抗猪流行性腹泻病毒变异株卵黄抗体的制备、纯化及其活性影响因素的研究

doi:10.16431/j.cnki.1671-7236.2018.08.031

Abstract

Abstract:

The purpose of the experiment was to prepare the yolk antibody (IgY) against variant of porcine epidemic diarrhea virus (PEDV) and explore methods that could efficiently,easily prepare and purify the prepared IgY.In this study,hens were immunized by variant PEDV (CH/JX01) and then the eggs were collected.Three different methods,water dilution method,polyethylene glycol 6000 (PEG-6000) method and water dilution-ammonium sulfate secondary precipitation method were introduced to purify the IgY targeted to PEDV,and the extraction quantity were evaluated by indirect ELISA and Western blotting.The results showed that the concentration of IgY extracted by water dilution method was the highest,reaching 6.204 mg/mL,followed by PEG-6000 method,4.673 mg/mL,and the concentration of IgY by water dilution-ammonium sulfate secondary precipitation method was 3.359 mg/mL.Based on the results of ELISA,the titer of IgY targeted PEDV from water dilution-ammonium sulfate secondary precipitation method was the highest with a titer of 1:12 800,while the titers of IgY from PEG-6000 and water dilution methods were 1:6 400 and 1:1 600,respectively.Compared the three different methods of purified IgY concentration and titer,it concluded that the purification effect of the water dilution-ammonium sulfate secondary precipitation method was better than the other two methods.The activity factors analysis results indicated that IgY purified by the water dilution-ammonium sulfate secondary precipitation method was stable at 37 to 60℃ and had good activity at pH 4.0 to pH 11.0.Under acidic conditions,20% sucralfate had a good protective effect on IgY activity.The results would provide an effective reference for the large-scale production and storage of the yolk antibody in the future.

Key words: egg yolk antibody; porcine epidemic diarrhea virus (PEDV); water dilution method; PEG-6000 method; water dilution-ammonium sulfate secondary precipitation method; activity

CLC Number:

S855.3

LEI Dan, LI Anqi, LUO Suxian, YE Yu, SONG Deping, TANG Yuxin. Preparation, Purification and Activity Factors Analysis of IgY Against Variant of Porcine Epidemic Diarrhea Virus[J]. , 2018, 45(8): 2293-2302.

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Preparation, Purification and Activity Factors Analysis of IgY Against Variant of Porcine Epidemic Diarrhea Virus

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