Scientists at Batavia Bioservices BV have developed and optimized DNA transfection protocols for CHO, Insect cells (i.e. SF9) and HEK293 cells. This technology allows for transient protein production in combination with commercially available, fully animal component free cell culture media.
The technology can be applied at small (10-20ml) scale in tube spin reactors all the way up to 50L scale, allowing the progression from DNA to delivery of up to multi-milligram quantities of material for screening or proof-of-concept studies within 3-6 weeks. This technology therefore provides clients with high quality research material for proof of concept studies or candidate selection studies within a matter of weeks thereby significantly reducing both time and cost. Hereunder three case studies that merit the spin tube technology are described.
The technology is applied in a high throughput mode at a very early stage of candidate selection. For example, multiple antibody variants against a single target have been expressed simultaneously by performing high throughput parallel transient transfections at 20ml scale. These variants have then been purified using small scale, single step purification technologies and analyzed for in-vitro activity by receptor binding studies. The process from DNA to analyzed functional data takes 3-4 weeks, resulting in the possibility for clients to rapidly perform early stage candidate selection.
The technology is also applied at a larger (1-2L) scale to screen multiple protein variants or muteins to determine for example in-vitro or in-vivo functionality such as efficacy or half-life. For example, multiple variants of multiple proteins including interferons, fertility hormones, fusion proteins and peptides have been expressed in parallel at 1-2L scale. Following purification, final material was release tested and supplied to the customer for proof-of-concept (POC) testing. Productivities are typically in the 1-10 mg/L range, enabling the production of mg quantities of purified protein for testing.
Should additional or larger POC studies be required, large scale transient production can be performed at up to 50L scale, which can deliver sufficient material to perform extensive efficacy testing within a period of 6-8 weeks.
Relevant publication:Adv Biochem Eng Biotechnol. 2010;115:33-53. Use of orbital shaken disposable bioreactors for Mammalian cell cultures from the milliliter-scale to the 1,000-liter scale. Zhang X, et al