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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dc.contributor.authorKhoo, Teng Kew
dc.date.accessioned2018-10-10T02:15:46Z
dc.date.available2018-10-10T02:15:46Z
dc.date.issued2011-07
dc.description.abstractpanLF 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.en_US
dc.identifier.urihttp://hdl.handle.net/123456789/6731
dc.language.isoenen_US
dc.publisherUniversiti Sains Malaysiaen_US
dc.subjectFermentation of recombinant E. colien_US
dc.subjectto achieve high yield of biomassen_US
dc.titleFermentation Of Recombinant E. coli TOP10F' / pPROEX tm HTa / BmSXP To Achieve High Yield Of Biomass And Recombinant Antigen For Diagnostic Applicationen_US
dc.typeThesisen_US
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