Microtiter miniature shaken bioreactor system as a scale-down model for process development of production of therapeutic alpha-interferon2b by recombinant Escherichia coli

Joo Shun Tan,Sahar Abbasiliasi,Arbakariya B Ariff,Yew Joon Tam,Yee-Ying Lee,Teck-Kim Tang,Saeid Kadkhodaei

doi:10.1186/s12866-017-1145-9

Joo Shun Tan, Sahar Abbasiliasi + Show 5 more

Open Access

https://doi.org/10.1186/s12866-017-1145-9

Copy DOI

Abstract

BackgroundDemand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development. This can be efficiently boosted by using microtiter plate (MTP) cultivation setup embedded into an automated liquid-handling system. The objective of this study was to establish an automated microscale method for upstream and downstream bioprocessing of α-IFN2b production by recombinant Escherichia coli. The extraction performance of α-IFN2b by osmotic shock using two different systems, automated microscale platform and manual extraction in MTP was compared.ResultsThe amount of α-IFN2b extracted using automated microscale platform (49.2 μg/L) was comparable to manual osmotic shock method (48.8 μg/L), but the standard deviation was 2 times lower as compared to manual osmotic shock method. Fermentation parameters in MTP involving inoculum size, agitation speed, working volume and induction profiling revealed that the fermentation conditions for the highest production of α-IFN2b (85.5 μg/L) was attained at inoculum size of 8%, working volume of 40% and agitation speed of 1000 rpm with induction at 4 h after the inoculation.ConclusionAlthough the findings at MTP scale did not show perfect scalable results as compared to shake flask culture, but microscale technique development would serve as a convenient and low-cost solution in process optimization for recombinant protein.

Highlights

Demand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development
The highest quantity of αIFN2b released by osmotic shock extraction using the automated microscale platform (49.2 μg/L) was comparable to that of α-IFN2b released by the manual extraction
At a constant agitation speed (800 rpm), working volume (80%) and induction time (4 h), growth of E. coli was increased from 2.83 g/L to 3.1 g/L with increasing inoculum size from 2% to 8% (v/v)

Summary

Introduction

Demand for high-throughput bioprocessing has dramatically increased especially in the biopharmaceutical industry because the technologies are of vital importance to process optimization and media development. This can be efficiently boosted by using microtiter plate (MTP) cultivation setup embedded into an automated liquid-handling system. Recent advances in life sciences and the establishment of high throughput screening technologies have led to the identification of large numbers of potentially important biopharmaceuticals Many of these products are in the later stages of product development and a priority is. The miniature bioreactor systems described are demonstrated in replacing the traditionally popular shake flask and manual osmotic shock treatment for parallel, high throughput operation, by optimizing the fermentation parameters with α-IFN2b production as a model. Report on the use of automated osmotic shock treatment is not available in the literature

Objectives

Methods

Results

Discussion

Conclusion