Mammalian cell production of a respiratory syncytial virus (RSV) candidate vaccine recovered using a product-specific affinity column
2001 (English)In: Biotechnology and applied biochemistry, ISSN 0885-4513, E-ISSN 1470-8744, Vol. 34, 25-32 p.Article in journal (Refereed) Published
The recombinant production of a respiratory syncytial virus (RSV) candidate vaccine BBG2Na in baby hamster kidney cells (BHK-21 cells) was investigated. BBG2Na consists of a serum-albumin-binding region (BB) fused to a 101-amino-acid fragment of the RSV G-protein. Semliki Forest virus-based expression vectors encoding both intracellular and secreted forms of BBG2Na were constructed and found to be functional. Affinity recovery of BBG2Na employing human serum albumin columns was found to be inefficient due to the abundance of BSA in the applied samples. Instead, a strategy using a tailor-made affinity ligand based on a combinatorially engineered Staphylococcus aureus protein A domain, showing specific binding to the G-protein part of the product, was evaluated. In conclusion, a strategy for production and successful recovery of BBG2Na in mammalian cells was created, through the development of a product-specific affinity column.
Place, publisher, year, edition, pages
2001. Vol. 34, 25-32 p.
affibody, affinity chromatography, mammalian cell expression, Semliki Forest virus, Staphylococcus aureus protein A, bacterial receptor domain, taq dna-polymerase, g-protein fragment, combinatorial libraries, protective immunity, expression systems, fusion protein, binding, replicon, vectors
IdentifiersURN: urn:nbn:se:kth:diva-20911ISI: 000170684000005OAI: oai:DiVA.org:kth-20911DiVA: diva2:339608
QC 201005252010-08-102010-08-10Bibliographically approved