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Chicken egg yolk immunoglobulin ( I g Y ) developed against fusion protein LTB – ST a– ST b neutralizes the toxicity of E scherichia coli heat‐stable enterotoxins
Author(s) -
You J.,
Xu Y.,
Li H.,
Wang L.,
Wu F.,
Xu F.,
Jin L.,
Li S.,
Li X.
Publication year - 2014
Publication title -
journal of applied microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.889
H-Index - 156
eISSN - 1365-2672
pISSN - 1364-5072
DOI - 10.1111/jam.12525
Subject(s) - enterotoxin , enterotoxigenic escherichia coli , microbiology and biotechnology , escherichia coli , immunogenicity , biology , heat stable enterotoxin , antibody , heat labile enterotoxin , fusion protein , recombinant dna , toxoid , immunogen , gene , monoclonal antibody , biochemistry , immunization , immunology
Aim To obtain a recombinant enterotoxigenic E scherichia coli ( ETEC ) fusion enterotoxin protein LTB – ST a– ST b ( B ab) that can express the immunogenicity of the haptens ST a and ST b and induce their corresponding neutralizing antibodies. Methods and Results The three important ETEC enterotoxin genes coding LTB , ST a and ST b were PCR ‐amplified, and the amplified products were fused to construct the trivalent enterotoxin expression vector p ET 30‐ B ab. SDS ‐ PAGE and Western blot were used to verify the expression of the fusion protein B ab by E . coli BL 21 carrying plasmid p ET 30‐ B ab. Laying hens immunized with B ab developed high egg yolk immunoglobulin ( I g Y ) titres specific to LTB , ST a and ST b, and all were significantly higher than those in the control group ( P < 0·01). A suckling mouse assay showed that anti‐ B ab I g Y can neutralize the natural toxicity of ST a and ST b with the highest dilution of 1/2 and 1/32, respectively. Conclusions Genetically constructed B ab induced significant antibody responses against ST a and ST b in chickens, and the resulting I g Y had the capacity to neutralize the toxicity of ST . Significance and Impact of the Study The recombinant B ab protein containing three important ETEC enterotoxins may serve as an effective and convenient polyvalent toxoid that can be used to produce multiple antitoxin I g Y s to prevent colibacillosis caused by ETEC with various fimbriae in young animals.