A New Electrochemiluminescence Immunoassay Based on Magnetic Microbeads as Carrier of Labels
Bing Liu, Zhaoyang Tong, Lan-Qun HAO, Wei Liu, Xihui Mu, Zhiwei Liu, Qibin Huang
Abstract
Bing Liu, Zhaoyang Tong, Lan-Qun HAO, Wei Liu, Xihui Mu, Zhiwei Liu, Qibin Huang
Abstract
An ultrasensitive elctrochemiluminescense( ECL) immunoassay was proposed by using magnetic microbeads as the carrier of ECL labels for ECL signal amplification.First,polyclonal antibody( pcAb) of abrin was coated on microplate.Then,abrin was bound to pcAb.Next,biotinylated monoclonal antibody( mcAb) was conjugated to abrin,yielding the sandwich immunocomplex on the plate.After that,Ru( bpy)2 +3labeled magnetic microbeads was conjugated on the plate through specific biotin-streptavidin interaction.Finally,Ru( bpy)2 +3labeled magnetic microbeads was released from the plate and ECL of Ru( bpy)2 +3was measured for abrin determination.A logarithmic linear relationship between ECL intensity and the concentration of abrin in the range of 2-200 μg/L was obtained,the linear regression equation was lgY=1.295lgX + 0.201( R=0.995,n=7,p 0.0001) and the detection limit was 2 μg/L.This new approach holds great promise for highly sensitive detection of target proteins in various fields such as biodiagnostics,environmental monitoring and biodefense.
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An ultrasensitive elctrochemiluminescense( ECL) immunoassay was proposed by using magnetic microbeads as the carrier of ECL labels for ECL signal amplification.First,polyclonal antibody( pcAb) of abrin was coated on microplate.Then,abrin was bound to pcAb.Next,biotinylated monoclonal antibody( mcAb) was conjugated to abrin,yielding the sandwich immunocomplex on the plate.After that,Ru( bpy)2 +3labeled magnetic microbeads was conjugated on the plate through specific biotin-streptavidin interaction.Finally,Ru( bpy)2 +3labeled magnetic microbeads was released from the plate and ECL of Ru( bpy)2 +3was measured for abrin determination.A logarithmic linear relationship between ECL intensity and the concentration of abrin in the range of 2-200 μg/L was obtained,the linear regression equation was lgY=1.295lgX + 0.201( R=0.995,n=7,p 0.0001) and the detection limit was 2 μg/L.This new approach holds great promise for highly sensitive detection of target proteins in various fields such as biodiagnostics,environmental monitoring and biodefense.
Key concepts: Electrochemiluminescence, Chemistry, Immunoassay, Luminescent Measurements, Chromatography, Nanotechnology, Optoelectronics, Detection limit