A new immunoassay method called specific analyte labeling and recapture assay (SALRA) to quantitatively measure protein abundance was developed, and the assay conditions were optimized. The key features of this method include labeling the antigen bound to the capture antibody, eluting the labeled antigen, and recapturing it by the same capture antibody on the detection plate. The reporter molecules on the labeled antigen provide a convenient and reliable means for signal detection. We demonstrated that the dose-response curve of SALRA was comparable to that of sandwich enzyme-linked immunosorbent assay (ELISA) and better than that of the antigen direct labeling method. In addition, multiple proteins can be measured simultaneously by SALRA. Using the SALRA method, the detection limit for most of the cytokines tested was approximately 0.01ng/ml. Further SALRA tests on interleukin 6 (IL-6) showed the linear dose-response was 3.3 to 0.01ng/ml, the accuracy of the test was 71 to 91%, the intraassay variation was 3.6 to 7.4%, and the interassay variation was 3.8 to 10.0%. The applications of SALRA include quantitatively measuring proteins for which there are no ELISA tools available and providing a new platform for protein microarrays.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1016/j.ab.2010.01.038 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Influences of age and sex on the diagnostic accuracy of human epididymis secretory protein 4 for malignant pleural effusion.
Sci Rep
January 2025
Department of Laboratory Medicine, the Affiliated Hospital of Inner Mongolia Medical University, Hohhot, China.
Previous studies have suggested that the presence of human epididymal protein 4 (HE4) in pleural fluid can be used to diagnose malignant pleural effusion (MPE) with moderate accuracy. However, the factors that affect the diagnostic accuracy of HE4 remain unknown. This study aimed to examine how age and sex influence the diagnostic accuracy of HE4.
View Article and Find Full Text PDFMUC1 and glycan probing of CA19-9 captured biomarkers from cyst fluids and serum provides enhanced recognition of ovarian cancer.
Sci Rep
January 2025
Department of Life Technologies, Division of Biotechnology, University of Turku, Medisiina D, 5th floor, Kiinamyllynkatu 10, 20520, Turku, Finland.
Glycosylation changes of circulating proteins carrying the CA19-9 antigen may offer new targets for detection methods to be explored for the diagnosis of epithelial ovarian cancer (EOC). Search for assay designs for targets initially captured by a CA19-9 antigen reactive antibody from human body fluids by probing with fluorescent nanoparticles coated with lectins or antibodies to known EOC associated proteins. CA19-9 antigens were immobilized from ascites fluids, ovarian cyst fluids or serum samples using monoclonal antibody C192 followed by probing of carrier proteins using anti-MUC16, anti-MUC1 and, anti STn antibodies and seven lectins, all separately coated on nanoparticles.
View Article and Find Full Text PDFImpact of SARS-CoV-2 spike antibody positivity on infection and hospitalisation rates in immunosuppressed populations during the omicron period: the MELODY study.
Lancet
January 2025
Centre for Inflammatory Disease, Department of Immunology and Inflammation, Imperial College London, London, UK; Imperial College Renal and Transplant Centre, Imperial College Healthcare NHS Trust, Hammersmith Hospital, London, UK. Electronic address:
Background: In the UK, booster COVID-19 vaccinations have been recommended biannually to people considered immune vulnerable. We investigated, at a population level, whether the absence of detectable anti-SARS-CoV-2 spike protein IgG antibody (anti-S Ab) following three or more vaccinations in immunosuppressed individuals was associated with greater risks of infection and severity of infection.
Methods: In this prospective cohort study using UK national disease registers, we recruited participants with solid organ transplants (SOTs), rare autoimmune rheumatic diseases (RAIRDs), and lymphoid malignancies.
Gold nanorod in silver tetrahedron: Cysteamine mediated synthesis of SERS probes with embedded internal markers for AFP detection.
Anal Chim Acta
February 2025
The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, No. 28 Xianning West Road, Xi'an, 710049, China. Electronic address:
Background: Plasmonic core-shell nanostructures with embedded internal markers used as Raman probes have attracted great attention in surface-enhanced Raman scattering (SERS) immunoassay for cancer biomarkers due to their excellent uniform enhancement. However, current core-shell nanostructures typically exhibit a spherical shape and are coated with a gold shell, resulting in constrained local field enhancement.
Results: In this work, we prepared a core-shell AuNR@BDT@Ag structure by depositing silver on the surface of Raman reporter-modified gold nanorods (AuNR).
An ultrasensitive ECL immunosensor with a dual signal amplification strategy using AuNPs@GO@SmMoSe and Gd(MoO) for estriol detection.
Anal Chim Acta
February 2025
School of Chemistry and Chemical Engineering, University of Jinan, Jinan, 250022, PR China; Department of Chemistry, Sungkyunkwan University, Suwon, 16419, Republic of Korea. Electronic address:
Background: Estriol (E3) is a common estrogen responsible for regulating the female reproductive system, but excessive amount can pose health risks to humans and wild life. Therefore, sensitive and accurate detection of estriol level is crucial. A novel competitive ECL immunosensor based on a dual signal amplification strategy of AuNPs@GO@SmMoSe and Gd(MoO) was fabricated for ultrasensitive detection of estriol.
View Article and Find Full Text PDFWant AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!