Towards the purification of IgY from egg yolk by centrifugal partition chromatography

Although their high potential as alternative biopharmaceuticals, less than 2% of the total polyclonal antibodies produced worldwide correspond to immunoglobulin Y (IgY) due difficulties in isolating them from egg yolk (complex biological matrix). In this work, water-soluble proteins fraction (WSPF) was first obtained and present identified by one-dimensional gel electrophoresis (SDS-PAGE) label-free quantitative nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS). The WSPF then applied create aqueous biphasic systems (ABS) composed polyethylene glycol 1000 g·mol−1 (PEG 1000) K2HPO4/ KH2PO4 buffer, followed centrifugal partition chromatography (CPC) purify IgY. characterization showed presence six major proteins: target antibody IgY, serum albumin (α-livetin), ovalbumin, ovotransferrin, vitellogenin 1 2. results ABS revealed a affinity all polymer-rich phase. However, changing PEG salt concentrations, higher selectivity observed for with remaining partitioning between two phases. best were CPC, finally allowing multi-stage technology scale-up. CPC operating conditions optimized, obtain IgY 50.6% purity.