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Studies on the Preparation of Immunoglobulin Y from Egg Yolk against Piglet Diarrhea

Author: JiaLi
Tutor: YangYanJun
School: Jiangnan University
Course: Of Food Science
Keywords: Piglets Diarrhea E. coli Immunity Adjuvant Yolk Immunoglobulin
CLC: TQ464.7
Type: Master's thesis
Year: 2006
Downloads: 321
Quote: 1
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Abstract


Enterotoxigenic Escherichia coli (ETEC) is one of the major bacteria that cause diarrhea, ETEC adhesion molecules have been isolated and identified K88, K99, 987P, and F41, etc.. K88-K99-987P and K88-K99-F41, respectively Immunization mixed with different adjuvants (Freund's adjuvant, white oil adjuvant, aluminum hydroxide latex and phosphate-buffered saline solution) to study the anti-diarrhea in piglets egg yolk immunoglobulin (IgY), preparation method, and the egg yolk IgY was separated and purified, a systematic study of the biological activity and its characteristics and IgY. In this paper, enable Escherichia coli of K88, K99, 987P and F41 strains producing high unit cilia antigen Minca medium to determine the the bacteria best collection time of 12h by the determination of the growth curve. Hens immune yolk antibody activity monitoring, found that the Freund adjuvant vaccine and White Oil seedlings can induce sustained, stable and highly active antibody. Determine the water dilution method combined flocculant extracted IgY process route, the water-soluble protein yield was 84.47%. SDS-PAGE showed a the IgY the relative molecular mass of about 188,000, its light and heavy chains of relative molecular mass of about 28400 and 66500, respectively. Clarification → affinity purified IgY was diluted with water → flocculation, yield 17.92%; diluted with water → flocculation → → pro ammonium sulfate precipitation and purified by chromatography the IgY, yield 16.43%. Extracted from an egg yolk IgY about the whole egg weight of 1.22 ‰. Immunization of laying hens with Freund's adjuvant the IgY-K88-K99-987P IgY was-of K88-K99-F41 activity than the control group increased by 5 titers; laying hens with white oil adjuvant vaccine get IgY-K88-K99-987P and IgY was-of K88-K99-F41 activity than the control group increased 4 titers. The IgY was heat experiments show that: the activity of the purified IgY and IgY purified little difference between 20 ° C to 50 ° C; when the temperature continues to rise, of IgY the unpurified heat resistance than the purification of IgY better. The IgY acid, alkali and experimental results show that: the higher the temperature of IgY acid, the poorer the ability of the base, the purified IgY to the change of pH at 60 ℃ than at 20 ° C and 37 ℃ sensitive. 37 ℃, when the pH is between 2.0 to 4.0 and 8.0 to 10.0, and its change on the activity of non-purified IgY impact than the effect on the activity of the purified IgY significant. Pepsin hydrolysis the IgY the experimental: IgY was tolerated protease hydrolysis capacity and pH. When the pH at 2.0 and 3.0, the pepsin hydrolysis of IgY significantly;, pepsin hydrolysis of IgY effect is not significant when the pH is 4.0 to 6.0. IgY vitro antibacterial experiments show: K88 growth and reproduction can be suppressed, when the the IgY added amount of between 1 ~ 10mg/mL, IgY may be in vitro binding and K88. Vivo animal experiments showed that: composite egg yolk antibody powder in the high-dose group better diarrhea treatment effect, a cure rate of 100%; yolk antibody powder low dose group composite effect of poor treatment of diarrhea, piglet mortality by 50 %; piglet mortality in the control group 75%. Began three days before of piglets challenged to its orally egg yolk antibody powder, can play a better protective effect of piglets, the protection rate of 100%.

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CLC: > Industrial Technology > Chemical Industry > Pharmaceutical chemical industry > Drug production of biological products > Amino acids,peptides,proteins
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