Protein Separations Using Affinity Ultrafiltration with Small Charged Ligands

使用小带电配体的亲和超滤分离蛋白质

基本信息

批准号：
0345528
负责人：
Andrew Zydney
金额：
$ 25万
依托单位：
Pennsylvania State Univ University Park
依托单位国家：
美国
项目类别：
Standard Grant
财政年份：
2004
资助国家：
美国
起止时间：
2004-03-01 至 2008-02-29
项目状态：
已结题

来源：
https://www.nsf.gov/awardsearch/showAward?AWD_ID=0345528&HistoricalAwards=false
关键词：
Protein Separations Using Affinity Ultrafiltration

项目摘要

AbstractProposal Title: Protein Adsorption and Transport in Chromatographic BioprocessingProposal Number: CTS-0345528Principal Investigator: Andrew ZydneyInstitution: Pennsylvania State UniversityThe objective of this project is to examine a novel approach for protein purification using small, charged, affinity ligands to achieve high-resolution separations by affinity ultrafiltration. Affinity ultrafiltration exploits the high selectivity that can be obtained by molecular recognition, but the product and ligand are allowed to interact in free solution, thereby eliminating the mass transfer limitations and concerns about ligand accessibility that often hinder applications of packed column affinity chromatography. The binding interaction causes a change in electrical charge of the protein-ligand complex, and this is used to obtain high product retention by exploiting electrostatic interactions during ultrafiltration. Initial experimental studies will be performed using Cibacron Blue as the charged affinity ligand and human serum albumin as the model protein. Preliminary theoretical calculations indicate that the change in protein charge will provide high selectivity for membrane separations between albumin and protein impurities that are unable to bind Cibacron Blue. These results will demonstrate the feasibility of this technique, while providing a quantitative framework for design and optimization of affinity ultrafiltration processes using small charged affinity ligands. In terms of the broader impacts, this project will provide graduate and undergraduate students with hands-on experience dealing with membrane processes and protein separations. The results of this work may aid in the development of new technologies that provide high-resolution protein purification at a price, scale, and quality appropriate for high-value products.

摘要提案标题：色谱生物加工中的蛋白质吸附和运输提案编号：CTS-0345528主要研究员：Andrew Zydney机构：宾夕法尼亚州立大学该项目的目的是研究一种使用小型带电亲和配体进行蛋白质纯化的新方法，通过以下方法实现高分辨率分离：亲和超滤。亲和超滤利用了通过分子识别可以获得的高选择性，但产物和配体可以在自由溶液中相互作用，从而消除了传质限制和对配体可及性的担忧，而这些限制常常阻碍填充柱亲和色谱的应用。结合相互作用引起蛋白质-配体复合物的电荷变化，这用于通过在超滤过程中利用静电相互作用来获得高产物保留。初步实验研究将使用 Cibacron Blue 作为带电亲和配体和人血清白蛋白作为模型蛋白进行。初步理论计算表明，蛋白质电荷的变化将为白蛋白和无法结合 Cibacron Blue 的蛋白质杂质之间的膜分离提供高选择性。这些结果将证明该技术的可行性，同时为使用小带电亲和配体的亲和超滤过程的设计和优化提供定量框架。就更广泛的影响而言，该项目将为研究生和本科生提供处理膜过程和蛋白质分离的实践经验。这项工作的结果可能有助于开发新技术，以适合高价值产品的价格、规模和质量提供高分辨率的蛋白质纯化。