Detection of attograms of antigen by a high-sensitivity enzyme-linked immunoabsorbent assay (HS-ELISA) using a fluorogenic substrate

Measurement of IgG-Blocking Antibodies by ELISA Using Monoclonal Antibody and Fluorogenic Substrate

“…Virtually no cross-reactivity with immunoglobulins of goat, rab bit, horse and sheep was defined. Our results are in agreement with the suggestion that by using McAb, assay specificity, sensitivity, precision and accuracy in the ELISA can be simultaneously improved [18,20,34], Use of a fluorogenic substrate with a time-resolved fluorometer has been suggested to provide a signifi cant advantage in the measurement of antibodies [22][23][24][25]. Our experience with an AP-G used with a flu orogenic substrate indicated that nonspecific binding was consistently low (<7.1% of top standard curve point).…”

“…We have employed a mouse monoclonal antihu man IgG antibody (McAb) in a solid-phase immu noassay system utilizing an enzyme-conjugated sec ond antibody [18][19][20][21] and a fluorogenic substrate [22][23][24][25] in an effort to increase sensitivity and assay performance in a model system to quantitate IgG antibodies to short ragweed pollen allergens. The performance of the assay was highly satisfactory, providing an immunoassay which is an attractive alternative to existing methods for IgG antibody determination.…”

Single-Molecule ELISA

“…First it must be noted that similar sensitivity claims were made for 2 methods described in reports published some 30 years ago (4,5 ). Both methods relied on enzymes as signal amplifiers.…”

Development, comparison and utilization of enhanced immunoassays for the detection of lettuce mosaic virus

“…Yolken and Stopa (1979) obtained about a hundred fold increase in sensitivity of detection of human rotavirus in stool specimens. Shalev et al (1980) found that when NPP was used in ELISA, the limit of detection was 10 nM but with MUP, sensitivity increased to 10 pM resulting in a thousand fold increase in detection sensitivity. Increased sensitivity was also obtained using the fluorogenic substrate, MUG, for the quantitation of serum ferritin (Konjin et al, 1982), and for the detection of viral hepatitis in humans and animals (Neurath and Strick, 1981).…”

“…The search for sensitivity enhancement of ELISA for antigen detection led to the employment of fluorogenic enzyme substrates such as MUP (Shalev et al, 1980;Torrance and Jones, 1982;Yolken and Stopa, 1979) or MUG (Konjin et al, 1982;Neurath and Strick, 1981). It has long been known that fluorometric assays are more sensitive than absorption assay techniques (Guilbault, 1973;Lowry, 1957 Neurath and Strick (1981) claimed that hepatitis B surface antigen could be detected at concentrations of 5 to 10 pg in blood samples when the other fluorogenic substrate, MUG, was used.…”