Proteolysis in microfluidic droplets: an approach to interface protein separation and peptide mass spectrometry.
A versatile microreactor protocol based on microfluidic droplets has been developed for on-line protein digestion. Proteins separated by liquid chromatography are fractionated in water-in-oil droplets and digested in sequence. The microfluidic reactor acts also as an electrospray ionization emitter for mass spectrometry analysis of the peptides produced in the individual droplets. Each droplet is an enzymatic micro-reaction unit with efficient proteolysis due to rapid mixing, enhanced mass transfer and automated handling. This droplet approach eliminates sample loss, cross-contamination, non-specific absorption and memory effect. A protein mixture was successfully identified using the droplet-based micro-reactor as interface between reverse phase liquid chromatography and mass spectrometry.
Department of Chemistry, Institute of Biomedical Sciences and State Key Laboratory of Molecular Engineering of Polymers, Fudan University, Shanghai, China.
Ji
J
Ji
Lei
L
Nie
Liang
L
Qiao
Yixin
Y
Li
Liping
L
Guo
Baohong
B
Liu
Pengyuan
P
Yang
Hubert H
HH
Girault
D000595:Amino Acid Sequence; D000818:Animals; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003574:Cytochrome c Group / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D004867:Equipment Design; D006801:Humans; D046210:Microfluidic Analytical Techniques / Q000295:instrumentation*; D008969:Molecular Sequence Data; D009113:Muramidase / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009211:Myoglobin / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism; D009821:Oils / Q000737:chemistry; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D059748:Proteolysis; D021241:Spectrometry, Mass, Electrospray Ionization / Q000295:instrumentation* / Q000379:methods; D014357:Trypsin / Q000378:metabolism; D014867:Water / Q000737:chemistry
D003574:Cytochrome c Group; D009211:Myoglobin; D009821:Oils; D010455:Peptides; D011506:Proteins; D014867:Water; D009113:Muramidase; D014357:Trypsin