Effect of pH change on size and nanomechanical behavior of whey protein microgels

Microgels specific structural and functional features are attracting high research interest in several applications such as bioactives and drug delivery or functional food ingredients. Whey protein microgels (WPM) are obtained by heat treatment of whey protein isolate (WPI) in order to promote intramolecular cross-linking. In the present work, atomic force microscopy (AFM) was used in contact mode and in liquid to investigate WPM particles topography and mechanical properties at the nanoscale at native pH (6.5) and acid pH (5.5 and 3.0). Prior to AFM, WPM particles were captured on a gold substrate via low energy interactions by means of specific monoclonal antibodies. AFM images clearly showed an increase in the size of WPM particles induced by pH decrease. AFM in force spectroscopy mode was employed to monitor the elasticity of WPMs. The obtained effective Young's modulus data showed a significant increase in stiffness at pH 5.5 and pH 3.0, over 15-fold compared to native pH. These findings indicate that the mechanical profile of the WPM network varied with the pH decrease. The WPM topographic and nanomechanical changes induced by acidification were most likely due to substantial changes in the shape and inner structure of WPM particles. Our results suggest that internally cross-linked structures, modified by acidification could display interesting functional properties when used as a food ingredient.