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A linear-polymer-based lactoferrin-selective recognition element for an ELISA mimic: A proof of concept
M A Goicolea1, A Gómez-Caballero1, M Saumell-Esnaola2
1Department of Analytical Chemistry, Faculty of Pharmacy, University of the Basque Country (UPV/EHU), 01006, Vitoria-Gasteiz, Spain.
Analytica Chimica Acta
|January 16, 2022
Summary
Researchers developed a novel linear polymer (LP) for selective lactoferrin (LF) detection, offering a potential antibody replacement in bioassays. This flexible polymer shows high specificity and applicability in disease diagnostics.
Area of Science:
- Polymer Chemistry
- Biomaterials Science
- Analytical Chemistry
Background:
- Developing synthetic polymers for specific macromolecule recognition, like proteins, remains a significant challenge.
- Traditional synthetic polymers for macromolecule imprinting often suffer from drawbacks associated with insolubility and inflexibility.
- Natural antibodies, while effective, can be expensive and have limited stability for widespread bioassay applications.
Purpose of the Study:
- To introduce a novel linear polymer (LP) format as a selective recognition element for the globular protein lactoferrin (LF).
- To demonstrate the proof-of-concept for using LPs as a viable alternative to natural antibodies in bioassays.
- To evaluate the performance of the LP in a lactoferrin-selective enzyme-linked immunosorbent assay (ELISA).
Main Methods:
- A solid-phase synthesis strategy employing reversible deactivation radical polymerization (RDRP) was utilized.
- Unlike classical imprinting, this method does not require a cross-linker, resulting in a soluble and flexible copolymer.
- The synthesized LP was employed as the recognition element in a LF-specific ELISA to quantify LF concentrations.
Main Results:
- The developed linear polymer demonstrated selective recognition of lactoferrin within a concentration range of 0.1 nM to 0.25 μM.
- The assay showed a significant difference between non-specific signals and those obtained with the polymeric material, with an EC50 value of 11.8 ± 1.4 nM.
- High selectivity was confirmed through competitive inhibition assays against related proteins (lysozyme, trypsin, albumin), with cross-reactivity values as low as 0.016%.
Conclusions:
- The novel linear polymer (LP) offers a promising, flexible, and soluble alternative to traditional synthetic polymers for macromolecule imprinting.
- This LP format has significant potential to replace natural antibodies in various bioassays, including ELISA, dot blot, and western blot.
- The method's applicability was validated by determining urine LF levels in patients with inflammatory and infectious urinary tract diseases.
