Method for radioiodination of proteins using N-succinimidyl 3-hydroxy-4-iodobenzoate.
A conjugation method has been developed for the radioiodination of proteins which should be adaptable to kit formulation. m-Hydroxybenzoic acid was converted to 3-hydroxy-4-[131I]iodobenzoic acid in 65% radiochemical yield using Chloramine-T as the oxidant. This intermediate was then converted to N-succinimidyl 3-hydroxy-4-[131I]iodobenzoate ([131I]mSHIB) in 75% yield by reaction with N-hydroxysuccinimide and dicyclohexylcarbodiimide in a reaction time of only 10 min. Monoclonal antibody (mAb) 81C6 was labeled in 40-60% yield by reaction with [131I]mSHIB. Performing purifications of radioiodinated compounds using cartridges instead of HPLC did not alter conjugation efficiency, mAb immunoreactivity, or tissue distribution. Thyroid uptake of labeled mAb was low but up to 2.4 times higher than that seen when the mAb was labeled with N-succinimidyl 3-[125I]-iodobenzoate. These results suggest that [131I]mSHIB may be a useful reagent for the radioiodination of proteins, particularly in contexts when less complicated purification methods would be advantageous.
Vaidyanathan
G
G
Department of Radiology, Duke University Medical Center, Durham, North Carolina 27710, USA. g405@acpub.duke.edu
Affleck
D J
DJ
Zalutsky
M R
MR
D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000493:pharmacokinetics; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes / Q000737:chemistry; D007463:Iodobenzoates / Q000737:chemistry*; D007553:Isotope Labeling / Q000379:methods*; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D011506:Proteins / Q000737:chemistry*; D014018:Tissue Distribution
D000911:Antibodies, Monoclonal; D007202:Indicators and Reagents; D007457:Iodine Radioisotopes; D007463:Iodobenzoates; D011506:Proteins; C062337:N-succinimidyl 3-iodobenzoate