Process Monitoring of Protein Microheterogeneity by Capillary Immunophoresis

通过毛细管免疫电泳对蛋白质微观异质性进行过程监测

• 批准号: 9525821
• 负责人: Cheng Lee
• 金额: $ 14万
• 依托单位: Iowa State University
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 1996
• 资助国家: 美国
• 起止时间: 1996-03-15 至 1999-02-28
• 项目状态: 已结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=9525821&HistoricalAwards=false
• 关键词: Process; Monitoring; Protein; Microheterogeneity; Capillary

9525821 Lee This proposal is on research to develop capillary immunophoresis as an analytical technique to assess microheterogeneity in pharmaceutical proteins, including variations in post- translational modifications and degradation products. The technique consists of coupling a capillary electrophoresis system with an immunoreagent reaction capillary post-system which incorporates laser-induced fluorescence detection. Specifically it is proposed to study: (1) the relationship between separation conditions and separation resolution of protein variants in capillary zone electrophoresis and capillary isoelectric focusing, (2) the quantitative dependence of applied electric field strengths, capillary dimensions, and solution ionic strengths on the effectiveness of protein transfer and mixing of protein variants with antibodies in a post-capillary immunoreactor, and (3) the direct sensing of the antigen (protein variants)-antibody reaction by measuring the induced electrostatic potentials and pH changes around the antigen- antibody complexes. ***

这个建议是研究发展毛细管免疫电泳作为一种分析技术来评估药物蛋白的微观异质性，包括翻译后修饰和降解产物的变化。该技术由毛细管电泳系统与免疫试剂反应毛细管后系统耦合组成，毛细管后系统包含激光诱导荧光检测。具体建议研究：(1)毛细管区带电泳和毛细管等电聚焦分离条件与蛋白质变异分离分辨率之间的关系；(2)外加电场强度、毛细管尺寸和溶液离子强度对蛋白质转移和蛋白质变异与抗体在毛细管后免疫反应器中混合的有效性的定量依赖；(3)通过测量抗原-抗体复合物周围的诱导静电电位和pH变化，直接感知抗原（蛋白质变体）-抗体反应。＊＊＊