蛋黃IgY免疫親和式層析法純化豬血漿中丙型球蛋白

Isolation of Pig γ-Globulin by Anti-γ-Globulin IgY Immunoaffinity Chromatography

杜艷櫻(Yann-Ying Tu) ； 陳昭誠(Chao-Cheng Chen)

《臺灣營養學會雜誌》 35卷3期 (2010/09) Pp. 109-114

https://doi.org/10.6691/NSJ.201009_35(3).0003

摘要

本研究主要以蛋黃抗丙型球蛋白IgY免疫親和式層析法純化豬血中丙型球蛋白。丙型球蛋白濃度約為(5 mg/mL)經抗丙型球蛋白IgY之免疫親和式層析法純化後之純化效益為15倍，回收率為81.1%。純化之抗丙型球蛋白IgY與丙型球蛋白，經SDS-PAGE梯度電泳及免疫轉印法分析，主要色帶分別在分子量66.4及55.6 kDa有抗丙型球蛋白IgY及丙型球蛋白的重鏈，此外無其他色帶，顯示純度頗高。丙型球蛋白與抗丙型球蛋白IgY免疫親和式膠體之親和性吸附作用力之Omax約0.27 mg/mL wet gel，Kd值約4.2×10^(-5)M。本研究發現免疫親和式層析法應用於純化高純度的豬血丙型球蛋白，具有醫藥量化生產潛力。

關鍵字

豬血丙型球蛋白 ； IgY免疫親和式層析法 ； 解離常數 ； 最大吸附量 ； SDS-PAGE梯度電泳 ； 免疫轉印法

並列摘要

The objective of this study was to investigate the isolation of pig γ-globulin by anti-γ-globulin IgY immunoaffinity chromatography. The purification efficiency of the γ-globulin concentration in pig blood (about 5 mg/mL) purified by anti-γ-globulin IgY immunoaffinity chromatography was 15 with a recovery of 81.1%. Electrophoresis of the gradient sodium dodecylsulfate polyacrylamide gel electrophoresis and Western blotting specific against anti-γ-globulin IgY and γ-globulin were demonstrated to be of high purity for only one band the heavy chain ) with a molecular weight of 66.4 kDa for the anti-γ-globulin IgY and 55.6 kDa for y-globulin, and the absence of detectable protein bands was confirmed. The dissociation constant (Kd) and binding capacity (Omax) of anti-γ-globulin IgY immunoaffinity chromatography for y-globulin were respectively calculated to be 4.2×10^(-6) M and 0.27 mg/mL wet gel. Immunoaffinity chromatography exhibits a potentially important advantage of scaling up for use with therapeutic agents.

並列關鍵字

pig γ-globulin ； IgY immunoaffinity chromatography ； dissociation constant Kd ； binding capacity Qmax ； SDS-PAGE gradient electrophoresis ； Western blot