Hi, we are in need of chromatography systems and looking at what is available at the market. Needless to say I haven't used this technique yet so starting from scratch. We need to purify viral vectors by anion exchange and affinity chromatography. Depending on the vector type we need to purify particles of 20-150 nm diameter which are in isotonic salt solutions and eluted with high salt or isotonic buffers like Sodium citrate. The flow paths need to be bio-inert and the machines certified for cGMP conditions if possible.
The vectors are highly shear sensitive. Does that mean FPLC is what we need?
Any input on machines to look into would be highly appreciated!
Hi Ulrike,
Can you share what the scale of your purification is (volume and mass)? Also, is this a two step purification (AX clean up followed directly by affinity)?
If the vectors are shear sensitive then a low to medium chrom system is most likely needed in order to avoid unnecessary physical stress on the vectors.
Bio-Rad has an instrument called the NGC which is a great medium pressure chrom system - lots of advantages for Multi-Dimension purifications. It is completely Bio-Inert (Titanium and PEEK) and has stringent standards for Regulatory Requirements. The software and licensing is free (and also intuitive and powerful for analysis).The support you get before and after purchase is also a notable benefit over what is seen from the alternative, more commonly seen FPLCs.
For a low pressure system with slightly fewer bells and whistles there is the BioLogic Low Pressure chrom system which is super easy to use, versatile and it is a workhorse.
Do you already know what columns you will be working with?
Good luck with your research!
Katie
GE's Akta system can do this and scales to large scale cGMP very well, here is a informative white paper that may be relevant:
https://promo.gelifesciences.com/gl/BP2/misc/29043549AC_03.pdf?extcmp=BP2_SU-GL-Brochures-Influenza
CIM monoliths from BIA Separations - ion exchangers or HIC or affinity-based - are very practical and efficient for virus purification using FPLC systems. Due to their special characteristics (convective-mass transfer) they are especially reccomended for shear sensitive molecules:
http://www.biaseparations.com/applications/viruses-vlps
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