Modeling the navigating forces behind BSA aggregation in a microfluidic chip.

Area of Science:

• Biophysics
• Materials Science
• Chemical Engineering

Background:

• Protein aggregation is implicated in various diseases and is influenced by physical and chemical factors.
• Investigating protein aggregation dynamics requires advanced methodologies to capture rapid processes.

Purpose of the Study:

• To dynamically investigate bovine serum albumin (BSA) aggregation using microfluidic chips.
• To elucidate the molecular mechanisms and driving forces behind BSA aggregation in a microfluidic system.
• To compare aggregation kinetics in static vial-based systems versus dynamic microfluidic systems.

Main Methods:

• Utilized microfluidic chip-based dynamic systems and vial-based static systems for BSA aggregation studies.
• Employed biophysical and microscopic experimental methods.
• Conducted computational simulations using MATLAB and COMSOL Multiphysics.

Main Results:

• BSA aggregation was successfully induced in microfluidic chips on a timescale of seconds.
• Brownian movement, advective mixing, and laminar flow were identified as key factors favoring amyloid-like aggregate formation.
• Heating initiated partial unfolding, while space restriction and electrostatic/van der Waals forces drove initial cluster formation.
• Hydrodynamic forces, hydrophobic interactions, and space restriction led to aggregate deposition and structural conversion to amyloid-like structures.

Conclusions:

• Microfluidic systems enable rapid, dynamic investigation of protein aggregation mechanisms.
• A combination of physical forces (Brownian motion, shear, space restriction) and chemical interactions drives BSA aggregation.
• The study elucidated the forces and molecular mechanisms underlying BSA aggregation in microfluidic environments within seconds.