Bioactive modification of microfluidic chip surface for detecting recombinant human bone morphogenetic protein-2
In order to develop a rapid, specific, and accurate detection method for the concentration of recombinant human bone morphogenetic protein-2 (rhBMP-2) through Escherichia coli-based expression systems, enzyme-linked immunosorbent assay (ELISA) was combined with microfluidic chip. The specific first...
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| Main Authors: | , , |
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| Format: | Article |
| Language: | English |
| Published: |
Zhejiang University Press
2020-08-01
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| Series: | 浙江大学学报. 农业与生命科学版 |
| Subjects: | |
| Online Access: | https://www.academax.com/doi/10.3785/j.issn.1008-9209.2019.08.071 |
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| Summary: | In order to develop a rapid, specific, and accurate detection method for the concentration of recombinant human bone morphogenetic protein-2 (rhBMP-2) through Escherichia coli-based expression systems, enzyme-linked immunosorbent assay (ELISA) was combined with microfluidic chip. The specific first antibody for rhBMP-2 was screened from several commercial products. Based on the regulation strategy of antibody orientation for enhancing detection signal, plasma-protein A method was then used to modify the detection microwells of the microfluidic chip. After tuning the conditions of plasma treatment on the detection microwells, adsorption efficiency of the first antibody and strength of the final detection signal were evaluated. It was found that the better capture efficiency of the first antibody could be obtained by using the higher power in the plasma treatment process. The best plasma condition was the power of 100 W and treatment time of 30 s. After the optimized modification conditions were applied for the microfluidic chip, the dilute concentrations of rhBMP-2 in a range of 0-2 000 pg/mL were achieved. In comparison with the standard assay carried out in the 96-well microtiter plate, the microwells of microfluidic chip exhibited a broader linear detection range (0-2 000 pg/mL vs. 0-250 pg/mL) and a much less reagent consumption (Each sample needed 600 μL reagent consumption in the standard assay, while about 16 μL in the microwell assay, which was 97.3% reduction in dosage). Clearly, this plasma-protein A immobilization strategy holds a great potential for polymeric microfluidic chip based assay in biomedical application, food safety, and environment monitoring. |
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| ISSN: | 1008-9209 2097-5155 |