Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode

Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode Publisher Pubmed

Karam S^{1, 2} ; Raigani M² ; Afshar SH^{1, 2} ; Talebkhan Y² ; Bayat E² ; Komijani S² ; Nematollahi L² ; Barkhordari F² ; Ardestani MS³ ; Davami F²

Show Affiliations

1. International Campus, School Of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran
2. Biotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran
3. Department of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

Source: Iranian Biomedical Journal Published:2021

Abstract

Background: Pseudomonas aeruginosa is one of the opportunistic pathogens causing frequent hospital-acquired life-threatening infections in mechanically ventilated patients. The most significant virulence factor of P. aeruginosa is T3SS. PcrV is an important structural protein of the T3SS. Methods: In the current investigation, a recombinant scFv mAb against the PcrV protein was expressed in EnBase® (fed-batch) cultivation mode. The pETiteTM N-His SUMO Kan vector, including anti-PcrV scFv gene, was transformed into Escherichia coli (BL21) cells. The expression and solubility of anti-PcrV scFv protein were investigated at two different temperatures (25 °C and 30 °C) and at different induction times (4, 6, 8, 12, and 24 hours). Results: Increased efficiency was achieved by EnBase® compared to LB broth; owing to the slow release of glucose, the maximum level of solubility and total protein expression was observed in EnBase® cultivation system at 30 °C and 24 h post induction. Furthermore, IC50 for anti-PcrV scFv protein was determined to be approximately 7 μg/mL. Conclusion: Anti-PcrV scFv produced in this study showed promising in vitro results, protecting RBC from lysis by P. aeruginosa (exoU+). © 2021, Pasteur Institute of Iran. All rights reserved.

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Experts (# of related papers)

Amirhasan Zarnani (1)

Style	Citing Format
MLA	Karam S, et al.. "Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode." Iranian Biomedical Journal, vol. 25, no. 6, 2021, pp. 390-398.
APA	Karam S, Raigani M, Afshar SH, Talebkhan Y, Bayat E, Komijani S, Nematollahi L, Barkhordari F, Ardestani MS, Davami F (2021). Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode. Iranian Biomedical Journal, 25(6), 390-398.
Chicago	Karam S, Raigani M, Afshar SH, Talebkhan Y, Bayat E, Komijani S, Nematollahi L, Barkhordari F, Ardestani MS, Davami F. "Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode." Iranian Biomedical Journal 25, no. 6 (2021): 390-398.
Harvard	Karam S et al. (2021) 'Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode', Iranian Biomedical Journal, 25(6), pp. 390-398.
Vancouver	Karam S, Raigani M, Afshar SH, Talebkhan Y, Bayat E, Komijani S, et al.. Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode. Iranian Biomedical Journal. 2021;25(6):390-398.
BibTex	@article{ author = {Karam S and Raigani M and Afshar SH and Talebkhan Y and Bayat E and Komijani S and Nematollahi L and Barkhordari F and Ardestani MS and Davami F}, title = {Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode}, journal = {Iranian Biomedical Journal}, volume = {25}, number = {6}, pages = {390-398}, year = {2021} }
RIS	TY - JOUR AU - Karam S AU - Raigani M AU - Afshar SH AU - Talebkhan Y AU - Bayat E AU - Komijani S AU - Nematollahi L AU - Barkhordari F AU - Ardestani MS AU - Davami F TI - Production of an Antibody Fragment (Scfv) Targeting Pcrv Protein of Pseudomonas Aeruginosa in Fed-Batch Cultivation Mode JO - Iranian Biomedical Journal VL - 25 IS - 6 SP - 390 EP - 398 PY - 2021 ER -

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