Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125
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...
Published in: | Applied Microbiology and Biotechnology |
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ftunivnapoliiris:oai:www.iris.unina.it:11588/587258 2024-09-09T19:08:32+00:00 Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 Maria Giuliani Filomena Sannino Gennaro Antonio Apuzzo Gennaro Marino PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA Maria, Giuliani Parrilli, Ermenegilda Filomena, Sannino Gennaro Antonio, Apuzzo Gennaro, Marino Tutino, MARIA LUISA 2014 http://hdl.handle.net/11588/587258 https://doi.org/10.1007/s00253-014-5582-1 eng eng info:eu-repo/semantics/altIdentifier/wos/WOS:000336268900010 volume:98 issue:11 firstpage:4887 lastpage:4895 numberofpages:9 journal:APPLIED MICROBIOLOGY AND BIOTECHNOLOGY http://hdl.handle.net/11588/587258 doi:10.1007/s00253-014-5582-1 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84901671977 info:eu-repo/semantics/restrictedAccess Pseudoalteromonas haloplanktis TAC125 Psychrophilic gene expression system scFvOx Single-chain antibody info:eu-repo/semantics/article 2014 ftunivnapoliiris https://doi.org/10.1007/s00253-014-5582-1 2024-06-17T15:19:31Z 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. Article in Journal/Newspaper Antarc* Antarctic IRIS Università degli Studi di Napoli Federico II Antarctic The Antarctic Applied Microbiology and Biotechnology 98 11 4887 4895 |
institution |
Open Polar |
collection |
IRIS Università degli Studi di Napoli Federico II |
op_collection_id |
ftunivnapoliiris |
language |
English |
topic |
Pseudoalteromonas haloplanktis TAC125 Psychrophilic gene expression system scFvOx Single-chain antibody |
spellingShingle |
Pseudoalteromonas haloplanktis TAC125 Psychrophilic gene expression system scFvOx Single-chain antibody Maria Giuliani Filomena Sannino Gennaro Antonio Apuzzo Gennaro Marino PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
topic_facet |
Pseudoalteromonas haloplanktis TAC125 Psychrophilic gene expression system scFvOx Single-chain antibody |
description |
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. |
author2 |
Maria, Giuliani Parrilli, Ermenegilda Filomena, Sannino Gennaro Antonio, Apuzzo Gennaro, Marino Tutino, MARIA LUISA |
format |
Article in Journal/Newspaper |
author |
Maria Giuliani Filomena Sannino Gennaro Antonio Apuzzo Gennaro Marino PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA |
author_facet |
Maria Giuliani Filomena Sannino Gennaro Antonio Apuzzo Gennaro Marino PARRILLI, ERMENEGILDA TUTINO, MARIA LUISA |
author_sort |
Maria Giuliani |
title |
Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
title_short |
Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
title_full |
Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
title_fullStr |
Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
title_full_unstemmed |
Recombinant production of a single-chain antibody fragment in Pseudoalteromonas haloplanktis TAC125 |
title_sort |
recombinant production of a single-chain antibody fragment in pseudoalteromonas haloplanktis tac125 |
publishDate |
2014 |
url |
http://hdl.handle.net/11588/587258 https://doi.org/10.1007/s00253-014-5582-1 |
geographic |
Antarctic The Antarctic |
geographic_facet |
Antarctic The Antarctic |
genre |
Antarc* Antarctic |
genre_facet |
Antarc* Antarctic |
op_relation |
info:eu-repo/semantics/altIdentifier/wos/WOS:000336268900010 volume:98 issue:11 firstpage:4887 lastpage:4895 numberofpages:9 journal:APPLIED MICROBIOLOGY AND BIOTECHNOLOGY http://hdl.handle.net/11588/587258 doi:10.1007/s00253-014-5582-1 info:eu-repo/semantics/altIdentifier/scopus/2-s2.0-84901671977 |
op_rights |
info:eu-repo/semantics/restrictedAccess |
op_doi |
https://doi.org/10.1007/s00253-014-5582-1 |
container_title |
Applied Microbiology and Biotechnology |
container_volume |
98 |
container_issue |
11 |
container_start_page |
4887 |
op_container_end_page |
4895 |
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1809822768076685312 |