Purification of allophycocyanin from Porphyra haitanensis and prepraration of its polyclonal antibody

In this study, a series of methods including grinding in the presence of liquid N₂, ultrasonic disruption, ammonium sulfate precipitation and gel filtration chromatography on DEAE-Sephadex-A-50 were applied to isolation of allophycocyanin, a crucial chlorophyll-protein in the photosynthesis of algae and an important marine bio-active substance, from Porphyra haitanensis. The purified allophycocyanin was injected into rabbits to prepare its polyclonal antibody, and specificity of polyclonal antibody was detected by Western blotting. The results indicated that the purified allophycocyanin producted two bands on SDS-PAGE, which denoted the a and β subunit of the allophycocyanin, and the purity and rate of recovery of allophycocyanin obtained were high, up to 3.5 (A₆₅₀/A₂₈₀) and 70.5% respectively. The result of indirect ELISA assay showed that the liter of the polyclonal antibody was nearly 1: 100000.