Abhishek Mishra , Sarah Steinbach , Ibrahim M. Tamer , Hector Budman
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引用次数: 0
Abstract
Whooping cough, also known as pertussis, is an infectious respiratory disease caused by Bordetella pertussis, a type of gram-negative, aerobic, pathogenic bacteria with a coccobacillus shape and an outer capsule. The acellular vaccine for this disease contains a combination of pertussis toxin, fimbriae, filamentous hemagglutinin, and pertactin. The main constraint in the manufacturing of the vaccine relates to pertactin productivity due to its low abundance in the fermentation broth. Being the most abundant nutrient in the media, variability in initial glutamate concentration is hypothesized to be a major cause of process variability. This study proposes the combined use of cytometry and chromatography based separation to study the impact of glutamate-induced oxidative stress on the growth and productivity of pertactin antigen. Surface expression of pertactin was observed using fluorescence microscopy and flow cytometry while extracellular concentration was quantified using affinity chromatography. Oxidative stress levels and secretion of NADPH, a crucial reactant in antioxidant reactions, were monitored in flask and bioreactor experiments. The results established clear correlations between oxidative stress with growth and productivity and differentiate between the reduction in productivity to growth and to synthesis rate of pertactin. These findings are industrially relevant for improving productivity and reducing process variability.
期刊介绍:
The Biochemical Engineering Journal aims to promote progress in the crucial chemical engineering aspects of the development of biological processes associated with everything from raw materials preparation to product recovery relevant to industries as diverse as medical/healthcare, industrial biotechnology, and environmental biotechnology.
The Journal welcomes full length original research papers, short communications, and review papers* in the following research fields:
Biocatalysis (enzyme or microbial) and biotransformations, including immobilized biocatalyst preparation and kinetics
Biosensors and Biodevices including biofabrication and novel fuel cell development
Bioseparations including scale-up and protein refolding/renaturation
Environmental Bioengineering including bioconversion, bioremediation, and microbial fuel cells
Bioreactor Systems including characterization, optimization and scale-up
Bioresources and Biorefinery Engineering including biomass conversion, biofuels, bioenergy, and optimization
Industrial Biotechnology including specialty chemicals, platform chemicals and neutraceuticals
Biomaterials and Tissue Engineering including bioartificial organs, cell encapsulation, and controlled release
Cell Culture Engineering (plant, animal or insect cells) including viral vectors, monoclonal antibodies, recombinant proteins, vaccines, and secondary metabolites
Cell Therapies and Stem Cells including pluripotent, mesenchymal and hematopoietic stem cells; immunotherapies; tissue-specific differentiation; and cryopreservation
Metabolic Engineering, Systems and Synthetic Biology including OMICS, bioinformatics, in silico biology, and metabolic flux analysis
Protein Engineering including enzyme engineering and directed evolution.