Concanavalin A Partial purification of RhoD Antigen

Concanvalin A (Con A) chromatography is an alternative method for the partial purification of RhoD from red blood cell membrane. To partially purify RhoD, white ghost was prepared, solubilized, passed through Con A column and glycoproteins were eluted with α-methyl-D-mannoside. Affinity chromatography by applying Human RhoD immunoglobulin bound to Cyanogen bromide-activated-Sepharose 4B was used along with Con A column to compare and evaluate Con A chromatography for partial purification of RhoD antigen. The results of two columns after SDS/PAGE was subjected for immunoblot and also used in hemagglutination inhibition test. Both types of column led to isolation of two specific bands of polypeptides detectable in immunoblotting with approximate molecular weights of 29 kDa and 60 kDa respectively. Furthermore Con A chromatography partially purified RhoD antigen, was able to inhibit hemagglutination which indicates partially purified protein is functionally active.