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Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography
Zhang, Yan1,2; Luo, Jian1; Bi, Jingxiu1,3; Wang, Jun1,2; Sun, Lijing1,2; Liu, Yongdong1; Zhang, Guifeng1; Ma, Guanghui1; Su, Zhiguo1; Su, ZG
2010-06-04
Source PublicationJOURNAL OF CHROMATOGRAPHY A
ISSN0021-9673
Volume1217Issue:23Pages:3668-3673
AbstractTransgenic 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.
KeywordSeparation Homologous Protein Human Alpha-lactalbumin Transgenic Bovine Milk
SubtypeArticle
WOS HeadingsScience & Technology ; Life Sciences & Biomedicine ; Physical Sciences
DOI10.1016/j.chroma.2010.03.060
Indexed BySCI
Language英语
WOS KeywordPROTEINS ; ACID ; CALCIUM ; BINDING ; SITE
WOS Research AreaBiochemistry & Molecular Biology ; Chemistry
WOS SubjectBiochemical Research Methods ; Chemistry, Analytical
WOS IDWOS:000278158600005
Citation statistics
Cited Times:10[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Version出版稿
Identifierhttp://ir.ipe.ac.cn/handle/122111/6131
Collection生化工程国家重点实验室
Corresponding AuthorSu, ZG
Affiliation1.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
Recommended Citation
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, ZG.(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.
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