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Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis
Published on: March 7, 2018
Gold nanoparticle-based immunoassay by using non-stripping chemiluminescence detection
Chun-Feng Duan1, Yu-Qi Yu, Hua Cui
1Department of Chemistry, University of Science & Technology of China, Hefei, Anhui 230026, People's Republic of China.
Insights
A new chemiluminescence immunoassay for human immunoglobulin G (IgG) uses gold nanoparticles for sensitive detection. This simplified method offers a faster, automated alternative for clinical diagnostics.
Area of Science:
- Analytical Chemistry
- Biochemistry
- Immunology
Background:
- Accurate quantification of human immunoglobulin G (IgG) is crucial for diagnosing various diseases.
- Existing chemiluminescence immunoassay (CLIA) methods can be complex, requiring stringent procedures or difficult synthesis.
Purpose of the Study:
- To develop a novel, microplate-compatible chemiluminescence immunoassay for human IgG determination.
- To utilize a luminol-AgNO(3)-gold nanoparticle CL system for enhanced sensitivity and simplicity.
Main Methods:
- Immobilization of goat-anti-human IgG primary antibody on polystyrene microwells.
- Formation of a sandwich-type immunocomplex with human IgG and a gold-labeled secondary antibody.
- Triggering chemiluminescence via the gold label's catalytic effect on the luminol-AgNO(3) reaction.
Main Results:
- The CL intensity showed linearity with the logarithm of human IgG concentration from 25 to 5000 ng/mL.
- A detection limit of 12.8 ng/mL (approx. 80 pM) was achieved at S/N = 3.
- The developed method demonstrated simplicity, time-saving advantages, and potential for automation.
Conclusions:
- The novel CL immunoassay provides a simple, sensitive, and automatable method for human IgG determination.
- This approach avoids complex stripping procedures or challenging synthesis steps common in other gold-labeled CLIA methods.
- The assay is promising for the routine analysis of clinically significant bioactive analytes.
Abstract:
A novel microplate-compatible chemiluminescence (CL) immunoassay has been developed for the determination of human immunoglobulin G (IgG) based on the luminol-AgNO(3)-gold nanoparticles CL system. Polystyrene microtiter plates were used for both immunoreactions and CL measurements. The primary antibody, goat-anti-human IgG, was first immobilized on polystyrene microwells. Then the antigen (human IgG) and the gold-labeled second antibody were connected to the microwells successively to form a sandwich-type immunocomplex. The gold label could trigger the reaction between luminol and AgNO(3), accompanied by light emission. Under the optimized conditions, the CL intensity of the system was linear with the logarithm of the concentration of human IgG in the range from 25 to 5000 ng mL(-1), with a detection limit of 12.8 ng mL(-1) ( approximately 80 pM) at a signal to noise ratio of three (S/N = 3). Compared with other reported CL immunoassay method based on gold labels, the proposed CL protocol avoids a strict stripping procedure or difficult to control synthesis processes, making the method more simple, time-saving and easily automated. The present CL method is promising for the determination of clinically important bioactive analytes.
