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  • Impact of conjugation strategies for targeting of antibodies in gold nanoparticles for ultrasensitive detection of 17β-estradiol.

Impact of conjugation strategies for targeting of antibodies in gold nanoparticles for ultrasensitive detection of 17β-estradiol.

Scientific reports (2019-09-27)
Jairo Pinto Oliveira, Adilson Ribeiro Prado, Wanderson Juvencio Keijok, Paulo Wagnner Pereira Antunes, Enrique Ronald Yapuchura, Marco Cesar Cunegundes Guimarães
ABSTRACT

Antibody-coated nanoparticles have recently attracted considerable attention, with the focus falling on diagnostics. Nevertheless, controlled antibody bioconjugation remains a challenge. Here, we present two strategies of bioconjugation with the aim of evaluating the best approach for the coupling of antibodies on the surface of nanomaterials in an oriented way. We employed electrostatic interaction (physical adsorption) and covalent conjugation in the orientation of antibodies on the metallic surface as coupling methods, and their influence on the detection of 17β-estradiol was addressed with localized surface plasmon resonance. The understanding of these mechanisms is fundamental for the development of reproducible inorganic bioconjugates with oriented surface as well sensibility of immunoassays.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
16-Mercaptohexadecanoic acid, 98%
Sigma-Aldrich
16-Mercaptohexadecanoic acid, 90%
Sigma-Aldrich
β-Estradiol, powder, γ-irradiated, suitable for cell culture
Sigma-Aldrich
Poly(ethylene glycol) methyl ether thiol, average Mn 6,000
Supelco
Sodium Citrate, Pharmaceutical Secondary Standard; Certified Reference Material
Sigma-Aldrich
N-(3-Dimethylaminopropyl)-N′-ethylcarbodiimide hydrochloride, crystalline
Sigma-Aldrich
N-Hydroxysuccinimide, 98%
Sigma-Aldrich
Gold(III) chloride trihydrate, ACS reagent, ≥49.0% Au basis