Nanocrystal Quantum Dot-Conjugated Anti-Myeloperoxidase Antibody as the Detector of Activated Neutrophils

Fluorescent nanocrystal quantum dots (QDs) have been applied to a wide range of biological studies by taking advantage of their fluorescence properties. Here we show that QDs conjugated with antibody against neutrophil peroxidase, myeloperoxidase (MPO). We designed a novel method to conjugate QDs to...

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Bibliographic Details
Published in:IEEE transactions on nanobioscience 2007-12, Vol.6 (4), p.341-345
Main Authors: Hoshino, A., Nagao, T., Nakasuga, A., Ishida-Okawara, A., Suzuki, K., Yasuhara, M., Yamamoto, K.
Format: Magazinearticle
Language:English
Subjects:
Activated
Activation analysis
Antibodies
Antibodies, Antineutrophil Cytoplasmic - administration & dosage
Antibodies, Antineutrophil Cytoplasmic - chemistry
Antigen-Antibody Complex
Antigens, Surface - analysis
Biochemistry
bioimaging
Cell Separation
Dentistry
Detectors
diagnosis of activated neutrophils
Diseases
Flow Cytometry
Fluorescence
Fluorescent Antibody Technique, Direct - methods
Humans
Immune system
Medical diagnostic imaging
Nanocrystals
Nanoparticles - chemistry
Nanostructure
Neutrophil Activation
Neutrophils - immunology
Peroxidase - analysis
Peroxidase - immunology
Probes
Quantum dot conjugated with antibody
Quantum Dots
Staining and Labeling
Surface chemistry
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Summary:Fluorescent nanocrystal quantum dots (QDs) have been applied to a wide range of biological studies by taking advantage of their fluorescence properties. Here we show that QDs conjugated with antibody against neutrophil peroxidase, myeloperoxidase (MPO). We designed a novel method to conjugate QDs to antibody without losing any antibody function including their antigen recognizing and Fc-receptor binding activities. When we applied anti-MPO antibody (Ab) with conventional organic probes in the case of immunostaining of living cells, the antibodies lost their fluorescence because of MPO enzymic activity to produce reactive oxygen species. Our QD-conjugated anti-MPO (alpha-MPO-QDs) can detect MPO on the surface of activated neutrophils. In addition, anti-MPO-QDs did not react to the inactivated neutrophils. In conclusion, we demonstrated that antibody visualized the expression of MPO on the neutrophil surface after stimulation with proinflammatory cytokines. Taken together, these techniques have the possibility that QDs can reveal the activation of neutrophils by immunostaining and flow cytometric analysis as a powerful tool for diagnosis of the neutrophil activation in vitro .
ISSN:1536-1241
1558-2639
DOI:10.1109/TNB.2007.909008