Phase change enzyme immunoassay |
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| Authors: | D E Mahan L Morrison L Watson L S Haugneland |
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| Affiliation: | 1. Department of Chemistry “U. Schiff”, University of Florence, Via U. Schiff 6, 50019, Sesto Fiorentino, Florence, Italy;2. Department of Chemistry “U. Schiff”, University of Florence, Via della Lastruccia 3-13, 50019 Sesto Fiorentino, Florence, Italy;3. Department of Neuroscience, Psychology, Drug Research and Child Health, Section of Pharmaceutical and Nutraceutical Sciences, University of Florence, Via U. Schiff 6, 50019 Sesto Fiorentino, Florence, Italy;1. Division of Immunology, Department of Biology, University of Konstanz, D-78457, Konstanz, Germany;2. Center for Integrated Protein Science Munich at the TUM Department of Chemistry, Technical University of Munich, D-85748, Garching, Germany;3. Biotechnology Institute Thurgau at the University of Konstanz (BITg), CH-8280, Kreuzlingen, Switzerland;1. Department of Clinical and Molecular Sciences, DISCLIMO, Università Politecnica delle Marche, Ancona, Italy;2. Scientific Direction, IRCCS INRCA, Ancona, Italy;3. Center of Clinical Pathology and Innovative Therapy, IRCCS INRCA, Ancona, Italy;1. Department of Gastroenterology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China;2. Department of Gastroenterology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China;1. School of Clinical Medicine, Xuzhou Medical University, 209 Tongshan Road, Xuzhou 221004, PR China;2. Jiangsu Cancer Hospital, Jiangsu Institute of Cancer Research, Nanjing Medical University Affiliated Cancer Hospital, 42 Baiziting, Nanjing 210009, PR China;3. Department of General Surgery, the First Affiliated Hospital with Nanjing Medical University, 300 Guangzhou Road, Nanjing 210029, PR China;4. Jiangsu Key Lab of Cancer Biomarkers, Prevention and Treatment, Nanjing Medical University, 101 Longmian Avenue, Nanjing 211166, PR China |
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| Abstract: | A novel enzyme-linked immunoassay employing a partitioning chromophore was developed. The assay system consisted of an aqueous phase and an immiscible organic solvent. Antigen-antibody interaction was indicated by transfer of a chromogenic indicator from the aqueous phase to an organic layer. The indicator employed was a water-soluble phosphate ester of phenylazophenol. Hydrolysis of the ester by acid or alkaline phosphatase produced a water-insoluble phenol that partitioned into toluene. The enzyme employed in this assay format can be covalently linked to antibody or a specific antibody for the phosphatase can be used. Phase change immunoassays were developed for the measurement of alkaline phosphatase, human IgG in whole blood, and the human tumor marker prostatic acid phosphatase. Solid supports of small polystyrene latex particles and Sephadex were employed. |
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