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Label Free and Multiplexed Immunosensor Chips for Detection of Macromolecular and Cellular Biomarkers

Han, Yu
http://orcid.org/0000-0002-1460-7511
http://rave.ohiolink.edu/etdc/view?acc_num=akron1447634222

Abstract Details

2015, Doctor of Philosophy, University of Akron, Mechanical Engineering.
Biomarker detection represents an important task for many scientific fields such as disease diagnosis, biodefense, environmental monitoring and biological research. In this research, micro resistive pulse immunoaggregation sensors were studied to quantitatively detect macromolecular and cellular biomarker concentrations. First, to prove the concept of the immunoaggregation assay and resistive pulse sensing, goat anti-rabbit IgG, used as a model biomarker, was detected by the micro resistive pulse immunoaggregation sensor. A detection range from 16.0 to 160 ng/mL was achieved. The human ferritin in 10% fetal bovine serum was then detected to prove the device’s performance in a complex media. A detection range from 0.104 to 208 ng/mL was achieved. In addition, the studies showed that the detection range can be shifted to lower and higher concentrations by decreasing and increasing MP concentrations, respectively. Second, the multiplexed immunoaggregation assay was also studied in this research to achieve multiple biomarker detection in a single test. A mixture consisting of multiple types of MPs functionalized by different antibody with different sizes and magnetic properties was used for testing the multiplexed assay. 2.00 µm non-magnetic MPs coated with rabbit anti-mouse IgG antibody and 2.80 µm magnetic MPs coated with goat anti-human ferritin antibody were used to detect the concentration of mouse anti-rabbit IgG and human ferritin, respectively. Detection ranges from 5.20 to 208 ng/mL and 3.10 to 51.2 × 103 ng/mL were achieved for human ferritin and mouse anti-rabbit IgG detection. Next, a two-stage resistive pulse sensor was invented to detect target cells in a mixed cell population. S. cerevisiae and Chlorella were used as target cells and control cells, respectively. The specific capture efficiency of S. cerevisiae was greater than 94.8%, while the nonspecific capture efficiency was less than 3.4%. S. cerevisiae ratio measurements in a mixture with Chlorella showed that for S. cerevisiae to Chlorella ratios ranging from 1.0 to 2.0, the measurement errors were less than 7%, while the errors were 20% to 32% for lower ratios, ranging from 0.1 to 0.5, caused by nonspecific attachment. The testing results demonstrated that the immunoaggregation biomarker sensor and MP cellular sensor enable reliable detection of target macromolecular and cellular biomarkers in samples at very low concentrations, without the fluorescence and enzyme labeling of biomarker. These innovative biosensors will lead to a fast and cost-effective bioassay instrument for disease diagnosis, environmental monitoring and homeland security.
Jiang Zhe (Advisor)
Gang Cheng (Committee Member)
Qin Liu (Committee Member)
Francis Loth (Committee Member)
Chang Ye (Committee Member)
138 p.
Mechanical Engineering

Recommended Citations

Citations

  • Han, Y. (2015). Label Free and Multiplexed Immunosensor Chips for Detection of Macromolecular and Cellular Biomarkers [Doctoral dissertation, University of Akron]. OhioLINK Electronic Theses and Dissertations Center. http://rave.ohiolink.edu/etdc/view?acc_num=akron1447634222

    APA Style (7th edition)

  • Han, Yu. Label Free and Multiplexed Immunosensor Chips for Detection of Macromolecular and Cellular Biomarkers. 2015. University of Akron, Doctoral dissertation. OhioLINK Electronic Theses and Dissertations Center, http://rave.ohiolink.edu/etdc/view?acc_num=akron1447634222.

    MLA Style (8th edition)

  • Han, Yu. "Label Free and Multiplexed Immunosensor Chips for Detection of Macromolecular and Cellular Biomarkers." Doctoral dissertation, University of Akron, 2015. http://rave.ohiolink.edu/etdc/view?acc_num=akron1447634222

    Chicago Manual of Style (17th edition)

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This open access ETD is published by University of Akron and OhioLINK.