Hydropericardium syndrome primarily affects the broilers between the ages of 2-7 weeks. 1he vaccine prepared from infected liver extract treated with formaldehyde is being used to protect the broilers from the disease. The current study was carried out to check the presence of immunogenic proteins in commercially available HPS vaccines Hyper immune serum was raised in 60 (3 weeks old) broiler birds using different commercially available HPS vaccine. The sera were subjected to AGPT for screening out HPS positive serum samples. The HPS infected livers were homogenized and loaded in alreadv prepared agar gel plates. The plates were incubated for 48 hours at room temperature in humid plastic box. Only 5 liver samples were found positive .The HPS autogenous vaccines,HPS positive liver samples normal liver samples were subjected to homogenization, sonication, chloroform treatment, ammonium sulphate precipitation, mixed with equal volume denaturing buffer and then boiled for 3 minutes to extract total proteins of Sample. . Large variations in the protein loading of samples in adjoining lanes lead to distortion.

Spectrophotometer determination of protein concentration assay were used to quantitate the known protein samples (bovine serum albumen is commonly used standard or this method) to standardize protein concentration. A28o were used to determine protein concentration and a calibration curve was created by plotting and performing regression analysis of A28o versus concentration of the standards and absorbance of the sample were used to determine the concentration from the calibration curve by using the formula. (concentration =Standard Factor* Dilution Factor* Optical Density.

Total protein analysis was carried out by using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and western blotting technique.

A typical gel of 9% acrylamide composition was used to nicely separate polypeptides of samples to analyze the entire profile of a fraction that contains heavy and light polypeptides. Best results were obtained when 30 pi of a 20-30 mg/mi final concentrations of denatured protein sample were loaded per sample well .Each sample was repeated twice on gel. The gel was cutted at the centre vertically. 0.1% Coomassie Blue dye in 50% methanol, 10% glacial acetic acid were used to stain half of the gel for detecting protein while half of the gel was subjected to western blotting. Relative molecular weight (MW) of each protein fraction was determined by plotting a standard curve. The western blot analysis of proteins of hydropericardium syndrome virus infected liver and HPS autogenous vaccine, separated on 9% gel showed one immunogenic protein, molecular weight 15-20 kDa. However, further studies are needed to establish its immunogenic nature and feasibility for its use as vaccine.