SBIR Phase II: Method for Isolating Catalytic Antibodies

* 9710660 One Cell Systems，Inc Trnovsky 这个小型企业创新研究第二阶段项目提出通过将细胞封装在凝胶微滴（GMD）中，然后使用流式细胞术对其进行分析和分选，来分离具有催化特性的可卡因特异性抗体。 I期研究证明了使用GMD技术筛选细胞的催化活性的可行性，使用四种不同的模型，每种模型代表催化抗体表达的不同位置。 将荧光产物保留在琼脂糖微球内，并回收感兴趣的细胞用于使用荧光激活细胞分选进行克隆或基因分析。 在限定的微环境中采用该测定允许基于产物形成和/或结合来鉴定和回收单细胞。 由于可卡因特异性抗体预计是罕见的和弱催化，微胶囊技术是特别适合这些调查。 这些研究的成功完成将导致开发一种快速、自动化的方法，用于分离、分析和回收产生感兴趣抗体的细胞。 催化抗体代表了几乎取之不尽的生物催化剂来源。 产生特定蛋白酶的能力将对用作治疗剂以及工业酶的生物技术和医学产生重大影响。 全球催化抗体市场总额估计超过10亿美元。 ***

*** 9710660 One Cell Systems, Inc Trnovsky This Small Business Innovation Research Phase II project proposes to isolate cocaine specific antibodies with catalytic properties by encapsulating cells in gel microdrops (GMD) and subsequently analyzing and sorting them using flow cytometry. Phase I research demonstrated the feasibility of using GMD technology to screen cells for catalytic activity using four different models each representing a distinct location for catalytic antibody expression. Fluorescent products are retained within the agarose microsphere and cells of interest recovered for cloning or gene analysis using fluorescent activated cell sorting. Employing this assay in defined microenvironments allows the identification and recovery of single cells based on product formation and/or binding. Since cocaine specific antibodies are expected to be rare and weakly catalytic, the microencapsulation technology is particularly suited for these investigations. Successful completion of these studies will lead to development of a rapid, automated method for isolating, analyzing and recovering cells producing antibodies of interest. Catalytic antibodies represent a virtually inexhaustible source of biocatalysts. The ability to generate specific proteases would have a significant impact on biotechnology and medicine for use as therapeutics as well as industrial enzymes. The total worldwide market for catalytic antibodies is estimated to be in excess of $1.0 billion. ***