Stability analysis on the radioactive iodine-labelled prostate cancer-specific recombinant oncolytic adenovirus

摘要

The aim of the present study was to construct the I-125-replication-selective oncolytic adenovirus (RSOAds) human telomerase reverse transcriptase (hTERT)/prostate specific antigen (PSA) nuclide-oncolytic virus marker by labelling the hTERT/PSA double-regulation replicative oncolytic adenovirus with I-125 nuclide, and investigate the influence of viral markers under various reaction conditions on labelling efficiency. N-bromosuccinimide (NBS) was used as the oxidizer for I-125 labelling, and the best conditions for labelling were identified through the reactions between oncolytic adenovirus at various concentrations and NBS. Dosage of I-125, reaction duration, pH values and reaction volume were respectively evaluated to determine their effects on the labelling efficiency of I-125-RSOAds-hTERT/PSA nuclide-oncolytic adenovirus markers. Purified nuclide-oncolytic adenovirus markers were isolated by gel-filtration chromatography; paper chromatography was performed to assay the radiochemical purity of I-125-RSOAds-hTERT/PSA markers at various time points. Radiochemical purity of I-125-RSOAds-hTERT/PSA was >95%, and could be maintained at 4 degrees C for 7 days. The best reaction conditions were set as follows: 0.5 mu l of I-125 (similar to 0.2 m Ci, 7.4 MBq); 25 qg of NBS; 100 mu l of 8x10(9) VP/ml I-125-RSOAds-hTERT/PSA virus solution; 30 min of reaction duration; pH 7.5; 120 mu l of PBS. Labelling hTERT/PSA double-regulation replicative oncolytic adenovirus with I-125 was identified to be available, and the radiochemical purity of acquired virus markers could be maintained under specific conditions.