title: Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis.
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.
authors.affiliation: Technology Research Laboratory, Shimadzu Corporation, Seika-cho, Soraku-gun, Kyoto, Japan.
authors.forename: Tatsuya
authors.initials: T
authors.lastname: Munaka
authors.forename: Hirohisa
authors.initials: H
authors.lastname: Abe
authors.forename: Masaki
authors.initials: M
authors.lastname: Kanai
authors.forename: Takashi
authors.initials: T
authors.lastname: Sakamoto
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakanishi
authors.forename: Tetsuji
authors.initials: T
authors.lastname: Yamaoka
authors.forename: Shuichi
authors.initials: S
authors.lastname: Shoji
authors.forename: Akira
authors.initials: A
authors.lastname: Murakami
mesh_terms: D000818:Animals; D016880:Anisotropy; D000921:Antibody-Producing Cells; D004867:Equipment Design; D006825:Hybridomas / Q000737:chemistry / Q000166:cytology / Q000378:metabolism*; D007074:Immunoglobulin G / Q000032:analysis* / Q000378:metabolism; D007202:Indicators and Reagents; D056656:Lab-On-A-Chip Devices; D008163:Luminescent Measurements / Q000379:methods*; D051379:Mice; D046208:Microchip Analytical Procedures / Q000379:methods*; D051381:Rats; D012428:Ruthenium / Q000737:chemistry; D013997:Time Factors
chemical_list: D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012428:Ruthenium