Phosphorylated-tyrosine based pseudobioaffinity adsorbent for the purification of immunoglobulin G
Pavan, Gisele Luiza
Lazzarotto Bresolin, Igor Tadeu [UNIFESP]
Alves Bueno, Sonia Maria
Is part ofJournal Of Chromatography B-Analytical Technologies In The Biomedical And Life Sciences
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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.
CitationJournal Of Chromatography B-Analytical Technologies In The Biomedical And Life Sciences. Amsterdam, v. 1052, p. 10-18, 2017.
SponsorshipFAPESP - 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)
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