Patent ID: 6649736
Filing Date: 2003-11-18
Classification: C07K

Abstract:
A method of labeling a percentage of the solvent-accessible, reduced carbon atoms in a peptide or protein with a heavy hydrogen, the method comprising:(i) generating hydroxyl radical in a solution for a time and under conditions effective to achieve a total equivalent concentration of the hydroxyl radical of at least 10 &mgr;M, wherein said solution comprises the peptide or protein, an electron scavenger source, a heavy hydrogen source, and a heavy hydrogen donor; (ii) adding an amount of a heavy hydrogen donor to the solution to replace heavy hydrogen donor that is depleted during step (i); and (iii) optionally repeating steps (i) and (ii) until a cumulative total equivalent concentration of hydroxyl radical generated in the solution is sufficient to produce carbon-centered radicals of said percentage of solvent-accessible, reduced carbon atoms in said peptide or protein; wherein, when the total equivalent concentration of hydroxyl radical is achieved in step (i), the concentration of O2 dissolved in the solution is sufficiently low that a substantial number of the carbon-centered radicals formed in step (i) are repaired by donation of a hydrogen isotope by the heavy hydrogen donor.