Recycling immunoaffinity chromatography for multiple analyte analysis in biological samples |
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| Institution: | 1. Department of Pathology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692, Japan;2. Department of Radiology, Faculty of Medicine, University of Miyazaki, 5200 Kihara, Kiyotake, Miyazaki 889-1692, Japan;3. Department of Biochemistry, Kyushu University of Health and Welfare, 1714-1 Yoshino, Nobeoka 882-0072, Japan;4. Department of Pathology, Miyazaki Prefectural Hospital, 5-30 Kitatakamatsu, Miyazaki 880-5810, Japan;1. Center for Computational Science, RIKEN, Kobe, Hyogo, 650-0047, Japan;2. Department of Physics, Graduate School of Science, Nagoya University, Aichi, 464-8602, Japan;3. Institute of Transformative Bio-Molecules (WPI-ITbM), Nagoya University, Aichi, 464-8601, Japan |
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| Abstract: | The ability to isolate and measure multiple complex analytes in a single biological sample holds great potential in many biomedical fields, especially immunology and diagnostic clinical chemistry. We have developed a procedure involving recycling immunoaffinity chromatography for the simultaneous measurement of a number of analytes in a single sample. The procedure is based on the passage of a fluorochrome-labelled sample through a battery of small immunoaffinity columns, each column extracting a single analyte. Detection is achieved by acid elution of the bound analytes and laser-induced fluorescence. We have applied this system to a number of different biological fluids and found that it is capable of reliably isolating and measuring up to ten different cytokines in a 25-μl sample of human body fluid. |
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