Artículos de revistas
Separation Of Human Igg Fragments Using Copper, Nickel, Zinc, And Cobalt Chelated To Cm-asp-agarose By Positive And Negative Chromatography
Registro en:
1873-376X
Journal Of Chromatography B. ELSEVIER SCIENCE BV, n. 1017-1018, p. 163 - 173.
1570-0232
WOS:000373547200020
10.1016/j.jchromb.2016.01.058
Autor
Mourao
CA; Carmignotto
GP; Bueno
SMA
Institución
Resumen
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) This study evaluated the feasibility of using immobilized metal-ion affinity chromatography (IMAC) for separation of human Fab fragments using four different transition metal ions copper, nickel, zinc, and cobalt chelated to CM-Asp (carboxymethylaspartate) immobilized on the agarose gel. The Fab and Fc fragments (from human IgG digested with papain) interacted differently with the chelates studied, depending on the adsorption buffer system. The interaction between chelate and Fc fragment is predominantly based on the coordination bonds using adsorption buffer containing NaCl. Negative chromatography was performed on Cu(II)-CM-Asp-agarose obtaining 2.9 mg of Fab per mL of adsorbent in nonretained fractions (Fc fragment-free without uncleaved IgG). The adsorption of Fab fragments is governed by electrostatic forces in the absence of NaCl in the adsorption buffer. High selectivity was achieved on Co(II)-CM-Asp-agarose and 5.7 mg of Fab per mL of adsorbent was obtained in eluted fractions without Fc fragments, although having uncleaved IgG. The results showed that chromatography on transition metal ions chetated to CM-Asp-agarose is a promising approach to separation of Fab fragments from papain-digested human IgG solution. (C) 2016 Elsevier B.V. All rights reserved. 1017-1018
163 173 CNPq CAPES CAPES/PROEX, Brazil Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)