Lomonosov Institute of Fine Chemical Technologies, MIREA-Russian Technological University, Vernadskogo Pr. 86, 119571 Moscow, Russia.
Federal State Autonomous Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of the Ministry of Health of the Russian Federation, 8-2 Trubetskaya Str., 119991 Moscow, Russia.
Biomolecules. 2024 Jul 14;14(7):849. doi: 10.3390/biom14070849.
Affinity chromatography is a widely used technique for antibody isolation. This article presents the successful synthesis of a novel affinity resin with a mutant form of protein A (BsrtA) immobilized on it as a ligand. The key aspect of the described process is the biocatalytic immobilization of the ligand onto the matrix using the sortase A enzyme. Moreover, we used a matrix with primary amino groups without modification, which greatly simplifies the synthesis process. The resulting resin shows a high dynamic binding capacity (up to 50 mg IgG per 1 mL of sorbent). It also demonstrates high tolerance to 0.1 M NaOH treatment and maintains its effectiveness even after 100 binding, elution, and sanitization cycles.
亲和层析是一种广泛应用于抗体分离的技术。本文介绍了一种新型亲和树脂的成功合成，该树脂将突变体形式的蛋白 A（BsrtA）固定在其上作为配体。所描述过程的关键方面是使用 sortase A 酶将配体生物催化固定在基质上。此外，我们使用了未经修饰的带有伯氨基的基质，这大大简化了合成过程。所得树脂具有高动态结合容量（每 1 毫升吸附剂可达 50 毫克 IgG）。它还表现出对 0.1 M NaOH 处理的高耐受性，并且即使在 100 次结合、洗脱和消毒循环后仍能保持其有效性。
