Practical Course Dr. Uwe Horn/ Prof Dr. Dirk Hoffmeister

From Strains to Biotechnological Processes

2 - 3 November 2010, HKI, Biotechnikum, meeting point 9:00 HKI main building, foyer

Limited number of  participants :10 * 

*In order to provide each participant a thorough hands on experience we are restring the number of participants to 10. 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.