PMID- 15595729
OWN - NLM
STAT- MEDLINE
DCOM- 20050419
LR  - 20131121
IS  - 1535-3893 (Print)
IS  - 1535-3893 (Linking)
VI  - 3
IP  - 6
DP  - 2004 Nov-Dec
TI  - Titania and alumina sol-gel-derived microfluidics enzymatic-reactors for peptide 
      mapping: design, characterization, and performance.
PG  - 1201-9
AB  - The design and characterization of titania-based and alumina-based 
      Poly(dimethylsiloxane) (PDMS) microfluidics enzymatic-reactors along with their 
      analytical features in coupling with MALDI-TOF and ESI-MS were reported. 
      Microfluidics with microchannel and stainless steel tubing (SST) were fabricated 
      using PDMS casting and O(2)-plasma techniques, and were used for the preparation 
      of an enzymatic-reactor. Plasma oxidation for the PDMS microfluidic system 
      enabled the channel wall of the microfluidics to present a layer of silanol 
      (SiOH) groups. These SiOH groups act as anchors onto the microchannel wall linked 
      covalently with the hydroxyl groups of trypsin-encapsulated sol matrix. As a 
      result, the trypsin-encapsulated gel matrix was anchored onto the wall of the 
      microchannel, and the leakage of gel matrix from the microchannel was effectively 
      prevented. A feature of the microfluidic enzymatic-reactors is the feasibility of 
      performing on-line protein analysis by attached SST electrode and replaceable 
      tip. The success of trypsin encapsulation was investigated by AFM imaging, assay 
      of enzymatic activity, CE detection, and MALDI-TOF and ESI-MS analysis. The 
      lab-made devices provide an excellent extent of digestion even at a fast flow 
      rate of 7.0 microL/min, which affords the very short residence time of ca. 2 s. 
      With the present device, the digestion time was significantly shortened compared 
      to conventional tryptic reaction schemes. In addition, the encapsulated trypsin 
      exhibits increased stability even after continuous use. These features are 
      required for high-throughput protein identification.
FAU - Wu, Huiling
AU  - Wu H
AD  - Department of Chemistry, Research Center for Proteome, and Institute of Genetics, 
      Fudan University, Shanghai 200433, People's Republic of China.
FAU - Tian, Yuping
AU  - Tian Y
FAU - Liu, Baohong
AU  - Liu B
FAU - Lu, Haojie
AU  - Lu H
FAU - Wang, Xiaoyan
AU  - Wang X
FAU - Zhai, Jianjun
AU  - Zhai J
FAU - Jin, Hong
AU  - Jin H
FAU - Yang, Pengyuang
AU  - Yang P
FAU - Xu, Yunmin
AU  - Xu Y
FAU - Wang, Honghai
AU  - Wang H
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - J Proteome Res
JT  - Journal of proteome research
JID - 101128775
RN  - 0 (Dimethylpolysiloxanes)
RN  - 0 (Enzymes, Immobilized)
RN  - 0 (Proteins)
RN  - 0 (Silicones)
RN  - 15FIX9V2JP (titanium dioxide)
RN  - 63148-62-9 (baysilon)
RN  - D1JT611TNE (Titanium)
RN  - EC 3.4.21.4 (Trypsin)
RN  - LMI26O6933 (Aluminum Oxide)
SB  - IM
MH  - Aluminum Oxide
MH  - Dimethylpolysiloxanes
MH  - Electrodes
MH  - Enzymes, Immobilized/*metabolism
MH  - Equipment Design
MH  - Mass Spectrometry
MH  - Microfluidic Analytical Techniques/*instrumentation/methods
MH  - Peptide Mapping/*methods
MH  - Phase Transition
MH  - Proteins/analysis
MH  - Silicones
MH  - Surface Properties
MH  - Titanium
MH  - Trypsin/*metabolism
EDAT- 2004/12/15 09:00
MHDA- 2005/04/20 09:00
CRDT- 2004/12/15 09:00
PHST- 2004/12/15 09:00 [pubmed]
PHST- 2005/04/20 09:00 [medline]
PHST- 2004/12/15 09:00 [entrez]
AID - 10.1021/pr049889z [doi]
PST - ppublish
SO  - J Proteome Res. 2004 Nov-Dec;3(6):1201-9. doi: 10.1021/pr049889z.