Polygalacturonase를 검출하기 위한 종이 기반의 효소결합 면역반응 센서 제작
Fabrication of a paper-based ELISA to detect polygalacturonase
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In this paper, we describe the fabrication of a paper-based enzyme-linked immunosorbent assay (ELISA) to detect polygalacturonase (PG), which is used as a biomarker to determine whether a plant is infected with a disease. The proposed paper-based ELISA can analyze the concentration of PG in a short time using a small sample compared to the traditional ELISA, which is generally performed using a well plate. To increase the resolution of the sensor, we optimized the dilution ratio of the HRP-conjugated goat anti-rabbit IgG antibodyand the dilution ratio of the anti-PG and HRP-conjugated goat anti-rabbit IgG antibodies. Furthermore, for quantitative analysis of PG concentration, Delta RGB analysis was conducted to detect color changes in the sensing window displayed by the PG samples at various concentrations. Based on the experiment, the fabricated paper-based ELISA could measure at least 0.25 µg of PG and the measurement range was 0.25–2 µg. Therefore, the paper-based ELISA for detecting PG is expected to be able to determine the presence or absence of disease in crops at the infection stage in the future.
Abstract
In this paper, we describe the fabrication of a paper-based enzyme-linked immunosorbent assay (ELISA) to detect polygalacturonase (PG), which is used as a biomarker to determine whether a plant is infected with a disease. The proposed paper-based ELISA can analyze the concentration of PG in a short time using a small sample compared to the traditional ELISA, which is generally performed using a well plate. To increase the resolution of the sensor, we optimized the dilution ratio of the HRP-conjugated goat anti-rabbit IgG antibodyand the dilution ratio of the anti-PG and HRP-conjugated goat anti-rabbit IgG antibodies. Furthermore, for quantitative analysis of PG concentration, Delta RGB analysis was conducted to detect color changes in the sensing window displayed by the PG samples at various concentrations. Based on the experiment, the fabricated paper-based ELISA could measure at least 0.25 µg of PG and the measurement range was 0.25–2 µg. Therefore, the paper-based ELISA for detecting PG is expected to be able to determine the presence or absence of disease in crops at the infection stage in the future.
참고문헌 (12)
- J. D. Walton, "Deconstructing the cell wall", Plant Physiol., 104(4), pp.1113~1118 (1994)
- R. D´ovidio, "Polygalacturonases, polygalacturonase-inhibiting proteins and pectic oligomers in plant-pathogen interactions", Biochim. Biophys. Acta, 1696(2), pp.237~244 (2004)
- D. F. Bateman, "Degradation of plant cell walls and membranes by microbial enzymes", Encycl. Plant Physiol., 4, pp.316~355 (1976)
- M. Li, "Pathogenicity of cell wall degrading enzymes produced by botryodiplodia theobromae Pat. Against mangoes", Agricultural Biotechnology, 1(6), pp.18~21 (2012)
- Y. Q. Zhao, "Activity pathogenic effect of cell wall degrading enzyme in tobacco target spot pathogen Rhizoctonia solani", Tobacco Science & Technology, 11, pp.84~88 (2014)
- J. Zhang, "Cell wall-degrading enzymes of didymella bryoniae in relation to fungal growth and virulence in cantaloupe fruit", Eur. J. Plant Pathol., 139(4), pp.749~761 (2014)
- A. K. Chaurasia, "In vitro and in vivo production of polygalacturonase, polymethylgalacturonase and cellulase enzymes by Alternaria solani at different incubation periods", Afr. J. Plant Sci., 8(5), pp.248~253 (2014)
- D. B. Gawade, "Extracellular enzymes activity determining the virulence of rhizoctonia bataticola, causing root rot in soybean", Physiol. Mol. Plant Pathol., 100, pp.49~56 (2017)
- T. Tu, "High-yield production of a low-temperature-active polygalacturonase for papaya juice clarification", Food Chem., 141(3), pp.2974~2981 (2013)
- G. L. Miller, "Use of dinitrosalicylic acid reagent for determination of reducing sugar", Anal. Chem., 31(3), pp.426~428 (1959)
- M. Somogyi, "A new reagent for the determination of sugars", J. Biol. Chem., 160(1), pp.61~68 (1945)
- S. Torres, "A colorimetric method to quantify endo-polygalacturonase activity", Enzyme Microb. Technol, 48(2), pp.123~128 (2011)