置换剂。合成工作将包括第二笔资助中确定的低分子量置换剂的结构变体以及我们实验室最近确定的其他有希望的候选物的变体。本研究中开发的置换剂将用于详细研究置换剂化学性质及其作为高亲和力或选择性置换剂解决各类分离问题的能力之间的相互作用。这项研究将评估与置换器设计相关的几个假设。本研究开发的低分子量置换剂将用于从复杂的生物混合物中纯化蛋白质和寡核苷酸。最后，这些高亲和力置换剂将用于开发通用置换技术，用于纯化一系列蛋白质，并可能应用于蛋白质组学。拟议的研究将为蛋白质和寡核苷酸纯化的高效低分子量置换剂的设计提供重要的见解，并将使各种生物制药的更有效纯化成为可能。此外，这项工作将促进医学研究人员在实验室规模上使用这种高分辨率/高通量技术，从稀释的复杂混合物中快速纯化新型生物分子。