Purification of human blood eosinophils by a combination method using anti-CD16 monoclonal antibody, immunobeads, and Nycodenz density gradient
| Autor: | Yoichiro Sasai, Shingo Tsuda, K. Shirouzu, Minoru Miyasato, Naohisa Kitamura, Takekuni Nakama |
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| Rok vydání: | 1995 |
| Předmět: |
Density gradient
medicine.drug_class Cell Separation Dermatology Biology Monoclonal antibody Biochemistry Peripheral blood mononuclear cell Dermatitis Atopic law.invention Mice Ribonucleases Reference Values law Centrifugation Density Gradient medicine Animals Humans Centrifugation Molecular Biology Chemiluminescence Blood Cells Eosinophil Granule Proteins Receptors IgG Antibodies Monoclonal Blood Proteins Eosinophil Molecular biology Microspheres Eosinophils medicine.anatomical_structure Luminescent Measurements Immunologic Techniques biology.protein Schistosoma Antibody Granulocytes |
| Zdroj: | Journal of Dermatological Science. 10:118-129 |
| ISSN: | 0923-1811 |
| DOI: | 10.1016/0923-1811(95)00394-8 |
| Popis: | A simple method is described for the procurement of human blood eosinophil phenotypes by combining an anti-CD16 monoclonal antibody, immunobeads, and a non-toxic and non-ionic density gradient medium, Nycodenz. The purification depends on the removal of mononuclear cells using a 1.076/1.102 g/ml Nycodenz density gradient, partial removal of neutrophils based on different binding to plastic dishes, interaction of residual neutrophils with immunobeads via an anti-CD16 monoclonal antibody and, finally, extraction of eosinophil phenotypes by sifting the immunobeads-loaded neutrophils through an 1.080/1.102 g/ml Nycodenz density gradient. This method permits simultaneous preparation of highly purified normodense (> 1.080 g/ml) and hypodense eosinophils (< 1.080 g/ml) with reasonable chemiluminescence responses to opsonized zymosans and helminthotoxic activity to opsonized schistosomula corresponding to their own immunocytological properties. |
| Databáze: | OpenAIRE |
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