Design and Development of a Field Applicable Gold Nanosensor for the Detection of Luteinizing Hormone


In this paper, we describe a novel strategy for the fabrication of a nanosensor for detecting luteinizing hormone (LH) of sheep using a gold nanoparticle-peptide conjugate. A new peptide sequence "CDHPPLPDILFL" (leutinizing hormone peptide, LHP) has been identified, using BLAST and Clustal W analysis, to detect antibody of LH (sheep). LHP has been synthesized and characterized, and their affinity toward anti-LH was established using enzyme linked immunosorbant assay (ELISA) technique. The thiol group in LHP directly binds with gold nanoparticles (AuNPs) to yield AuNP-LHP construct. Detailed physicochemical analysis of AuNP-LHP construct was determined using various analytical techniques. Nanosensor using gold nanoparticle peptide conjugate was developed on the basis of competitive binding of AuNP-LHP and LH toward anti-LH. Nitrocellulose membrane, precoated with anti-LH, was soaked in the mixture of AuNP-LHP and sample of analysis (LH). In the absence of LH (sheep), anti-LH coated on the membrane binds with AuNP-LHP, leading to a distinctive red color, while in the presence of LH, no color appeared in the membrane due to the interaction of anti-LH with LH thereby preventing the binding of AuNP-LHP with membrane bound anti-LH. The sensor assay developed in this study can detect LH (sheep) up to a minimal concentration of ~50 ppm with a high degree of reproducibility and selectivity. The gold-nanoparticle-peptide based nanosensor would be a simple, portable, effective, and low cost technique for infield applications.



Keywords and Phrases

Competitive binding; Design and Development; Gold nanoparticle; Gold Nanoparticles; Immunosorbant assay; Low costs; Membrane-bound; Nitrocellulose membranes; Novel strategies; Peptide sequences; Physico-chemical analysis; Reproducibilities; Thiol groups, Antibodies; Metal nanoparticles; Nanosensors; Nitrocellulose; Peptides, Gold, gold; luteinizing hormone; metal nanoparticle; oligopeptide, amino acid sequence; animal; article; blood; chemistry; enzyme linked immunosorbent assay; equipment; equipment design; genetic procedures; surface property, Amino Acid Sequence; Animals; Biosensing Techniques; Enzyme-Linked Immunosorbent Assay; Equipment Design; Gold; Luteinizing Hormone; Metal Nanoparticles; Oligopeptides; Surface Properties

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Article - Journal

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© 2012 American Chemical Society (ACS), All rights reserved.

Publication Date

01 Nov 2012