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Photoimmobilized array of panel cells for assay of antibodies
Yoshihiro Ito1, Tetsuya Yamauchi, Makoto Uchikawa
1Kanagawa Academy of Science and Technology, KSP East 309, 3-2-1 Sakado, Takatsu-ku, Kawasaki, Kanagawa 213-0012, Japan.
Antibodies in blood are checked with panel blood cells before blood transfusion. In this investigation, for the first time, a panel cell-microarray was prepared by using a photoimmobilization method. Different types of red blood cells were microarrayed on a plate. A water-soluble photoreactive polymer as a matrix was synthesized by the coupling reaction of azidoaniline with poly(2-methacryloyloxyethylphosphorylcholine-co-methacrylic acid). The polymer was mixed with cells and the mixtures were microspotted on substrate and photoirradiated after drying in air. For the antibody assay, monoclonal antibodies or human serum was added to the cell-arrayed plate and adsorbed antibodies were detected by horseradish peroxidase-labeled secondary antibody, which recognized the adsorbed antibodies. Antibodies specifically adsorbed on the immobilized cells as expected. The aggregation method has been available for this type of assay, but extensive experience was needed to apply it correctly. The method using a cell array will be useful for antibody detection.
Antibodies in blood are checked with panel blood cells before blood transfusion. In this investigation, for the first time, a panel cell-microarray was prepared by using a photoimmobilization method. Different types of red blood cells were microarrayed on a plate. A water-soluble photoreactive polymer as a matrix was synthesized by the coupling reaction of azidoaniline with poly(2-methacryloyloxyethylphosphorylcholine-co-methacrylic acid). The polymer was mixed with cells and the mixtures were microspotted on substrate and photoirradiated after drying in air. For the antibody assay, monoclonal antibodies or human serum was added to the cell-arrayed plate and adsorbed antibodies were detected by horseradish peroxidase-labeled secondary antibody, which recognized the adsorbed antibodies. Antibodies specifically adsorbed on the immobilized cells as expected. The aggregation method has been available for this type of assay, but extensive experience was needed to apply it correctly. The method using a cell array will be useful for antibody detection.
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