A high cell density fermentation strategy to produce recombinant malarial antigen in E. coli

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Abstract
A high cell density cultivation method was developed to produce recombinant PvRII, a malaria vaccine candidate, in E. coli for use in vaccine studies. Cells were grown in completely defined media and glucose was fed to achieve a specific growth rate of 0.12 h−1 until cells reached 55 g dry wt l−1. Culture was then induced with 1 mm IPTG and cells were further grown for 4 h to reach 85 g dry wt l−1 at 0.1 h−1. Recombinant PvRII was purified from inclusion bodies under denaturing conditions using metal affinity chromatography which yielded 10 mg PvRII g−1 dry wt. After refolding, PvRII was greater than 98% pure, homogeneous and functionally active in that it specifically bound Duffy positive human red cells.