Obtention of β- phycoerythrin and its use in the conjugation of specific antibodies

Abstract

The objective of this work was to obtain pure preparations of β-phycoerythrin and its conjugation with specific antibodies. For this, a process of purification of biomass obtained from the alga Porphyridium cruentum, 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 β- 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.

Keywords: phycoerythrin, conjugated, flow cytometry, diagnoses.