A rational optimization approach for the development of a multiplexed lateral flow immunoassay: detection of nonepithelial ovarian cancer markers in human serum
Author(s)
Type
Journal Article
Abstract
With the rise of non-communicable diseases, lateral flow immunoassays (LFIAs) are well-positioned to address the demand for disease monitoring. We present the use of peroxidase-mimicking platinum nanozyme conjugates targeting three ovarian germ cell tumor markers (alpha-fetoprotein (AFP), human chorionic gonadotropin (HCG), and cancer antigen 125 (CA125)) in LFIA. A “design of experiments” (DoE) approach was used to optimize antibody–nanozyme conjugation, superseding conventional “one-factor-at-a-time” optimization strategies, which neglect factor-to-factor interactions obscuring identification of optimal conditions. Crucial to disease monitoring, assays must demonstrate limits of detection (LoD) within clinically defined ranges and produce quantitative readouts. We address limitations of typical LoD calculations, which assume homoscedastic variance across marker concentrations, presenting an alternative model and applying it to assess LFIA performance, subsequently demonstrating clinically relevant LoDs in human serum. To robustly derive marker concentrations from LFIA readouts, the model was coupled with the resolution molecular concentration and, using patient samples, validated against laboratory gold standard values. AFP and HCG assays align well with gold standard measurements, with sensitivities of 87.5% and 100%, and specificities of 98.3% and 100%, respectively. This work outlines a development pipeline of a patient sample validated semiquantitative LFIA, utilizing DoE for streamlined optimization and improving modeling approaches to quantify performance in a manner representative of assay function.
Date Issued
2026-06-04
Date Acceptance
2026-01-28
Citation
Advanced Science, 2026, 13 (31)
ISSN
2198-3844
Publisher
Wiley
Journal / Book Title
Advanced Science
Volume
13
Issue
31
Copyright Statement
© 2026 The Author(s). Advanced Science published by Wiley-VCH GmbH This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
License URL
Identifier
https://www.ncbi.nlm.nih.gov/pubmed/41739995
Subjects
design of experiments
lateral flow immunoassays
limits of detection
nanozymes
Humans
Female
Ovarian Neoplasms
Immunoassay
Biomarkers, Tumor
Chorionic Gonadotropin
alpha-Fetoproteins
CA-125 Antigen
Limit of Detection
Publication Status
Published
Coverage Spatial
Germany
Article Number
e23192
Date Publish Online
2026-02-25