Purification and radioimmunoassay of human alpha-1-fetoprotein: the effect of aggregates on the radioimmunoassay.
A simple method for the extraction of alpha1-fetoprotein from fetal serum in high yield and of sufficient purity for use as radio-iodinated tracer in a radioimmunoassay is described. The purified alpha1-fetoprotein formed aggregates during storage. The presence of 125I-labelled aggregates affected the double antibody radioimmunoassay by increasing the non-specific bound, resulting in decreases assay sensitivity. Freshly prepared 125I-labelled monomer alpha1-fetoprotein is prepared for each assay. The radioimmunoassay described is sensitive to between 100 and 200 pg per tube. Normal adult serum levels were found to be in the range from less than 1 mug/l to 16 mug/l.
D004589:Electrophoresis, Disc; D005260:Female; D005326:Fetal Proteins / Q000032:analysis*; D005333:Fetus; D006801:Humans; D046911:Macromolecular Substances; D008832:Microchemistry; D008970:Molecular Weight; D011247:Pregnancy; D011485:Protein Binding; D011863:Radioimmunoassay / Q000379:methods; D000509:alpha-Fetoproteins / Q000032:analysis* / Q000302:isolation & purification
D005326:Fetal Proteins; D046911:Macromolecular Substances; D000509:alpha-Fetoproteins