[Soluble expression and target study to brain of anti-TfR ScFv].
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.
Yan
Bing
B
Beijing Institute of Biotechnology, Beijing 100071, China. yanbing72@hotmail.com
Zhu
Heng-Qi
HQ
Huang
Pei-Tang
PT
D000818:Animals; D000906:Antibodies / Q000008:administration & dosage* / Q000235:genetics* / Q000378:metabolism; D000917:Antibody Formation; D001483:Base Sequence; D001812:Blood-Brain Barrier / Q000187:drug effects*; D016503:Drug Delivery Systems; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D051381:Rats; D017207:Rats, Sprague-Dawley; D011990:Receptors, Transferrin / Q000276:immunology*
D000906:Antibodies; D011990:Receptors, Transferrin