Cloning and expression of a single human immunoglobulin heavy-chain variable domain with vascular endothelial growth factor binding activity / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 1555-1562, 2010.
Artículo
en Chino
| WPRIM
| ID: wpr-351562
ABSTRACT
In the application of therapeutic antibodies, large molecular weight of antibodies is always a problem that prevents them from penetrating into tissues or binding to antigenic determinants. To overcome this problem, we investigated the function of the heavy chain variable domain of a monoclonal anti-VEGF human IgM antibody derived from the Five-Feature Translocus Mice. We cloned the cDNA of the heavy chain variable domain, which was then inserted into pET28a vector and expressed in Escherichia coli. After purification and renaturation of the denatured recombinant protein, we obtained a 16 kDa antibody fragment, which is named as rhVVH. By immunoassaying its VEGF-binding capability in vitro, we proved that rhVVH retains this activity as the complete IgM. Importantly, rhVVH is shown to inhibit the HUVEC cell proliferation in a concentration-dependent manner. Our results indicate that the single heavy chain variable domain might inherit part of the biological function of the complete IgM antibody, which provided a valuable potential in further research on antibody miniaturisation.
Texto completo:
Disponible
Índice:
WPRIM (Pacífico Occidental)
Asunto principal:
Proteínas Recombinantes
/
Región Variable de Inmunoglobulina
/
Datos de Secuencia Molecular
/
Secuencia de Bases
/
Secuencia de Aminoácidos
/
Clonación Molecular
/
Cadenas Pesadas de Inmunoglobulina
/
Factor A de Crecimiento Endotelial Vascular
/
Escherichia coli
/
Anticuerpos de Cadena Única
Límite:
Animales
/
Humanos
Idioma:
Chino
Revista:
Chinese Journal of Biotechnology
Año:
2010
Tipo del documento:
Artículo
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