Kaupbayeva Bibifatima, Murata Hironobu, Matyjaszewski Krzysztof, Russell Alan J, Boye Susanne, Lederer Albena
Department of Biological Sciences, Carnegie Mellon University 4400 Fifth Avenue Pittsburgh PA 15213 USA.
Center for Polymer-Based Protein Engineering, Carnegie Mellon University 5000 Forbes Avenue Pittsburgh PA 15213 USA.
Chem Sci. 2021 Sep 27;12(41):13848-13856. doi: 10.1039/d1sc03033g. eCollection 2021 Oct 27.
Polymer-based protein engineering has enabled the synthesis of a variety of protein-polymer conjugates that are widely applicable in therapeutic, diagnostic and biotechnological industries. Accurate characterizations of physical-chemical properties, in particular, molar masses, sizes, composition and their dispersities are critical parameters that determine the functionality and conformation of protein-polymer conjugates and are important for creating reproducible manufacturing processes. Most of the current characterization techniques suffer from fundamental limitations and do not provide an accurate understanding of a sample's true nature. In this paper, we demonstrate the advantage of asymmetrical flow field-flow fractionation (AF4) coupled with multiple detectors for the characterization of a library of complex, zwitterionic and neutral protein-polymer conjugates. This method allows for determination of intrinsic physical properties of protein-polymer chimeras from a single, rapid measurement.
基于聚合物的蛋白质工程技术已能够合成多种蛋白质-聚合物共轭物，这些共轭物在治疗、诊断和生物技术行业中具有广泛的应用。准确表征其物理化学性质，特别是摩尔质量、尺寸、组成及其分散性，是决定蛋白质-聚合物共轭物功能和构象的关键参数，对于建立可重复的制造工艺也很重要。目前大多数表征技术都存在根本局限性，无法准确了解样品的真实性质。在本文中，我们展示了不对称流场-流分级分离法（AF4）与多个检测器联用，用于表征复杂的两性离子和中性蛋白质-聚合物共轭物文库的优势。该方法能够通过单次快速测量确定蛋白质-聚合物嵌合体的固有物理性质。
