| Registro completo |
| Provedor de dados: |
BJMBR
|
| 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
|
| Direitos: |
info:eu-repo/semantics/openAccess
|
