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KMID : 0390320180280020011
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Chungbuk Medical Journal
2018 Volume.28 No. 2 p.11 ~ p.16
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Nonspecific reaction of fluorescent conjugated antibody
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Seon Seong-Gyu
Park Bo-Reum
Lim Yeong-Hyeon
Song Hyung-Geun
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Abstract
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Purpose: Detection of cancer-specific biological markers is mostly carried out using antibodies. Diagnosis of cancer-specific markers mainly through immunohistochemistry is time-consuming and costly, and can result in false negatives due to paraffin. On the other hand, flow cytometry has the advantage of analyzing the antigen in the living body as it is. In this paper, we investigated the nonspecific reactions that can occur due to fluorescent conjugated antibody in the diagnosis of biomarkers using flow cytometry.
Materials and Methods: Using a mouse 1E1 antibody and a chimeric 1E1 antibody that recognize ICAM-4 (CD242), antibody binding was analyzed by direct and indirect staining in K562 cells expressing ICAM-4 and 786-O, CEM-CM3 cells not expressing it
Results: When direct staining was used, fluorescence-linked chimeric antibodies showed nonspecific results. In the indirect staining method, neither mouse nor chimeric antibody showed nonspecific results.
Conclusion: When fluorescence-linked chimeric antibodies are used as direct staining for the detection of cancer-specific antigens, false - positive reactions may occur.
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KEYWORD
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Non-specific reactions, recombinant chimeric antibodies, flow cytometry
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