A generalized kinetic model for amine modification of proteins with application to phage display

蛋白质胺修饰的广义动力学模型及其在噬菌体展示中的应用

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作者:Xiaofang Jin, Jessica Rose Newton, Stephen Montgomery-Smith, George Smith

Abstract

Amine modification of filamentous virions (phage particles) is widely used in phage display technology to couple small groups such as biotin or fluorescent dyes to the major coat protein pVIII. We have developed a generalized kinetic model for protein amine modification and applied it to the modification of pVIII with biotin and the near-infrared fluorophor Alexa Fluor 680. Empirically optimized kinetic parameters for the two modification reactions allow the modification level to be predicted for a wide range of virions and modifying reagent concentrations. Virions with 0.03 biotins per pVIII subunit have 50% of the maximal binding capacity for a streptavidin conjugate.

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