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Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125

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Abstract

Recombinant protein production in cold-adapted bacteria has proved to be a valuable option to overcome solubility concerns often came up in conventional expression hosts. ScFvs are examples of “difficult proteins” due to their tendency to form inclusion bodies when expressed in Escherichia coli. In this paper, the recombinant production of a single-chain antibody (ScFvOx) in the psychrophilic bacterium Pseudoalteromonas haloplanktis TAC125 is reported. The expression vector for the ScFvOx production was designed to address the recombinant protein in the periplasmic space and to allow the formation of the antibody disulphide bonds. For periplasmic export, two different export mechanisms were evaluated. By combining the genetic tools available for recombinant protein expression in psychrophilic hosts with an ad hoc medium and fermentation modality and optimised expression conditions at low temperatures, we obtained the highest yield of soluble and epitope-binding ScFvOx reported so far by conventional prokaryotic expression. The observed proficiency of the Antarctic bacterium to produce recombinant antibody fragments was related to the unusually high number of genes encoding peptidyl prolyl cis-trans isomerases found in P. haloplanktis TAC125 genome, making this bacterium the host of choice for the recombinant production of this protein class.

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Acknowledgments

This work was supported by Programma Nazionale di Ricerche in Antartide PDR 2010/A1.05.

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Author notes

  1. Maria Giuliani

    Present address: Novartis Vaccines and Diagnostics, Via Fiorentina, 1, 53100, Siena, Italy

Authors and Affiliations

  1. Dipartimento di Scienze Chimiche, Università di Napoli “Federico II”, Via Cinthia, 4, 80126, Naples, Italy

    Maria Giuliani, Ermenegilda Parrilli, Filomena Sannino, Gennaro Antonio Apuzzo, Gennaro Marino & Maria Luisa Tutino

  2. Consiglio Nazionale delle Ricerche, Istituto di Biochimica delle Proteine, Via P. Castellino, 111, 80134, Naples, Italy

    Filomena Sannino

  3. Dipartimento di Scienze Chimiche, Università di Napoli “Federico II”, Complesso universitario Monte Sant’Angelo, Via Cintia, 80126, Naples, Italy

    Maria Luisa Tutino

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  1. Maria Giuliani
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  2. Ermenegilda Parrilli
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Correspondence to Maria Luisa Tutino.

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Giuliani, M., Parrilli, E., Sannino, F. et al. Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125. Appl Microbiol Biotechnol 98, 4887–4895 (2014). https://doi.org/10.1007/s00253-014-5582-1

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  • DOI: https://doi.org/10.1007/s00253-014-5582-1

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