title: A high cell density fermentation strategy to produce recombinant malarial antigen in E. coli.
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.
authors.affiliation: Malaria Research Group, International Centre for Genetic Engineering and Biotechnology (ICGEB), Aruna Asaf Ali Marg, New Delhi 110067, India. shams@icgeb.res.in
authors.forename: Syed Shams
authors.initials: SS
authors.lastname: Yazdani
authors.forename: Ahmad Rushdi
authors.initials: AR
authors.lastname: Shakri
authors.forename: Chetan E
authors.initials: CE
authors.lastname: Chitnis
mesh_terms: D000953:Antigens, Protozoan / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D002452:Cell Count; D018929:Cell Culture Techniques / Q000379:methods*; D049109:Cell Proliferation; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D005285:Fermentation / Q000502:physiology; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D024861:Genetic Enhancement / Q000379:methods; D015202:Protein Engineering / Q000379:methods*; D015800:Protozoan Proteins / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D011956:Receptors, Cell Surface / Q000032:analysis / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D011994:Recombinant Proteins / Q000096:biosynthesis
chemical_list: D000953:Antigens, Protozoan; C066001:Duffy antigen binding protein, Plasmodium; D015800:Protozoan Proteins; D011956:Receptors, Cell Surface; D011994:Recombinant Proteins