Fermentation Of Recombinant E. coli TOP10F' / pPROEX tm HTa / BmSXP To Achieve High Yield Of Biomass And Recombinant Antigen For Diagnostic Application
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Date
2011-07
Authors
Khoo, Teng Kew
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
panLF Rapid™ is a rapid IgG4 antibody detection test which is based on the
detection of anti-filarial IgG4 antibodies that react with recombinant B. malayi
antigens, BmR1 and BmSXP. This diagnostic kit is very useful for the detection of
lymphatic filariasis (LF), especially in assisting the WHO on its certification and
surveillance activities of post-mass drug administration that is in relation to its
‘Global Programme to Eliminate Lymphatic Filariasis’ effort. The production of this
test kit has received a significant demand in the market, hence there is a need to upscale
the production of the recombinant antigens and increase the purification
efficiency in order to increase the production rate and also reduce the cost of
production. In this study the yield of BmSXP recombinant antigen was maximized by
achieving high biomass yield using fed-batch culture in a bioreactor, and the
recovery rate of the protein of interest was optimized in the downstream purification
process. The cultivation of the recombinant bacteria
(TOP10F’/pPROEX™HTa/BmSXP) was initially optimized in small-scale
fermentation using shake flask culture where it yielded 4.2 g.L-1 and
0.576 mg.g DCW-1 of BmSXP. The up-scaling process was then performed using fedbatch
fermentation where cells were grown in modified Terrific broth medium and
glucose was fed exponentially at a controlled rate using Multifermenter Control
Software (MFCS) for automated feeding. Varying an assortment of feeding
strategies, specific growth rate (μ) and induction strategies, biomass concentration of
19.43 g.L-1 and 11.16 mg.g DCW-1 of BmSXP were obtained based on exponential
feeding strategy at μ of 0.20 h-1, and with 1 mM single pulse IPTG induction at the
late-log phase of the bacterial growth curve. It was also observed that at this feeding
rate, the fermentation yield coefficients of overall specific productivity (YP/X),
biomass yield (YX/S) and product yield (YP/S) were high.
Description
Keywords
Fermentation of recombinant E. coli , to achieve high yield of biomass