Developing a polyepitope vaccine a chimeric protein which contains diverse types of antigenic epitopes is a promising strategy to prevent malaria Previously we had constructed a library of polyeptitope chimeric genes against Plasmodium falciparum without any protein tags In an attempt to develop an efficient and universal procedure for purification of polyepitope chimeric proteins we assembled an immunoaffinity chromatography (IAC) column with affinity-purified specific polyclonal IgY (mplgY) antibodies that recognized the same C-terminal epitope tag of chimeric proteins in the library A single-step and universal protocol was established and successfully applied for the purification of chimeric proteins Using this protocol chimeric proteins were specifically purified from an Escherichia colt expression system and the purity and authenticity were verified by gel electrophoresis and Western blot analysis Moreover the comparison between this IAC method and the conventional chromatography using two anion exchange columns followed by a step of gel filtration showed that the new method was more efficient with an 8-fold greater yield The results suggest that this IAC method will be an efficient approach for the purification of polyepitope vaccine candidates against P falciparum in our future study and also be valuable for other similar applications Crown Copyright (C) 2010 Published by Elsevier Inc All rights reserved