Obtaining a anti-CD45 leukocyte marker for flow cytometry
The reactive anti-CD45 is useful for the diagnostic and the study of T or B cells immunodeficiency. In this works were assayed three protocols with the purpose to obtain this reagent for the production, each one modifying the relation among the fluorochrome and the antibody (F:P) to conjugate, referred to the proposal of 1 : 2 recommended. The antibody was purified by affinity chromatography with 99,3 % of purity; was evaluated the relation FITC/Protein to 495 nm and 280 nm; and also, was obtained the leukocyte recognition percent through cytometry of flow by three tried protocols which evidence good biological activity (90,7 % - 96,8 %). The best protocol was P-2, with the lowest production cost and highest profits compared with product imported usually used in health system. This study permitted establishes the process to obtain anti-CD45 - FITC conjugated in LABEX laboratories.
Keywords: monoclonal antibodies, leukocyte marker, Fluorescein isotiocyanate (FITC), flow cytometry.
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