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I'm not sure of your exact situation, but the first step is to carefully reread the method. You can either restart the method and you next breakpoint to quickly get you back where you need to be. The other option with out restarting the method is to use the "block" instruction to just re-read in the elution block where the air sensor prematurely triggered. Gook luck! btw I haven't covered the "block" instruction yet.

Nice! I have those systems too.

I'm glad you liked the video!

Yes and no. I would say you want to do a pump wash every time you start a new run, but not every time you switch to a new inlet position. So say you start a run by washing inlets 1 and 2 you don't have to rewash them later in the middle of the run to switch from one inlet to the next. UNLESS the solution in one of the inlets would totally muck up the run some how, like in affinity chromatography, for example you don't want to have Ni anywhere near the column when loading you sample with a His-tag (HisTrap) column.

Glad you liked it!

I'm glad you liked this video! I worked really hard on it and to find that someone found it useful is great!

Thanks for watching!

I have a whole series of videos about navigating modules so you can get the chromatogram you are looking for. They were some of the first videos I did, if you haven't seen them I'd recommend checking them out, czcams.com/video/m5kp9wM63ZQ/video.html

It's hard to know what's going on with the information you provided. Tell me more or message me a snap shot of your chromatogram. Here are some guesses about what I think is going on. Did you integrate the peak? The area integrated needs to just be the peak of interest. Be sure the baseline is properly drawn/calculated, you can really only adjust the baseline in the classic evaluation module (so far as I know). Sorry I didn't answer quickly...

Good question. Ideally you should do it before, the slurry will settle very quickly if it is fully equilibrated, but if you don't mind waiting for it to settle then you can do it while it is on the column.

Thanks for the idea!

Click on the text tab in the middle of the screen, go to 2:43 mark in this video to see what I'm talking about. You'll have to read through the method to find the setting you want. Once you find the setting be sure to press the change button, then save. Good luck.

Thanks, I'm glad you liked the video! You can change the unit for results. In any of the modules go to Tools in the menu bar and select Options. Then in the window that pops up select the View tab, towards the bottom of the tab there is an option for Pressure Unit. It doesn't look like you can change the units in the Method Editor or System Control, only for results. It's a good question though.

Glad it was helpful! I don't know why this video was such a challenge to make, probably because everyone is going to have something different, so I'm glad you liked it enough to comment.

I hope to soon. I have so many ideas and each video takes a bit to make. Is there a kind of column in particular you want to see packed?

HIC

Done! I did one, I should have some more up sometime in the near future. Not making any promises. czcams.com/video/z02_8-yjmOk/video.html

Yeah you could, you'd either need two valves plumbed like the outlet valve or one like the column valve. There's no reason you couldn't have a second column valve you'd just have you use your imagination when it came to understanding the flow path.

Glad it was helpful! Really means a lot! I'm trying to gear this channel towards people, such as yourself, that are new to the Avant and other ÄKTA systems.

☺@@UnicornStuff Wish your channel gets more views and more people learn from here. Surprisingly, when I search for Äkta, the recommendation did not come directly; it only came up when I searched with Unicorn. Such a nice channel.❤

What really! I never imagined someone would actually use my content. So excited my videos actually fill a need. Cheers!

Glad it was helpful! It can be really helpful to get the hang of Unicorn and the Avant at first.

Good question. There could be a variety of reasons all of which could be true in this case. There could be impurities in the sample that elute later. There could one target protein, but it is heterogeneous because of post transnational modifications. I believe the best answer however is the concentration of salt wasn't strong enough to get all the protein off for that step. The solution would be to increase the step length or concentration a little bit. Sorry for the late reply.

You're welcome! I'm planning on making more videos.

Glad to hear that!

Great question. Most protein A type resins will foul if you use >10 mM NaOH because the ligand is unstable in higher concentrations. There are some NaOH resistant resins MabSelect (Cytiva) and Toyopearl (Tosoh) are my two favorite.

No it is not, one of the check valves is always open. It is necessary to pump wash the system afterwards regardless. Thanks for your question.

Glad you liked it!

Glad it was helpful!

Thanks for your feedback. I'm working on using the audio editor more. I'd like to have background music to cover up some of the breathing noises I make. Hopefully we find something we both like.

@@UnicornStuff I see. Understand! Thank you so much! I

Thanks! Yes I'm making more & super excited you found my channel!

@@UnicornStuff looking forward to seeing more about how to use the unicorn software , methods editor and results evaluation 🙏👍