The application of the biotin/avidin system to the von Willebrand factor antigen immunoassay. 1988

L D Taylor
Department of Clinical Haematology and Oncology, Royal Children's Hospital, Melbourne, Australia.

A vWf: Ag ELISA which uses biotinylated anti-vWf: Ag IgG and an avidin/biotinylated peroxidase (ABP) complex detection system has been developed. The sensitivity of the ELISA for vWf: Ag was 4 X 10(-5) U/ml which is approximately 10-fold lower than previously reported vWf: Ag ELISA's. The avidin used for the preparation of the ABP complex was glutaraldehyde-treated. This treatment increased the reactivity 2-3-fold as compared to ABP complex prepared using untreated avidin. The reactivity of two commercial detection reagents was also tested. The effect of the reduction of the total assay time to 3.5 h is also described. In this form the detection limit for vWf: Ag was 2 X 10(-4) U/ml, the intra-assay precision at vWf: Ag concentrations of 0.01 and 0.0025 U/ml was 3.8% and 4.9% respectively and ELISA results for normal individuals still correlated well with those obtained using electro-immunodiffusion (r = 0.93).

UI MeSH Term Description Entries
D004797 Enzyme-Linked Immunosorbent Assay An immunoassay utilizing an antibody labeled with an enzyme marker such as horseradish peroxidase. While either the enzyme or the antibody is bound to an immunosorbent substrate, they both retain their biologic activity; the change in enzyme activity as a result of the enzyme-antibody-antigen reaction is proportional to the concentration of the antigen and can be measured spectrophotometrically or with the naked eye. Many variations of the method have been developed. ELISA,Assay, Enzyme-Linked Immunosorbent,Assays, Enzyme-Linked Immunosorbent,Enzyme Linked Immunosorbent Assay,Enzyme-Linked Immunosorbent Assays,Immunosorbent Assay, Enzyme-Linked,Immunosorbent Assays, Enzyme-Linked
D006801 Humans Members of the species Homo sapiens. Homo sapiens,Man (Taxonomy),Human,Man, Modern,Modern Man
D000941 Antigens Substances that are recognized by the immune system and induce an immune reaction. Antigen
D001360 Avidin A specific protein in egg albumin that interacts with BIOTIN to render it unavailable to mammals, thereby producing biotin deficiency.
D001710 Biotin A water-soluble, enzyme co-factor present in minute amounts in every living cell. It occurs mainly bound to proteins or polypeptides and is abundant in liver, kidney, pancreas, yeast, and milk. Vitamin H,Biodermatin,Biokur,Biotin Gelfert,Biotin Hermes,Biotin-Ratiopharm,Biotine Roche,Deacura,Gabunat,Medebiotin,Medobiotin,Rombellin,Biotin Ratiopharm,Gelfert, Biotin,Hermes, Biotin,Roche, Biotine
D014841 von Willebrand Factor A high-molecular-weight plasma protein, produced by endothelial cells and megakaryocytes, that is part of the factor VIII/von Willebrand factor complex. The von Willebrand factor has receptors for collagen, platelets, and ristocetin activity as well as the immunologically distinct antigenic determinants. It functions in adhesion of platelets to collagen and hemostatic plug formation. The prolonged bleeding time in VON WILLEBRAND DISEASES is due to the deficiency of this factor. Factor VIII-Related Antigen,Factor VIIIR-Ag,Factor VIIIR-RCo,Plasma Factor VIII Complex,Ristocetin Cofactor,Ristocetin-Willebrand Factor,von Willebrand Protein,Factor VIII Related Antigen,Factor VIIIR Ag,Factor VIIIR RCo,Ristocetin Willebrand Factor

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