Department of Food Science, University of Wisconsin, Madison, WI 53706, USA.
J Chromatogr A. 2011 Apr 29;1218(17):2445-50. doi: 10.1016/j.chroma.2011.01.025. Epub 2011 Jan 18.
Proteins conjugated to neutral biopolymers are of keen interest to the food and pharmaceutical industries. Conjugated proteins are larger and more charge shielded than un-reacted proteins, making purification difficult using conventional beaded chromatographic supports because of slow mass transfer rates, weak binding, and viscous solutions. Past methods developed for pharmaceuticals are unsuitable for foods. In this work, a food-grade whey protein-dextran conjugate was purified from a feed solution also containing un-reacted protein and dextran using either a column packed with 800 mL of a beaded support that was specifically designed for purification of conjugated proteins or an 8 mL tube monolith. The monolith gave a similar dynamic binding capacity as the beaded support (4-6 g/L), at a 42-fold greater mass productivity, and 48-fold higher flow rate, albeit at somewhat lower conjugate purity. Performance of the monolith did not depend on flow rate. In conclusion, monoliths were found to be well suited for the purification of whey protein-dextran conjugates.
与中性生物聚合物偶联的蛋白质是食品和制药行业非常关注的对象。与未反应的蛋白质相比，偶联蛋白质更大，电荷屏蔽更强，因此使用传统的珠状色谱支持物进行纯化非常困难，因为传质速率慢、结合力弱和溶液粘性大。过去为药物开发的方法不适合食品。在这项工作中，从含有未反应的蛋白质和葡聚糖的进料溶液中，使用专门设计用于纯化偶联蛋白质的 800 毫升珠状载体填充柱或 8 毫升管整体柱，从乳清蛋白-葡聚糖缀合物中进行纯化。整体柱的动态结合容量与珠状载体（4-6 g/L）相似，但质量生产率高 42 倍，流速高 48 倍，尽管缀合物纯度略低。整体柱的性能不依赖于流速。总之，整体柱非常适合乳清蛋白-葡聚糖缀合物的纯化。
