A modified sandwich ELISA for accurate measurement of HbF in α-thalassemia carriers containing Hb Bart's and Hb Portland 1. Issue 3 (4th May 2018)

• Record Type: Journal Article
• Authors: Kerdpoo, Sasiwan; Laopajon, Witida; Kasinrerk, Watchara; Pata, Supansa; Tatu, Thanusak
• Is Part Of: Journal of immunoassay & immunochemistry. Volume 39:Issue 3(2018)
• Page Start: 323
• Page End: 336
• Publication Date: 2018-05-04
• Subjects: Hemoglobin F -- hemoglobin Bart's -- antibody -- α-thalassemia -- sandwich ELISA; Immunoassay -- Periodicals; Immunochemistry -- Periodicals; Immunoassay -- periodicals; Immunochemistry -- periodicals; 571.96405
• DOI: 10.1080/15321819.2018.1488726
• Languages: English
• ISSNs: 1532-1819

Abstract

ABSTRACT: Hemoglobin F (HbF) in blood lysate can be accurately measured by various methods, including immunoassay. In this study, we have produced polyclonal antibody (pAb) against HbF and established a modified sandwich-type ELISA for HbF quantification in blood lysates. The modified sandwich ELISA utilized anti-γ-globin monoclonal antibody clones Thal N/B as the capture antibody (Ab) coated on solid-phase, fluorescein isothiocyanate (FITC)-labeled pAb as the detecting Ab, and HPR-labeled anti-FITC Ab as the signal-generating Ab. By using an optimized blood lysate dilution, the HbF could be measured with no interference from hemoglobin Bart's (Hb Bart's) and hemoglobin Portland (Hb Portland 1) presented in α-thalassemia carriers. HbF levels measured by the modified sandwich ELISA were comparable to those quantified by the standard cation-exchange high performance liquid chromatography. We suggested that this modified sandwich ELISA was able to accurately measure HbF levels even in α-thalassemia carriers containing Hb Bart's and Hb Portland 1 and be an alternative method for HbF measurement.