Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis

Tatsuya Munaka, Hirohisa Abe, Masaki Kanai, Takashi Sakamoto, Hiroaki Nakanishi, Tetsuji Yamaoka, Shuichi Shoji, Akira Murakami

    Research output: Contribution to journalArticle

    5 Citations (Scopus)

    Abstract

    This article presents a real-time monitoring system for cellular analysis using micro total analysis systems technology. Time-resolved luminescence anisotropy analysis was adopted for real-time detection of small amounts of a target protein produced by a small number of cells. The system was tested by real-time monitoring of the antibody secretion by hybridomas. The cells were successfully cultivated in a micro-incubation chamber (240 nl) fabricated on a microchip. The quantification of the antibody was achieved using the Ru(II) complex-labeled Staphylococcus aureus protein A probe, which can bind specifically to the Fc region of the antibody. Using this system, we detected as little as 24 fmol of immunoglobulin G under physiological conditions without the bound/free separation protocol. We successfully achieved real-time and quantitative monitoring of small amounts of antibody production by approximately 200 hybridoma cells. This method could be applied to various cellular analyses using small numbers of cells.

    Original languageEnglish
    Pages (from-to)1-6
    Number of pages6
    JournalAnalytical Biochemistry
    Volume353
    Issue number1
    DOIs
    Publication statusPublished - 2006 Jun 1

    Fingerprint

    Anisotropy
    Hybridomas
    Luminescence
    Antibodies
    Monitoring
    Cell Count
    Computer Systems
    Staphylococcal Protein A
    Antibody Formation
    Staphylococcus aureus
    Immunoglobulin G
    Technology
    Proteins

    Keywords

    • μTAS
    • Cellular analysis
    • Real-time monitoring
    • Time-resolved luminescence anisotropy analysis

    ASJC Scopus subject areas

    • Biochemistry
    • Biophysics
    • Molecular Biology

    Cite this

    Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis. / Munaka, Tatsuya; Abe, Hirohisa; Kanai, Masaki; Sakamoto, Takashi; Nakanishi, Hiroaki; Yamaoka, Tetsuji; Shoji, Shuichi; Murakami, Akira.

    In: Analytical Biochemistry, Vol. 353, No. 1, 01.06.2006, p. 1-6.

    Research output: Contribution to journalArticle

    Munaka, Tatsuya ; Abe, Hirohisa ; Kanai, Masaki ; Sakamoto, Takashi ; Nakanishi, Hiroaki ; Yamaoka, Tetsuji ; Shoji, Shuichi ; Murakami, Akira. / Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis. In: Analytical Biochemistry. 2006 ; Vol. 353, No. 1. pp. 1-6.
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    AU - Nakanishi, Hiroaki

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