Phosphorylated-tyrosine based pseudobioaffinity adsorbent for the purification of immunoglobulin G

Phosphorylated-tyrosine based pseudobioaffinity adsorbent for the purification of immunoglobulin G

Author Pavan, Gisele Luiza Google Scholar
Lazzarotto Bresolin, Igor Tadeu Autor UNIFESP Google Scholar
Grespan, Angelica Google Scholar
Alves Bueno, Sonia Maria Google Scholar
Abstract The present study evaluated the phosphorylated-tyrosine (P-Tyr) based pseudobioaffinity adsorbent for the purification of human immunoglobulin G (IgG). P-Tyr was selected as a ligand to mimic the natural interactions that occur between the immunoreceptor tyrosine-based activation motif and the IgG. The ligand was coupled to bisoxirane-activated agarose gel and the effect of buffer system, pH, and conductivity was performed to elucidate the nature of IgG-P-Tyr interactions. P-Tyr-agarose was able to purify IgG from human plasma solution in HEPES buffer at pH 7.0 exhibiting a purification factor of 9.1 with IgG purity of 91% (based on ELISA analysis of albumin, transferrin, and immunoglobulins A, G, and M). The evaluation of different functional groups of P-Tyr on the adsorption of human IgG indicated the predominance of electrostatic interactions with phosphate groups, although the contributions of aromatic and carboxylic groups also play a role. The thermodynamic parameters (Delta H degrees, Delta S degrees, Delta G degrees) for IgG adsorption onto P-Tyr-agarose were determined from the temperature dependence. The maximum IgG binding capacity at 20 degrees C was 273.51 +/- 12.63 mg g(-1) and the dissociation constant value of the complex IgG-P-Tyr was in the order of 10(-5) mol L-1 indicating low-affinity. (C) 2017 Published by Elsevier B.V.
Keywords O-Phospho-tyrosine
Human immunoglobulin G
Adsorption
Purification
Human plasma
xmlui.dri2xhtml.METS-1.0.item-coverage Amsterdam
Language English
Sponsor FAPESP - Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (Brazil)
CAPES - Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (Brazil)
CNPq - Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (Brazil)
Date 2017
Published in Journal Of Chromatography B-Analytical Technologies In The Biomedical And Life Sciences. Amsterdam, v. 1052, p. 10-18, 2017.
ISSN 1570-0232 (Sherpa/Romeo, impact factor)
Publisher Elsevier Science Bv
Extent 10-18
Origin http://dx.doi.org/10.1016/j.jchromb.2017.03.018
Access rights Closed access
Type Article
Web of Science ID WOS:000402218900002
URI https://repositorio.unifesp.br/handle/11600/54578

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