| RRC ID |
10909
|
| Author |
Munaka T, Abe H, Kanai M, Sakamoto T, Nakanishi H, Yamaoka T, Shoji S, Murakami A.
|
| Title |
Real-time monitoring of antibody secretion from hybridomas on a microchip by time-resolved luminescence anisotropy analysis.
|
| Journal |
Anal Biochem
|
| 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.
|
| Volume |
353(1)
|
| Pages |
1-6
|
| Published |
2006-6-1
|
| DOI |
10.1016/j.ab.2006.03.009
|
| PII |
S0003-2697(06)00174-6
|
| PMID |
16620756
|
| MeSH |
Animals
Anisotropy
Antibody-Producing Cells
Equipment Design
Hybridomas / chemistry
Hybridomas / cytology
Hybridomas / metabolism*
Immunoglobulin G / analysis*
Immunoglobulin G / metabolism
Indicators and Reagents
Lab-On-A-Chip Devices
Luminescent Measurements / methods*
Mice
Microchip Analytical Procedures / methods*
Rats
Ruthenium / chemistry
Time Factors
|
| IF |
2.877
|
| Times Cited |
6
|
|
WOS Category
|
CHEMISTRY, ANALYTICAL
BIOCHEMICAL RESEARCH METHODS
BIOCHEMISTRY & MOLECULAR BIOLOGY
|
| Resource |
| Human and Animal Cells |
12-1B6(RCB0870) |
