IgG purification by negative chromatography in amine-based ligands: A comparison of L-lysine and poly-L-lysine

IgG purification by negative chromatography in amine-based ligands: A comparison of L-lysine and poly-L-lysine

Author Lazzarotto Bresolin, Igor Tadeu Autor UNIFESP Google Scholar
Fioritti, Renato Rodrigues Google Scholar
Alves Buenoa, Sonia Maria Google Scholar
Institution Universidade Estadual de Campinas (UNICAMP)
Universidade Federal de São Paulo (UNIFESP)
Abstract Agarose gels with immobilized amine-based ligands L-lysine (Lys) and poly-L-lysine (PLL) exhibited the ability to adsorb human serum proteins and to separate immunoglobulin G (IgG) in nonretained chromatographic fractions (negative chromatography). the effect of the buffer system and pH on IgG purification on Lys-agarose and PLL-agarose showed that PLL was the most selective ligand. PLL-agarose was able to purify IgG from human serum solution diluted in Bis-Tris buffer at pH 6.0 with 79% yield and 88.7% purity (based on total protein concentration and nephelometric analysis of albumin, transferrin, and immunoglobulins A, G, and M). Human serum albumin (HSA) and IgG adsorption data showed that the first followed the Langmuir model whereas the second followed the Langmuir-Freundlich model, due to the presence of a positive cooperativity effect (n = 1.60), with the maximum adsorption capacities of 76.4 mg mL(-1) for HSA and 23.6 mg mL(-1) for IgG. the Delta G(max) values obtained by non-linear regression of HSA and IgG adsorption data of Temkin model were similar (-7.0 kcal mol(-1)), indicating that the process is spontaneous in nature. the joint analysis of these data shows that PLL-agarose can be considered an alternative adsorbent for the isolation of IgG from human serum with the potential to be integrated to a purification process on a large scale. (C) 2011 Elsevier B.V. All rights reserved.
Keywords IgG purification
Human serum
Negative chromatography
Electrostatic interactions
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Date 2011-12-01
Published in Process Biochemistry. Oxford: Elsevier B.V., v. 46, n. 12, p. 2277-2285, 2011.
ISSN 1359-5113 (Sherpa/Romeo, impact factor)
Publisher Elsevier B.V.
Extent 2277-2285
Origin http://dx.doi.org/10.1016/j.procbio.2011.09.005
Access rights Closed access
Type Article
Web of Science ID WOS:000297832000009
URI http://repositorio.unifesp.br/handle/11600/34303

Show full item record


File Size Format View

There are no files associated with this item.

This item appears in the following Collection(s)




My Account