Soluble expression and target study to brain of anti-TfR ScFv / 生物工程学报
Chinese Journal of Biotechnology
;
(12): 342-347, 2004.
Article
Dans Chinois
| WPRIM
| ID: wpr-249985
ABSTRACT
The single-chain antibody gene (ox26-scFv) to transferrin receptor (TfR) was synthesized and amplified by three-step PCR. After sequencing, the gene was cloned into prokaryotic expression vector pTIG-Trx which carried thioredoxin (Trx) gene and a C-terminal His.tag. The Ox26-scFv proteins achieved 31% yields of total bacteria proteins at 20 degrees C, after 0.02mM IPTG induction using the strain E. coli BL21 (DE3). The soluble scFv proteins in cytoplasm suspension were about 35% and the inclusion bodies were about 65%. The soluble products were purified by immobilized metal chelation affinity chromatography (Ni-NTA), a single band with molecular weight 29 kD appeared on SDS-PAGE gel. Rat GH3 cell immunocytochemistry staining showed that Ox26-scFv protein could recognize and bind to transferrin receptor. Injected SD rats with Ox26-scFv proteins by tail veins, the antibodies were detected from brain tissues specially on the brain capillaries 4 h later which indicate that Ox26-scFv proteins have a good target function to brain capillaries and can permeate the blood-brain barrier mediated by the transferrin receptors.
Texte intégral:
Disponible
Indice:
WPRIM (Pacifique occidental)
Sujet Principal:
Récepteurs à la transferrine
/
Données de séquences moléculaires
/
Séquence nucléotidique
/
Barrière hémato-encéphalique
/
Systèmes de délivrance de médicaments
/
Rat Sprague-Dawley
/
Allergie et immunologie
/
Vecteurs génétiques
/
Génétique
/
Métabolisme
Limites du sujet:
Animaux
langue:
Chinois
Texte intégral:
Chinese Journal of Biotechnology
Année:
2004
Type:
Article
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