Preparation of R-phycoerythrin from red alga and its application for antibody labeling and detection

R-phycoerythrin (R-PE) was highly purified from red alga (Bangia fusco-purpurea) by ammonium sulfate fractionation and DEAE-Sepharose anion-exchange column chromatography, the purity rate (A 565 /A280) of R-PE was over 6.0.SDS-PAGE demonstrated that relative molecular masses of R-PE subunits were about 19 and 21ku, respectively.Optimum concentration of heterobifunctional reagent N-succinimidyl-3-2-pyridyldithio propionate (SPDP) was used to polymerize goat anti-mouse IgG antibody and the cross-linking was identified using SDS-PAGE and fluorescence analysis.R-PE was successfully crosslinked with goat anti-mouse IgG as secondary antibody.Detection of major allergen parvalbumin in fish using R-PE labelled IgG were carried out by Dot blot and Western blot together with mouse anti-silver carp parvalbumin as primary antibody.The results demonstrated that R-PE labelled IgG could effectively detect the existence of parvalbumin and revealed good immunological specificity.The application of R-phycoerythrin as fluorescent probe is expected to shorten the detection time of immune hybridization, and simplify the operation processes.