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Universal affinity membrane chromatography for rapid, one-step purification of proteins

用于快速、一步纯化蛋白质的通用亲和膜层析

基本信息

批准号：
10250634
负责人：
Scott M Husson
金额：
$ 25.66万
依托单位：
TRIALTUS BIOSCIENCE, LLC
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-09-15 至 2023-05-14
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10250634
关键词：
Address Affinity Alabama Benchmarking Binding Binding Proteins Biological Products Biological Sciences COVID-19 CRISPR/Cas technology Cells Chimeric Proteins Chromatography Clustered Regularly Interspaced Short Palindromic Repeats Column Chromatography Complex Cystic Fibrosis Data Development Diagnostic Reagent Kits Disease Drops Engineering Enzymes Escherichia coli Genes Goals Guide RNA Health Heart Diseases Human Immobilization Industry Ligands Malignant Neoplasms Measures Membrane Mendelian disorder Pathology Performance Pharmaceutical Preparations Phase Plant Resins Play Polymers Process Production Productivity Proteins Protocols documentation Public Health Race Recovery Research Ribonucleoproteins Role SARS-CoV-2 spike protein Sales Scientist Sickle Cell Anemia Small Business Technology Transfer Research Speed Structure System Technology Testing Time Universities Validation Variant Work antibody test base commercial application commercialization cost effective density experimental study gene therapy genome editing improved innovation interest new technology phase 1 study prevent prospective protein E protein purification prototype public health relevance research and development residence screening vaccine evaluation

项目摘要

Project Summary The goal of this STTR Phase I project is to demonstrate the feasibility of developing the first universal affinity membrane for the rapid, selective, one-step purification of difficult-to-purify proteins. Despite the urgency for expanding the application of gene editing systems and accelerating vaccine and antibody test development, the rapid purification of critical proteins such as CRISPR-Cas9 and the SARS-COV2 spike protein with high yield, high purity, and high activity remains a difficult challenge. The proposed innovation will benefit public health and have a significant impact on the biopharmaceutical industry by addressing this challenge. The products of this innovation will be first-in-market universal affinity membrane chromatography columns based on TriAltus’ ultrahigh affinity CL7/Im7 purification system. Research will be done in partnership with Clemson University and the University of Alabama at Birmingham. Purification of CRISPR-Cas9 protein from E. coli lysate will be used as a demonstration case for evaluating the performance of the affinity membrane innovation. The Specific Aims of this Phase I study are to (1) Synthesize and characterize Im7-activated affinity membranes and (2) Test prototype affinity membrane columns for capture purification of CRISPR-Cas9. In Specific Aim 1, we will establish the feasibility of synthesizing the affinity membranes by immobilizing four Im7 ligand variants at high density on polymer-grafted macroporous membranes. We will measure protein binding capacity, recovery, and column reusability in screening experiments using purified, CL7-tagged CRISPR-Cas9. In Specific Aim 2, we will quantify and benchmark performance for capture step purification of CRISPR-Cas9 from E. Coli lysate. We will measure Cas9 binding capacity, recovery, purity, and activity. In Phase II, the team plans to comprehensively evaluate the roles that synthesis conditions and membrane support structure play on binding capacity; delineate operating ranges that maximize productivity, recovery, purity, and activity; collaborate with a membrane technology company on research to establish a scalable process to produce membrane columns for external validation; and conduct field research with external partners and prospective customers. Commercialization will bring the first universal affinity membrane chromatography column to the market. Market entry for the new column products will be to research and development scientists. The columns will provide a universal platform for the rapid, cost-effective purification of tag-free proteins with high yield, purity, and activity. By simplifying purification protocols and reducing purification times, products derived from this membrane innovation will have a major impact in the race to develop new protein drugs, gene therapies, vaccines, and antibody test kits.

项目总结