Vimer Shay, Ben-Nissan Gili, Marty Michael, Fleishman Sarel J, Sharon Michal
Department of Biomolecular Sciences, Weizmann Institute of Science, Rehovot, Israel.
Department of Chemistry & Biochemistry, University of Arizona, Tucson, Arizona, USA.
Proteomics. 2021 Nov;21(21-22):e2000300. doi: 10.1002/pmic.202000300. Epub 2021 Aug 9.
In recent decades, antibodies (Abs) have attracted the attention of academia and the biopharmaceutical industry due to their therapeutic properties and versatility in binding a vast spectrum of antigens. Different engineering strategies have been developed for optimizing Ab specificity, efficacy, affinity, stability and production, enabling systematic screening and analysis procedures for selecting lead candidates. This quality assessment is critical but usually demands time-consuming and labor-intensive purification procedures. Here, we harnessed the direct-mass spectrometry (direct-MS) approach, in which the analysis is carried out directly from the crude growth media, for the rapid, structural characterization of designed Abs. We demonstrate that properties such as stability, specificity and interactions with antigens can be defined, without the need for prior purification.
近几十年来,抗体(Abs)因其治疗特性以及在结合广泛抗原方面的多功能性而受到学术界和生物制药行业的关注。为了优化抗体的特异性、功效、亲和力、稳定性和产量,人们开发了不同的工程策略,从而实现了用于筛选先导候选物的系统筛选和分析程序。这种质量评估至关重要,但通常需要耗时且费力的纯化程序。在这里,我们采用了直接质谱(direct-MS)方法,即直接从粗生长培养基中进行分析,以对设计的抗体进行快速的结构表征。我们证明,无需事先纯化即可定义诸如稳定性、特异性以及与抗原的相互作用等特性。