High yield and secretion of recombinant human apolipoprotein AI in Pichia pastoris

Protein Expression and Purification
Mei-qing FengPei Zhou

Abstract

Apolipoprotein AI (ApoAI) is an important apolipoprotein in plasma and is known to have various physiological functions suitable for pharmaceutical applications. Human blood has been the only source of this protein for research and large-scale applications. To obtain large amounts of ApoAI a Pichia pastoris expression system was first used to obtain a high level of expression of secreted, recombinant protein. The human gene encoding ApoAI was inserted into the secretion vector pPIC9K and used to transform P. pastoris GS115. AP16, a high expression transformant with high G418 resistance, was obtained. After induction with methanol, the expression level of rhApoAI (recombinant human ApoAI) was 160 mg/L in a 14L fermentor. RhApoAI was purified by cold acetone precipitation followed by Q-Sepharose Fast Flow ion exchange column chromatography with 60% recovery. The N-terminal amino acid sequence and molecular weight (mass spec.) of rhApoAI are identical to native human ApoAI. Purified rhApoAI has specific binding activity with liver cells SMC7721 and binding can be inhibited by native human ApoAI.

References

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Citations

Jul 30, 2010·Journal of Lipid Research·Rachel J BraceSally P A McCormick
Oct 8, 2013·Journal of Drug Targeting·Baolong WangMeiqing Feng
Dec 3, 2015·Biotechnology Journal·Vignesh Narasimhan JanakiramanXavier Santarelli
Aug 2, 2011·Protein Expression and Purification·Sarah FiddymentAngel-Luis García-Otín
May 27, 2008·Protein Expression and Purification·Wang Jian-qiuYan Wei-qun
Apr 6, 2007·Protein Expression and Purification·Xun-Long ShiPei Zhou
Feb 26, 2008·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Xindu Geng, Lili Wang
Nov 28, 2015·Bulletin of Experimental Biology and Medicine·A V RyabchenkoL M Polyakov
Mar 25, 2017·Protein Expression and Purification·Nikita IkonRobert O Ryan

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