Zeolite nanoparticle modified microchip reactor for efficient protein digestion.
An enzymatic microreactor has been fabricated based on the poly(methyl methacrylate) (PMMA) microchchip surface-modified with zeolite nanoparticles. By introducing the silanol functional groups, the surface of PMMA microchannel has been successfully modified with silicalite-1 nanoparticle for the first time due to its large external surface area and high dispersibility in solutions. Trypsin can be stably immobilized in the microchannel to form a bioreactor using silica sol-gel matrix. The immobilization of enzyme can be realized with a stable gel network through a silicon-oxygen-silicon bridge via tethering to those silanol groups, which has been investigated by scanning electron microscopy and microchip capillary electrophoresis with laser-induced fluorescence detection. The maximum proteolytic rate constant of the immobilized trypsin is measured to be about 6.6 mM s(-1). Using matrix assisted laser desorption and ionization time-of-flight mass spectrometry, the proposed microreactor provides an efficient digestion of cytochrome c and bovine serum albumin at a fast flow rate of 4.0 microL min(-1), which affords a very short reaction time of less than 5 s.
Department of Chemistry, Fudan University, Shanghai 200433, People's Republic of China.
Yi
Y
Huang
Wei
W
Shan
Baohong
B
Liu
Yun
Y
Liu
Yahong
Y
Zhang
Yue
Y
Zhao
Haojie
H
Lu
Yi
Y
Tang
Pengyuan
P
Yang
D000595:Amino Acid Sequence; D045304:Cytochromes c / Q000032:analysis / Q000737:chemistry; D046209:Electrophoresis, Microchip; D004800:Enzymes, Immobilized / Q000737:chemistry; D007700:Kinetics; D008855:Microscopy, Electron, Scanning; D008969:Molecular Sequence Data; D053758:Nanoparticles / Q000737:chemistry*; D019904:Polymethyl Methacrylate / Q000737:chemistry; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000032:analysis / Q000737:chemistry; D012821:Silanes / Q000737:chemistry; D017640:Silicates / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014357:Trypsin / Q000737:chemistry; D017641:Zeolites / Q000737:chemistry*
D004800:Enzymes, Immobilized; D011506:Proteins; D012821:Silanes; D017640:Silicates; C082343:silanol; D017641:Zeolites; D012710:Serum Albumin, Bovine; D045304:Cytochromes c; D019904:Polymethyl Methacrylate; D014357:Trypsin