Genetic engineering approach to develop next-generation reagents for endotoxin quantification:

Innate Immunity

HikaruMizumura¹ 

关键词: Amoebocyte lysate reagent;    endotoxin;    horseshoe crab;    mammalian cell;    recombinant cascade reagent;   

DOI  :  10.1177/1753425916681074

学科分类：生物科学（综合）

来源: Sage Journals

【 摘 要 】

The bacterial endotoxin test, which uses amebocyte lysate reagents of horseshoe crab origin, is a sensitive, reproducible and simple assay to measure endotoxin concentration. To develop sustainable raw materials for lysate reagents that do not require horseshoe crabs, three recombinant protease zymogens (factor C, derived from mammalian cells; factor B; and the proclotting enzyme derived from insect cells) were prepared using a genetic engineering technique. Recombinant cascade reagents (RCRs) were then prepared to reconstruct the reaction cascade in the amebocyte lysate reagent. The protease activity of the RCR containing recombinant factor C was much greater than that of recombinant factor C alone, indicating the efficiency of signal amplification in the cascade. Compared with the RCR containing the insect cell-derived factor C, those containing mammalian cell-derived factor C, which features different glycosylation patterns, were less susceptible to interference by the injectable drug components. The s...

【 授权许可】

CC BY