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Tey Beng Ti,Al-Rubeai Mohamed The Korean Society for Biotechnology and Bioengine 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3
The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The ${\mu}_{max}$ and $K_s$ for the Bcl-2 cell line is $0.927day^{-1}\;and\;0.947\%(v/v)$ respectively, which are $21\%$ greate and $7\%$ lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a $17\%$ decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EM suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.
Optimal Conditions for Hepatitis B Cove Antigen Production in Shaked Flask Fermentation
Tey Beng Ti,Yong Kok Hoe,Ong Hong Puay,Ling Tau Chuan,Ong Swee Tin,Tan Yan Peng,Ariff Avbakariya,Tan Wen Siang The Korean Society for Biotechnology and Bioengine 2004 Biotechnology and Bioprocess Engineering Vol.9 No.5
The effects of various environmental factors such as pH (5, 6, 7, 8 and 9), temperature (30, 37 and $40^{\circ}C$) and rotational speed (150, 200 and 250 rpm) on the growth and the hepatitis B core antigen (HBcAg) production of Escherichia coli W3110IQ were examined in the present Study. The highest growth rate is achieved at pH 7, $37^{\circ}C$ and at a rotational Speed of 250 rpm which is 0.927 $h^{-1}$. The effect of pH on cell growth is more substantial compared to other parameters; it recorded a $123\%$ different between the highest growth rate (0.927 $h^{-1}$) at pH 7 and lowest growth at pH 5. The highest protein yield is achieved at pH 9, rotational speed of 250 rpm and $40^{\circ}C$. The yield of protein at pH 7 is $154\%$ higher compared to the lowest yield achieved at pH 5. There is about $28\%$ different of the protein yield for the E. coli cultivated at 250 rpm compared to that at 150 rpm which has the lowest HBcAg yield. The yield of protein at $40^{\circ}C$ is $38\%$ higher compared to the lowest yield achieved at $30^{\circ}C$.
Beng Ti Tey,Mung Ing Chua,Ghee Sung Chua,Michelle Yeen Tan Ng,Dayang Radiah Awang Biak,Wen Siang Tan,Tau Chuan Ling 한국생물공학회 2006 Biotechnology and Bioprocess Engineering Vol.11 No.2
The influence of temperature and agitation on the growth of Escherichia coli expressing hepatitis B core antigen (HBcAg) in stirred tank bioreactor were investigated. The highest specific growth rate for E. coli (0.844 h-1) was achieved at a temperature of 37°C and an agitation speed of 250 rpm. The activation energy for the growth of the E. coli strain W3110IQ in the stirred tank bioreactor was estimated to be 11 kcal/mol. The highest protein yield was achieved at a temperature of 44°C and an agitation speed of 250 rpm. The relative protein concentration at 44°C is 30 and 6% higher compared to that at 30 and 37°C, respectively.
Beng Ti Tey,Yen Mei Chow,Mohd Nordin Ibrahim,Arbakariya Ariff,Tau Chuan Ling 한국생물공학회 2006 Biotechnology and Bioprocess Engineering Vol.11 No.5
The bed stability of Streamline DEAE (ρ = 1.2 g/mL) in a 20 mm (i.d.) glass expanded bed contactor, and its performance on the recovery of glucose 6-phosphate dehydrogenase (G6PDH) from unclarified yeast homogenate were investigated. A residence time distribution study showed that a stable expanded bed was achieved. The theoretical plate and Bodenstein numbers determined were 25 and 53, respectively. A recovery yield of 87% and purification factor of 4.1 were achieved in the operation using 5% (w/v) biomass concentration feedstock. The performance of the anion exchange EBAC was still considerable good at a biomass concentration as high as 15% (w/v).
Tey, Beng Ti,Chua, Mung Ing,Chua, Ghee Sung,Ng, Michelle Yeen Tan,Biak, Dayang Radiah Awang,Tan, Wen Siang,Ling, Tau Chuan The Korean Society for Biotechnology and Bioengine 2006 Biotechnology and Bioprocess Engineering Vol.11 No.2
The influence of temperature and agitation on the growth of Escherichia coli expressing hepatitis B core antigen (HBcAg) in stirred tank bioreactor were investigated. The highest specific growth rate for E. coli$(0.844 h^{-1})$ was achieved at a temperature of $37^{\circ}C$ and an agitation speed of 250 rpm. The activation energy for the growth of the E. coli strain W3110lQ in the stirred tank bioreactor was estimated to be 11 kcal/mol. The highest protein yield was achieved at a temperature of $44^{\circ}C$ and an agitation speed of 250 rpm. The relative protein concentration at $44^{\circ}C$ is 30 and 6% higher compared to that at 30 and $37^{\circ}C$, respectively.
Optimal Conditions for Hepatitis B Core Antigen Production in Shaked Flask Fermentation
Beng Ti Tey,Tau Chuan Ling,Yan Peng Tan,Swee Tin Ong,Arbakariya Ariff,Wen Siang Tan,Hong Puay Ong,Kok Hoe Yong 한국생물공학회 2004 Biotechnology and Bioprocess Engineering Vol.9 No.5
The effects of various environmental factors such as pH (5, 6, 7, 8 and 9), temperature (30, 37 and 40C) and rotational speed (150, 200 and 250 rpm) on the growth and the hepatitis B core antigen (HBcAg) production of Escherichia coli W3110IQ were examined in the present study. The highest growth rate is achieved at pH 7, 37C and at a rotational speed of 250 rpm which is 0.927 h-1. The effect of pH on cell growth is more substantial compared to other parameters; it recorded a 123% different between the highest growth rate (0.927 h-1) at pH 7 and lowest growth at pH 5. The highest protein yield is achieved at pH 9, rotational speed of 250 rpm and 40C. The yield of protein at pH 7 is 154% higher compared to the lowest yield achieved at pH 5. There is about 28% different of the protein yield for the E. coli cultivated at 250 rpm compared to that at 150 rpm which has the lowest HBcAg yield. The yield of protein at 40C is 38% higher compared to the lowest yield achieved at 30C.
Beng Ti Tey,Mohamed Al-Rubeai 한국생물공학회 2005 Biotechnology and Bioprocess Engineering Vol.10 No.3
The over-expression of Bcl-2 has greatly improved the culture period, specific growth rate, and maximum viable cell density of NS0 cells culture under low serum condition. Further analysis of these data suggests that a saturation model of the Monod type can be used to represent the relationships of specific growth rate and initial serum concentration. The max and Ks for the Bcl-2 cell line is 0.927 day-1 and 0.947% (v/v) respectively, which are 21% greater and 7% lower respectively than its control counterpart. Study on the amino acid supplementation revealed that Bcl-2 cell lines possess greater improvement in the specific growth rate and maximum viable cell density compared to the control cell lines. A further increase in the amino acid supplementation has resulted a 17% decrease in specific growth rate and no improvement in maximum viable cell density in the control culture. However, the Bcl-2 cell line exhibited a better growth characteristic in this culture condition compared to that of control cell lines. The higher specific growth rate and maximum viable cell density of the Bcl-2 cell line in medium fortified with serum and MEM EAA suggested a more efficient nutrient metabolism compared to that in the control cell line. The low serum and amino acid utilisation rate and the higher cell yield may prove to be important in the development of serum/protein free culture.
Shamala Salvamani,Beng Ti Tey,Wen Cheng Ng,Wen Siang Tan 한국생물공학회 2011 Biotechnology and Bioprocess Engineering Vol.16 No.6
Nipah Virus (NiV) is an emerging zoonotic paramyxovirus that can be fatal in humans and various types of animals. The phospho (P) protein of NiV plays an important role in RNA synthesis, replication, and genome synthesis. In this study, the NiV P gene was cloned into a pTrcHis2-TOPO vector and the recombinant protein containing a His-tag was produced in Escherichia coli. SDSPAGE and Western blot analysis using the anti-His antibody confirmed the protein expression. An optimization study of E. coli fermentation showed that the optimal cultivation temperature was 37℃, while the optimal induction time for P protein expression was at 9 h with 1 mM IPTG. Solubility analysis showed that E. coli cultivated at 37℃produced the highest fraction (70%) of soluble P protein. The recombinant P protein was purified from clarified E. coli lysate using an immobilized metal affinity chromatography (IMAC) technique to a purity of 92.67%, with a purification factor of 11.58. The purified P protein strongly reacted with the anti-NiV swine sera collected during a NiV outbreak, suggesting its potential as a diagnostic reagent.
Chow, Yen Mei,Tey, Beng Ti,Ibrahim, Mohd Nordin,Ariff, Arbakariya,Ling, Tae Chuan The Korean Society for Biotechnology and Bioengine 2006 Biotechnology and Bioprocess Engineering Vol.11 No.3
A dense, pellicular UpFront adsorbent ($p=1.5 g/cm^3$, UpFront Chromatography, Cophenhagen, Denmark) was characterized in terms of hydrodynamic properties and protein adsorption performance in expanded bed chromatography. Cibacron Blue 3GA was immobilised into the adsorbent and protein adsorption of bovine serum albumin (BSA) was selected to test the setup. The Bodenstein number and axial dispersion coefficient estimated for this dense pellicular adsorbent was 54 and $1.63{\times}10^{-5}m^2/s$, respectively, indicating a stable expanded bed. It could be shown that the BSA protein was captured by the adsorbent in the presence of 30% (w/v) of whole-yeast cells with an estimated dynamic binding capacity $(C/C_o = 0.01)$ of approximately 6.5 mg/mL adsorbent.