Repository hosted by TU Delft Library

Home · Contact · About · Disclaimer ·
 

Expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori

Publication files not online:

Author: Joosten, V. · Gouka, R.J. · Hondel, C.A.M.J.J. van den · Verrips, C.T. · Lokman, B.C.
Type:article
Date:2005
Institution: TNO Kwaliteit van Leven TNO Voeding
Source:Applied Microbiology and Biotechnology, 4, 66, 384-392
Identifier: 238288
doi: doi:10.1007/s00253-004-1689-0
Keywords: Biology · Biotechnology · Cloning · Degradation · Functions · Fungi · Proteins · RNA · Vanadium compounds · Aspergillus expression · Heavy-chain antibody fragments · mRNA levels · Antibodies · Antibody · Messenger RNA · Protein · Antibody · Antibody production · Aspergillus awamori · Culture medium · Expression vector · Extracellular space · Molecular cloning · Nonhuman · Nucleotide sequence · Protein degradation · Protein expression · Amino Acid Sequence · Animals · Aspergillus · Camelids, New World · Cloning, Molecular · Gene Expression · Immunoglobulin Heavy Chains · Immunoglobulin Variable Region · Molecular Sequence Data · Recombinant Proteins · Sequence Homology, Amino Acid · Transformation, Genetic · Aspergillus · Aspergillus awamori · Lama glama · vectors

Abstract

We report the expression and production of llama variable heavy-chain antibody fragments (VHHs) by Aspergillus awamori. Fragments encoding VHHs were cloned in a suitable Aspergillus expression vector and transformants secreting VHH fragments were analysed for integrated gene copy-numbers, mRNA levels and protein production. Functional V HHs were detected in the culture medium, indicating the feasibility of producing this type of protein in a fungal expression system. Secreted V HHs were subjected to (extracellular) degradation, which could be partially prevented by the addition of BSA to the culture medium. © Springer-Verlag 2004.