Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities
| Autor: | Carmen I. Tobos, David M. Rissin, David C. Duffy, Antony J Sheehan |
|---|---|
| Rok vydání: | 2020 |
| Předmět: |
Analyte
Antibody microarray Optical Phenomena Cross Reactions 010402 general chemistry 01 natural sciences Antibodies Analytical Chemistry Microtiter plate Limit of Detection medicine Humans Multiplex Interleukin 5 Immunoassay Chromatography biology medicine.diagnostic_test Chemistry Tumor Necrosis Factor-alpha Interleukins 010401 analytical chemistry 0104 chemical sciences Protein profiling biology.protein Antibody |
| Zdroj: | Analytical chemistry. 92(7) |
| ISSN: | 1520-6882 |
| Popis: | We have developed a customizable contact printed multiplex immunoassay capable of simultaneously measuring up to five analytes with attomolar sensitivities. This enzyme-linked immunosorbent assay (ELISA) was based on spotting different antibodies in a circular pattern at the bottom of a microtiter plate well. Unlike traditional antibody printing for ELISA that prints a capture antibody specific to a target of interest, in this ELISA we printed unique "anchor" antibodies at the well surface, each having a high affinity for a specific peptide target. By coupling each peptide to a unique assay capture antibody, this array of anchor antibodies enabled a customizable contact printed multiplex immunoassay workflow. As a proof of concept, we developed a 5-plex assay measuring interleukin 5 (IL-5), interleukin 6 (IL-6), interleukin 10 (IL-10), interleukin 22 (IL-22), and tumor necrosis factor alpha (TNF-α). Measurements of these five analytes in serum and plasma correlated well between the method utilizing the anchor antibodies and peptides and the traditional capture antibody printing approach, with r2 values of 0.99, 0.93, 0.99, 0.96, and 0.75 for IL-5, IL-6, IL-10, IL-22, and TNFα, respectively. This approach makes customizable multiplex ultrasensitive ELISA available to laboratories without access to the precision printing instrumentation and will be useful for antibody screening, custom assay development, biomarker detection, and protein profiling for diagnostic applications. |
| Databáze: | OpenAIRE |
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