GOALI: Understanding the effect of sequence on expression of recombinant monoclonal antibodies

0854099Sharfstein拟议项目的智力价值是通过以下方式对基因和蛋白质序列与重组单克隆抗体表达水平之间的关系形成基本理解：* 建立重组治疗性单克隆抗体的序列变体文库 * 在中国卵巢（CHO）细胞中表达这些变体 * 分析mRNA水平和细胞内和细胞外抗体水平，并将表达与基因和蛋白质序列相关联 * 将本研究的结果与UCB Celltech（斯劳，英国）先前对其他抗体进行的研究进行比较，以确定是否始终受蛋白质稳定性控制或其他机制是否也起作用。 拟议工作的更广泛影响是：* 为化学工程的女博士生提供独特的教育经验，结合工业实习和国际经验，可以作为国内研究生国际经验的典范 * 为美国的生物技术和生物制药行业提供关键知识 * 发展伦斯勒理工学院和UCB之间的持续关系，这将在邀请IGERT提出的分子生物加工建议中发挥重要作用

0854099SharfsteinThe intellectual merit of the proposed project is to develop a fundamental understanding of the relationships between gene and protein sequence and expression level of recombinant monoclonal antibodies by:* Creating a library of sequence variants for a recombinant therapeutic monoclonal antibody* Expressing these variants in Chinese Hamster Ovary (CHO) cells* Analyzing the mRNA levels and intra- and extra-cellular antibody levels and correlating the expression with gene and protein sequence* Comparing the results of this study with previous studies of other antibodies performed by UCB Celltech (Slough, UK) to determine whether always controlled by protein stability or if other mechanisms also play a role. The broader impacts of the proposed work are to: * Provide a unique educational experience for a female doctoral student in Chemical Engineering, combining an industrial internship and an international experience that can serve as a model for international experiences for domestic graduate students* Provide critical knowledge to the biotechnology and biopharmaceutical industry in the United States* Develop an ongoing relationship between Rensselaer Polytechnic Institute and UCB that will play an important role in an invited IGERT proposal in molecular bioprocessing