I like to ask for people's opions on the best method to concentrate tissue culture (TC) supernatants that contain antibody. My efforts are aimed at 1 to 5 liters of either hybridoma or CHO cell supernatants with murine or human IgG.
I started using a Vivaflow 50 PES with a 100k MWCO, as suggested by Vivasccience: http://www.vivascience.com/en/ultrafiltration/vivaflow_50.shtml
However, I've found that recovery has been absolutely horrible. I have more activity in the flow through than in the concentrate. We did not push the pressure beyond recommendation either.
I've also had poblems with the Vivaspin concentrators from this company, I have a lost antibody after spinning. Once it was clearly due to the plastic support holding the membranes breaking during the spin. Again, we're spinning these things at less than the recomended g-force. All in all this company is not on my recommend list.
I' ve got a pellicon tangential flow device on order with a 50,000 MWCO. Hopefully that will work, because this really should be quite simple. I've also switched to Millipore concnetrators (50 ml tubes-15ml supernatant size) and I'm having better luck with these.
I would love to hear what others are using to concentrate supernatants for subsequent protein A or G column purification.