Hi everyone, apologies if this is not in the right forum.
I was wondering if there’s any vendors that sell pre-packed disposable gravity columns for HIS TAG protein purification from mammalian cells.
We have a working purification protocol on a liquid handler (Biomek) that transfers 30-120 mL of supernatant to flow through a 6 mL column packed with 0.5 mL of resin. The limitation is that we pack the columns in house, which leads to different flow rates between each column. Because of it, the protocol time is dependent on the slowest flow rate.
We would like to optimize this so that each column has the same flow rate. Therefore, we believe that finding a vendor that sells it pre-packed would solve the issue. We are open to other suggestions too!
We have looked into those types of tips before as we do have Hamilton instruments as well. However, it is not compatible for large scale volume that we are dealing with But it does hold value for pilot scale purification that we are also doing.
Unfortunately you’re looking at a scale that is too large of a scale typically handled by high-throughput lab automation. You’re either going to need to look at doing your own custom solution by packing your own columns using something like these Bio-Rad columns: https://www.bio-rad.com/en-us/category/empty-gravity-flow-columns?ID=66878000-baac-4a58-8a3c-9887c3825dd1. Or you’ll have to step up to something lower throughput but more highly controlled on an AKTA purification system. You might be able to ask Biotage about their PhyPrep (Biotage PhyPrep Maxi Mega Giga) typically used for lage scale DNA purification and see if they offer a His-Resin version.
I’ve make my own workflow in the past using gravity fed desalting columns (PD-10) and a custom jig on a Tecan Evo, but since its gravity fed you will definitely not have control over flowrate like a typical packed column. You can always work out a fixed volume and fixed timing situation where you might not care about maximally efficient purification.
One other place to check is DPX Technologies (https://dpxtechnologies.com/). They have an open tip format where you may be able to fill some empty tips with your own resin or purchase them pre-filled. But I don’t think they have a 6mL tip format. Doesn’t hurt to ask though. Good luck!
But it does hold value for pilot scale purification that we are also doing.