We are using amicon centrifugal filter units (15ml) to concentrate cell culture medium containing a monoclonal antibody. Since we have a large quantity of medium to concentrate (3L) and the units are very expensive, I was wondering if anybody can suggest if they can be used multiple times. In theory it shouldn't be a problem, but it would be good to hear from someone who has done this successfully already.
For such high volumes you should rather try stirred cells from Millipore (https://www.millipore.com/catalogue/module/c3259) or other companies especially if you plan to concentrate antibodies on a regular basis. The centrifugal filters may be washed and reused since you plan to concentrate the same supernate but with time they will clog and besides they are not very practical for such volumes
For such high volumes you should rather try stirred cells from Millipore (https://www.millipore.com/catalogue/module/c3259) or other companies especially if you plan to concentrate antibodies on a regular basis. The centrifugal filters may be washed and reused since you plan to concentrate the same supernate but with time they will clog and besides they are not very practical for such volumes
Of course it's not ideal, but in theory i can't see any problem with it so long as the filter doesn't clog. Millipore stirred cells is not an option for us as we are only doing this as a once off and are not planning on isolating more antibodies in the future. I have given this a try and was able to re-use each filtering unit up until 3 times without compromising recovery and with a good held overall.
Sorry I missed this part of the question. When I was using centrifugal filters for protein concentration, I was incubating them before with 1% BSA (in the same buffer of the protein extract). Subsequently I was washing them with buffer with 0.1% SDS (a couple of times) and I was reusing as Francesco suggested with no particular losses in the ultrafiltrate
I was told by a tutor that such centrifugal filters could be reused continuously, a critical step, so said that tutor, was to not let the membrane dry itself, for that reason he added a solution of PBS 1X with sodium azide to the 0.02% (the sodium azide to avoid contamination) , I did re used the centrifugal filters several times, and got good results for practical terms, i did not had however time to check how many re-uses where possible and how was the concentration efficiency of the membrane affected by re-using it
Instead of clogging, I found that Amicon centrifugal filtration columns get leaky after reused for 5-6 times. I lost quite a lot of gold nanoparticles when I tried to filter them.
Hi George,
when you say you incubate the column with 1% BSA do you mean you run 1% BSA through the column or just let this solution sit in the column? If the answer is latter, how long do you incubate? Also what is the purpose of 0.1% SDS when you wash your columns?
Hi Sujeet,
Yes, you can. I used to reuse them. I used to store them as described by Eduardo.
Hi Sook,
From your experience, how many times do you suggest the filter membrane be used maximally before any leakage can occur? Secondly, how and what did you use for cleaning before using again?
I am waiting for your response. Thanks
I would not reuse them if working with mutants. It is difficult to clean them completely and the leftover may cause problems.
Mubashshir Rasheed do you know how long you were able to store them for?
Kate Secombe I used to store them 4-6 months depending on the usage. With the frequent use the efficiency to concentrate goes down, therefore, I will suggest not to reuse them more than 5-6 times.
Dear all,
Is it okay to reuse the same protein concentrator for different proteins, after all the cleaning is done?
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