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
in Chinese
| WPRIM (Western Pacific)
| ID: wpr-249985
Responsible library:
WPRO
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.
Full text:
Available
Database:
WPRIM (Western Pacific)
Main subject:
Receptors, Transferrin
/
Molecular Sequence Data
/
Base Sequence
/
Blood-Brain Barrier
/
Drug Delivery Systems
/
Rats, Sprague-Dawley
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Allergy and Immunology
/
Genetic Vectors
/
Genetics
/
Metabolism
Limits:
Animals
Language:
Chinese
Journal:
Chinese Journal of Biotechnology
Year:
2004
Document type:
Article