An efficient direct competitive nano-ELISA for residual BSA determination in vaccines
| Autor: | Li-Sheng Ding, Jie Li, Qian-Long Wang, Lin-Sen Qing, Pei Luo, Xing-De Li, Wan-Jun Tao |
|---|---|
| Rok vydání: | 2017 |
| Předmět: |
Silicon dioxide
Nanoparticle Enzyme-Linked Immunosorbent Assay 02 engineering and technology Residual 01 natural sciences Biochemistry Horseradish peroxidase Analytical Chemistry chemistry.chemical_compound Microscopy Electron Transmission Limit of Detection Nanotechnology Bovine serum albumin Detection limit Vaccines Chromatography biology Amidogen 010401 analytical chemistry Reproducibility of Results Serum Albumin Bovine Silicon Dioxide 021001 nanoscience & nanotechnology 0104 chemical sciences Linear range chemistry biology.protein 0210 nano-technology |
| Zdroj: | Analytical and Bioanalytical Chemistry. 409:4607-4614 |
| ISSN: | 1618-2650 1618-2642 |
| Popis: | A simple, fast, and highly sensitive direct competitive enzyme-linked immunosorbent assay (ELISA) based on bovine serum albumin (BSA) antigen labeled amine-terminated silicon dioxide (SiO2–NH–BSA) nanoparticles was developed to determine residual BSA in vaccines. As nano-ELISA using nanomaterials with a very high surface-to-volume ratio has emerged as a promising strategy, SiO2–NH–BSA nanoparticles were prepared in this study by the coupling of BSA to SiO2 nanoparticles modified with amidogen, followed by the quantification of BSA via a direct competitive binding of BSA-antigen-labeled SiO2 nanoparticles to anti-BSA antibody conjugated with horseradish peroxidase. The validation study showed that the linear range of this method was from 1 to 90 ng/mL (r = 0.998) and the limit of detection was 0.67 ng/mL. The intra-assay and interassay coefficients of variation were less than 10% at three concentrations (10, 40, and 70 ng/mL), and the recovery was 92.4%, indicating good specificity. As a proof of principle, this new method was applied in the analysis of residual BSA in five different vaccines. Bland–Altman plots revealed that there was no significant difference in the accuracy and precision between our new method and the most commonly used sandwich ELISA. From the results taken together, the new method developed in this study is more sensitive and facile with lower cost and thus demonstrated potential to be applied in the quality control of biological products. |
| Databáze: | OpenAIRE |
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