PMID- 17321125
OWN - NLM
STAT- MEDLINE
DCOM- 20070726
LR  - 20090714
IS  - 0956-5663 (Print)
IS  - 0956-5663 (Linking)
VI  - 22
IP  - 12
DP  - 2007 Jun 15
TI  - Enzyme immunosensing of pepsinogens 1 and 2 by scanning electrochemical 
      microscopy.
PG  - 3099-104
AB  - Scanning electrochemical microscopy (SECM) was applied to a dual enzyme 
      immunoassay for the detection of pepsinogen 1 (PG1) and pepsinogen 2 (PG2). 
      Sandwich-type immunocomplexes labeled with horseradish peroxidase (HRP) were 
      constructed on microspots consisting of anti-PG1 IgG antibody and anti-PG2 IgG 
      antibody. These microspots were fabricated on a hydrophobic glass substrate using 
      a capillary microspotting technique. In the presence of H(2)O(2) and 
      ferrocenemethanol (FcOH; used as an electron mediator), the labeled HRP catalyzed 
      the oxidation of FcOH by H(2)O(2) to generate the oxidized form of FcOH (Fc(+)OH) 
      at localized areas corresponding to microspots containing both immunocomplexes. 
      The enzymatically generated Fc(+)OH was reduced and detected with a SECM probe 
      (0.05 V versus Ag/AgCl), and the substrate surface was mapped to generate SECM 
      images of the PG1 and PG2 spots. Relationships between the reduction current in 
      the SECM images and the concentrations of PG1 and PG2 were obtained in the range 
      1.6-60.3 ng/ml protein. Dual imaging of PG1 and PG2 was achieved using microspots 
      containing PG1 and PG2 immunocomplexes separated by a 200 microm physical barrier 
      on the substrate. Pyramidal hole arrays with 100 microm x 100 microm openings on 
      the silicon wafer were utilized to fabricate spots using antibodies on 
      poly(dimethylsiloxane) (PDMS) membranes. Current responses obtained from 
      microspots fabricated with pyramidal holes are significantly sharper compared to 
      the responses obtained from spots fabricated using the capillary method.
FAU - Yasukawa, Tomoyuki
AU  - Yasukawa T
AD  - Graduate School of Environmental Studies, Tohoku University, 6-6-11 Aoba, 
      Aramaki, Aoba, Sendai 980-8579, Japan. yasu@bioinfo.che.tohoku.ac.jp
FAU - Hirano, Yu
AU  - Hirano Y
FAU - Motochi, Naomi
AU  - Motochi N
FAU - Shiku, Hitoshi
AU  - Shiku H
FAU - Matsue, Tomokazu
AU  - Matsue T
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20070128
PL  - England
TA  - Biosens Bioelectron
JT  - Biosensors & bioelectronics
JID - 9001289
RN  - 0 (Dimethylpolysiloxanes)
RN  - 0 (Membranes, Artificial)
RN  - 0 (Silicones)
RN  - 61536-72-9 (Pepsinogen C)
RN  - 63148-62-9 (baysilon)
RN  - 9001-10-9 (Pepsinogen A)
SB  - IM
MH  - Biosensing Techniques/*instrumentation/methods
MH  - Dimethylpolysiloxanes
MH  - Electrochemistry/*instrumentation
MH  - Immunoenzyme Techniques/*instrumentation/methods
MH  - Membranes, Artificial
MH  - Microscopy/*methods
MH  - Pepsinogen A/*analysis
MH  - Pepsinogen C/*analysis
MH  - Silicones
EDAT- 2007/02/27 09:00
MHDA- 2007/07/27 09:00
CRDT- 2007/02/27 09:00
PHST- 2006/10/03 00:00 [received]
PHST- 2006/12/22 00:00 [revised]
PHST- 2007/01/23 00:00 [accepted]
PHST- 2007/02/27 09:00 [pubmed]
PHST- 2007/07/27 09:00 [medline]
PHST- 2007/02/27 09:00 [entrez]
AID - S0956-5663(07)00036-X [pii]
AID - 10.1016/j.bios.2007.01.015 [doi]
PST - ppublish
SO  - Biosens Bioelectron. 2007 Jun 15;22(12):3099-104. doi: 
      10.1016/j.bios.2007.01.015. Epub 2007 Jan 28.