徳島大学 教育・研究者情報データベース(EDB)

1. 阿部 佳織

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2. (英) Hashimoto Y

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3. (英) Yatsushiro S

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4. (英) Yamamura S

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5. 板東 美香([徳島大学.大学院医歯薬学研究部.歯学域.口腔科学部門.臨床歯学系.歯周歯内治療学]/[徳島大学.病院.診療科.歯科.歯周病科(第二保存科)])

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6. 廣島 佑香([徳島大学.大学院医歯薬学研究部.歯学域.口腔科学部門.基礎歯学系.口腔微生物学])

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7. 木戸 淳一

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8. (英) Tanaka M

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9. 篠原 康雄([徳島大学.先端酵素学研究所.基幹研究部門])

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10. 大家 利彦(産業技術総合研究所健康工学研究センター)

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11. 馬場 嘉信

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12. 片岡 正俊

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(英) Simultaneous immunoassay analysis of plasma IL-6 and TNF-α on a microchip.

(英) Sandwich enzyme-linked immunosorbant assay (ELISA) using a 96-well plate is frequently employed for clinical diagnosis, but is time-and sample-consuming. To overcome these drawbacks, we performed a sandwich ELISA on a microchip. The microchip was made of cyclic olefin copolymer with 4 straight microchannels. For the construction of the sandwich ELISA for interleukin-6 (IL-6) or tumor necrosis factor-α (TNF-α), we used a piezoelectric inkjet printing system for the deposition and fixation of the 1st anti-IL-6 antibody or 1st anti-TNF-α antibody on the surface of the each microchannel. After the infusion of 2 µl of sample to the microchannel and a 20 min incubation, 2 µl of biotinylated 2nd antibody for either antigen was infused and a 10 min incubation. Then 2 µl of avidin-horseradish peroxidase was infused; and after a 5 min incubation, the substrate for peroxidase was infused, and the luminescence intensity was measured. Calibration curves were obtained between the concentration and luminescence intensity over the range of 0 to 32 pg/ml (IL-6: R(2) = 0.9994, TNF-α: R(2) = 0.9977), and the detection limit for each protein was 0.28 pg/ml and 0.46 pg/ml, respectively. Blood IL-6 and TNF-α concentrations of 5 subjects estimated from the microchip data were compared with results obtained by the conventional method, good correlations were observed between the methods according to linear regression analysis (IL-6: R(2) = 0.9954, TNF-α: R(2) = 0.9928). The reproducibility of the presented assay for the determination of the blood IL-6 and TNF-α concentration was comparable to that obtained with the 96-well plate. Simultaneous detection of blood IL-6 and TNF-α was possible by the deposition and fixation of each 1st antibody on the surface of a separate microchannel. This assay enabled us to determine simultaneously blood IL-6 and TNF-α with accuracy, satisfactory sensitivity, time saving ability, and low consumption of sample and reagents, and will be applicable to clinic diagnosis.

1. (英) Enzyme-Linked Immunosorbent Assay
2. (英) Humans
3. (英) Immunoassay
4. (英) Interleukin-6
5. (英) Lab-On-A-Chip Devices
6. (英) Luminescent Measurements
7. (英) Reproducibility of Results
8. (英) Tumor Necrosis Factor-alpha

1. (英) Article.ELocationID: 10.1371/journal.pone.0053620

2. (英) Article.PublicationTypeList.PublicationType: Journal Article

3. (英) Article.PublicationTypeList.PublicationType: Research Support, Non-U.S. Gov't

4. (英) OtherID: PMC3541141