abstract
Antibodies obtained from egg yolk of immunized hens, immunoglobulinY (IgY), are an alternative to the most focused mammal antibodies, because they can be obtained in higher titers by less invasive approaches. However, the production cost of high-quality IgY for large-scale applications remains higher than that of other drug therapies due to the lack of efficient purification methods. The search for new purification platforms is thus vital. The solution could be liquid-liquid extraction by using aqueous biphasic systems (ABS). Herein, we report the extraction and attempted purification of IgY from chicken egg yolk by using a new ABS composed of polymers and Good's buffer ionic liquids (GB-ILs). New self-buffering and biocompatible ILs based on the cholinium cation and anions derived from Good's buffers were synthesized and the self-buffering characteristics and toxicity were characterized. Moreover, when these GB-ILs are combined with PPG 400 (poly(propylene) glycol with a molecular weight of 400gmol(-1)) to form ABS, extraction efficiencies, of the water-soluble fraction of proteins, ranging between 79 and 94% were achieved in a single step. Based on computational investigations, we also demonstrate that the preferential partitioning of IgY for the GB-IL-rich phase is dominated by hydrogen-bonding and van der Waals interactions.
keywords
AQUEOUS 2-PHASE SYSTEMS; EGG-YOLK IMMUNOGLOBULINS; BIOLOGICAL-RESEARCH; BIPHASIC SYSTEMS; GLASS-ELECTRODE; CYTOCHROME-C; EXTRACTION; PURIFICATION; PROTEINS; IGY
subject category
Chemistry
authors
Taha, M; Almeida, MR; Silva, FAE; Domingues, P; Ventura, SPM; Coutinho, JAP; Freire, MG
our authors
Ana Francisca Osório de Almeida Coelho e Silva
Junior Researcher
Ana Mafalda Rodrigues Almeida Rocha
Researcher
Mara Guadalupe Freire Martins
Coordinating Researcher
Mohamed Taha Ahmed Amin
Post-doc Fellowship
Sónia Patrícia Marques Ventura
Assistant ProfessorGroups
G4 - Renewable Materials and Circular Economy
G5 - Biomimetic, Biological and Living Materials
Projects
Igy Technology: A Purication Platform using Ionic-Liquid-Based Aqueous Biphasic Systems (IGYPURTECH)
acknowledgements
The authors are grateful for financial support from FEDER funds through the program COMPETE and from the national fund through the Portuguese Foundation for Science and Technology (FCT) under the scope of the projects Pest-C/CTM/LA0011/2013 and PTDC/AAC-AMB/119172/2010. The authors are also thankful to FCT for financial support through the doctoral and post-doctoral grants of SFRH/BD/94901/2013, SFRH/BPD/78441/2011 and SFRH/BPD/79263/2011 of F.A.eS., M.T. and S.P.M.V., respectively. M.G.F. acknowledges the European Research Council (ERC) for the Grant ERC-2013-StG-337753.