Covalent conjugation of cysteine-engineered scFv to PEGylated magnetic nanoprobes for immunotargeting of breast cancer cells

被引：20

作者：

Alric, Christophe ^{[1
]}

Aubrey, Nicolas ^{[2
]}

Allard-Vannier, Emilie ^{[1
]}

di Tommaso, Anne ^{[2
]}

Blondy, Thibaut ^{[1
]}

Dimier-Poisson, Isabelle ^{[2
]}

Chourpa, Igor ^{[1
]}

Herve-Aubert, Katel ^{[1
]}

机构：

[1] Univ Tours, Nanomedicaments & Nanosondes EA6295, F-37200 Tours, France

[2] Univ Tours, INRA Infectiol & Sante Publ UMR1282, F-37000 Tours, France

来源：

RSC ADVANCES | 2016年 / 6卷 / 43期

关键词：

IRON-OXIDE NANOPARTICLES; GOLD NANOPARTICLES; PROTEIN-L; IN-VITRO; ANTIBODY; BIODISTRIBUTION; DELIVERY; PURIFICATION; DESIGN; MRI;

D O I：

10.1039/c6ra06076e

中图分类号：

O6 [化学];

学科分类号：

0703 ;

摘要：

In the present study, we describe the synthesis and characterization of new generation of cancer-targeting magnetic nanoprobes: superparamagnetic iron oxide nanoparticles (SPIONs) coated with polyethylene glycol (PEG) shell functionalized with recombinant anti-HER2 single chain fragment variable (scFv) of Trastuzumab antibody. An anti-HER2 scFv with terminal cysteine (scFv 4D5-Cys) has been rationally engineered in order to favor its orientation-and site-directed covalent conjugation to the polymeric surface of PEGylated SPIONs. Optimization of scFv and nanoparticles production allowed to obtain well-characterized SPIONs-PEG-scFv nanoparticles carrying similar to 7 fragments per nanoparticle, having a hydrodynamic diameter of ca. 86 nm and nearly neutral surface. The nanoprobes-scFv capability to recognize the HER2 protein has been confirmed by enzyme-linked immunosorbent assay (ELISA). Compared to non-targeted PEGylated SPIONs, the SPIONs-PEG-scFv nanoprobes showed an enhanced binding to HER2-overexpressing cells (SK-BR3) in vitro as it was shown by immunofluorescence. Finally, ICP-AES measurements shown that in 1 hour the uptake of SPIONs-PEG-scFv in HER2-overexpressing cells is 2.1 times greater than non-targeted PEGylated SPIONs. Therefore, both due to their physico-chemical characteristics and the immunotargeting of HER2-positive breast cancer cells, the SPIONs-PEG-scFv appear as promising nanoplatforms for future applications in theranostic treatment of cancers.

引用

页码：37099 / 37109

页数：11

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