Chemiluminescence-imaging detection of DNA on a solid-phase membrane by using a peroxidase-labeled macromolecular probe
| Autor: | Tsutomu Kabashima, Md. Golam Azam, Mutsumi Yamasuji, Takayuki Shibata, Masaaki Kai, Tomasz Krawczyk |
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| Rok vydání: | 2014 |
| Předmět: |
Streptavidin
Adult Macromolecular Substances Biotin Horseradish peroxidase Analytical Chemistry law.invention chemistry.chemical_compound law Complementary DNA Image Processing Computer-Assisted Humans Cells Cultured Horseradish Peroxidase Chemiluminescence biology Mouth Mucosa Nucleic Acid Hybridization Dextrans DNA Telomere Avidin Blotting Southern Biochemistry chemistry Biotinylation Luminescent Measurements biology.protein Biophysics Macromolecule |
| Zdroj: | Talanta. 139 |
| ISSN: | 1873-3573 |
| Popis: | We have developed a novel method for sensitive chemiluminescence (CL)-imaging detection of DNA by using a macromolecular probe synthesized by attaching multiple molecules of horseradish peroxidase (HRP) and biotin in dextran backbone. The probe formed a macromolecular assembly by binding to streptavidin which specifically recognized biotinylated complementary DNA, which was hybridized to a target DNA on a solid-phase membrane. This methodology was applied to CL-imaging detection of a synthetic telomere DNA (TTAGGG)10 and human telomere DNA by using the CL probe comprising of dextranT2000 (MW=ca. 2000 kDa) bonded to approximately 42 molecules of HRP and 210 molecules of biotin. The human telomere DNA in a small number of buccal mucous cells (ca. 70 cell numbers) of cheek tissue was quantitatively determined by the proposed CL detection method that afforded approximately 10 times higher sensitivity than that of the conventional CL method using commercially available HRP–avidin probe. |
| Databáze: | OpenAIRE |
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