Large-Scale Production of

Recombinant Proteins
Lecture 6

ISAT 451, Fall 2003

College of Integrated Science
and Technology
James Madison University
 Large-Scale Protein
Production

 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

 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
 Batch Fermentation

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

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 can
last up to 1,000
hours.
 Why do Cells Stop
Growing?

 Depletion of nutrients
 Lack of oxygen

 Change in pH

 Growth inhibition from

metabolic end products

 Stirred Tank Bioreactors

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

Introduction of
sterile air or
pure gases is
used for
agitation and
oxygenation.
 Internal-Loop
Airlift Bioreactor

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

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