A recombinant vaccine against hydatidosis: production of the antigen in Escherichia coli.
A commercial process was developed for producing a recombinant vaccine against hydatidosis in farm animals. The vaccine antigen consisting of a surface protein of the oncospheres of the hydatid worm (Echinococcus granulosus), was produced as inclusion bodies in Escherichia coli. Fed-batch cultures of E. coli using Terrific broth in stirred bioreactors at 37 degrees C, pH 7.0, and a dissolved oxygen level of 30% of air saturation produced the highest volumetric concentrations of the final solubilized antigen. An exponential feeding strategy proved distinctly superior to feeding based on pH-stat and DO-stat methods. The plasmid coding for the antigen was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG) at 4 h after initiation of the culture. The minimum IPTG concentration for full induction was 0.1 mM.
Manderson
Daniel
D
Institute of Technology and Engineering PN456, Massey University, Palmerston North, New Zealand.
Dempster
Robert
R
Chisti
Yusuf
Y
D000818:Animals; D000947:Antigens, Helminth / Q000096:biosynthesis* / Q000235:genetics; D003470:Culture Media; D004443:Echinococcosis / Q000517:prevention & control*; D048209:Echinococcus granulosus / Q000235:genetics / Q000276:immunology*; D004926:Escherichia coli / Q000235:genetics; D005786:Gene Expression Regulation; D006863:Hydrogen-Ion Concentration; D002479:Inclusion Bodies / Q000378:metabolism; D007544:Isopropyl Thiogalactoside / Q000494:pharmacology; D010273:Parasitic Diseases, Animal / Q000517:prevention & control; D010957:Plasmids; D013696:Temperature; D014614:Vaccines, Synthetic / Q000096:biosynthesis / Q000235:genetics
D000947:Antigens, Helminth; D003470:Culture Media; D014614:Vaccines, Synthetic; D007544:Isopropyl Thiogalactoside