Positive response to surfactants on the interfacial behavior and aggregation stability of Fab fragments from yolk immunoglobulin.

The antigen binding fragment (Fab) is pepsin-digested product from egg yolk immunoglobulin (IgY), which shows lower immunogenicity and higher antibacterial activity. However, it limited the application of Fab due to the spontaneous adsorption and aggregation at the air-liquid interface. The present work is to investigate the effect of surfactants polysorbate 20 (PS20), poloxamer 188 (P188), and polyethylene glycol (PEG) on the aggregation stability of Fab of IgY. The results confirmed the positive role of surfactants in improving Fab stability. PS20 could effectively prevent the generation of Fab aggregates (DLS and light-obscuration analysis). It could also distinctly increase the internal hydrophobicity level, fortify the surface charge by altering the molecular conformational characteristics of Fab. The results of CLSM and surface tension demonstrated that P188 and PEG were co-adsorbed with Fab at the air-liquid interface and inhibited the formation of aggregation. PS20 competitively adsorbed in the gap between Fab molecules to inhibit the formation of aggregates. These findings would give an in-depth understanding of protein aggregation behavior influenced by surfactants and provide a theoretical basis for the development of functional food based on Fab active fragments.