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Isolation of lactoferrin from bovine colostrum by ultrafiltration coupled with strong cation exchange chromatography on a production scale

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Title: Isolation of lactoferrin from bovine colostrum by ultrafiltration coupled with strong cation exchange chromatography on a production scale
Authors: Lu, Rong Rong lurr@sytu.edu.cn, Xu, Shi Ying1, Wang, Zhang1, Yang, Rui Jin1
Source: Journal of Membrane Science. Jul2007, Vol. 297 Issue 1/2, p152-161. 10p.
Subjects: Lactoferrin, Colostrum, Cations, Chromatographic analysis
Abstract: Abstract: Lactoferrin (LF) was isolated from bovine colostrum by ultrafiltration and then purified with a fast flow strong cation exchange chromatography system in a production scale. A two-step ultrafiltration process was performed with membranes of nominal molecular weight cut-offs of 100kDa for ultrafiltration (UF) step 1 (UF-1) and 10kDa in the UF step 2 (UF-2). The UF-1 process was performed at fixed transmembrane pressure (TMP), tangential flow velocity and temperature equivalent to 200kPa, 5m/s, and 25°C, respectively. The optimum operating parameters for the UF-2 were a tangential flow velocity of 4m/s, limiting TMP of 150kPa, and an operating temperature of 50°C. The predicted permeate flux based on a resistance mathematical model was not significantly (p >0.05) different from those of the actual experiment. The LF concentrated in the UF-2 retentate reached a purity of 30.88% (w/w) and a recovery of 94.04%. A stepwise procedure for purification of the crude LF was conducted using a preparative-scale strong cation exchange chromatography. The LF eluted with 1.0M NaCl aqueous buffer showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular weight of 80,400Da. The purity and the recovery of the final LF product were 94.20% and 82.46%, respectively. The process developed in this work is a significant improvement over the commercial practice for the fractionation of LF from bovine colostrum. [Copyright &y& Elsevier]
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Abstract:Abstract: Lactoferrin (LF) was isolated from bovine colostrum by ultrafiltration and then purified with a fast flow strong cation exchange chromatography system in a production scale. A two-step ultrafiltration process was performed with membranes of nominal molecular weight cut-offs of 100kDa for ultrafiltration (UF) step 1 (UF-1) and 10kDa in the UF step 2 (UF-2). The UF-1 process was performed at fixed transmembrane pressure (TMP), tangential flow velocity and temperature equivalent to 200kPa, 5m/s, and 25°C, respectively. The optimum operating parameters for the UF-2 were a tangential flow velocity of 4m/s, limiting TMP of 150kPa, and an operating temperature of 50°C. The predicted permeate flux based on a resistance mathematical model was not significantly (p >0.05) different from those of the actual experiment. The LF concentrated in the UF-2 retentate reached a purity of 30.88% (w/w) and a recovery of 94.04%. A stepwise procedure for purification of the crude LF was conducted using a preparative-scale strong cation exchange chromatography. The LF eluted with 1.0M NaCl aqueous buffer showed a single band in sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) with a molecular weight of 80,400Da. The purity and the recovery of the final LF product were 94.20% and 82.46%, respectively. The process developed in this work is a significant improvement over the commercial practice for the fractionation of LF from bovine colostrum. [Copyright &y& Elsevier]
ISSN:03767388
DOI:10.1016/j.memsci.2007.03.039