Effect of CZE Buffer Components on Nanobody Separation

This technical note presents a study on the impact of capillary zone electrophoresis (CZE) buffer components on the separation of charge variants from Nanobody samples. The objective is to evaluate the SCIEX CZE kit for reducing testing variability and to assist users in implementing CZE methods with improved reproducibility. The document outlines the optimization of CZE buffers and buffer pH to achieve optimal separation, detailing the design of experiments (DoE) approach used in the study. It describes the preparation of Nanobody samples, the CZE separation method employed, and the analysis of results. The findings indicate that the concentration of TETA and the choice of polymer in the buffer significantly affect separation quality. The study concludes that while in-house buffer preparations can yield reproducible results, the SCIEX CZE Rapid Charge Variant Analysis kit provides better consistency and separation performance, making it a recommended starting point for Nanobody analysis.