HT-production of Abs using Interaction Trap in Yeast

描述(申请人提供)：Affomix希望将抗体芯片商业化，使研究人员能够研究人类蛋白质组。目前，研究人员使用基因芯片作为替代手段来估计细胞和组织中各种蛋白质的浓度。但如果可以选择，大多数研究人员倾向于直接研究蛋白质，因为：i)基因表达与蛋白质浓度之间的相关性较差，ii)使用基因芯片时缺乏任何关于翻译后修饰的信息。问题在于，研究细胞和组织中蛋白质水平和翻译后修饰的台架水平方法是不可用的。抗体芯片可以解决这个问题，但到目前为止，还没有一种廉价而简便的方法来分离针对人类蛋白质组中近25,000种蛋白质的抗体。 我们的战略是创造一种高通量(HT)分离高亲和力抗体的新方法。总体计划是开发和使用一种自动化形式的酵母双杂交(Y2H)系统，以提供一条商业上可行的分子生物学“流水线”，通过该流水线生产数千种不同的抗体。 Affomix有三个主要的商业目标：10向研究人员提供高质量的抗体芯片，2)提供针对特定目标的人类抗体识别的定制服务，3)开发我们自己的抗体疗法。成功完成这一第一阶段提案将使我们能够完成前两个目标。这一特定赠款申请的重要短期目标是建立一种可靠的方法来评估给定抗体的强度：抗原相互作用。成功完成四个州的具体目标将为今后的扩大努力提高效率和进行分析奠定基础。完成我们的第二阶段特定目标将使我们能够为研究人员提供重要的新试剂，用于研究细胞的蛋白质组组成和多样性，并促进识别、跟踪、量化、控制、捕获和成像生命系统中的单个蛋白质和分子机制。

DESCRIPTION (provided by applicant): Affomix would like to commercialize antibody chips to enable researchers to study the human proteome. Currently, researchers use gene chips as a surrogate means to estimate the concentration of the vast variety of proteins in cells and tissues. But given the choice, most researchers would prefer to study the proteins directly because of the; i) poor correlation between gene expression and protein concentration, and ii) lack of any information regarding post-translational modification when using gene chips. The problem has been that bench-level methods for studying protein levels and post-translational modification occurring in cells and tissues are not available. Antibody chips would address this problem but until now there has not been a cheap and facile method to isolate antibodies against the nearly 25,000 proteins in the human proteome. Our strategy is to create a new approach for the high-throughput (HT) isolation of high-affinity antibodies. The overall plan is to develop and use an automated format of the yeast two-hybrid (Y2H) system to provide a commercially viable molecular biology 'assembly line' by which to generate thousands of different antibodies. Affomix has three primary commercial objectives; 10 supply researchers with high-quality antibody chips, 2) perform custom services for the identification of human antibodies against specified targets, and, 3) develop our own antibody therapeutics. Successful completion of this Phase I proposal will allow us to complete these first 2 objectives. The significant short-term objective of this specific grant application is to establish a robust assay for estimating the strength of a given antibody:antigen interaction. Successful completion of the four state specific aims will set the state for the increased efficiencies and analysis in a future scaled-up effort. Completion of our Phase II specific aims will allow us to provide researchers with significant new reagents with which to study the proteomic makeup and diversity of cells, and facilitate efforts to identify, track, quantify, control, capture, and image both individual proteins and molecular mechanisms in living systems.