In an ion-exchange chromatography, if 14cm length and 5cm diameter constitute 290mL resin can pass 800mL sample then how to select column if we want to run 2500mL sample. Here, the resin should bind DNA/proteins or other impurities.
Your any kind of explanation would be highly appreciated. Thank you.
The capacity of an ion exchange column is determined by the number of ionic sites per gram of column packing.
You can just proportion the column.
(290 mL column)/ (800 mL sample) = x/((2500 mL sample) x= ~906 mL column packing.
Scale up can be estimated using simple math. We have a general rule in chromatography. If you double the diameter of the column and keep the length and particle size (media) the same, then your capacity (loading) will increase 4x. *That should be enough for your example and would result in the same back-pressure at the same flow rate.
What you really need to do first is conduct a loading study with your existing set-up to determine how much material you can load onto the column before separation is lost. Once you determine the maximum load, you can also determine how much column you need. Isn't science (or perhaps math in this case) wonderful?
Dear Sir. Concerning your issue, you can use the van Deemter equation to derive a rule scaling. The scaling is done by the concept of time scales. The time scales are kept identical for all the columns by scaling the flow rate to the total void volume and the load to the amount of the media. The gradient retention behaviour of protein is independent of the column aspect ratio, but it's a function of the initial salt concentration and the gradient slop G′. For more information you can visit the link:
http://www.sciencedirect.com/science/article/pii/S0021967306005917
I hope that would helpful.
Thanks.
concerning the scaling up for coulmn chromatography, start first with batch experimnet and then start build up the coulumn which you need to do some experiment for the right way of extraction
Dear Professor Sayed,
Thank you for your response. I did several experiments regrading this issue. In my conclusion, a 800 mL viral sample (that not much viscous) can easily run through a 274 ml packed column. Now, I want to scale-up it with 8000mL sample then how I can maintain my column length and diameter? Suppose, I have experimented with a column length 240mm and diameter 60mm.
http://images.alfresco.advanstar.com/alfresco_images/pharma/2014/08/22/4d954911-b358-4e43-963e-ee16f20497d5/article-43408.pdf
red the following it will help
Dear Van So Le,
The diamter is important for ion-exchange but If I intend to increase the column height rather than diameter in case of unavailability of widen column then how possibility to get a purify product. Suppose, I want to use 50cm length and 50cm diamter column for 2500mL sample run where the total resin volume same as 28 cm length, 6.42 cm diameter column. Thank you.
Dear Bill Letter,
Thanks for your explanation regarding my query. According to your information I need to double the column diameter to increase 4X my loading capacity but how I can resolve the problem if this type of column is unavailable. I have also a query that, will I need to adjust the resin volume by increasing the column length while column diameter is crucial for ion-exchange.
I was trying to convey to you that you can use simple math to understand the relationship that the surface area of the media has relative to the loading capacity. You do not have the use the exact sizes in my example. *Strongly suggest you do a loading study first so you can determine the exact capacity needed first.
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