Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography

CORC > 过程工程研究所 > 中国科学院过程工程研究所 > 生化工程国家重点实验室

	Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography
	Zhang, Yan 1,2; Luo, Jian 1; Bi, Jingxiu 1,3; Wang, Jun 1,2; Sun, Lijing 1,2; Liu, Yongdong 1; Zhang, Guifeng 1; Ma, Guanghui 1; Su, Zhiguo 1
刊名	JOURNAL OF CHROMATOGRAPHY A
	2010-06-04
卷号	1217 期号:23 页码:3668-3673
关键词	Separation Homologous protein Human alpha-lactalbumin Transgenic bovine milk
ISSN号	0021-9673
通讯作者	Su, ZG
英文摘要	Transgenic bovine milk could be a rich source of recombinant human proteins. However, the co-presence of bovine and human homologous proteins can be a challenge for product purification. In this study, the average surface hydrophobicity and electric potential of human alpha-lactalbumin (HLA) and bovine alpha-lactalbumin (BLA) were analyzed and compared through the exposure area calculation of different amino acids. Based on the analysis, calcium independent hydrophobic interaction chromatography was selected for separation of recombinant human alpha-lactalbumin (rHLA) from BLA in transgenic bovine milk. The operating conditions for the best separation of two proteins were predicted by fluorescence data. Three commercially available HIC resins (Butyl Sepharose 4 FF, Octyl Sepharose 4 FF, Phenyl Sepharose 6 FF) were compared. The transgenic milk was skimmed and treated by pH adjustment to remove a large quantity of casein protein. The supernatant was loaded on the hydrophobic interaction chromatographic matrix. The correct elution fraction was further treated with gel filtration chromatography. The overall recovery of rHLA was up to 67.1% with the purity greater than 95%. Circular dichroism spectroscopy (CD) and mass spectrogram (MS) confirmed the native state and glycosylated form of the purified rHLA. (C) 2010 Elsevier B.V. All rights reserved.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine ; Physical Sciences
类目[WOS]	Biochemical Research Methods ; Chemistry, Analytical
研究领域[WOS]	Biochemistry & Molecular Biology ; Chemistry
关键词[WOS]	PROTEINS ; ACID ; CALCIUM ; BINDING ; SITE
收录类别	SCI
语种	英语
WOS记录号	WOS:000278158600005
公开日期	2013-11-18
内容类型	期刊论文
版本	出版稿
源URL	[http://ir.ipe.ac.cn/handle/122111/6131]
专题	过程工程研究所_生化工程国家重点实验室
作者单位	1.Chinese Acad Sci, Inst Proc Engn, Natl Key Lab Biochem Engn, Beijing 100190, Peoples R China 2.Chinese Acad Sci, Grad Sch, Beijing 100049, Peoples R China 3.Univ Adelaide, Sch Chem Engn, Adelaide, SA 5005, Australia
推荐引用方式 GB/T 7714	Zhang, Yan,Luo, Jian,Bi, Jingxiu,et al. Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography[J]. JOURNAL OF CHROMATOGRAPHY A,2010,1217(23):3668-3673.
APA	Zhang, Yan.,Luo, Jian.,Bi, Jingxiu.,Wang, Jun.,Sun, Lijing.,...&Su, Zhiguo.(2010).Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography.JOURNAL OF CHROMATOGRAPHY A,1217(23),3668-3673.
MLA	Zhang, Yan,et al."Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography".JOURNAL OF CHROMATOGRAPHY A 1217.23(2010):3668-3673.