Next-gen diagnostics: CRISPR-chip for monoclonal antibodies quantification at the point-of-care
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Author
Del Giovane S., Migliorelli D., Porchetta A., Altug H., Burr L., Stefano
Migliorelli, Davide
Porchetta, Alessandro
Altug, Hatice
Burr, Loïc
DOI
Abstract
Monoclonal antibodies (mAbs) are used to treat numerous cancers, offering specific therapy that improves patient outcomes. Measurements of mAbs in the bloodstream during cancer treatment are essential for monitoring the therapeutic antibody dose, assuring treatment efficacy, reducing side effects, and tracking drug resistance.
Enzyme-Linked Immunosorbent Assay (ELISA) and High-Pressure Liquid Chromatography tandem Mass Spectrometry (HPLC-MS) are the gold standard for quantitative mAbs analysis in clinical studies. Both methods involve expensive, bulky, and complicated instrumentation, making them unsuitable for point-of-care (POC) use.
To address this need, we introduce a POC platform for quantitative mAb detection, which leverages a recently developed quantitative mAb detection method, based on the use of a DNA circuit for the recognition of the mouse anti-hemagglutinin (antiHA) from influenza, and the activation of the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) and CRISPR-associated protein (Cas). CRISPR systems' adaptability, low cost, high sensitivity and repeatability make our approach a great candidate for POC applications. Though promising, the conventional protocols run in test tubes, require four incubation steps and consecutive reagent additions. Thus, our study focuses on developing a simplified on-chip CRISPR-based assay without scarifying on the performance.
For this purpose, a two-step protocol is developed, including sample dilution and injection, requiring only 10 µL of volume, and providing results within a 60 minutes sample-to-answer timeframe. The assay is integrated on a cost-effective chip, less than 1 € for chemicals, and additionally it is verified that the dispensed reagents can be stored at -20°C for at least 1 month without degradation.
In conclusion, we demonstrate a proof-of-concept CRISPR-chip, which has the potential to impact modern diagnostics and the clinical market by introducing a new tool for protein quantification at the POC.
Publication Reference
Euroanalysis 2025, Barcelona, Spain
Year
2025-09-03