Development of a microfluidics-based gel protein recovery system

✍ Scribed by Trust Tariro Razunguzwa; April Biddle; Heather Anderson; Dongliang Zhan; Matthew Powell

• Publisher: John Wiley and Sons
• Year: 2009
• Tongue: English
• Weight: 761 KB
• Volume: 30
• Category: Article
• ISSN: 0173-0835
• DOI: 10.1002/elps.200900485

✦ Synopsis

Development of a microfluidics-based gel protein recovery system

A PMMA microfluidic chip, in which fluid is manipulated to transport protein from a PAGE gel piece to a collection reservoir via a microfluidic channel, has been developed. The protein sample is mobilized out of the gel (loaded in a chip access hole) into a low EOF-CE microfluidic channel under the influence of an electric field. Simultaneously, hydrostatic pressure from the filled buffer reservoirs is used to direct the protein sample to a third reservoir, through a field-free channel connected to the electrophoresis channel. Using this novel process of protein transport from a gel sample, proteins from Coomassie-stained gels have been transferred into solution in 15-30 min, with good sample recovery, using a run buffer containing an anionic acid-labile surfactant. A variety of small-and medium-sized proteins were successfully recovered and detected using both electrospray and MALDI MS over gel loads of 0.1-10 mg. This technological tool is very important for extracting quality intact protein samples from polyacrylamide gels, from which accurate protein molecular weights and protein sequences can be obtained using intact molecules.