Determination of fetal hemoglobin by time-resolved immunofluorometric assay.
We have developed a "sandwich"-type time-resolved immunofluorometric assay (IFMA) for fetal hemoglobin (HbF) in hemolysates from adults and newborns, amniotic fluid, and plasma, based on a polyclonal and a monoclonal antibody against human fetal hemoglobin. Microtiter wells are coated with polyclonal capture antibody, and the gamma-chain-specific monoclonal tracer antibody is labeled with a europium chelate. In a simple and fast assay procedure, prediluted hemolysates are incubated in the microtiter wells first with capture antibody for 1 h and, after washing, for 1 h with tracer antibody. The wells are washed and the fluorescence of europium is measured. The mean analytical recovery is 102% and results by IFMA agreed well with values obtained by high-performance liquid chromatography. The analytical range of IFMA is large and well suited for clinical purposes. The detection limit of the assay is 0.2 microgram/L and the measuring range extends to 500 micrograms/L.
Turpeinen
U
U
Department of Obstetrics and Gynecology, Helsinki University Central Hospital, Finland.
Stenman
U H
UH
D000653:Amniotic Fluid / Q000737:chemistry; D000911:Antibodies, Monoclonal; D001663:Bilirubin / Q000097:blood; D002851:Chromatography, High Pressure Liquid; D004912:Erythrocytes / Q000737:chemistry; D005063:Europium; D005260:Female; D005319:Fetal Hemoglobin / Q000032:analysis*; D015200:Fluoroimmunoassay / Q000379:methods* / Q000706:statistics & numerical data; D006801:Humans; D007231:Infant, Newborn; D011786:Quality Control; D012016:Reference Values
D000911:Antibodies, Monoclonal; D005063:Europium; D005319:Fetal Hemoglobin; D001663:Bilirubin