Evidence map›Paper›PMID 34641643›Full record

ArticleIranian biomedical journal2021

Production of an Antibody Fragment (scFv) Targeting PcrV Protein of Pseudomonas aeruginosa in Fed-Batch Cultivation Mode

Saba Karam, Mozhgan Raigani, Sahar Hassani Afshar, Yeganeh Talebkhan, Elham Bayat, Samira Komijani, Leila Nematollahi, Farzaneh Barkhordari, Mehdi Shafiee Ardestani, Fatemeh Davami

Abstract read
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Article in Iranian biomedical journal, 2021. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.

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1 · What the graph read from it

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2 · The registry

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3 · Its place in the literature

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4 · The record

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5 · Who and what money

Authors and funding

10 authors.

Saba KaramInternational Campus, School Of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Mozhgan RaiganiBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Sahar Hassani AfsharInternational Campus, School Of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Yeganeh TalebkhanBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Elham BayatBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Samira KomijaniBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Leila NematollahiBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Farzaneh BarkhordariBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.
Mehdi Shafiee ArdestaniDepartment of Radiopharmacy, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran.
Fatemeh DavamiBiotechnology Research Center, Pasteur Institute of Iran, Pasteur Avenue, Tehran, Iran.

Funding

No grant is acknowledged in the PubMed record.

6 · The paper itself

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 type III secretion system (T3SS). PcrV is an important structural protein of the T3SS. Methods: In the current investigation, a recombinant single-chain fragment variable (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 Luria–Bertani 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+).

Indexed as

Pseudomonas aeruginosaAntibodies, BacterialAntigens, BacterialBacterial ToxinsPore Forming Cytotoxic ProteinsSingle-Chain AntibodiesTemperatureTime FactorsAntibodies, BacterialAntigens, Bacterialantigen V, PseudomonasBacterial ToxinsPore Forming Cytotoxic ProteinsSingle-Chain AntibodiesFed BatchPseudomonas aeruginosarecombinant proteinscFv

Identifiers

PMID34641643
PMCPMC8744694

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