Evans Leslie, Hughes Miranda, Waters Joanne, Cameron Jason, Dodsworth Neil, Tooth David, Greenfield Anthony, Sleep Darrell
Novozymes Biopharma UK Ltd., Castle Court, 59 Castle Boulevard, Nottingham NG7 1FD, United Kingdom.
Protein Expr Purif. 2010 Oct;73(2):113-24. doi: 10.1016/j.pep.2010.05.009. Epub 2010 May 27.
An expression system is described for the production of monomeric scFvs and scFv antibody fragments genetically fused to human albumin (at either the N- or C-terminus or both). Based upon strains of Saccharomyces cerevisiae originally developed for the production of a recombinant human albumin (Recombumin) this system has delivered high levels of secreted product into the supernatant of shake flask and high cell density fed-batch fermentations. Specific binding to the corresponding ligand was demonstrated for each of the scFvs and scFv-albumin fusions and pharmacokinetic studies showed that the fusion products had greatly extended circulatory half-lives. The system described provides an attractive alternative to other microbial systems for the manufacture of this type of product.
描述了一种表达系统,用于生产与人类白蛋白(在N端或C端或两端)基因融合的单体单链抗体可变区片段(scFv)和scFv抗体片段。基于最初为生产重组人白蛋白(重组白蛋白)而开发的酿酒酵母菌株,该系统已在摇瓶上清液和高细胞密度补料分批发酵中产生了高水平的分泌产物。每个scFv和scFv-白蛋白融合物都证明了与相应配体的特异性结合,药代动力学研究表明融合产物的循环半衰期大大延长。所描述的系统为制造这类产品提供了一种有吸引力的替代其他微生物系统的方法。