Abstract
A new electrochemical assay for the detection of secreted alkaline phosphatase (SEAP) from transfectant HeLa cells is proposed using a microarray device and scanning electrochemical microscopy (SECM). The assay consists of two steps: the first is the incubation of a transfected cell in a microarray culture device covered with a substrate modified with anti-SEAP under physiological conditions without any additives. The array device consists of a 4×4 array of microwells having a size of 100μm×100μm (diameter×depth). The second step is SECM measurement of secreted SEAP at the antibody-immobilized substrate. This assay ensures accuracy and intactness because the undesired influence of endogeneous ALP is eliminated and the transfected cells are incubated in a culture device under suitable conditions. We successfully detected the expression of SEAP from intact cells at the single-cell level using this assay. The system is useful as a cell-based gene-expression assay.
| Original language | English |
|---|---|
| Pages (from-to) | 2163-2167 |
| Number of pages | 5 |
| Journal | Biotechnology and Bioengineering |
| Volume | 109 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - 2012 Aug |
Keywords
- Biosensor
- Cell array
- Electrochemistry
- Lab on a chip
- Reporter gene assay