Reference : Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine
Scientific journals : Article
Human health sciences : Multidisciplinary, general & others
Human health sciences : Anesthesia & intensive care
Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine
Deby, Ginette [Université de Liège - ULiège > > Centre de l'oxygène : Recherche et développement (C.O.R.D.) >]
Pincemail, Joël mailto [Centre Hospitalier Universitaire de Liège - CHU > > Chirurgie cardio-vasculaire >]
Thirion, A. [ > > ]
Deby, C. [ > > ]
Lamy, Maurice mailto [Université de Liège - ULiège > Département des sciences cliniques > Anesthésie et réanimation]
Franchimont, P. [> > > >]
Yes (verified by ORBi)
[en] Human ; Leucocytes ; Myeloperoxidase-iodination-radioimmunoassay
[en] In order to obtain a radioimmunoassay (RIA) technique for the measurement of human plasma myeloperoxidase (MPO), we purified the enzyme from polymorphonuclear granulocytes (neutrophils), and compared three methods of labeling it with 125Iodine:chloramine T, lactoperoxidase, and an original technique of 'self labeling' based on the ability of the enzyme to oxidize and bind 125I in the presence of H2O2. The chloramine T technique produced a degraded protein, as well shown by a high non-specific binding of tracer to antibody. The lactoperoxidase technique did not succeed in labeling MPO with an adequate specific activity. In contrast, the self-labeling method gave a stable tracer with a specific activity of 23 microCi/micrograms MPO (85 MBq), a satisfactory level of immunoreactivity, and a low-specific binding (less than or equal to 3%). After labeling, purification of tracer was achieved by gel filtration chromatography in phosphate buffer (0.05 M; pH7) to which 0.1% poly-L-lysine was added. The labeled molecule remained stable for 40 days and could be used for RIA with a polyclonal antibody raised in rabbits.

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