Affinophoresis in two-dimensional agarose gel electrophoresis: specific separation of biomolecules by a moving affinity ligand |
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| Authors: | K Shimura K Kasai |
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| Affiliation: | 1. Department of Neurology, Mayo Clinic, 4500 San Pablo Road, Jacksonville, FL 32224, USA;2. Department of Neuroscience, Mayo Clinic, 4500 San Pablo Road, Jacksonville, FL 32224, USA;3. Lund University, Department of Clinical Sciences, Lund, Neurology, Getingevägen 4, 22185 Lund, Sweden;4. Department of Neuropathology, Mayo Clinic, 4500 San Pablo Road, Jacksonville, FL 32224, USA;1. National and Local United Engineering Lab of Metabolic Control Fermentation Technology, Tianjin University of Science and Technology, Tianjin 300457, China;2. College of Biotechnology, Tianjin University of Science and Technology, Tianjin 300457, China;3. The Institute of Seawater Desalination and Multipurpose Utilization, SOA, Tianjin 300192, China;1. State Key Laboratory of Microbial Technology, National Glycoengineering Research Center, Shandong University, Jinan 250100, PR China;2. Marine Biology Institute, Shantou University, Shantou 515063, PR China;3. CAS Key Lab of Biobased Materials, Qingdao Institute of Bioenergy and Bioprocess Technology, Chinese Academy of Sciences, Qingdao, PR China;1. Molecular Biotechnology and Health Sciences Department, University of Torino, Quarello 15, 10135 Torino, Italy;2. Molecular Neuropathobiology Laboratory, Department of Molecular Life Sciences, Tokai University School of Medicine, 143 Shimokasuya, Isehara, Kanagawa 259-1193, Japan;1. Chemical and Biological Engineering, and Clinical & Translational Research Center, State University of New York, Buffalo, NY 14260, USA;2. Department of Chemistry, Biomedical Chemistry Institute, New York University, New York, NY 10003, USA |
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| Abstract: | Affinophoresis is an electrophoretic separation technique for biomolecules which uses an affinophore. An affinophore is a macromolecular polyelectrolyte bearing affinity ligands. It migrates rapidly in an electric field, and consequently the electrophoretic mobility of molecules having affinity for the ligand is specifically changed. This technique has now been incorporated in two-dimensional agarose gel electrophoresis in a procedure which utilizes normal electrophoresis in the first dimension and affinophoresis in the second dimension. Proteins which do not have affinity for the ligand migrate to locations along a diagonal line passing through the origin, whereas proteins which have affinity are carried away from the line by the affinophore. Accordingly, molecules having affinity for the ligand can be readily assigned. Trypsins contained in Pronase and pancreatin were separated by this procedure using an affinophore bearing a competitive inhibitor for trypsin, benzamidine, on a polyanionic molecule (a polyacrylic acid derivative). |
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