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Lectin Affinity Chromatography: An Efficient Method to Purify Horse IgG3.
De-Simone, Salvatore G; Provance, David W.
Afiliação
  • De-Simone SG; FIOCRUZ, Center for Technological Development in Health (CDTS)/National Institute of Science and Technology for Innovation on Neglected of Population Diseases (INCT-INDP), Rio de Janeiro, RJ, Brazil. dsimone@cdts.fiocruz.br.
  • Provance DW; FIOCRUZ, Oswaldo Cruz Institute, Laboratory of Experimental and Computational Biochemistry of Pharmaceuticals, Rio de Janeiro, RJ, Brazil. dsimone@cdts.fiocruz.br.
Methods Mol Biol ; 2178: 301-310, 2021.
Article em En | MEDLINE | ID: mdl-33128757
Affinity chromatography is a separation method based on a specific binding interaction between an immobilized ligand and its binding partner. An important class of ligands for the effective separation and purification of biotechnologically important substances is lectins, a group of naturally occurring molecules widely found in plants that display a range of specificities to bind different sugars. As sugars are often added to proteins through the process of glycosylation, ∼1/3 of all genetically encoded proteins are glycosylated, numerous cognate pairs of lectins with glycosylation groups have been discovered. Their specific binding interactions have not only allowed the development of numerous methodological strategies involving immobilized lectins to isolate molecules of interests but also for understanding the intermolecular interactions and alterations in glycosylation during a diverse set of biological phenomena, including tumor cell metastasis, intracellular communication, and inflammation. In this chapter, we describe a basic procedure for the separation of horse antibody classes by affinity chromatography based on differences in their glycosylation patterns. This procedure has been utilized for the purification of horse IgG3 (hoIgG3) from other six Ig from equine sera in a single step by using an Artocarpus integrifolia Jacalin column. This class of antibody comprises the therapeutic fraction generated in equine for passive antibody therapy and can serve as a biomarker for patient hypersensitivity. During the course of developing the protocol, the affinity interaction constant between the huIgE-hypersensitive immunoglobulin and the purified hoIgG3 was also determined.
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Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Imunoglobulina E / Imunoglobulina G / Cromatografia de Afinidade / Lectinas de Plantas / Cavalos Tipo de estudo: Guideline Limite: Animals / Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos
Texto completo
Adicionar na Minha BVS
Imprimir
XML
PubMed Links
Buscar no Google

Texto completo: 1 Coleções: 01-internacional Base de dados: MEDLINE Assunto principal: Imunoglobulina E / Imunoglobulina G / Cromatografia de Afinidade / Lectinas de Plantas / Cavalos Tipo de estudo: Guideline Limite: Animals / Humans Idioma: En Revista: Methods Mol Biol Assunto da revista: BIOLOGIA MOLECULAR Ano de publicação: 2021 Tipo de documento: Article País de afiliação: Brasil País de publicação: Estados Unidos