Practical Course Dr. Uwe Horn
From Strains to Biotechnological
Processes
November 21-24, 2011,
HKI, Bio Pilot Plant, meeting point 9:00 HKI main building, foyer
Limited number of participants:
8 *;
minimum number of participants: 3
- Course is fully
booked! -
For the waiting list, please
email
christine.vogler@hki-jena.de
until November 15
*In
order to provide each participant a thorough hands on experience
we are restring the number of participants to 8. Should there
be more interested participants we would be happy to repeat the
course next year.
Wine, sauerkraut or yogurt; since time immemorable the art of
fermentation has been known to the man-kind. Metamorphosis of
this art to science and then to technology began in the last
century with the production of bio-active substances from
micro-organisms at the industrial scale.
In the coursework a specific application of high cell density
fermentation technology would be covered:
1. Periplasmic expression of camelid VHH antibody fragment and
antibody purification from the biomass
In contrast to the conventional mammalian antibodies, camelid
antibodies bear only single variable heavy domain (VHH) and lack
the CH1 domain and the variable light chain. Because of the
small size ~15kDa, VHH domains are very useful for recombinant
antibody libraries. Specific camelid antibodies have been
generated in our department, screened by phage display technique
and engineered for periplasmic expression in E.coli. Herein,
E.coli would be cultivated by high cell density fed-batch
fermentation in a defined glucose mineral salt media followed by
downstream procedure of biomass accumulation, cell lysis and
antibody purification.
In addition we give an introduction to the possibilities of our
biopilot plant around the cultivation of different MO and their
downstream processing.
References:
1. Habicht G et al. : Directed selection of a conformational
antibody domain that prevents mature amyloid fibril formation by
stabilizing Abeta protofibrils. Proc Natl Acad Sci U S A. 2007
Dec 4;104(49): 19232-7.
2. Horn U et al.: High volumetric yields of functional dimeric
miniantibodies in Escherichia coli, using an optimized
expression vector and high-cell-density fermentation under non
-limited growth conditions. Appl Microbiol Biotechnol. 1996
Dec;46(5-6):524-32.
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