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Revista Cubana de Química
versão On-line ISSN 2224-5421
Resumo
FERNANDEZ-DUARTE, George; GUTIERREZ-CALZADO, Esteban e GARCIA-REYES, Maibis L. Obtention of ß-phycoerythrin and its use in the conjugation of specific antibodies. Rev Cub Quim [online]. 2017, vol.29, n.2, pp. 212-228. ISSN 2224-5421.
The objective of this work was to obtain pure preparations of ß-phycoerythrin and its conjugation with specific antibodies. For this, a process of purification of biomass obtained from the Porphyridium cruentum alga, from which an initial filtration was obtained, was submitted to a molecular exclusion chromatographic process with Sephadex G-25, and later to ion exchange with DEAE cellulose. It was then conjugated to antibodies by the Hardy protocol, which involves three steps, with derivatizing reagents, culminating in a covalent phycoerythrin-antibody binding. The quality of the conjugate obtained was evaluated by Flow Cytometry. The results obtained show a yield between 30 and 51 %, with purity levels of ß- phycoerythrin molecule greater than 90%, and the interpretation of the diagrams obtained by Flow Cytometry, testifying the quality of the conjugate obtained, for later use for diagnostic purposes.
Palavras-chave : phycoerythrin; conjugated; flow cytometry; diagnoses.