Article (Scientific journals)
Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine
Deby, Ginette; Pincemail, Joël; Thirion, A. et al.
1991In Experientia, 47 (9), p. 952-7
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Keywords :
Human; Leucocytes; Myeloperoxidase-iodination-radioimmunoassay
Abstract :
[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.
Disciplines :
Human health sciences: Multidisciplinary, general & others
Anesthesia & intensive care
Author, co-author :
Deby, Ginette ;  Université de Liège - ULiège > Centre de l'oxygène : Recherche et développement (C.O.R.D.)
Pincemail, Joël ;  Centre Hospitalier Universitaire de Liège - CHU > Chirurgie cardio-vasculaire
Thirion, A.
Deby, C.
Lamy, Maurice ;  Université de Liège - ULiège > Département des sciences cliniques > Anesthésie et réanimation
Franchimont, P.
Language :
English
Title :
Self-Labeling of Human Polymorphonuclear Leucocyte Myeloperoxidase with 125iodine
Publication date :
15 September 1991
Journal title :
Experientia
ISSN :
0014-4754
Publisher :
Birkhauser Verlag, Switzerland
Volume :
47
Issue :
9
Pages :
952-7
Peer reviewed :
Peer Reviewed verified by ORBi
Available on ORBi :
since 25 March 2009

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