Alpha-Fetoprotein (AFP)
Lab area
Clinical Biochemistry - General
Method and equipment
Equipment : Roche Cobas Pro e801
Method : The electrochemiluminescence immunoassay “ECLIA” is intended for use
on cobas e immunoassay analyzers.
on cobas e immunoassay analyzers.
Sandwich principle:
▪ 1st incubation a biotinylated monoclonal AFP‑specific antibody, and a monoclonal AFP‑specific antibody labeled with a ruthenium complexa) react to form a sandwich complex.
▪ 2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.
▪ The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell II M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
▪ Results are determined via a calibration curve which is instrument specifically generated by 2‑point calibration and a master curve provided via the cobas link
▪ 1st incubation a biotinylated monoclonal AFP‑specific antibody, and a monoclonal AFP‑specific antibody labeled with a ruthenium complexa) react to form a sandwich complex.
▪ 2nd incubation: After addition of streptavidin-coated microparticles, the complex becomes bound to the solid phase via interaction of biotin and streptavidin.
▪ The reaction mixture is aspirated into the measuring cell where the microparticles are magnetically captured onto the surface of the electrode. Unbound substances are then removed with ProCell II M. Application of a voltage to the electrode then induces chemiluminescent emission which is measured by a photomultiplier.
▪ Results are determined via a calibration curve which is instrument specifically generated by 2‑point calibration and a master curve provided via the cobas link
Expected turn-around time
STAT/ Urgent: 6 hours
Routine: 24 hours
Specimen type
Serum, Sodium/Lithium Heparin plasma, Amniotic fluid
Specimen requirements
150 uL
Storage and transportation
Shipping information
The Hospital for Sick Children
Rapid Response Laboratory
555 University Avenue, Room 3642
Toronto, ON
Canada
M5G 1X8
Phone: 416-813-7200
Toll Free: 1-855-381-3212
Hours: 7 days/week, 24 hours/day
Background and clinical significance
The discovery of alpha-fetoprotein (AFP) in fetal serum was first recorded by Bergstrand and Czar in 1956.
Alpha-fetoprotein is a single polypeptide chain glycoprotein with a molecular weight of approximately 70,000 daltons. It is secreted into fetal serum, reaching a peak at about 13 weeks gestation and gradually declining thereafter. Elevated serum AFP levels subsequently reappear during pregnancy and in conjunction with several malignant diseases
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