High selective purification of IgY from quail egg: Process design and quantification of deep eutectic solvent based ultrasound assisted liquid phase microextraction coupled with preparative chromatography.

The present research investigates on task-specific deep eutectic solvents (TDES) based aqueous two-phase extraction and purification of immunoglobulins by chromatography from quail egg. The synthesis of TDES was accomplished with quaternary ammonium salt as hydrogen bond acceptor (HBA) and glycerol as hydrogen bond donor (HBD). Aqueous two-phase (ATPS) formation of TDESs with various salts was established and phase partitioning ability was evaluated using standard bovine immunoglobulin. Ultrasound-assisted liquid-liquid microextraction (UA-LLME) was performed with ATPS of better partitioning ability for quail egg yolk. Optimization of influential variables with response surface methodology was accomplished and maximum yield was achieved for 85% (v/v) of NADES for feed of 18 μg/ml of egg yolk isolate with ultrasound temperature of 35 °C and 12 min contact time respectively. The quantification of recovery was accomplished by gel filtration process with the determination of retention time, volume and resolution of separation using marker protein. Ultrapure immunoglobulin-Y (IgY) is achieved with anion exchange chromatography and yield was calculated. The results revealed that the selective concentration of immunoglobulin-Y from quail egg yolk could be performed using environment-friendly TDES based UA-LLME coupled with chromatography.