Preclinical evaluation of a humanized NR-LU-10 antibody-streptavidin fusion protein for pretargeted cancer therapy

被引：33

作者：

Goshorn, S ^{[1
]}

Sanderson, J ^{[1
]}

Axworthy, D ^{[1
]}

Lin, YK ^{[1
]}

Hylarides, M ^{[1
]}

Schultz, J ^{[1
]}

机构：

[1] NEORX Corp, Seattle, WA 98119 USA

来源：

CANCER BIOTHERAPY AND RADIOPHARMACEUTICALS | 2001年 / 16卷 / 02期

关键词：

multivalent; pretargeted radioimmunotherapy; single-chain Fv; streptavidin; tumor targeting; E. coli expression;

D O I：

10.1089/108497801300189209

中图分类号：

R73 [肿瘤学];

学科分类号：

100214 ;

摘要：

A humanized single chain F nu antibody fragment specific to the EGP40 antigen was genetically engineered as a streptavidin fusion (scF nu SA) for use in pretargeted radioimmunotherapy. The scF nu SA construct was expresses as a soluble, tetrameric species in the Escherichia coli periplasm at 110-140 mg/liter. The fusion protein was purified from crude lysates by iminobiotin affinity chromatography with an overall yield of 50-60%. Characterization of the purified protein by SDS-PAGE, light scattering, and size exclusion chromatography demonstrated that the fusion protein was tetrameric with a molecular weight of similar to 172,000. Competitive immunoreactivity assays showed a two-fold greater binding to the antigen than the comparable whole antibody. The purified protein had a biotin disassociation rate identical to recomdinant streptavidin and bound an average of three of four possible biotins per molecules. The radiolabeled fusion protein showed a faster blood clearance rate in normal mince than the corresponding whole antibody-streptavidin molecule was demonstrated in nude bearing SW1222 human colon carcinoma xenografts. A single dose of 800 mu Ci of Y-90-DOTA-biotin produced cures in mice with established subcutaneous human small cell lung or colon cancer xenografts.

引用

页码：109 / 123

页数：15

共 37 条

[1] MOLECULAR-CLONING AND NUCLEOTIDE-SEQUENCE OF THE STREPTAVIDIN GENE [J].