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Biochim Biophys Acta. 2014 Feb;1840(2):722-9. doi: 10.1016/j.bbagen.2013.04.039. Epub 2013 May 2.

Immuno-spin trapping from biochemistry to medicine: advances, challenges, and pitfalls. Focus on protein-centered radicals.

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Laboratory of Experimental Medicine and Therapeutics, Institute Multidisciplinary of Biological Investigations-San Luis (IMIBIO-SL), National Bureau of Science and Technology (CONICET) and National University of San Luis, San Luis, 5700 San Luis, Argentina.



Immuno-spin trapping (IST) is based on the reaction of a spin trap with a free radical to form a stable nitrone adduct, followed by the use of antibodies, rather than traditional electron paramagnetic resonance spectroscopy, to detect the nitrone adduct. IST has been successfully applied to mechanistic in vitro studies, and recently, macromolecule-centered radicals have been detected in models of drug-induced agranulocytosis, hepatotoxicity, cardiotoxicity, and ischemia/reperfusion, as well as in models of neurological, metabolic and immunological diseases.


To critically evaluate advances, challenges, and pitfalls as well as the scientific opportunities of IST as applied to the study of protein-centered free radicals generated in stressed organelles, cells, tissues and animal models of disease and exposure.


Because the spin trap has to be present at high enough concentrations in the microenvironment where the radical is formed, the possible effects of the spin trap on gene expression, metabolism and cell physiology have to be considered in the use of IST and in the interpretation of results. These factors have not yet been thoroughly dealt with in the literature.


The identification of radicalized proteins during cell/tissue response to stressors will help define their role in the complex cellular response to stressors and pathogenesis; however, the fidelity of spin trapping/immuno-detection and the effects of the spin trap on the biological system should be considered. This article is part of a Special Issue entitled Current methods to study reactive oxygen species - pros and cons and biophysics of membrane proteins. Guest Editor: Christine Winterbourn.


5,5-dimethyl-1-pyrroline N-oxide; AG; Anti-DMPO; BSA; DMPO; Disease/exposure model; ESR; HPLC; I/R; IST; Immuno-spin trapping; LC; LPS; MRI; MS; OA; Protein radical; Reactive chemical species; SOD; Spin trap; TPO; aminoglutethimide; bovine serum albumin; electron spin resonance; high performance liquid chromatography; hoMb; horse myglobin; huHb/Mb; human hemoglobin/myoglobin; immuno-spin trapping; ischemia/reperfusion; lipopolysaccharide; liquid chromatography; magnetic resonance imaging; mass spectrometry; octanoic acid; superoxide dismutase; thyroid peroxidase

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