Serum Institute of India Private Limited is developing a recombinant circumsporozoite protein R21 based pre-erythrocytic vaccine, against Plasmodium falciparum, in collaboration with Jenner Institute, Oxford, UK. Based on the gene sequence developed by Jenner Institute, the Hansenula polymorpha production strain was developed by ARTES Biotechnology GmbH. The production strain was transformed with an expression plasmid designed for expression of the fusion protein CS-HBsAg (R21). On receipt of Pre-MCB from ARTES, in-house research cell bank were prepared which were used in experimental batches at SIIPL. Based on our extensive fermentation experience on Hansenula platform, we optimized the fermentation and expression of R21. Further a mixed approach of SIIPL purification and Oxford process was designed and optimized for purification of the Recombinant R21 antigen. The VLP purification was assessed for purity, content, Size and DNA throughout the purification process.