Highly Effective Synthetic Polymer-Based Blockers of Non-Specific Interactions in Immunochemical Analyses

Abstract

In vitro diagnostic methods face non-specific interactions increasing their background level and influencing the efficacy and reproducibility. Currently, the most important and employed blocker of non-specific interactions is bovine serum albumin (BSA), an animal product with some disadvantages like its batch-to-batch variability and contamination with RNases. Herein, we developed amphiphilic water-soluble synthetic copolymers based on the highly biocompatible, non-immunogenic and nontoxic N-2-(hydroxypropyl)methacrylamide (HPMA)-based copolymers or poly(oxazoline)s as highly effective synthetic blockers of non-specific interactions and an effective BSA alternative. The highest blocking capacity was observed for HPMA-based polymers containing two hydrophobic anchors taking advantage of the combination of two structurally different hydrophobic molecules. Polymers prepared by free radical polymerisation with broader dispersity were slightly better in terms of surface covering. The sandwich ELISA evaluating human thyroid-stimulating Hormone in patient samples revealed that the designed polymers can fully replace BSA without compromising the assay results. Importantly, as a fully synthetic material, the developed polymers are fully animal pathogen-free; thus, they are highly important materials for further development.

Keywords

• HPMA copolymer
• blocker
• non-specific interactions
• immunochemical analyses
• bovine serum albumin
• TSH ELISA kit