Labeling peptides with technetium-99m using a bifunctional chelator of a N-hydroxysuccinimide ester of mercaptoacetyltriglycine.
A modified mercaptoacetyltriglycine (MAG3) chelator, which has acetyl S-protection and which is derivitized with N-hydroxysuccinimide (NHS) ester for conjugation, has been used to radiolabel four small (approximately 6- to 7-kDa) peptides, bovine pancreatic trypsin inhibitor, epidermal growth factor, human neutrophil elastase inhibitor and plasmin inhibitor, with 99mTc.


METHODS
Each peptide was specifically labeled at the MAG3 chelation sites at ambient temperature and neutral pH. Specific activities of 100-150 mCi/mg were achieved at labeling efficiencies of about 50%, but specific activities of 3500 mCi/micromol could be attained.


RESULTS
By a variety of assays, protein activity was unimpaired by the conjugation and labeling for two of the four peptides. The activities for plasmin of the plasmin inhibitor and bovine pancreatic trypsin inhibitor were reduced by conjugation, presumably because of a sensitive lysine residue in the structure of each of these two peptides. Multiple peaks were present in the high-performance liquid chromatography radiochromatograms, especially of human neutrophil elastase inhibitor; however, most peaks could be shown to be labeled active peptide. Stability during cysteine challenge at modest cysteine-to-peptide molar ratios and during incubation in serum was observed in each case. Large differences among the labeled peptides were apparent in the 3-hr biodistributions of 99mTc in normal mice.


CONCLUSIONS
The use of NHS-S-acetyl-MAG3 may be a convenient method of radiolabeling peptides with 99mTc.
Hnatowich
D J
DJ
Department of Nuclear Medicine, University of Massachusetts Medical Center, Worcester 01655, USA.
Qu
T
T
Chang
F
F
Ley
A C
AC
Ladner
R C
RC
Rusckowski
M
M
D000818:Animals; D000933:Antifibrinolytic Agents / Q000493:pharmacokinetics; D007611:Aprotinin / Q000493:pharmacokinetics; D002417:Cattle; D004815:Epidermal Growth Factor / Q000493:pharmacokinetics; D006801:Humans; D007553:Isotope Labeling; D008297:Male; D051379:Mice; D019275:Radiopharmaceuticals / Q000493:pharmacokinetics; D015842:Serine Proteinase Inhibitors / Q000493:pharmacokinetics; D013667:Technetium / Q000493:pharmacokinetics; D014018:Tissue Distribution
D000933:Antifibrinolytic Agents; D019275:Radiopharmaceuticals; D015842:Serine Proteinase Inhibitors; D004815:Epidermal Growth Factor; D013667:Technetium; D007611:Aprotinin