An inhibition enzyme immunoassay for myelin basic protein |
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Authors: | Nigel P Groome Janice Harland Jonathan Appleby |
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Institution: | Biology Department, Oxford Polytechnic, Headington, Oxford OX3 0BP, U.K. |
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Abstract: | An improved Enzyme Immunoassay for Myelin Basic Protein is described. Myelin Basic Protein covalently attached to glass balls, and Myelin Basic Protein in samples compete with each other for binding of a peroxidase conjugated anti Myelin Basic Protein antibody. The peroxidase activity on the balls is then inversely proportional to the amount of Myelin Basic Protein in the sample. A detection limit of 0.6 ng/ml is demonstrated for diluent or spinal fluid. For plasma a dilution step increases this to 1.8 ng/ml. Both the coated balls and the peroxidase conjugate are stable for long periods. The assay requires no expensive equipment. Although the assay appears to be valid for subcellular fractions spinal fluid and plasma, successful detection of Myelin Basic Protection peptides in clinical samples may require careful selection of suitable antisera. The assay would be very suitable for eventual use with an appropriate monoclonal antibody. |
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Keywords: | MBP Myelin Basic Protein CSF Cerebrospinal fluid ELISA Enzyme linked immunoadsorbent assay ABTS 2 2′ Azinodi-(3 ethylbenzthiazoline-6-sulphonic acid) HRP horse radish peroxidase |
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