基于串联亲和层析法纯化并鉴定猪初乳中SIgA

doi:10.11843/j.issn.0366-6964.2023.09.027

Abstract

Abstract: SIgA is the primary effector molecule of mucosal immunity, and it is often used as a target for early diagnosis of diseases in clinic. In addition, more and more attention has been paid for therapeutic SIgA mAbs and mucosal vaccines targeting SIgA producing. Isolation and purification of intact and active SIgA is a prerequisite for product development. In order to improve the purification efficiency of SIgA from swine colostrum, in this study, a new way was established,including using tandem affinity chromatography, strong anion column and superpose 6 gel filtration. By using this way, 3 mg of active SIgA was abtained from 10 mL of colostrum. Western blot immunoblotting was performed with anti-pig IgA heavy chain MAb and anti-SC MAb, and the results showed that both of the MAbs had good reactivity with purified SIgA here. ELISA method was used to test the reactivity of SIgA purified here and anti-pig IgA heavy chain MAb, it showed that the purified SIgA in our study had higher reactivity than SIgA purified by the old method (ammonium sulphate precipitation, DEAE52 weak anion chromatography, SephadexG-200 gel chromatography and multiple Protein A affinity chromatography). Finally, purified SIgA was further checked by mass spectrometry. Our research established an efficient method for the purification of SIgA from swine colostrum, and also provided a reference for the purification of SIgA from other animals.

Key words: porcine colostrum, SIgA, purification, identification

CLC Number:

S852.4

YANG Yue, WANG Rui, GAN Yuan, HAO Fei, XIE Xing, ZHANG Lei, SHAO Guoqing, MENG Qingguo, CHEN Rong, FENG Zhixin. Purification and Identification of SIgA in Porcine Colostrum based on Tandem Affinity Chromatography[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(9): 3884-3894.

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Purification and Identification of SIgA in Porcine Colostrum based on Tandem Affinity Chromatography

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