Protein Stability and Functionality Enhancement Method

描述（由申请人提供）：本研究的长期目标是开发定向进化平台技术，称为插入，删除，替换随机突变（ids.PCR），用于商业应用于诊断。我们建议开发诊断报告酶，用于诊断和治疗监测，需要对复杂混合物中存在的靶蛋白进行敏感，特异性，定量和多路检测。我们将生成β -内酰胺酶变体，其中包含插入的氨基酸序列，与特定疾病相关的靶蛋白具有结合亲和力。这些功能增强的“诊断报告酶”将结合β -内酰胺酶的高效催化活性和特异性抗体样结合亲和力，从而快速、特异性、选择性和敏感地检测与疾病相关的生物标志物。在这项II期研究中，将使用两个商业上相关的蛋白质靶点来筛选和表征进化的酶，以对单独的靶点和复杂的患者样本做出反应。

DESCRIPTION (provided by applicant): The long range goal of this research is to develop the directed evolution platform technology termed Insertion, Deletion, Substitution random mutagenesis (ids.PCR) for commercial applications in diagnostics. We propose to develop diagnostic reporter enzymes for diagnosis and therapeutic monitoring that require sensitive, specific, quantitative and multiplexed detection of target proteins present in complex mixtures. We will generate beta-lactamase variants that contain inserted amino acid sequences with binding affinity for specific disease-associated target proteins. These functionally enhanced "Diagnostic Reporter Enzymes" will combine the highly efficient catalytic activity of beta-lactamase with specific antibody-like binding affinities, resulting in rapid, specific, selective, and sensitive detection of biological markers associated with disease. For this Phase II research, two commercially relevant protein targets will be used to screen and characterize the evolved enzymes for response to the target alone and in complex patient samples. PROPOSED COMMERCIAL APPLICATIONS: Ids.PCR is a platform evolution technology that has far-reaching potential commercial applications. The demonstration of diagnostic reporter enzymes evolved using this technology would have widespread application in diagnostics and therapeutic monitoring. The goal is to ultimately use these low-cost single molecule reagents to screen large numbers of biological markers for analysis of human diseases in complex patient samples.