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

• Published in: Applied microbiology and biotechnology Vol. 66; no. 4; pp. 384 - 392
• Main Authors: Joosten, Vivi, Gouka, Robin J, van den Hondel, Cees A M J J, Verrips, C Theo, Lokman, B Christien
• Format: Journal Article
• Language: English
• Published: Germany 01-01-2005
• Subjects: Amino Acid Sequence; Animals; Antibodies; Aspergillus - genetics; Aspergillus awamori; Biodegradation; Camelids, New World - genetics; Camelids, New World - immunology; Cloning, Molecular; Expression vectors; Gene Expression; Immunoglobulin Heavy Chains - biosynthesis; Immunoglobulin Heavy Chains - genetics; Immunoglobulin Variable Region - biosynthesis; Immunoglobulin Variable Region - genetics; Molecular Sequence Data; mRNA; Q1; Q2; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; Sequence Homology, Amino Acid; Transformation, Genetic
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-004-1689-0

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