HUBEI AGRICULTURAL SCIENCES ›› 2020, Vol. 59 ›› Issue (15): 108-113.doi: 10.14088/j.cnki.issn0439-8114.2020.15.021

• Animal Husbandry & Veterinary Medicine • Previous Articles     Next Articles

Comparison and preliminary application of five extraction methods of egg yolk antibody

XIANG Dong-mei1,2, YI Si-yu1,3, TANG Qing-hai1, WU Si-wen1, WANG Shu-shu1, LI Yan1, LI Lu1, HAN Tao-tao1   

  1. 1. College of Life Sciences and Environment/Institute of Bio-technology for Livestock, Hengyang Normal University, Hengyang 421008, Hunan, China;
    2. College of Life Sciences, Qingdao Agricultural University, Qingdao 266109, Shandong, China;
    3. College of Animal Science and Technology, Guangxi University, Nanning 530004, China
  • Received:2019-11-26 Online:2020-08-10 Published:2020-09-24

Abstract: In order to obtain a relatively good extraction process of egg yolk antibody, five methods including octanoic acid method, water dilution method, chloroform method, ammonium sulfate precipitation method and polyethylene glycol precipitation method were improved in this experiment. Expression of recombinant porcine pathogenic Escherichia coli toxin STxB(rSTxB) was induced by IPGT, and then the immunogen was prepared by emulsifying the rSTxB with the adjuvant 71VG, and the hens were immunized, IgY was extracted by modified polyethylene glycol method. The purity of the IgY antibody was detected by SDS-PAGE electrophoresis, and the immunoactivity and titers of yolk antibody and serum antibody titer were detected by Western blot. The results showed that the extraction efficiencies of octanoic acid method, water dilution method, chloroform method, ammonium sulfate precipitation method and polyethylene glycol precipitation method were 96.00,81.99,155.50,65.34,46.00 mg/mL,respectively, and the extraction purity of polyethylene glycol precipitation was the highest. The molecular weight of rSTxB protein was about 25 kDa, the antibody titer in serum was up to 64 000 times, and the highest dilution factor of yolk antibody was 16 000 times. For the comprehensive evaluation, the extraction method of polyethylene glycol precipitation was relatively superior, and the extracted yolk antibody showed good immunological activity.

Key words: egg yolk antibody, extraction process, polyethylene glycol precipitation, STxB protein, adjuvant

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