You are on page 1of 12

Large-Scale Production of Recombinant Proteins

Lecture 6
Lecture 6

ISAT 451, Fall 2003 College of Integrated Science and Technology James Madison University

Large-Scale Protein Production

Large-Scale Protein Production  Most commercially valuable proteins (industrial enzymes or therapeutic proteins) require living cells

Most commercially valuable proteins (industrial enzymes or therapeutic proteins) require living cells to make the product Although cells can be readily grown in a standard 200 ml laboratory flask, commercial production may require containers in excess of 10,000 liters What are the limitations of large-

Microbial Growth Conditions

Microbial Growth Conditions  Sterile culture medium containing a carbon source and essential salts or an

Sterile culture medium containing a carbon source and essential salts or an enriched medium

Pure culture for inoculation oxygenation and mixing Regulation of temperature Regulation of pH

Microbial Cell Growth

Microbial Cell Growth

Batch Fermentation

Batch Fermentation Sterile medium is inoculated and fermentation proceeds without the addition of fresh growth medium

Sterile medium is inoculated and fermentation proceeds without the addition of fresh growth medium.

Fed Batch

Fed Batch Nutrients are added at intervals during fermentation. No growth medium is removed.

Nutrients are added at intervals during fermentation. No growth medium is removed.

Continuous Fermentation Fresh growth medium is added continuously during fermentation and spent medium is removed. Fermentation
Continuous Fermentation
Continuous Fermentation

Fresh growth medium is added continuously during fermentation and spent medium is removed.

Fermentation can last up to 1,000 hours.

Why do Cells Stop Growing?

Why do Cells Stop Growing?  Depletion of nutrients  Lack of oxygen  Change in

Depletion of nutrients Lack of oxygen Change in pH Growth inhibition from metabolic end products

Stirred Tank Bioreactors

Stirred Tank Bioreactors An external motor is used to agitate the growth medium with impellers. Sterile

An external motor is used to agitate the growth medium with impellers.

Sterile air or combinations of gases can be introduced (sparging)

Bubble Column Bioreactor

Bubble Column Bioreactor Introduction of sterile air or pure gases is used for agitation and oxygenation.

Introduction of sterile air or pure gases is used for agitation and oxygenation.

Bubble Column Bioreactor Introduction of sterile air or pure gases is used for agitation and oxygenation.

Internal-Loop Airlift Bioreactor

Internal-Loop Airlift Bioreactor Sterile air or pure gases are introduced to create circulation internal to the
Internal-Loop Airlift Bioreactor Sterile air or pure gases are introduced to create circulation internal to the

Sterile air or pure gases are introduced to create circulation internal to the reactor.

External-Loop Airlift Bioreactor

External-Loop Airlift Bioreactor Sterile air or pure gases are introduced to drive circulation through an external
External-Loop Airlift Bioreactor Sterile air or pure gases are introduced to drive circulation through an external

Sterile air or pure gases are introduced to drive circulation through an external loop.