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Kinetics of immunoassays with particles as labels: Effect of antibody coupling using dendrimers as linkers

Gubala, V., Crean, C., Nooney, R., Hearty, S., McDonnell, B., Heydon, K., O'Kennedy, R., MacCraith, B.D., Williams, D.E. (2011) Kinetics of immunoassays with particles as labels: Effect of antibody coupling using dendrimers as linkers. Analyst, 136 (12). pp. 2533-2541. ISSN 0003-2654. (doi:10.1039/c1an15017k) (KAR id:45229)

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In this article, we report on poly(amidoamine) dendrimers (PAMAM) as coupling agents for recombinant single-chain (ScFv) antibodies to nanoparticle (NP) labels, for use in immunoassay. We present a simple theory for the kinetics of particle capture onto a surface by means of an antibody-antigen reaction, in which the important parameter is the fraction of the particle surface that is active for reaction. We describe how increasing the generation number of the linking dendrimers significantly increased the fraction of the NP surface that is active for antigen binding and consequently also increased the assay kinetic rates. Use of dendrimers for conjugation of the NP to the antibody resulted in a significantly higher surface coverage of active antibody, in comparison with mono-valent linker chemistry. As a direct consequence, the increase in effective avidity significantly out-weighed any effect of a decreased diffusion coefficient due to the NP, when compared to that of a molecular dye-labelled antibody. The signal to noise ratio of the G4.5 dendrimer-sensitised nanoparticles out-performed the dye-labelled antibody by approximately four-fold. Particle aggregation experiments with the multi-valent antigen CRP demonstrated reaction-limited aggregation whose rate increased significantly with increasing generation number of the dendrimer linker. © 2011 The Royal Society of Chemistry.

Item Type: Article
DOI/Identification number: 10.1039/c1an15017k
Additional information: Unmapped bibliographic data: LA - English [Field not mapped to EPrints] J2 - Analyst [Field not mapped to EPrints] C2 - 21541412 [Field not mapped to EPrints] AD - Biomedical Diagnostics Institute (BDI), Dublin City University, Collins Avenue, Glasnevin, Dublin 9, Ireland [Field not mapped to EPrints] AD - School of Biotechnology, Dublin City University, Dublin 9, Ireland [Field not mapped to EPrints] AD - MacDiarmid Institute for Advanced Materials and Nanotechnology, Department of Chemistry, University of Auckland, Private Bag 92019, Auckland 1142, New Zealand [Field not mapped to EPrints] DB - Scopus [Field not mapped to EPrints]
Uncontrolled keywords: dendrimer, nanoparticle, PAMAM Starburst, recombinant protein, single chain fragment variable antibody, article, chemistry, genetics, immunoassay, immunology, kinetics, metabolism, methodology, Dendrimers, Immunoassay, Kinetics, Nanoparticles, Recombinant Proteins, Single-Chain Antibodies
Divisions: Divisions > Division of Natural Sciences > Medway School of Pharmacy
Depositing User: Vladimir Gubala
Date Deposited: 14 Dec 2017 20:16 UTC
Last Modified: 16 Nov 2021 10:18 UTC
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