Mammalian cell production of a respiratory syncytial virus (RSV) candidate vaccine recovered using a product-specific affinity column.
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.
Andersson
C
C
Department of Biotechnology, Royal Institute of Technology (KTH), SE-100 44 Stockholm, Sweden.
Hansson
M
M
Power
U
U
Nygren Pk
Ståhl
S
S
D000818:Animals; D015153:Blotting, Western; D002460:Cell Line; D002846:Chromatography, Affinity; D006224:Cricetinae; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D015151:Immunoblotting; D008856:Microscopy, Fluorescence; D008956:Models, Chemical; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000378:metabolism; D022261:Respiratory Syncytial Virus Vaccines / Q000737:chemistry*; D012136:Respiratory Syncytial Viruses / Q000378:metabolism*; D012709:Serum Albumin / Q000737:chemistry; D013211:Staphylococcus aureus / Q000737:chemistry; D014158:Transcription, Genetic; D014162:Transfection
D011993:Recombinant Fusion Proteins; D022261:Respiratory Syncytial Virus Vaccines; D012709:Serum Albumin