Pessela Benevides C C, Torres Rodrigo, Batalla Pilar, Fuentes Manuel, Mateo Cesar, Fernández-Lafuente Roberto, Guisán José M
Departamento de Biocatalisis, Instituto de Catalisis-CSIC, Campus UAM, Cantoblanco, 28049 Madrid, Spain.
Biotechnol Prog. 2006 Mar-Apr;22(2):590-4. doi: 10.1021/bp050223z.
We have developed a new protocol with only two steps for purification of immunoglobulins (Ig) from a protein concentrate of whey. Following this protocol, we have an 80% recovery of immunoglobulins, fairly pure. The purification was achieved by eliminating the BSA, via a strong adsorption on DEAE-agarose. Full desoprtion of the other serum proteins could be achieved without contamination with BSA. Thus, a protein solution containing only Ig and very small proteins (e.g., beta-lactoglobulins and alpha-lactalbumin) was obtained. Offering this protein mixture to a lowly activated aminated support, only Ig adsorbed on the support. It has been shown that BSA is able to interact with other proteins (including Ig and lactalbumins). This ability to form complexes with other proteins prevented the success of the direct adsorption of Ig on this mildly activated support, even although Ig should be the largest protein presented in dairy whey.
我们开发了一种新的方案,只需两步即可从乳清蛋白浓缩物中纯化免疫球蛋白(Ig)。按照此方案,我们可回收80%的免疫球蛋白,纯度相当高。通过在DEAE-琼脂糖上强烈吸附来去除牛血清白蛋白(BSA),从而实现了纯化。可以在不被BSA污染的情况下完全洗脱其他血清蛋白。因此,得到了一种仅含有Ig和非常小的蛋白质(如β-乳球蛋白和α-乳白蛋白)的蛋白质溶液。将这种蛋白质混合物提供给轻度活化的胺化支持物时,只有Ig吸附在支持物上。已表明BSA能够与其他蛋白质(包括Ig和乳白蛋白)相互作用。这种与其他蛋白质形成复合物的能力阻碍了Ig直接吸附到这种轻度活化支持物上的成功,尽管Ig应该是乳清中最大的蛋白质。