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光照辅助干燥法用于稳定蛋白质。

Light-assisted drying for protein stabilization.

机构信息

The Univ. of North Carolina at Charlotte, United States.

出版信息

J Biomed Opt. 2018 Jul;23(7):1-8. doi: 10.1117/1.JBO.23.7.075007.

Abstract

A light-based processing method to create an amorphous trehalose matrix for the stabilization of proteins is discussed. This method has potential applications in the stabilization of protein-based therapeutics and diagnostics. During light-assisted drying (LAD), proteins suspended in a trehalose solution are dehydrated using near-infrared (NIR) laser light. The goal of this study was to determine processing parameters that resulted in fast processing times and low end moisture contents (EMC), while maintaining the functionality of embedded proteins. We compared the effect of changing processing wavelength, power and resulting sample temperature, and substrate material on the EMC for two NIR laser sources (1064 and 1850 nm). The 1850-nm laser resulted in the lowest EMC (0.03  ±  0.01  gH2O  /  gDryWeight) after 20 min of processing on glass microfiber paper. This suggests a storage temperature of 68.3°C. We also tested the functionality of a model protein, lysozyme, after LAD processing using a standard assay. LAD showed no significant effect on the functionality of lysozyme when processed at a maximum temperature of ∼44  °  C to an EMC of 0.17  ±  0.06  gH2O  /  gDryWeight. LAD is a promising technique for forming amorphous trehalose solids that could stabilize proteins at ambient temperatures.

摘要

一种基于光的处理方法,用于创建用于稳定蛋白质的无定形海藻糖基质,用于稳定蛋白质的无定形海藻糖基质。这种方法在蛋白质基治疗剂和诊断剂的稳定化方面具有潜在的应用。在光辅助干燥(LAD)期间,悬浮在海藻糖溶液中的蛋白质使用近红外(NIR)激光进行脱水。本研究的目的是确定导致快速处理时间和低最终水分含量(EMC)的处理参数,同时保持嵌入式蛋白质的功能。我们比较了改变处理波长、功率和样品温度以及基底材料对两种近红外激光源(1064nm 和 1850nm)的 EMC 的影响。1850nm 激光在玻璃微纤维纸上处理 20min 后,导致 EMC(0.03±0.01gH2O/g干重)最低。这表明储存温度为 68.3°C。我们还使用标准测定法测试了 LAD 处理后模型蛋白溶菌酶的功能。当在最大温度约 44°C 至 EMC 为 0.17±0.06gH2O/g干重的条件下进行 LAD 处理时,LAD 对溶菌酶的功能没有显著影响。LAD 是一种很有前途的形成无定形海藻糖固体的技术,可在环境温度下稳定蛋白质。

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