Slip Flow Chromatography

描述(由申请人提供)：普渡大学和BioVidria，Inc.的这项STTR第二阶段合作的目标是将用于蛋白质层析的颠覆性新技术商业化，然后通过服务于制药行业增长最快的部分：蛋白质药物来最大限度地发挥其影响。该产品是一种反相色谱柱，使用直径仅为500纳米的二氧化硅颗粒，从而显著提高了蛋白质分离的分辨率和灵敏度。滑移流动使得如此小的颗粒能够获得高的流速。主要的市场机会是分析基于单抗的生物工程药物的异质性。这些药物具有很高的靶向性，例如，针对肿瘤的毒性。需要解决的问题是，单抗在储存过程中会发生分子内和分子间的二硫键争抢，从而导致患者具有免疫原性。拟议的滑流柱将独特地解析和分离这些扰乱的版本，使配方能够合理设计，以降低免疫原性。第一阶段的研究集中在填充毛细管中的蛋白质和多肽的分离，展示了十倍窄的区域和十倍于滑流的流动增强。第二阶段的研究将集中于扩大规模，开发直径2.1毫米的填充不锈钢柱。扩大规模将允许客户将这些色谱柱与当前的商业仪器一起使用，这对于服务于制药行业是必不可少的。第二阶段建议的具体目标是：1)优化将500 nm颗粒填充到不锈钢柱中的工艺；2)开发一种可扩展的工艺，以使键合相的硅醇活性可以忽略不计；以及3)开发用于所有三种类型抗体平台的单抗二硫键重排产物的分离方法。我们聘请了三家大公司提供样本：基因泰克(IgG1)、辉瑞(IgG2)和礼来公司(IgG4)。

DESCRIPTION (provided by applicant): The goal of this STTR Phase II collaboration between Purdue University and bioVidria, Inc. is to commercialize disruptive new technology for protein chromatography and then maximize its impact by serving the fastest growing segment of the pharmaceutical industry: protein drugs. The product is a reversed phase chromatography column that uses silica particles of only 500 nm in diameter, thereby significantly increasing resolution and sensitivity in protein separations. Slip flow is what enables high flow rates with such small particles. The primary market opportunity is the analysis of the heterogeneity of bioengineered drugs based on monoclonal antibodies. These drugs offer high target specificity, e.g., toxicity directed at the tumor. The problem to be solved is that monoclonal antibodies can undergo intra- and inter-molecular disulfide scrambling during storage, which causes immunogenicity in patients. The proposed slip-flow column will uniquely resolve and isolate these scrambled versions, enabling rational design of formulations to reduce immunogenicity. The Phase I research focused on protein and peptide separations in packed capillaries, demonstrating ten-fold narrower zones and a ten-fold flow enhancement from slip flow. The Phase II research will focus on scaling up to develop packed stainless steel columns of 2.1 mm in diameter. The scale-up will allow customers to use these columns with current commercial instruments, which is essential for serving the pharmaceutical industry. The Specific Aims of the Phase II proposal are to 1) optimize the process for packing 500 nm particles into stainless steel columns, 2) develop a scalable process to make bonded phases with negligible silanol activity, and 3) develop separation methods for resolving products of disulfide rearrangement of monoclonal antibodies for all three types of antibody platforms. We enlist three major companies to provide samples: Genentech (IgG1), Pfizer (IgG2) and Eli Lilly (IgG4).