Department of Analytical Chemistry, University of Pannonia, Egyetem u. 10., H-8200 Veszprém, Hungary.
Institute of Pharmaceutical Sciences of Western Switzerland, University of Geneva, CMU-Rue Michel Servet 1, 1211, Geneva 4, Switzerland.
J Pharm Biomed Anal. 2020 Jun 5;185:113207. doi: 10.1016/j.jpba.2020.113207. Epub 2020 Feb 25.
The hyphenation of non-denaturing liquid chromatographic (LC) modes (ion exchange (IEX), size exclusion (SEC) and hydrophobic interaction chromatography (HIC)) with mass spectrometry (MS) has attracted significant attention in the last few years. The inherent problem of these couplings is that non-denaturing LC separations have tended to use non-volatile mobile phase additives. Indeed, classical methods have not been directly compatible with MS. Therefore two approaches can be used to address this challenge: (1) finding innovative volatile mobile phases or (2) adding a desalting step prior to MS detection via the use of multidimensional LC. These two possibilities have been applied to the characterization of charge-, size- and hydrophobic variants of various monoclonal antibodies (mAbs) and related products and have been reviewed in this paper.
近年来，非变性液相色谱（LC）模式（离子交换（IEX）、尺寸排阻（SEC）和疏水相互作用色谱（HIC））与质谱（MS）的键合引起了广泛关注。这些偶联物存在一个固有问题，即非变性 LC 分离往往使用非挥发性流动相添加剂。实际上，传统方法与 MS 并不直接兼容。因此，可以采用两种方法来解决这一挑战：（1）寻找创新的挥发性流动相；或（2）在通过多维 LC 进行 MS 检测之前添加脱盐步骤。这两种可能性已应用于各种单克隆抗体（mAb）和相关产品的电荷、大小和疏水性变体的表征，并在本文中进行了综述。
