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Coiled-Coil-Based Biofunctionalization of 100 nm Gold Nanoparticles with the Trastuzumab Antibody for the Detection of HER2-Positive Cancer Cells
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DOI：10.1021/acs.langmuir.3c01621.png)
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Abstract
En 中文
Wecompared different biofunctionalization strategiesfor immobilizingtrastuzumab, an IgG targeting the HER2 biomarker, onto 100 nm sphericalgold nanoparticles because of the E/K coiled-coil peptide heterodimer.First, Kcoil peptides were grafted onto the gold surface while theirEcoil partners were genetically encoded at the C-terminus of trastuzumab'sFc region, allowing for a strong and specific interaction betweenthe antibodies and the nanoparticles. Gold nanoparticles with no Kcoilpeptides on their surface were also produced to immobilize Ecoil-taggedtrastuzumab antibodies via the specific adsorption of their negativelycharged Ecoil tags on the positively charged gold surface. Finally,the nonspecific adsorption of wild-type trastuzumab on the gold surfacewas also assessed, with and without Kcoil peptides grafted on it beforehand.We developed a thorough workflow to systematically compare the immobilizationstrategies regarding the stability of nanoparticles, antibody coverage,and ability to specifically bind to HER2-positive breast cancer cells.All nanoparticles were highly monodisperse and retained their localizedsurface plasmon resonance properties after biofunctionalization. Asignificant increase in the amount of immobilized antibodies was observedwith the two oriented coil-based strategies compared to nonspecificadsorption. Finally, all biofunctionalization strategies allowed forthe detection of HER2-positive breast cancer cells, but among theinvestigated approaches, we recommend using the E/K coiled-coil-basedstrategy for gold nanoparticle biofunctionalization because it allowsfor the qualitative and quantitative detection of HER2-positive cellswith a higher contrast compared to HER2-negative cells.
Keywords:
PROTEIN-G
ORIENTED IMMOBILIZATION
MONOCLONAL-ANTIBODIES
LIGHT-SCATTERING
DRUG-DELIVERY
IGG
STRATEGY
QUANTIFICATION
ADSORPTION
SURFACE
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