Department of Chemical and Biological Engineering, Zhejiang University, Hangzhou, Zhejiang Province, 310027, China.
Int J Environ Res Public Health. 2011 Sep;8(9):3764-76. doi: 10.3390/ijerph8093764. Epub 2011 Sep 21.
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.
本工作研究了一步阳离子交换色谱法，使用 SPEC 70 SLS 离子交换树脂从脱脂牛初乳中同时分离乳铁蛋白（Lf）和乳过氧化物酶（Lp）。建立并优化了用于 Lf 和 Lp 测定的 RP-HPLC 方法，条件如下：检测波长 220nm，流速 1mL/min，乙腈浓度在 20min 内从 25%升至 75%。使用 Lf 标准品作为底物优化了 Lf 在 SPEC 70 SLS 树脂上的吸附过程。在 15°C、pH7.0 和吸附时间 3h 的条件下，SPEC 70 SLS 树脂的最大静态结合容量为 22.0mg/g 树脂。Lf 的吸附过程可以很好地用朗缪尔吸附等温线模型描述，在 15°C 时的最大吸附容量为 21.73mg/g 树脂。在脱脂初乳的分批分级中，在上述条件下，SPEC 70 SLS 树脂同时吸附 Lf 和 Lp 的结合容量分别为 7.60 和 6.89mg/g 树脂，均优于相同条件下的 CM Sepharose F.F.或 SP Sepharose F.F.树脂。因此，SPEC 70 SLS 树脂被认为是从脱脂初乳中直接、经济地纯化 Lf 和 Lp 的成功候选者。
