Inhibiting amyloid-like aggregation through bio-conjugation of proteins with polymer surfactant

TL;DR

This study demonstrates that conjugating proteins with polymer surfactants can effectively inhibit amyloid-like aggregation, providing a potential strategy for protein stabilization with implications in biotechnology and medicine.

Contribution

The paper introduces a novel approach using bio-conjugation of proteins with polymer surfactants to prevent amyloid aggregation, showing enhanced stability compared to native proteins.

Findings

Polymer surfactant conjugates remain stable for 5 days under aggregation conditions.

Native BSA forms amyloid fibrils, unlike conjugates.

Heat-treated BSA forms self-standing films with amyloid structures.

Abstract

Prevention of protein aggregation and thus stabilization of proteins has large biological and biotechnological implications. Here, we show that inhibition of amyloid-like aggregates is possible in stoichiometric conjugates of polymer surfactant and bovine serum albumin (BSA) chosen as a model protein. We investigate using a combination of Thioflavin-T fluorescence spectroscopy, dynamic light scattering and FTIR spectroscopy the aggregation behavior in polymer surfactant modified and unmodified (native) BSA solutions. The BSA-polymer surfactant conjugates are stable up to 5 days under aggregation conditions, while native BSA forms amyloid fibrillar structures. Further, DLS-based micro-rheology studies performed with heat-treated 100 to 200 {\mu}M native BSA aggregates provided understanding of the equilibrium elastic and viscous moduli over a very large frequency range, reaching MHz, which are inaccessible using bulk rheology. Our results indicate that after 6 days of aggregation conditions, elastic moduli showed values between 1.2 to 3.6 Pa corresponding to an entanglement length ({\xi}) of 105 nm. Interestingly, heating 200 {\mu}M native BSA solution at 65 degree C for 2 days in a plastic Eppendorf resulted in self-standing films. These films exhibited strong ThT-fluorescence intensity and a predominant \b{eta}-sheet secondary structure from the FTIR studies, suggesting that self-standing microstructure resulted from hierarchical self-assembly of amyloid fibrils.