Process Monitoring of Protein Microheterogeneity by Capillary Immunophoresis

9525821 Lee该提案是关于开发毛细血管免疫电泳作为一种分析技术的研究，以评估药物蛋白质的微观异质性，包括翻译后修饰和降解产物的变化。 该技术包括耦合毛细管电泳系统与免疫试剂反应毛细管后系统，其中包括激光诱导荧光检测。 具体而言，建议研究：（1）在毛细管区带电泳和毛细管等电聚焦中蛋白质变体的分离条件和分离分辨率之间的关系，（2）所施加的电场强度，毛细管尺寸，和溶液离子强度对蛋白质转移和蛋白质变体与抗体在毛细管后免疫反应器中混合的有效性的影响，以及（3）通过测量抗原-抗体复合物周围的诱导静电势和pH变化来直接感测抗原（蛋白质变体）-抗体反应。 ***

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. ***