High-throughput protein concentration and buffer exchange: comparison of ultrafiltration and ammonium sulfate precipitation.
High-throughput protein purification is a complex, multi-step process. There are several technical challenges in the course of this process that are not experienced when purifying a single protein. Among the most challenging are the high-throughput protein concentration and buffer exchange, which are not only labor-intensive but can also result in significant losses of purified proteins. We describe two methods of high-throughput protein concentration and buffer exchange: one using ammonium sulfate precipitation and one using micro-concentrating devices based on membrane ultrafiltration. We evaluated the efficiency of both methods on a set of 18 randomly selected purified proteins from Shewanella oneidensis. While both methods provide similar yield and efficiency, the ammonium sulfate precipitation is much less labor intensive and time consuming than the ultrafiltration.
Moore
Priscilla A
PA
Molecular Biosciences, Pacific Northwest National Laboratory, Richland, WA, USA.
Kery
Vladimir
V
D000645:Ammonium Sulfate / Q000737:chemistry*; D000818:Animals; D001426:Bacterial Proteins / Q000737:chemistry*; D002021:Buffers; D011232:Chemical Precipitation; D008567:Membranes, Artificial; D020592:Shewanella / Q000737:chemistry*; D014462:Ultrafiltration / Q000379:methods*
D001426:Bacterial Proteins; D002021:Buffers; D008567:Membranes, Artificial; D000645:Ammonium Sulfate