Department of Chemistry, The College of William & Mary, Williamsburg, VA 23187, USA.
Department of Chemistry, The College of William & Mary, Williamsburg, VA 23187, USA.
Bioorg Chem. 2018 Feb;76:326-331. doi: 10.1016/j.bioorg.2017.11.020. Epub 2017 Dec 5.
The efficient preparation of protein bioconjugates represents a route to novel materials, diagnostics, and therapeutics. We previously reported a novel bioorthogonal Glaser-Hay reaction for the preparation of covalent linkages between proteins and a reaction partner; however, deleterious protein degradation was observed under extended reaction conditions. Herein, we describe the systematic optimization of the reaction to increase coupling efficiency and decrease protein degradation. Two optimized conditions were identified varying either the pH of the reaction or the bidentate ligand employed, allowing for more rapid conjugations and/or less protein oxidation.
高效制备蛋白质生物缀合物是一种制备新型材料、诊断试剂和治疗剂的途径。我们之前报道了一种新的生物正交 Glaser-Hay 反应,用于制备蛋白质和反应伙伴之间的共价键;然而,在延长的反应条件下观察到蛋白质降解。本文描述了对该反应的系统优化,以提高偶联效率并减少蛋白质降解。通过改变反应的 pH 值或使用双齿配体,确定了两种优化条件,使偶联反应更快或蛋白质氧化程度更低。