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|Method & Line||Sample & Target||Product||Package Info|
|MicroELISA||Plasma,Serum,Amniotic fluid||EIAgen AFP Kit||Tests per Package: 96|
|EIAgen||The EIAgen AFP Kit has been designed for the quantitative determination of Alpha-fetoprotein in human serum, plasma or amniotic fluid. The standards are calibrated against the WHO 1st International Reference Preparation (IRP) for human AFP (72/225).||Code: LI4017K||Package: 1 Microplate|
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The EIAgen AFP Kit has been designed for the quantitative determination of Alpha-fetoprotein in human serum, plasma or amniotic fluid. The standards are calibrated against the WHO 1st International Reference Preparation (IRP) for human AFP (72/225).
Alpha-fetoprotein (AFP) is a fetal serum protein which circulates in trace amounts in healthy adults (2). AFP is produced mainly by the fetal yolk sac and fetal liver and to a lesser extent by the fetal gastrointestinal tract and kidneys (3). It is a single chain glycoprotein with a molecular weight of approximately 65-70 kilodaltons. The physico-chemical properties and amino acid composition of AFP are similar to those of serum albumin (4), from which it is antigenically distinct (5).
Clinical interest in AFP is in oncology where serum AFP is used for the diagnosis and monitoring of tumours (6, 7, 8) and in prenatal screening where amniotic fluid AFP is used to screen for foetal abnormalities such as spina bifida and Down’s Syndrome (9).
Fetal plasma AFP diffuses into the fetal urine and is excreted into the amniotic fluid from where it diffuses into the maternal circulation. The concentration of AFP in the fetal plasma peaks (2-3 mg/mL) at 12 - 14 weeks and then rapidly falls (9). The levels in the amniotic fluid are parallel to those in the fetal plasma, but are about two orders of magnitude lower (µg range) than fetal plasma concentration. In contrast, maternal serum AFP concentrations increase geometrically until 30 weeks gestation and then decline (10). Following birth, AFP levels in both mother and newborn return rapidly to basal levels (<10 ng/mL).
Approximately 70% of patients with primary hepatocellular carcinoma show elevated levels of AFP (7). Levels of 1,000 - 10,000 ng/mL have been reported (11). In the case of testicular teratoma a direct relationship has been observed between incidence of elevated AFP levels and the stage of disease (12). No increased AFP levels are found in testicular seminomas (13). The application of AFP measurement to the management of carcinoma patients has been well documented (14). Failure of AFP levels to return to normal post-operatively suggests the presence of residual tumour.
Increased serum AFP levels have also been observed in benign conditions e.g. acute viral infections, chronic active hepatitis and cirrhosis. Elevated amniotic fluid AFP has also been found in other serious conditions e.g. fetal death, omphalocele and Turner’s syndrome.
The EIAgen AFP assay is based on the two steps immunoenzymatic sandwich principle, in conjunction with the Biotine-Streptavidin technology.
Two monoclonal anti-AFP of high affinity and specificity are used: one is labelled with Horse Radish Peroxidase (HRP) and the other with Biotin, while the microplate wells are coated with Streptavidin.
Samples, calibrators and controls are dispensed into the wells, followed by the Biotin anti-AFP conjugate.
During the incubation the monoclonal bind the AFP molecule to one specific sites, and contemporaneously, the Streptavidin immobilizes the forming immunological complex to the wells through the binding to the biotin moiety of the biotinilated antibody.
After washing to eliminate the not reacted species the second monoclonal conjugated to HRP is added.
After the second incubation the wells are washed to remove the unbound labelled antibodies and the mixture of chromogen/substrate is added.
The reaction is then blocked by adding the Stop Solution and the developed colour is measured photometrically. The intensity of the colour is directly proportional, within the working range of the assay, to the concentration of AFP in the sample. The concentration of AFP in a patient sample or controls is then determined by interpolation on the calibration curve.
Each kit contains sufficient reagents to perform 96 tests (code LI4017K).
Bring reagents to room temperature before use.
AFP Sample Diluent
Wash Buffer Conc. 10x
Number of tests