Download LAB SESSION 1: Bioprocessing

LAB SESSION 1: Bioprocessing
Explore Bioprocessing through a hands-on learning lab.
Bioprocessing is used in many pharmaceutical
applications in the
development
of
new drugs and
technologies.
Using
genetic
modification,
bacteria can be
manipulated
to
produce a wide
range of proteins.
Named after German physician
Drug development,
Theodor
Escherich (1857-1911)
diagnostic kits and
E.
coli
bacterium
is used for
aiding
in
overexpression of many
understanding the
interactions
important recombinant proteins
between cells and
(source: Kent School District;
their
environment
photo: Dennis Knunkle).
are
among
the
many, constantly increasing applications of
genetically modified (recombinant) proteins.
A key issue in the production of recombinant
proteins by bacteria is the investigator’s ability to
identify, purify and quantify the proteins of
interest. A wide variety of techniques are
available providing the investigator with many
possible outcomes of purity and concentration.
During this lab session, we will
investigate one of the most
widely used techniques in
protein purification, gel column
chromatography (GCC). Using
GCC, the investigator can purify
a mixture of many different
proteins or chemicals.
The
mechanism behind the process
relies on the characteristics of
the chemical or in the case of
proteins,
molecular
weight,
thereby
supplying
the
investigator
with
additional
pertinent information.
GCC Column using colored molecular weight
markers to demonstrate technique
During GCC, a liquid sample is applied to a
“solid” column consisting of porous beads.
Fractions collected as the solution passes
through the column will contain differing
chemicals or proteins based on their
characteristics, effectively separating the
different components.
Our Bioprocessing lab
session utilizes one of
the most widely used
proteins in research
today,
the
green
fluorescent
protein
(GFP).
Originally
isolated from the jelly
fish
Aequorea
Victoria, GFP is now
used
in
many
pharmacological
areas
of
protein
research. Used as a
marker
gene,
its
Aequorea Victoria from
glowing green color
http://www.accessexcellence.org:8080/RC/AB/
can easily be seen
BA/drug_testers/aequorea_win.html
with a simple UV light
source.
Attaching
GFP to other proteins of interests makes it
possible to determine the production or even
uptake of proteins being produced.
Within the gel column, students
can visually watch as the GFP is
separated from the other proteins.
They will learn techniques used to
quantify the amount of protein as
well as the purification technique
used. Students will gain an
understanding of the usefulness
of both the techniques and the
unique protein, GFP.
Gel column containing
GFP isolated from E. coli