Isolation, purification, and immuno biochemical characterization of IgG from dog (Canis lupus familiaris)

Abstract

Background: Immunoglobulin G (IgG) plays a pivotal role for secondary immune response in animals which trigger passive immunization against harmful pathogens in dogs. Purpose: The purpose of the present study was to isolate, purify, and immuno biochemically characterize IgG isolated from serum of dog (Canis lupus familiaris). Methods: The serum was isolated from dog which had undergone ammonium sulphate precipitation, which was dialysed overnight in a refrigerator followed by Lowry Method from where the protein concentration was estimated to be 122.22 mg/ml and the Total Protein Concentration was estimated to be 6.147 g/dl that is 61.47 mg/ml. This study continued through Column Chromatography estimation by obtaining a bell shaped graph (Absorbance vs Number of Fractions) followed by SDS-PAGE from where Rf values of different bands were obtained. Marker (Known) and samples P1, P2, P3, P4 (Unknown) graphs were obtained. The marker graph was estimated by Log of Molecular Weight of Protein Ladder Marker and the unknown graphs were estimated by Anti-Log of Molecular Weight of Protein Ladder Marker. After that DID Test was done where a single precipitin line of purified IgG of Canine dog was observed against antiserum developed in rabbit.Results: A single precipitin line of purified IgG of Canine dog observed against antiserum developed in rabbit and was found to be immune-reactive by DID Test and was fully confirmed by SDS-PAGE. Discussion: The dog serum was isolated by Ammonium Sulphate, purified by Column Chromatography, followed by Immunobiochemical Characterization which was confirmed by SDS-PAGE by obtaining a single precipitin line of IgG by DID Test.