Miho Suzuki

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Organization: Saitama University , Japan
Department: Department of Functional Materials and Science
Title: Research Fellow(PhD)

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Co-reporter:Miho Suzuki, Hikari Udaka, Takeshi Fukuda
Journal of Pharmaceutical and Biomedical Analysis 2017 Volume 143(Volume 143) pp:
Publication Date(Web):5 September 2017
DOI:10.1016/j.jpba.2017.05.014
•Antibody-attached quantum dots were used to establish an ELISA-like assay, QLISA.•Antibody was reduced to expose thiol groups for directed linkage to quantum dots.•Efficiently labeled antibodies with bright quantum dots could amplify QLISA signal.•QLISA was proven to be a general time- and effort-saving assay.An approach similar to the enzyme-linked immunosorbent assay (ELISA), with the advantage of saving time and effort but exhibiting high performance, was developed using orientation-directed half-part antibodies immobilized on CdSe/ZnS quantum dots. ELISA is a widely accepted assay used to detect the presence of a target substance. However, it takes time to quantify the target with specificity and sensitivity owing to signal amplification. In this study, CdSe/ZnS quantum dots are introduced as bright and photobleaching-tolerant fluorescent materials. Since hydrophilic surface coating of quantum dots rendered biocompatibility and functional groups for chemical reactions, the quantum dots were modified with half-sized antibodies after partial reduction. The half-sized antibody could be bound to a quantum dot through a unique thiol site to properly display the recognition domain for the core process of ELISA, which is an antigen-antibody interaction. The reducing conditions were investigated to generate efficient conjugates of quantum dots and half-sized antibodies. This was applied to IL-6 detection, as the quantification of IL-6 is significant owing to its close relationships with various biomedical phenomena that cause different diseases. An ELISA-like assay with CdSe/ZnS quantum dot institution (QLISA; Quantum dot-linked immunosorbent assay) was developed to detect 0.05 ng/mL IL-6, which makes it sufficiently sensitive as an immunosorbent assay.Download high-res image (85KB)Download full-size image
Co-reporter:Miho Suzuki, Ichiro Sakata, Takafumi Sakai, Hiroaki Tomioka, Koichi Nishigaki, Marc Tramier, Maïté Coppey-Moisan
Analytical Biochemistry 2015 Volume 491() pp:10-17
Publication Date(Web):15 December 2015
DOI:10.1016/j.ab.2015.08.022

Abstract

Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis.

Dibenz[a,e]aceanthrylene
5H-Phenanthro[4,5-bcd]pyran-5-one
PUROMYCIN
Benzo(c)fluorene
5'-Adenylic acid,2'-deoxy-, homopolymer
9-Methylphenanthrene
Dibenzo[def,mno]chrysene
Dibenzo[def,p]chrysene
Benzo[rst]pentaphene
Streptavidin