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

Gubala, Vladimir, Crean, Carol, Nooney, Robert, Hearty, Stephen, McDonnell, Barry, Heydon, Katherine, O'Kennedy, Richard, MacCraith, Brian D, Williams, David 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) (The full text of this publication is not currently available from this repository. You may be able to access a copy if URLs are provided)

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Official URL
http://dx.doi.org/10.1039/c1an15017k

Abstract

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.

Item Type: Article
DOI/Identification number: 10.1039/c1an15017k
Additional information: number of additional authors: 8;
Subjects: Q Science
R Medicine > RS Pharmacy and materia medica
Divisions: Faculties > Sciences > Medway School of Pharmacy
Depositing User: Stewart Brownrigg
Date Deposited: 07 Mar 2014 00:05 UTC
Last Modified: 29 May 2019 12:24 UTC
Resource URI: https://kar.kent.ac.uk/id/eprint/40398 (The current URI for this page, for reference purposes)
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