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J Nucl Med. 2008; 49 (Supplement 1):309P
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Radiopharmaceutical Chemistry: Radiopharmacy

Radiopharmacy Posters

Synthesis and in vitro characterization of a dendrimer-MORF conjugate for amplification pretargeting

Xiangji Chen1, Shuping Dou1, Guozheng Liu1, Xinrong Liu1, Yi Wang1, Ling Chen1, Mary Rusckowski1 and Donald Hnatowich1

1 Radiology, University of Massachusetts Medical School, Worcester, Massachusetts

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Objectives: Multiple step amplification pretargeting can greatly improve target localization of radiolabeled oligomer effectors through the intermediate use of polymers conjugated with multiple copies of the complementary oligomer. A generation 3 (G3) PAMAM carboxyl terminal dendrimer was conjugated via carbodiimides with multiple copies of an 18-mer MORF and used in combination with the cMORF conjugated CC49 antibody and the 99mTc-cMORF for amplification pretargeting.

Methods: Characterization by native-PAGE and size-exclusion HPLC included both the average MORFs per G3-MORF (gpm) and the number accessible to the 99mTc labeled cMORF. After purification, the G3-MORF was investigated for its ability to link the CC49-cMORF and 99mTc-cMORF in solution. Thereafter amplification pretargeting was performed on protein G and protein L coated plates to which the CC49 antibody was immobilized via its Fc and VLK regions respectively. Amplification was measured in the same wells with respect to conventional pretargeting with MORF conjugated CC49 and 99mTc labeled cMORF but without the polymer.

Results: Nine of the 32 carboxyl groups were modified with MORF, of which 90% were accessible in solution to 99mTc-cMORF. The 99mTc-labeled cMORF/G3-MORF complex when hybridized with CC49-cMORF demonstrated a single peak on SE HPLC as evidence of complete hybridization between 99mTc-labeled cMORF/G3-MORF and CC49-cMORF. On protein G and L coated plate, the amplification factors were 6 and 14 respectively, showing that the polymer participated in amplifying the signal.

Conclusions: A G3-MORF dendrimer was synthesized and used successfully for in vitro amplification pretargeting.





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