A new method for broadening the detection range of immunoassay and its application in β-hCG quantitative detection

Zhang, Bei; Guo, Miao; Zhang, Tianjie; Liu, Dandan; Tan, Xin; Li, Xue; Yu, Yang; Li, Huiqiang

doi:10.1039/d2ay00220e

Cited by 3 publications

(2 citation statements)

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“…Coupling of E2-BSA and E3-BSA to chemibeads was performed according to the procedure in previous studies. 18 The general steps are as follows: first, 2 mg of chemibeads was washed with distilled water and resuspended in 0.05 M carbonate buffer (pH 9.6). A certain volume of E2-BSA or E3-BSA antigen solution was added, and the mixture was shaken at 37 °C overnight.…”

Section: Methodsmentioning

confidence: 99%

“…The simultaneous use of two competitive antigens with different affinities based on light-initiated chemiluminescence immunoassay (LiCA) provides a new way to solve the abovementioned problems. LiCA is the latest generation of chemiluminescence technology. , It is based on the antigen–antibody reaction to bring sensibeads and chemibeads close to each other to achieve the delivery of high-energy reactive oxygen species and induce a photo-excited chemiluminescence process to detect the light signal in a homogeneous system . The nanospheres have small diameters and good suspension in the liquid phase, making the antigen–antibody more likely to collide and interact with each other, which greatly improves the detection sensitivity and shortens the detection time.…”

Section: Introductionmentioning

confidence: 99%

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Improved Sensitivity and Wide Range Detection of Small Analytes Using a Two-Antigen-Combined Competitive Immunoassay

Zhang

Lv²,

Qiang

et al. 2022

ACS Omega

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Competitive immunoassays have unique advantages in the detection of small molecules and are widely used in clinical practice. However, the concentrations of some analytes usually vary greatly among different populations, which makes it difficult to balance the sensitivity and detection range of competitive immunoassays. Studies have shown that using haptens with weaker affinity for specific antibodies as competitive antigens can help improve the sensitivity of the method. Here, we developed a competitive light initiated chemiluminescence assay based on the combination of antigens with different affinities, which has high sensitivity and wide detection range. As a proof of concept, estradiol was used as the analyte. After the mixing ratio was optimized, the two labeled haptens played different competitive roles due to the different concentrations of estradiol to be tested, which improved the sensitivity of estradiol detection, while ensuring a certain detection range. The limit of detection of this method was 5.30 pg/mL, which is lower than most current estradiol immunoassay kits. Good linearity (R 2 = 0.9902) was obtained between estradiol concentrations of 17.07−2376.22 pg/mL. This study provides a new solution for the detection of small molecule biomarkers with a large concentration span, which also has considerable potential in other immunological detection methods.

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Section: Methodsmentioning

confidence: 99%