Refine
Has Fulltext
- no (2)
Document Type
- Article (2)
Language
- English (2)
Is part of the Bibliography
- yes (2)
Keywords
- Cancer cells (1)
- Cell detection (1)
- Nanospots (1)
- Nanostructured surfaces (1)
- SAW sensor (1)
- sam (R) 5 (1)
Institute
This work describes a cell-based assay that does not depend on radioactivity or laboratory animals for the detection of ligands of angiotensin II type 1 receptor (AT(1)R). The assay makes use of stable transfected Chinese hamster ovary cells (CHO-AT(1)R) expressing the AT(1)R. A sequential saturation assay principle was used in which receptor binding sites of the CHO-AT(1)R cells are blocked by the analyte in a concentration-dependent manner. Afterwards, TAMRA-angiotensin II, a fluorescence-labeled ligand, was added to bind to the remaining free binding sites of the receptor. In consequence, the fluorescence signal determined is inversely proportional to the concentration of the analyte.
A nanostructured chip surface was fabricated enabling binding via spaced antibodies specifically targeting surface proteins of cancer cells and detection of extremely low numbers of circulating tumor cells (CTC) without labeling using a sam (R) 5 biosensor. The antibody surfaces mostly were generated by self assembly of antibodies to gold nanospots on the sensitive SiO2-surface of a sam (R) 5 chip. Compared with a complete gold surface, only 40% of the amount of antibodies was bound to the nanospot surface, but structured such that 15-fold higher sensitivity to vital cancer cells was achieved. Human cancer cell lines JEG-3 (lmphoblastic leukemia) and MOLT-17 (placental choriocarcinoma) from cell cultures were successfully detected. The sensor showed significant responses on less than 10 cells injected in a single run. The extreme increase in sensitivity and its simple regeneration emphasizes the usefulness of its introduction in biomedical applications. (C) 2011 Elsevier B.V. All rights reserved.