An Air-well sparging minifermenter system for high-throughput protein production

Microbial Cell Factories

Research

Cecilia Deantonio¹  Diego Cotella¹  Claudio Santoro¹  Francesca Persichetti¹  Daniele Sblattero¹  Valentina Sedini¹  Fabio Quasso²  Patrizia Cesaro² 

[1] Department of Health Sciences and Interdisciplinary Research Center on Autoimmune Diseases (IRCAD), University of Eastern Piedmont Amedeo Avogadro, Via Solaroli 17, 28100, Novara, Italy;Department of Sciences and Innovation, University of Eastern Piedmont Amedeo Avogadro, Alessandria, Italy;

关键词: High-Throughput Protein Production;    Shake-flask;    E.coli;    Air-sparging;    Minifermenter;   

DOI  :  10.1186/s12934-014-0132-1

 received in 2014-04-30, accepted in 2014-08-27,  发布年份 2014

来源: Springer

【 摘 要 】

BackgroundOver the last few years High-Throughput Protein Production (HTPP) has played a crucial role for functional proteomics. High-quality, high yield and fast recombinant protein production are critical for new HTPP technologies. Escherichia coli is usually the expression system of choice in protein production thanks to its fast growth, ease of handling and high yields of protein produced. Even though shake-flask cultures are widely used, there is an increasing need for easy to handle, lab scale, high throughput systems.ResultsIn this article we described a novel minifermenter system suitable for HTPP. The Air-Well minifermenter system is made by a homogeneous air sparging device that includes an air diffusion system, and a stainless steel 96 needle plate integrated with a 96 deep well plate where cultures take place. This system provides aeration to achieve higher optical density growth compared to classical shaking growth without the decrease in pH value and bacterial viability. Moreover the yield of recombinant protein is up to 3-fold higher with a considerable improvement in the amount of full length proteins.ConclusionsHigh throughput production of hundreds of proteins in parallel can be obtained sparging air in a continuous and controlled manner. The system used is modular and can be easily modified and scaled up to meet the demands for HTPP.

【 授权许可】

Unknown   
© Deantonio et al.; licensee BioMed Central Ltd. 2014. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.