Combination of polyacrylamide gel electrophoresis with enzyme-linked immunosorbent assay: A simple method for determination of antibody specificity and detection of antigens in complex mixtures |
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| Authors: | H Lutz S Bruggmann L Corboz R Müller W Limacher H Weber K Wissler GH Theilen |
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| Institution: | 1. Department of Surgery, Clinical Oncology Section, School of Veterinary Medicine, University of California, Davis, CA 95616 USA;2. Clinic of Veterinary Medicine, University of Zuerich, CH-8057 Zuerich Switzerland;3. Clinic of Gynaecology, University of Zuerich, CH-8057 Zuerich Switzerland;4. Institute of Veterinary Hygiene, University of Zuerich, CH-8057 Zuerich Switzerland;5. Institute of Anatomy, University of Zuerich, CH-8057 Zuerich Switzerland;6. Institute of Parasitology, University of Zuerich, CH-8057 Zuerich Switzerland;7. Pharmacology and Biochemistry, Faculty of Veterinary Medicine, University of Zuerich, CH-8057 Zuerich Switzerland |
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| Abstract: | Complex antigenic mixtures were separated on polyacrylamide gels in the presence of SDS. After electrophoresis, the gels were cut into equal fractions. Antigens were eluted from the fractions and could be bound to different solid phases on which a conventional enzyme-linked immunosorbent assay was performed. Antibody binding fractions could then be related with the bands of a stained gel run in parallel. This technique proved to be fast and sensitive. Antigens present in nanogram amounts in individual fractions were sufficient for the detection of specific antibody. We describe here the application of this technique to antibodies specific of parasite, viral, bacterial and mycoplasmal antigens and to antibodies against hormones and a muscle protein. |
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