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Microbioreactor Continuous Culture Applications

Batch and Perfusion culture for mammalian process development

Cell densities greater than 70 million cells per mL and perfusion flow rates as high as 2 volumes per day are achievable with the Pharyx microbioreactor. By using a 2 mL working volume and a fully disposable fluid system, process development can be performed faster and with negligible materials cost.

Perfusion culture for antibiotics PK/PD

Dosage and clearance of a compound can be simulated using a microbioreactor perfusion culture. After a period of batch growth to an initial microbial cell density, a drug compound can be introduced through one of the fluid channels. After the initial drug compound concentration is reached, dilution of the compound through the perfusion flow is programmed to match the expected clearance rate. During dilution, microbial cell samples and perfusate samples are taken to measure the microbial cell viability and compound concentration. Automated refrigerated sampling is available along with integration with standard liquid handling robotics for automated cell viability measurement.

Chemostat culture for antibiotic resistance frequency studies

The frequency of mutations that impart antibiotic resistance is influenced by the environment. Stress from nutrient limitations can be induced in a controlled manner using microchemostat culture. Cell samples are automatically collected and chilled for resistance frequency determination.

Example Automated Experiment

Growth rate vs. Temperature
  • E. coli turbidostat
  • 1mL working volume
  • M9 + glucose
  • Automated temperature steps
Turbidostat culture: dilution rate adjusts to maintain fixed optical density. Dilution rate equals maximum growth rate for the experimental conditions. Doubling time decreases as temperature increases from 31C to 37C.

General Applications

Through microfluidic integration and automation, a variety of applications are possible with Pharyx cell culture technology.

  • Analysis of overflow metabolites through turbidostat operation
  • Automated growth media optimization by varying media composition
  • Metabolic Pathway Analysis through dynamic media flow
  • Exponential Growth Profiling
  • Quorom Sensing effects under different growth rates
  • Evolution Studies
  • Optimizing recombinant protein production
  • Strain Enrichment and Conditioning
  • Environmental condition dependent Genomic Studies (steady state or transient)
  • Metabolic Flux Analysis
  • Cell Line/Plasmid Stability
  • Screening and Clone Selection
  • Continuous or Batch Microproduction for potent chemicals

References:

Synthetic biology and microbioreactor platforms for programmable production of biologics at the point-of-care
Pablo Perez-Pinera, Ningren Han, Sara Cleto, Jicong Cao, Oliver Purcell, Kartik A. Shah, Kevin Lee, Rajeev Ram, and Timothy K. Lu, 2016.

A perfusion-capable microfluidic bioreactor for assessing microbial heterologous protein production.
Nicholas J. Mozdzierz, Kerry R. Love, Kevin S. Lee, Harry L. T. Lee, Kartik A. Shah, Rajeev J. Ram, and J. Christopher Love, 2015.

Microfluidics for Control in Synthetic Biology.
Ningren Han, Oliver Purcell, Fahim Farzadfard, Kevin S. Lee, Timothy K. Lu, and Rajeev J. Ram, 2014.

Fed-batch microbioreactor platform for scale down and analysis of a plasmid DNA production process.
Diana M. Bower, Kevin S. Lee, Rajeev J. Ram, and Kristala L.J. Prathers, 2012.

Microfluidic chemostat and turbidostat with flow rate, oxygen, and temperature control for dynamic continuous culture.
Kevin S. Lee, Paolo Boccazzi, Anthony J. Sinskey, and Rajeev J. Ram, 2011.