Production of rHyC antigen
The expression of rHyC protein was induced using 1 mM IPTG and culture was allowed to grow at 37 °C with shaking at 200 rpm for 6 hr. Protein was purified under denaturing condition using 8 M urea in phosphate buffer. The homogeneity of purified fusion protein with molecular weight of about 45 kDa was analysed in SDS-PAGE (Fig. 1 ). The estimated concentration of purified rHyC protein was 94 µg/mL of elutes after dialysis.