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使用Plackett-Burman和中心复合设计优化培养基成分以提高重组人干扰素-β在SE1中的表达

Optimization of medium composition to increase the expression of recombinant human interferon-β using the Plackett-Burman and central composite design in SE1.

作者信息

Pal Dharam, Patel Gopal, Dobariya Prakashkumar, Nile Shivraj Hariram, Pande Abhay H, Banerjee Uttam Chand

机构信息

Department of Biotechnology, National Institute of Pharmaceutical Education and Research (NIPER), S.A.S. Nagar (Mohali), Sector 67, 160062 Punjab, India.

Department of Pharmaceutical Technology (Biotechnology), National Institute of Pharmaceutical Education and Research (NIPER), S.A.S. Nagar (Mohali), Sector 67, 160062 Punjab, India.

出版信息

3 Biotech. 2021 May;11(5):226. doi: 10.1007/s13205-021-02772-1. Epub 2021 Apr 19.

Abstract

Recombinant human interferon-β (rhIFN-β) is therapeutically important and new commercially viable approaches are needed for its increased production. In this study, a codon-optimized gene encoding for rhIFN-β protein was designed and expressed in SE1. As a first step of medium optimization, growth of as a function of different media components was studied. Subsequently, to optimize the media composition, a response surface methodology (RSM) was used. Our results show that optimized medium (15.0 g/L tryptone, 12.3 g/L meat extract, 1.0 g/L MgSO and 0.5 g/L thiamine along with minimal medium) obtained in this study provide better growth of recombinant cells and the expression level of recombinant protein was ~ 1.7-fold more than Luria-Bertani medium. The optimized medium may be utilized for the large-scale production of rhIFN-β.

摘要

重组人干扰素-β(rhIFN-β)具有重要的治疗意义,需要新的商业可行方法来提高其产量。在本研究中,设计了编码rhIFN-β蛋白的密码子优化基因,并在SE1中进行表达。作为培养基优化的第一步,研究了不同培养基成分对其生长的影响。随后,为了优化培养基组成,使用了响应面法(RSM)。我们的结果表明,本研究获得的优化培养基(15.0 g/L胰蛋白胨、12.3 g/L肉提取物、1.0 g/L MgSO和0.5 g/L硫胺素以及基本培养基)能使重组细胞更好地生长,重组蛋白的表达水平比Luria-Bertani培养基高约1.7倍。优化后的培养基可用于rhIFN-β的大规模生产。

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