Environmental stimuli on the soluble expression of anti-human ovarian carcinoma × anti-human CD3 single-chain bispecific antibody in recombinant Escherichia coli

Zhu, Yi-Qi; Tong, Wang-Yu; Wei, Dongzhi; Zhang, Feng; Zhao, Jun

doi:10.1016/j.bej.2007.04.010

Cited by 3 publications

(4 citation statements)

References 24 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In spite of the uncontrolled-pH nature of flask cultures, these results indicate that neither the higher pH nor lower pH were beneficial to cell growth and antibiotic production. Similar results appeared in the previous work [ 16 , 17 ], in which it was reported that the antibiotic production by X. nematophila BJ and Xenorhabdus sp. D43 were affected by the initial pH within the range of 4.5–8.5 and optimal initial pH for cell growth and antibiotic production occurred at 6.0–8.0.…”

Section: Resultssupporting

confidence: 92%

“…RIO showed growth in vitro similar to that of other Xenorhabdus species grown in TSB, the culture broth pH of RIO decline from 7.07 to pH 6.89 during the first few hours of growth, and then increased throughout the course of the experiment reaching 8.79 at 168 h [ 23 ]. The culture broth pH of X. nematophila BJ varied from 6.5 to 8.5 [ 16 ]. Although microbial cells have the ability to respond to environment alterations and a remarkable ability to maintain the intracellular pH at a constant level even with large variations in the pH of the extracellular medium, the pH of the environment has a natural tendency to change along with the bioprocess.…”

Section: Discussionmentioning

confidence: 99%

“…Well-directed process parameters shift representing a valuable control strategy will be beneficial to the cell growth and metabolite biosynthesis. pH-shift operation has been proven to be an effectual strategy for the production of bioactive metabolites in other microorganism [ 11 – 16 ].…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Manipulation of pH Shift to Enhance the Growth and Antibiotic Activity of Xenorhabdus nematophila

Wang

Fang

Cheng

et al. 2011

BioMed Research International

View full text Add to dashboard Cite

To evaluate the effects of pH control strategy on cell growth and the production of antibiotic (cyclo(2-Me-BABA-Gly)) by Xenorhabdus nematophila and enhance the antibiotic activity. The effects of uncontrolled- (different initial pH) and controlled-pH (different constant pH and pH-shift) operations on cell growth and antibiotic activity of X. nematophila YL00I were examined. Experiments showed that the optimal initial pH for cell growth and antibiotic production of X. nematophila YL001 occurred at 7.0. Under different constant pH, a pH level of 7.5 was found to be optimal for biomass and antibiotic activity at 23.71 g/L and 100.0 U/mL, respectively. Based on the kinetic information relating to the different constant pH effects on the fermentation of X. nematophila YL001, a two-stage pH control strategy in which pH 6.5 was maintained for the first 24 h, and then switched to 7.5 after 24 h, was established to improve biomass production and antibiotic activity. By applying this pH-shift strategy, the maximal antibiotic activity and productivity were significantly improved and reaching 185.0 U/mL and 4.41 U/mL/h, respectively, compared to values obtained from constant pH operation (100.0 U/mL and 1.39 U/mL/h).

show abstract

Section: Resultssupporting

confidence: 92%

Section: Discussionmentioning

confidence: 99%

See 1 more Smart Citation

Manipulation of pH Shift to Enhance the Growth and Antibiotic Activity of Xenorhabdus nematophila

Wang

Fang

Cheng

et al. 2011

BioMed Research International

View full text Add to dashboard Cite

show abstract

“…The most soluble to insoluble ratio of fusion protein is achieved at 25 °C in three strains. Several studies have showed that cultivating recombinant E. coli cells at lower temperature increases solubility and stability of recombinant proteins by preventing their aggregation (Zhu et al 2007) Several studies have showed that recombinant proteins are mostly appeared in their soluble form when the hosts are induced at lower temperature (Akbari et al 2014;Santala and Lamminmäki 2004;Sonoda et al 2010;Zhu et al 2007).…”

Section: Discussionmentioning

confidence: 99%

Cloning, Expression and One-Step Purification of a Novel IP-10-(anti-HER2 scFv) Fusion Protein in Escherichia coli

Ahmadzadeh

Farshdari

Behdani

et al. 2020

Int J Pept Res Ther

View full text Add to dashboard Cite

Breast cancer is the most common cancer in women, worldwide. The correlation between breast cancer malignancy and human epidermal growth factor 2 (HER2) expression leads to application of monoclonal antibodies against HER2 in HER2overexpressing breast cancers. Variable fragments of light and heavy chains of monoclonal antibodies are linked in single chain variable fragment (scFv). Herein, IP-10 chemokine is conjugated to scFv against HER2 and thus CD8 + T cells can chemoattract to HER2-overexpressing tumors. IP-10-(anti-HER2 scFv) gene was cloned in pET22-b (+) and successful expression of the fusion protein in E. coli host was confirmed by detecting a 39 kDa band in Western blotting. The highest fusion protein expression in Origami (DE3), BL21 (DE3) and SHuffle ® were obtained 4 h after 0.1, 0.5 and 0.1 mM IPTG induction at 37, 37 and 30 °C, respectively. It was also demonstrated that the most solubility of the fusion protein is obtained at 25 °C in all the three examined strains. The highest soluble form of the fusion protein was expressed in SHuffle ® . However, due to low amount of the fusion protein expressed in SHuffle ® , we focused on its expression in Origami (DE3). Finally, purification of the fusion protein using Ni-NTA affinity chromatography under native, denaturing and hybrid conditions was investigated. Purification under hybrid condition caused the most purity of the fusion protein. This study opens up the avenue of exploring the use of IP-10-(anti-HER2 scFv) as a potential treatment for HER2-overexpresing tumors or as an adjuvant in combination with HER2-based vaccine.

show abstract

Anti-HER2 scFv Expression in Escherichia coli SHuffle®T7 Express Cells: Effects on Solubility and Biological Activity

et al. 2019

View full text Add to dashboard Cite

Environmental stimuli on the soluble expression of anti-human ovarian carcinoma × anti-human CD3 single-chain bispecific antibody in recombinant Escherichia coli

Cited by 3 publications

References 24 publications

Manipulation of pH Shift to Enhance the Growth and Antibiotic Activity of Xenorhabdus nematophila

Manipulation of pH Shift to Enhance the Growth and Antibiotic Activity of Xenorhabdus nematophila

Cloning, Expression and One-Step Purification of a Novel IP-10-(anti-HER2 scFv) Fusion Protein in Escherichia coli

Anti-HER2 scFv Expression in Escherichia coli SHuffle®T7 Express Cells: Effects on Solubility and Biological Activity

Contact Info

Product

Resources

About