Perturbation of epidermal growth factor clearance after radioiodination and its implications.

The clearance of human epidermal growth factor (hEGF1-53) has been thought to be mediated mainly by a high-capacity receptor system, yet relatively low in vivo clearance rates (<10 mL/min/kg) and long terminal elimination half-lives (>120 min) have been observed in rats receiving the peptide that was iodinated by the oxidative chloramine-T (CT) method. We investigated if a mild, less oxidative iodination by the lactoperoxidase (Enzymobeads, EB) method, which is known to yield an iodinated peptide with receptor-binding equivalence, could produce a labeled peptide that behaves pharmacokinetically similar to the native material. For comparison, a parallel study was also conducted with EB-125I-hEGF1-48, which in its native form has a much reduced receptor binding activity due to the loss of the C-terminal pentapeptide. Plasma radioactivity concentrations were determined by trichloroacetic acid (TCA) precipitation and immunoprecipitation. Rats cleared unlabeled hEGF1-53 and hEGF1-48 markedly faster (CL(tot) > 120 mL/min/kg) than their radiolabeled counterparts. Approximately 96% of the hEGF1-53 dose was cleared during the initial phase (0-4 min), as opposed to only 5-14% for the iodinated peptide. Similar change was also observed for EB-125I-hEGF1-48 and CT-125I-hEGF1-53. The pharmacokinetic behavior of EB-125I-hEGF1-53 was, in fact, comparable to that of CT-125I-hEGF1-53. These observations indicate that receptor-binding equivalence does not have direct relationship with in vivo EGF clearance. Both iodination methods (oxidative CT and less oxidative EB) might have perturbed one or more steps in the cascade of ligand-receptor internalization and intracellular procession, which in turn modified the disposition of the peptides. In addition, the two independent precipitation techniques for the same peptide generated different kinetic outcomes. The overall experimental results suggest that it is unacceptable to use an iodinated form to characterize the disposition of peptides/proteins like EGF with a specific receptor system mediating its clearance.