Tags

Type your tag names separated by a space and hit enter

Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities.
Anal Chem. 2020 04 07; 92(7):5613-5619.AC

Abstract

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.

Authors+Show Affiliations

Quanterix Corporation, 900 Middlesex Turnpike, Building 1, Billerica, Massachusetts 01821, United States.TGR BioSciences, 31 Dalgleish Street, Thebarton, South Australia 5031, Australia.Quanterix Corporation, 900 Middlesex Turnpike, Building 1, Billerica, Massachusetts 01821, United States.Quanterix Corporation, 900 Middlesex Turnpike, Building 1, Billerica, Massachusetts 01821, United States.

Pub Type(s)

Journal Article

Language

eng

PubMed ID

32122115

Citation

Tobos, Carmen I., et al. "Customizable Multiplex Antibody Array Immunoassays With Attomolar Sensitivities." Analytical Chemistry, vol. 92, no. 7, 2020, pp. 5613-5619.
Tobos CI, Sheehan AJ, Duffy DC, et al. Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities. Anal Chem. 2020;92(7):5613-5619.
Tobos, C. I., Sheehan, A. J., Duffy, D. C., & Rissin, D. M. (2020). Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities. Analytical Chemistry, 92(7), 5613-5619. https://doi.org/10.1021/acs.analchem.0c00631
Tobos CI, et al. Customizable Multiplex Antibody Array Immunoassays With Attomolar Sensitivities. Anal Chem. 2020 04 7;92(7):5613-5619. PubMed PMID: 32122115.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Customizable Multiplex Antibody Array Immunoassays with Attomolar Sensitivities. AU - Tobos,Carmen I, AU - Sheehan,Antony J, AU - Duffy,David C, AU - Rissin,David M, Y1 - 2020/03/12/ PY - 2020/3/4/pubmed PY - 2020/3/4/medline PY - 2020/3/4/entrez SP - 5613 EP - 5619 JF - Analytical chemistry JO - Anal. Chem. VL - 92 IS - 7 N2 - 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. SN - 1520-6882 UR - https://www.unboundmedicine.com/medline/citation/32122115/Customizable_Multiplex_Antibody_Array_Immunoassays_with_Attomolar_Sensitivities_ L2 - https://doi.org/10.1021/acs.analchem.0c00631 DB - PRIME DP - Unbound Medicine ER -
Try the Free App:
Prime PubMed app for iOS iPhone iPad
Prime PubMed app for Android
Prime PubMed is provided
free to individuals by:
Unbound Medicine.