Application
Competitive ELISA
We developed a quantitative and sensitive immunoassay to determine Human IgE in human serum using gold nanoclusters (AuNCs) as fluorescent labels. The synthesized AuNCs have a particle size of 2.7 ± 0.1 nm, a maximum fluorescence emission at 710 nm and high stability. The covalent bioconjugation of these AuNCs with the antibody was carried out by the carbodiimide reaction. A molar ratio 1:3 (antibody:AuNCs) was selected.
Human IgE from Abcam (ab65866) was used as standard to prepare the calibration curve in PBS and in human serum matrix (attached figure). Two immunoassay configurations were assayed (competitive and sandwich format), obtaining better detection limit with the competitive format (0.2 ng/mL). For both formats the detection limits were better than those obtained with current commercial ELISA kits (12 ng/mL).
Moreover, the developed fluorescent competitive immunoassay offers other interesting advantages such as fewer reagents and simpler procedures.
The applicability of this new AuNCs immunoprobe was successfully demonstrated for the determination of IgE in human serum, using the competitive configuration.
This research has been published in Biosensors and Bioelectronics,
http://dx.doi.org/10.1016/j.bios.2015.08.011
Authors: María Cruz Alonso, Laura Trapiella-Alfonso, José M Costa Fernández, Rosario Pereiro, Alfredo Sanz-Medel
Title: “Functionalized gold nanoclusters as fluorescent labels for immunoassays: Application to Human serum immunoglobulin E determination”
Human IgE from Abcam (ab65866) was used as standard to prepare the calibration curve in PBS and in human serum matrix (attached figure). Two immunoassay configurations were assayed (competitive and sandwich format), obtaining better detection limit with the competitive format (0.2 ng/mL). For both formats the detection limits were better than those obtained with current commercial ELISA kits (12 ng/mL).
Moreover, the developed fluorescent competitive immunoassay offers other interesting advantages such as fewer reagents and simpler procedures.
The applicability of this new AuNCs immunoprobe was successfully demonstrated for the determination of IgE in human serum, using the competitive configuration.
This research has been published in Biosensors and Bioelectronics,
http://dx.doi.org/10.1016/j.bios.2015.08.011
Authors: María Cruz Alonso, Laura Trapiella-Alfonso, José M Costa Fernández, Rosario Pereiro, Alfredo Sanz-Medel
Title: “Functionalized gold nanoclusters as fluorescent labels for immunoassays: Application to Human serum immunoglobulin E determination”
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.
María Cruz
Verified customer
提交于 Sep 29 2015