Utilization Of Fermentation And Purification Strategies To Enhance The Yield Of Bmr1 Antigen
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Date
2010-03
Authors
Arifin, Norsyahida
Journal Title
Journal ISSN
Volume Title
Publisher
Universiti Sains Malaysia
Abstract
Brugia Rapid TM is an established diagnostic test that is commercially available to detect infection to both B. malayi and B.timori infections (brugian filariasis). The test is a dipstick lateral flow test format that utilizes BmR1 recombinant antigen expressed by the clone Bm17DIII/pPROEXTMHT/TOP 10. Due to a significant demand for the test in the market; there is a need to scale up the production of the recombinant antigen and increase the purification efficiency to reduce the production cost of the test.
The cultivation of the recombinant bacteria was performed using fed-batch fermentation where cells were grown in a modified Terrific broth medium and glucose was fed exponentially at a controlled rate using Multifermenter Control Software (MFCS) for automated feeding during pre-induction and post-induction feeding. Varying the specific growth rate (μ) prior to induction showed that a final cell concentration of 24.3 g/L was obtained at specific growth rate of 0.15 h-1 with feeding at high rate during post-induction. However, further increase of the specific growth rate during pre-induction feeding does not produce a higher cell mass, in fact it decreased the BmR1 recombinant protein production. It was found that the highest BmR1 production of 9.82 g/L was obtained when the feeding was carried out at low constant rate (1.9 ml/min), combined with the specific growth rate controlled at 0.075 h-1. It was also observed that at this feeding rate, the overall specific productivity, the fermentation yield coefficients [biomass yield (Yx/s) and product yield (Yp/s)] was the highest amongst all the runs tested. This strategy has successfully controlled the accumulation of acetic acid by-inhibitory product below the reported growth inhibitory level of 1.26 g/L.
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Keywords
Utilization of fermentation and purification strategies , to enhance the yield of Bmr1 antigen