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Abstract

GENERATION AND CHARACTERIZATION OF CHICKEN EGG YOLK ANTIBODIES (IGY) AGAINST CLOSTRIDIUM DIFFICILE AND EVALUATING ITS INVITRO NEUTRALIZATION EFFICACY

Sue Elizabeth Shajan*, Antonysamy Michael, Mahenthiran Ramasamy

ABSTRACT

Passive immunization by oral administration of specific antibodies has been an attractive approach against gastrointestinal tract (GIT) pathogens in both humans and animals. Recently, laying chickens have attracted considerable attention as an alternative source of antibodies for the prevention and treatment of infectious GIT diseases. Chicken egg yolk antibodies (IgY) were raised in twenty two weeks old white leghorn chicken against formalin killed whole cell antigen of Clostridium difficile (ATCC 9689). The level of the antibody in serum was monitored and booster doses were given wherever necessary. The antibodies were purified from the egg yolk of immunized chicken using PEG and Ammonium Sulphate precipitation method and further purified by dialysis method. The protein profile of anti Clostridium difficile IgY antibodies were analyzed by SDS-PAGEand Quantitative Titration of IgYantibodies by ELISA. High titer of specific antibody was found to be 1: 100000 on 91stday detected by modified ELISA and the titer were maintained with booster doses. The protein concentration of the egg yolk was 39.99 ± 0.79 mg/ml and the total IgY concentration in egg yolk was increased during the immunization period and reached maximum of 15.26± 0.57 mg/ml. In-vitro efficacy of anti-C.difficile IgY was determined by growth inhibition assay. A significant reduction in the growth was observed after 16 hours of incubation. These findings may open the door for significant advances in IgY technology and passive immunotherapy with an alternative high specific nature and low cost effective.

Keywords: Gastrointestinal tract (GIT), Chicken egg yolk antibodies (IgY), polyethylene glycol (PEG), Sodium Dodecyl Sulphate Polyacrylamide Gel Electrophoresis (SDS-PAGE), Enzyme Linked Immuno Assay (ELISA).


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