We are buying new protein purification equipment for our company and need to decide if we will go for the more expensive Akta Avant or for the less expensive Akta FPLC. The plan with the equipment is to develop protein purification protocols that we can use during the protein production phase for FDA preclinical and clinical trials.
We will not be using the equipment for the actual production (this part will be outsourced to cGMP compliant facility), the equipment will only be used for the development of the protocol in as short a time as possible.
The protocol that we develop should also naturally be upscalable.
Which of these 2 pieces of equipment would you recommend and why? Or maybe you would recommend something else? Please advise.
I am in the same situation as you, albeit in academia, so I look forward to the responses here as well. I was looking at the AKTA Purifier, as well as the Avant. The GE rep is coming by tomorrow, I can pass along any info I get after his visit.
We use the AKTA Purifier in our lab, and along with Superdex columns, it does a fantastic job. It even performs well as a makeshift HPLC using a high-pressure attachment. We primarily do isolation of monomeric, oligomeric, or fibrillar amyloidogenic proteins, and have found that it provides great resolution and an excellent feature set. I'm not sure what extras you would get with the Avant, but (with the right setup), the Purifier provides support for multiple wavelength UV monitoring, automated fractionation, multiple pumps, etc.
Matthew, how easy is it to run back-to-back purifications on the Purifier? For example if I want to perform a crude purification and collection (e.g. from a His column) and immediately turn around and run that sample through a "polishing" step with the Superdex, is this easy to do? Also, is there a specification for how much volume can be purified during a run? In my case I'd like to purify a protein that is secreted into the media, so after collecting supernatants from a few T175 flasks, you can imagine I have a large volume to purify. Thanks for the input!
Back-to-back purifications are quite easy if you're using two separate columns. The process of changing a column can be done in less than a minute, and from there it's just a matter of loading your sample to be purified and starting a run. Run parameters can be preprogrammed to make starting one quick and easy. Within the run parameters you can specify things like gradients, maximum column pressure, flow rate, fraction volume, etc. Doing back to back purifications on the same column can be done, but I'd recommend re-equillibrating the column in buffer first, which will take some time.
I have not done large-volume purifications myself. We usually resuspend HFIP protein films or synthetic protein in 500 uL and load on a 1 mL loop. However, GE also sells "Superloops," which come in 10, 50, and 150 mL varieties for large-volume purifications. Harvard has a nice guide on using them (and a description of why the "partial filling" method is better than loading the entire loop volume) here: http://labs.idi.harvard.edu/springer/uploads/Protocols/Equipment%20Manuals/aktapurifierinstructionalmaterial.pdf
If you want to run back to back column with two different columns then AKTA Explorer is better option. Even in this system you don't need to change the column after 1st run. There are 8 different column positions and you can attach 8 different column to the system and can run 8 different columns back to back. Avant has advantage of having temperature control system in it. If you want to run your protein in room temperature condition then I think explorer is better option.
Please let us know if you have some new ideas now.
Regards.
You can purify as much amount of protein you want by changing the size of the column.
Does anybody have experience using the Akta FPLC specifically? Akta purifier is being discontinued and replaced by the Akta Pure, which is also much more expensive than the AKTA fplc. We would like to go for this one unless there are some very specific reasons to rather go for the more expensive models. Akta explorer is also discontinued and was replaced by the Avant.
I am also on the same boat. Trying to decide between the Pure and the FPLC. The problem is that my budget is more like FPLC but what we need is more on the Pure side.
Either one, will be a great asset to our proteomics facility, helping on the sample prep and purification of target proteins.
Same issue here, what we really would like to buy is the Avant, but trying to figure out if we can get away with the fplc, much better on the budget :)
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