牛免疫球蛋白G的体外人源唾液酸化及Fc片段的制备

摘要/Abstract

摘要： 探讨了将牛免疫球蛋白G（bIgG）糖链末端N-羟乙酰神经氨酸（Neu5Gc）酶切并连接人源N-乙酰神经氨酸（Neu5Ac）的方法，并在实现bIgG转化为人源IgG（hIgG）的基础上研究了hIgG可结晶片段（Fc）的制备。结果表明，以170 U/mL神经氨酸酶酶切bIgG（4.0 mg/mL）后，通过?-1,4-半乳糖基转移酶（B4GALT1）和?-2,6-唾液酸转移酶（ST6GAL1）分别转移半乳糖（Gal）和Neu5Ac残基，可制备增加了8.4个Gal和42个Neu5Ac残基的hIgG分子。此外，在10.0 mg/mL hIgG、0.05 mg木瓜蛋白酶/mg hIgG、10.0 mmol/L半胱氨酸激活剂、2.0 mmol/L EDTA溶液和pH 7.0的条件下，酶解3 h可制得hIgG的Fc片段。最终所得hIgG得率为71.7%，Fc片段得率为20.8%。本研究为bIgG的产品开发和营养价值评价提供科学依据。

关键词: 牛免疫球蛋白G（bIgG）, Fc片段, 体外糖基化, 唾液酸

Abstract: In the present study, the methods of digesting bovine N-glycolylneuraminic acid (Neu5Gc) from bovine immunoglobulin G (bIgG), ligating IgG with human N-acetylneuraminic acid (Neu5Ac) to realize the transform of bIgG to human IgG (hIgG), and the preparation conditions of the crystallizable fragment (Fc) of hIgG were explored. Firstly, bovine Neu5Gc in bIgG (4 mg/mL) was digested under the action of 170 units/mL neuraminidase, 8.4 galactose (Gal) and 42 Neu5Ac were then transferred into prepared hIgG with ?-1,4-galactosyltransferase (B4GALT1) and ?-2,6-sialyltransferase (ST6GAL1), respectively. Secondly, the Fc fragment of hIgG was prepared under the conditions of 10 mg/mL hIgG, 0.05 mg papain/mg hIgG, 10.0 mmol/L cysteine, 2.0 mmol/L EDTA solution, pH 7.0, and enzymatic hydrolysis time 3 h. The yields of glycosylated hIgG and Fc fragment were 71.7% and 20.8%, respectively. The present study provides a role in laying the scientific basis for the product development and nutritional value evaluation of bIgG.

Key words: Bovine immunoglobulin G (bIgG), Fc fragment, in vitro glycosylation, sialic acid

中图分类号:

S896.4

李天慧陈春旭陈贵杰孙怡曾晓雄. 牛免疫球蛋白G的体外人源唾液酸化及Fc片段的制备[J]. 食品科学, 0, (): 0-0.

Tian-Hui LI Chunxu CHEN. Human sialylation of bovine immunoglobulin G and preparation of its Fc fragment in vitro[J]. FOOD SCIENCE, 0, (): 0-0.

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