Optimisation of insect cell growth in deep-well blocks: development of a high-throughput insect cell expression screen.
This report describes a method to culture insects cells in 24 deep-well blocks for the routine small-scale optimisation of baculovirus-mediated protein expression experiments. Miniaturisation of this process provides the necessary reduction in terms of resource allocation, reagents, and labour to allow extensive and rapid optimisation of expression conditions, with the concomitant reduction in lead-time before commencement of large-scale bioreactor experiments. This therefore greatly simplifies the optimisation process and allows the use of liquid handling robotics in much of the initial optimisation stages of the process, thereby greatly increasing the throughput of the laboratory. We present several examples of the use of deep-well block expression studies in the optimisation of therapeutically relevant protein targets. We also discuss how the enhanced throughput offered by this approach can be adapted to robotic handling systems and the implications this has on the capacity to conduct multi-parallel protein expression studies.
Bahia
Daljit
D
Respiratory Disease Area, Novartis Institutes for Biomedical Research, Wimblehurst Road, Horsham, West Sussex, United Kingdom.
Cheung
Robert
R
Buchs
Mirjam
M
Geisse
Sabine
S
Hunt
Ian
I
D000818:Animals; D016367:Baculoviridae; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D019411:Clinical Laboratory Techniques / Q000379:methods*; D005822:Genetic Vectors; D007700:Kinetics; D019869:Phosphatidylinositol 3-Kinases / Q000096:biosynthesis / Q000235:genetics; D020033:Protein Isoforms / Q000096:biosynthesis / Q000235:genetics; D043705:Receptor, Adenosine A2A / Q000096:biosynthesis / Q000235:genetics; D018411:Spodoptera; D013997:Time Factors
D020033:Protein Isoforms; D043705:Receptor, Adenosine A2A; D019869:Phosphatidylinositol 3-Kinases