Simultaneous isolation of lactoferrin and lactoperoxidase from bovine colostrum by SPEC 70 SLS cation exchange resin

In this work, simultaneous isolation of lactoferrin (Lf) and lactoperoxidase (Lp) from defatted bovine colostrum by one-step cation exchange chromatography with SPEC 70 SLS ion-exchange resin was investigated. A RP-HPLC method for Lf and Lp determination was developed and optimized as the following...

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Bibliographic Details
Published in:International journal of environmental research and public health 2011-09, Vol.8 (9), p.3764-3776
Main Authors: Liang, Yafei, Wang, Xuewan, Wu, Mianbin, Zhu, Wanping
Format: Article
Language:English
Subjects:
Adsorption
Animals
Cation Exchange Resins - chemistry
Cattle
Chromatography
Chromatography, High Pressure Liquid
Colostrum - chemistry
Electrophoresis, Polyacrylamide Gel
Female
Hydrogen-Ion Concentration
Lactoferrin - chemistry
Lactoferrin - isolation & purification
Lactoperoxidase - chemistry
Lactoperoxidase - isolation & purification
Milk
Pollution
Resins
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Summary:In this work, simultaneous isolation of lactoferrin (Lf) and lactoperoxidase (Lp) from defatted bovine colostrum by one-step cation exchange chromatography with SPEC 70 SLS ion-exchange resin was investigated. A RP-HPLC method for Lf and Lp determination was developed and optimized as the following conditions: detection wavelength of 220 nm, flow rate of 1 mL/min and acetonitrile concentration from 25% to 75% within 20 min. The adsorption process of Lf on SPEC 70 SLS resin was optimized using Lf standard as substrate. The maximum static binding capacity of SPEC 70 SLS resin was of 22.0 mg/g resin at 15 °C, pH 7.0 and adsorption time 3 h. The Lf adsorption process could be well described by the Langmuir adsorption isotherm model, with a maximum adsorption capacity of 21.73 mg/g resin at 15 °C. In batch fractionation of defatted colostrum, the binding capacities of SPEC 70 SLS resin for adsorbing Lf and Lp simultaneously under the abovementioned conditions were 7.60 and 6.89 mg/g resin, respectively, both of which were superior to those of CM Sepharose F.F. or SP Sepharose F.F. resins under the same conditions. As a result, SPEC 70 SLS resin was considered as a successful candidate for direct and economic purification of Lf and Lp from defatted colostrum.
ISSN:1660-4601
1661-7827
1660-4601
DOI:10.3390/ijerph8093764