Data/hora: 06/06/2024 01:22:50

Provedor de dados: | 56
País: | Brazil
Título: | Expression, production, and renaturation of a functional single-chain variable antibody fragment (scFv) against human ICAM-1
Autores: | Sun,H.; Wu,G.M.; Chen,Y.Y.; Tian,Y.; Yue,Y.H.; Zhang,G.L.
Data: | 2014-07-01
Ano: | 2014
Palavras-chave: | Intercellular adhesion molecule-1; Single-chain variable antibody fragment; Expression; Purification; Renaturation; Biological activity.
Resumo: | Intercellular adhesion molecule-1 (ICAM-1) is an important factor in the progression of inflammatory responses in vivo. To develop a new anti-inflammatory drug to block the biological activity of ICAM-1, we produced a monoclonal antibody (Ka=4.19×10−8 M) against human ICAM-1. The anti-ICAM-1 single-chain variable antibody fragment (scFv) was expressed at a high level as inclusion bodies in Escherichia coli. We refolded the scFv (Ka=2.35×10−7 M) by ion-exchange chromatography, dialysis, and dilution. The results showed that column chromatography refolding by high-performance Q Sepharose had remarkable advantages over conventional dilution and dialysis methods. Furthermore, the anti-ICAM-1 scFv yield of about 60 mg/L was higher with this method. The purity of the final product was greater than 90%, as shown by denaturing gel electrophoresis. Enzyme-linked immunosorbent assay, cell culture, and animal experiments were used to assess the immunological properties and biological activities of the renatured scFv.
Tipo: | Info:eu-repo/semantics/article
Idioma: | Inglês
Identificador: | http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0100-879X2014000700540
Editor: | Associação Brasileira de Divulgação Científica
Relação: | 10.1590/1414-431X20143276
Formato: | text/html
Fonte: | Brazilian Journal of Medical and Biological Research  v.47 n.7 2014
Fonte: | info:eu-repo/semantics/openAccess