Studies On Base And Application Of Chicken Egg Yolk Antibodies

IgY is the primite serum immunolgolubin(Ig) of chicken and provides the passively immune protection through transporting maternal IgY to neonate via the yolk .IgY is the founctional equivalent of mammalian IgG in bird ,reptiles and amphibia.IgY is applicable on immunoassay and immunotheray, due to possessing the physical and chemic characteristics Now , there is no available evidence studied in the functional protein to prove exist of the highly concerved domain of immunolgolubin in the context of evolution To understand this concerved domain , the anti-IgY monoclonal and polyclonal antibodies from Balb/c mouse immunized with chicken yolk antibody were used recognising the common epitope derived from various animal Ig shared with IgY.IgY antiserum cross-reacted with 4 yolk antibody species (pigeon, quail, duck,goose) were studied by dot-ELISA assay,as well as the immunologic cross-reactions of these same specific were presented by indirect-ELISA assay, but all of 13 hybridoma cell lines did not identify immunolgolubins upwards of 3 different species.This results suggest that, it is difficult to screen this McAb through general immunization and hybridoma protocal.Using chicken IgY and various Ig species (shrimp, fish, turtle, pigeon, duck, rabbit, horse, human) carried out basic and booster immunizations respectively produced the mouse antiserum ,showed good cross-reactivity in different species according to ELISA assay.It was suggested that our designed shift immunization scheme can enrich the common epitope of shared with IgY,enhance excreting this antibody specially and revealed the possibility that the common antigenic determinant was existent in measured species,at least among avian species markedly. Studing on yolk antibodies,adopting hybridoma technic, screen two mouse anti-IgY McAbs,named DFF and DFA.The McAb additivity test and Western-Blot assay presented DFF and DFA recongized the different epitope of IgY heavy chain, respec-tively .Thereinto the high-affinity DFF was conjugated HRP ,so firstly the indirect ELISA was establish for the detection system of chicken yolk antibodies specific in our country.In the immunotherapy of infective diarrhoea ,hens were immunized with genetically engineered vaccine of V. cholera and fatal enterotoxigenie escherichia coli strain produced the high titer antibodies detected by self- product ELISA reagent, thereby we set down the chicken immune regula- tion . Special yolk antibodies appeared the capability of recognising, com-bining with recombine cholera enterotoxin(CT-B).The essential technology was eatablished for clinical application.