Title of article :
Production of Brugia malayi BmSXP recombinant protein expressed in Escherichia coli
Author/Authors :
T. K.، Khoo نويسنده Institute for Research in Molecular Medicine (INFORMM) , , A.، Santhanam نويسنده Institute for Research in Molecular Medicine (INFORMM) , , R.، Noordin نويسنده Institute for Research in Molecular Medicine (INFORMM) , , N.، Arifin نويسنده Institute for Research in Molecular Medicine (INFORMM) ,
Issue Information :
دوفصلنامه با شماره پیاپی - سال 2010
Abstract :
A rapid antibody detection test is very useful for detection of lymphatic filariasis, especially for certification and
surveillance of post-mass drug administration. One such kit, panLF RapidTM (commercialized by Malaysian Bio-
Diagnostic Research Sdn. Bhd.) had been developed in our laboratory for the detection of all species of filarial
infections. It is based on the detection of anti-filarial IgG4 antibodies that react with recombinant Brugia malayi antigens, BmR1 and BmSXP. In this study, the growth of recombinant bacteria that produce BmSXP was optimized under shake flask fermentation for high yield of the recombinant antigen. The optimizations involved selection of suitable growth medium, IPTG concentration and induction time. The medium that yielded the highest biomass as well as total protein was Terrific Broth (TB) medium, which is an undefined medium. Initiation of induction of protein expression was found to be best at mid-log phase (OD600 = 1.5), with IPTG concentration of 1.0 mM, and harvest time at 9 h post-induction. This study showed that under the optimized conditions, the shake flask culture produced 4 g/L biomass (dry cell weight) of recombinant Escherichia coli BmSXP/pPROEXHTa/TOP10F’, which yielded 2.42 mg/L of purified BmSXP recombinant antigen. The purified antigen was analyzed by SDS-PAGE and the antigenicity of protein was confirmed by Western blot.
Journal title :
Malaysian Journal of Microbiology
Journal title :
Malaysian Journal of Microbiology