Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis.
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.
Munaka
Tatsuya
T
Technology Research Laboratory, Shimadzu Corporation, Seika-cho, Soraku-gun, Kyoto, Japan.
Abe
Hirohisa
H
Kanai
Masaki
M
Sakamoto
Takashi
T
Nakanishi
Hiroaki
H
Yamaoka
Tetsuji
T
Shoji
Shuichi
S
Murakami
Akira
A
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
D007074:Immunoglobulin G; D007202:Indicators and Reagents; D012428:Ruthenium