Biologics Development, Bristol-Myers Squibb Company , 38 Jackson Road, Devens, Massachusetts 01434, United States.
Biophysical Characterization Group, Bristol-Myers Squibb Company , 311 Pennington Rocky Hill Road, Pennington, New Jersey 08534, United States.
Anal Chem. 2018 Feb 20;90(4):2542-2547. doi: 10.1021/acs.analchem.7b03831. Epub 2018 Feb 5.
Capillary gel electrophoresis using sodium dodecyl sulfate (CE-SDS) is used commercially to provide quantitative purity data for therapeutic protein characterization and release. In CE-SDS, proteins are denatured under reducing or nonreducing conditions in the presence of SDS and electrophoretically separated by molecular weight and hydrodynamic radius through a sieving polymer matrix. Acceptable performance of this method would yield protein peaks that are baseline resolved and symmetrical. Nominal CE-SDS conditions and parameters are not optimal for all therapeutic proteins, specifically for Recombinant Therapeutic Protein-1 (RTP-1), where acceptable resolution and peak symmetry were not achieved. The application of longer alkyl chain detergents in the running buffer matrix substantially improved assay performance. Matrix running buffer containing sodium hexadecyl sulfate (SHS) increased peak resolution and plate count 3- and 8-fold, respectively, compared to a traditional SDS-based running gel matrix. At Bristol-Myers Squibb (BMS), we developed and qualified a viable method for the characterization and release of RTP-1 using an SHS-containing running buffer matrix. This work underscores the potential of detergents other than SDS to enhance the resolution and separation power of CE-based separation methods.
十二烷基硫酸钠胶束毛细管电泳（CE-SDS）被广泛应用于商业领域，用于提供治疗性蛋白的定量纯度数据，从而实现其特征和放行检测。在 CE-SDS 中，蛋白在还原或非还原条件下于 SDS 存在的情况下变性，通过筛分聚合物基质，按分子量和流体力学半径大小进行电泳分离。该方法的良好性能可得到基线分辨且对称的蛋白峰。并非所有治疗性蛋白的 CE-SDS 条件和参数都是最佳的，具体来说，对于重组治疗蛋白 1（RTP-1），其无法达到可接受的分辨率和峰对称性。在运行缓冲基质中应用长链烷基清洁剂可极大地改善分析性能。与传统基于 SDS 的运行凝胶基质相比，含有十六烷基磺酸钠（SHS）的基质运行缓冲液可分别将峰分辨率和板计数提高 3 倍和 8 倍。在 Bristol-Myers Squibb（BMS），我们开发并验证了一种基于 SHS 运行缓冲基质的 RTP-1 特性和放行检测的可行方法。这项工作强调了除 SDS 以外的清洁剂具有增强基于 CE 的分离方法的分辨率和分离能力的潜力。
