Procedure for the preparation and Quality Control of peroxidase isoenzyme A2 isolated and Purified from Roots of Turnip Rape (Brassica napus L.) for use in Biotechnology

摘要

A procedure was designed for the purification of acidic peroxidase isoenzyme called A2 from Roots of Turnip that once processed to extract Proteins constitutes a Crude extract. This extract was dialyzed and then Purified by Ion Exchange Chromatography using a column of DEAE sephacell.These Proteins adsorbed on the column with a gradient of Sodium Chloride are eluted isoenzyme A2 presents a Band Called homogeneous Anionic Polyacrylamide electrophoresis gels and by IEF with a PL 3.6 and a molecular mass of 42 kDa.This Enzyme was carried out Biochemical Characterizations and preservation in time and ensured its quality in relation to other peroxidases that are commonly employed for Commercial Uses of Biotechnology. Some advantages of Affinity and behavior with the peroxide substrate when the same acts as a single substrate.Also proved its Efficiency when used in clinical diagnosis of reactions, as well as tests of the same Enzyme immunoassay, being very satisfactory. Even those tests can be reproduced without Variations after six months of Cold Preservation of the conjugate.It is concluded that this isoenzyme Purified is highly recommended to use Biotechnology in clinical Diagnostic Assays as well as in determinations of Enzyme immunoassay or detoxification, Laying Claim the process of isolation, purification and quality control.