PMID- 18942150
OWN - NLM
STAT- MEDLINE
DCOM- 20090806
LR  - 20131121
IS  - 1097-0290 (Electronic)
IS  - 0006-3592 (Linking)
VI  - 102
IP  - 3
DP  - 2009 Feb 15
TI  - Protein A-based antibody immobilization onto polymeric microdevices for enhanced 
      sensitivity of enzyme-linked immunosorbent assay.
PG  - 891-901
LID - 10.1002/bit.22136 [doi]
AB  - Highly efficient antibody immobilization is extremely crucial for the development 
      of high-performance polymeric microdevices for enzyme-linked immunosorbent assay 
      (ELISA). In this article, a site-selective tyrosinase (TR)-catalyzed protein A 
      strategy for antibody immobilization was developed to enhance the sensitivity of 
      ELISA in poly-(methyl methacrylate) (PMMA) microchannels for interferon-gamma 
      (IFN-gamma) assay. To effectively immobilize the target antibodies, oxygen plasma 
      was first used to activate the inert PMMA. This is followed by 
      poly(ethyleneimine) (PEI) coating, an amine-containing functional polymer. For 
      comparison, protein A was also immobilized through the commonly used 
      amine-glutaraldehyde (GA) chemistry. Oxygen plasma treatment effectively 
      increased the amount of PEI attachment and subsequent binding efficiency of the 
      primary antibody. The antibody immobilized via TR-catalyzed protein A was able to 
      provide much better specific antigen capture efficiency than GA chemistry due to 
      the optimal spacing and orientation. Consequently, by using this new method, the 
      detection signal and the signal-to-noise ratio of the ELISA immunoassay in 
      microdevices were all significantly improved. In comparison to the standard assay 
      carried out in the 96-well microtiter plate, the treated microchannels exhibited 
      a broader detection range and a shorter detection time. And the detection limit 
      was also decreased to 20 pg/mL, much lower than that obtained in other 
      microdevices.
FAU - Yuan, Yuan
AU  - Yuan Y
AD  - Department of Chemical and Biomolecular Engineering, NSF Nanoscale Science and 
      Engineering Center for Affordable Nanoengineering of Polymeric Biomedical 
      Devices, The Ohio State University, 140 West 19th Avenue, Columbus, Ohio 43210, 
      USA.
FAU - He, Hongyan
AU  - He H
FAU - Lee, L James
AU  - Lee LJ
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
PL  - United States
TA  - Biotechnol Bioeng
JT  - Biotechnology and bioengineering
JID - 7502021
RN  - 0 (Antibodies, Immobilized)
RN  - 0 (Imines)
RN  - 0 (Polyethylenes)
RN  - 0 (Staphylococcal Protein A)
RN  - 0 (poly(ethylene imine))
RN  - 82115-62-6 (Interferon-gamma)
RN  - 9011-14-7 (Polymethyl Methacrylate)
RN  - EC 1.14.18.1 (Monophenol Monooxygenase)
RN  - S88TT14065 (Oxygen)
SB  - IM
MH  - *Antibodies, Immobilized
MH  - Enzyme-Linked Immunosorbent Assay/*methods
MH  - Humans
MH  - Imines/chemistry
MH  - Interferon-gamma/*analysis/immunology
MH  - Microchemistry/*methods
MH  - Microchip Analytical Procedures/methods
MH  - Microscopy, Atomic Force
MH  - Monophenol Monooxygenase/metabolism
MH  - Oxygen/metabolism
MH  - Polyethylenes/chemistry
MH  - Polymethyl Methacrylate/chemistry
MH  - Protein Binding
MH  - Reproducibility of Results
MH  - Sensitivity and Specificity
MH  - *Staphylococcal Protein A
EDAT- 2008/10/23 09:00
MHDA- 2009/08/07 09:00
CRDT- 2008/10/23 09:00
PHST- 2008/10/23 09:00 [pubmed]
PHST- 2009/08/07 09:00 [medline]
PHST- 2008/10/23 09:00 [entrez]
AID - 10.1002/bit.22136 [doi]
PST - ppublish
SO  - Biotechnol Bioeng. 2009 Feb 15;102(3):891-901. doi: 10.1002/bit.22136.