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Type: Journal article
Title: Efficient separation of homologous α-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography
Other Titles: Efficient separation of homologous alpha-lactalbumin from transgenic bovine milk using optimized hydrophobic interaction chromatography
Author: Zhang, Y.
Luo, J.
Bi, J.
Wang, J.
Sun, L.
Liu, Y.
Zhang, G.
Ma, G.
Su, Z.
Citation: Journal of Chromatography A, 2010; 1217(23):3668-3673
Publisher: Elsevier Science BV
Issue Date: 2010
ISSN: 0021-9673
Statement of
Yan Zhang, Jian Luo, Jingxiu Bi, Jun Wang, Lijing Sun, Yongdong Liu, Guifeng Zhang, Guanghui Ma and Zhiguo Su
Abstract: 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 α-lactalbumin (HLA) and bovine α-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 α-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.
Keywords: Separation; homologous protein; human α-lactalbumin; transgenic bovine milk
Rights: © 2010 Elsevier B.V.
RMID: 0020097970
DOI: 10.1016/j.chroma.2010.03.060
Appears in Collections:Chemical Engineering publications

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