Development, optimization and use of an enzyme linked immunosorbent assay (ELISA) to measure factor VIII antigen utilizing monoclonal antibodies.
An enzyme linked immunosorbent assay (ELISA) has been developed to measure VIII:Ag in plasma and concentrates. The assay utilizes two commercially available monoclonal antibodies to VIII:Ag and provides an alternative to the established immunoradiometric assay (IRMA). It has the advantage of not requiring the use of radioactive material and human antibodies. The assay sensitivity is 0.006 u/ml and the interassay coefficient of variation is 6.3%. Forty-eight samples with VIII:Ag levels ranging from 0.006 to 1.5 u/ml were assayed by both ELISA and IRMA. The coefficient of correlation between the two assays was 0.89. In addition to measuring human VIII:Ag, it is also possible to detect antigen in several animal plasma and sera.
Hornsey
V S
VS
Scottish National Blood Transfusion Service, National Science Laboratory, Edinburgh.
Drummond
O
O
Eaglesfield
P
P
Pepper
D S
DS
Prowse
C V
CV
D000911:Antibodies, Monoclonal; D000941:Antigens / Q000097:blood*; D002850:Chromatography, Gel; D004797:Enzyme-Linked Immunosorbent Assay; D005169:Factor VIII / Q000276:immunology*; D006801:Humans; D015592:Immunoradiometric Assay; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D013997:Time Factors
D000911:Antibodies, Monoclonal; D000941:Antigens; D005169:Factor VIII