The use of avidin-biotin interaction in immunoenzymatic techniques

Biotin was covalently attached to antibodies, antigens and enzymes, and the effects of this labeling on the antigen and antibody binding capacity and on enzymatic activity were tested. Based on avidin-biotin interaction, the labeled proteins were used in quantitative enzyme-immunoassay and enzyme-im...

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Bibliographic Details
Published in:The journal of histochemistry and cytochemistry 1979-08, Vol.27 (8), p.1131-1139
Main Authors: Guesdon, JL, Ternynck, T, Avrameas, S
Format: Article
Language:English
Subjects:
Animals
Antigen-Antibody Reactions
Avidin
Binding Sites, Antibody
Biotin
Cattle
Immunoenzyme Techniques
Ovalbumin - analogs & derivatives
Rabbits
Sheep
Staining and Labeling
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Summary:Biotin was covalently attached to antibodies, antigens and enzymes, and the effects of this labeling on the antigen and antibody binding capacity and on enzymatic activity were tested. Based on avidin-biotin interaction, the labeled proteins were used in quantitative enzyme-immunoassay and enzyme-immunohistochemical staining procedures. Two procedures were developed. In the first procedure, named the Bridged Avidin-Biotin (BRAB) technique four steps were used sequentially in order to quantify or detect an immobilized antigen: 1) incubation with biotin-labeled antibody; 2) incubation with avidin; 3) incubation with biotin-labeled enzyme; 4) measurement or histochemical staining of the enzyme. The technique is based on the observation that avidin possesses four active sites. In the second procedure, named the Labeled Avidin-Biotin (LAB) technique, biotin-labeled antibody and enzyme-labeled avidin are used sequentially. Enzyme-associated antigen is then quantified or revealed immunohistochemically. The optimal conditions for enzyme-immunoassay and enzyme-immunohistochemical staining using BRAB and LAB procedures were established.
ISSN:0022-1554
1551-5044
DOI:10.1177/27.8.90074