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对新型重组布基胆酶医疗对策的分子设计,表达和生物活性进行有效并行评估

Joanne L Allard¹, Miguel Aguirre², Ruchi Gupta³

¹Chemical, Biological, Radiological and Nuclear Defence Branch, Defence Science and Technology Group (DSTG), Victoria, 3027, Australia; Protein Expression Facility, The University of Queensland, Brisbane, 4072, Australia.

Chemico-biological interactions

|September 2, 2024

概括

开发新的复合人本甲基胆酶 (rBChE) 生物清除剂为神经毒剂中毒的对策提供了一种具有成本效益的方法. 融合策略,特别是与人血清白蛋白 (hSA),显著增强酶活性和表达,改善治疗潜力.

科学领域:

生物化学生物化学
生物技术是生物技术.
药理学药理学是指药理学的学科.

背景情况:

目前用于神经毒剂中毒的医疗对策 (MCM) 的有效性和副作用有限,需要新的广泛疗法.
人体血衍生的丁胆酶 (huBChE) 是一种潜在的生物救生剂,但其大规模制造成本极高.

研究的目的:

解决哺乳动物细胞中生物活性,长效复合huBChE (rBChE) 的经济生产挑战.
直接比较新的rBChE设计策略,包括与蛋白质合作伙伴的普罗林丰富附着域 (PRAD) 共表达和融合.

主要方法:

开发一种预净化选方法,用于并行比较十种新型rBChE设计.
对表达效率,活性和广泛神经毒剂结合的评估.
使用了ExpiCHOTM Max和Expi293TM表达系统.

主要成果:

所有设计的rBChE变种都显示出作为广泛的MCM对抗G,V和A系列神经毒剂的功能.
与Expi293TM相比,ExpiCHOTM Max产生了更高的表达水平和活性.
与Fc或hSA融合的rBChE结构显著优于PRAD-BChE,其中BChE-hSA表现出最高的表达和活性.

结论:

与hSA等蛋白质伙伴的融合是增强rBChE活动和表达的有效策略.
开发的选方法简化了最佳rBChE构造的成本效益选择.
BChE-hSA具有与huBChE相似的功能,支持其作为下一代MCM的潜力.

关键词:

生物救援者生物救援者甲基胆化酶是一种甲基胆化酶.医疗反制措施医疗反制措施神经毒剂是一种神经毒剂.有机酸盐有机酸盐再组合蛋白质是一种重组蛋白质.

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