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首页> 外文期刊>Bioprocess and Biosystems Engineering >High-level production of a single chain antibody against anthrax toxin in Escherichia coli by high cell density cultivation
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High-level production of a single chain antibody against anthrax toxin in Escherichia coli by high cell density cultivation

机译：通过高细胞密度培养在大肠杆菌中大量生产抗炭疽毒素单链抗体

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Previously, we isolated the M18 scFv, which is an affinity matured antibody against the anthrax toxin PA, and observed that its single chain antibody (scAb) form (Ml8 scAb) exhibited superior stability compared to the scFv. Here, we report high cell density cultivations for preparative scale production of Ml8 scAb in a 3.5 L fer-menter. Briefly, a pH-stat feeding strategy was employed in fed-batch cultivation, and four different cell densities (OD_(600) of 40, 80, 120, and 150) were examined for the induction of scAb gene expression. Among the four cell densities investigated, lower cell densities (OD_(600)) of 40) showed higher post-induction cell growth and production yields (665 mg/L of scAb). Even though lower solubility (51 %) of scAb was achieved at lower cell density (OD_(600) of 40), monomeric scAb could be purified with high purity (>95%) using simple purification procedures. The purified scAb from high cell density cultures showed biological activity equivalent to that of scAb purified from shake flask cultivation.
机译：以前，我们分离了M18 scFv，这是一种针对炭疽毒素PA的亲和力成熟抗体，并观察到其单链抗体（scAb）形式（M18 scAb）与scFv相比显示出更高的稳定性。在这里，我们报道了在3.5 L发酵罐中进行规模化生产Ml8 scAb的高细胞密度培养。简而言之，在分批补料培养中采用了pH调节补料策略，并检查了四种不同的细胞密度（OD_（600）为40、80、120和150）来诱导scAb基因表达。在所研究的四种细胞密度中，较低的细胞密度（OD_（600）为40）显示较高的诱导后细胞生长和产量（665 mg / L scAb）。即使在较低的细胞密度（OD_（600）为40）下，scAb的溶解度较低（51％），也可以使用简单的纯化程序以高纯度（> 95％）纯化单体scAb。从高细胞密度培养物中纯化的scAb的生物学活性与从摇瓶培养中纯化的scAb相同。

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