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http://hdl.handle.net/10773/34417
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DC Field | Value | Language |
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dc.contributor.author | Almeida, Mafalda R. | pt_PT |
dc.contributor.author | Ferreira, Filipe | pt_PT |
dc.contributor.author | Domingues, Pedro dimas | pt_PT |
dc.contributor.author | Coutinho, João A. P. | pt_PT |
dc.contributor.author | Freire, Mara G. | pt_PT |
dc.date.accessioned | 2022-08-05T16:54:09Z | - |
dc.date.available | 2022-08-05T16:54:09Z | - |
dc.date.issued | 2022-10-15 | - |
dc.identifier.issn | 1383-5866 | pt_PT |
dc.identifier.uri | http://hdl.handle.net/10773/34417 | - |
dc.description.abstract | Although their high potential as alternative biopharmaceuticals, less than 2% of the total polyclonal antibodies produced worldwide correspond to immunoglobulin Y (IgY) due to the difficulties in isolating them from egg yolk (complex biological matrix). In this work, the water-soluble proteins fraction (WSPF) of egg yolk was first obtained and the proteins present identified by one-dimensional gel electrophoresis (SDS-PAGE) and label-free quantitative nano-liquid chromatography-tandem mass spectrometry (nano-LC-MS/MS). The egg yolk WSPF was then applied to create aqueous biphasic systems (ABS) composed of polyethylene glycol 1000 g·mol−1 (PEG 1000) and K2HPO4/ KH2PO4 buffer, followed by centrifugal partition chromatography (CPC) to purify IgY. The characterization of the WSPF showed the presence of six major proteins: the target antibody IgY, serum albumin (α-livetin), ovalbumin, ovotransferrin, vitellogenin 1 and vitellogenin 2. The results obtained by ABS revealed a high affinity of all proteins to the polymer-rich phase. However, by changing the PEG and salt concentrations, a higher selectivity was observed for IgY, with the remaining proteins partitioning between the two phases. The best ABS were applied in CPC, finally allowing a multi-stage partition and to the technology scale-up. The CPC operating conditions were optimized, allowing to obtain IgY with 50.6% of purity. | pt_PT |
dc.language.iso | eng | pt_PT |
dc.publisher | Elsevier | pt_PT |
dc.relation | info:eu-repo/grantAgreement/EC/H2020/899921/EU | pt_PT |
dc.relation | info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDB%2F50011%2F2020/PT | pt_PT |
dc.relation | info:eu-repo/grantAgreement/FCT/6817 - DCRRNI ID/UIDP%2F50011%2F2020/PT | pt_PT |
dc.relation | LA/P/0006/2020 | pt_PT |
dc.rights | openAccess | pt_PT |
dc.rights.uri | https://creativecommons.org/licenses/by/4.0/ | pt_PT |
dc.subject | Immunoglobulin Y | pt_PT |
dc.subject | Purification | pt_PT |
dc.subject | Aqueous biphasic systems | pt_PT |
dc.subject | Centrifugal partition chromatography | pt_PT |
dc.title | Towards the purification of IgY from egg yolk by centrifugal partition chromatography | pt_PT |
dc.type | article | pt_PT |
dc.description.version | published | pt_PT |
dc.peerreviewed | yes | pt_PT |
degois.publication.title | Separation and Purification Technology | pt_PT |
degois.publication.volume | 299 | pt_PT |
dc.identifier.doi | 10.1016/j.seppur.2022.121697 | pt_PT |
dc.identifier.articlenumber | 121697 | pt_PT |
Appears in Collections: | CICECO - Artigos DQ - Artigos REQUIMTE - Artigos |
Files in This Item:
File | Description | Size | Format | |
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Manuscript_Almeida et al _submitted.pdf | 734.8 kB | Adobe PDF | View/Open |
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