CHOM-CT-SiO_2亲和介质的制备及胰蛋白酶的纯化

Affinity matrix preparation of CHOM and initial exploration for trypsin purification

目的为提高胰蛋白酶的纯度、收率,尝试制备一种新的亲和介质用于其分离纯化。方法用壳聚糖包裹硅胶制得CTS-SiO2基质,再与从鸡蛋中分离出的鸡卵黏蛋白(CHOM)偶联成CHOM-CT-SiO2亲和介质;运用介质吸附,一步洗脱后得到胰蛋白酶纯品。结果与结论鸡卵黏蛋白能高效地与CTS-SiO2基质偶联,亲和介质选择性好、非特异吸附小,该介质可有效特异地亲和吸附胰蛋白酶,比活性5.1×103U.mg-1,收率94%。

OBJECTIVE A new technology of separation and purification was developed for increasing the purity and yield of trypsin.METHODS An affinity matrix was prepared by conjugation silica gel to chitosan and CHOM which is separated from egg.Trypsin was directly adsorbed on the affinity matrix,and then eluted by one step to obtain pure trypsin.RESULTS and CONCLUSION CHOM can be efficiently immobilized on CTS-SiO2.The affinity matrix has the property of selectivity,and less non-specific adsorption.The matrix can selectively adsorb trypsin,and the specific activity was more than 5.1×10^3 U·mg^-1 with 94% yield.