The antibody cleared from the blood pool in a typical pattern for a whole antibody. was confirmed by the 48 hour biodistribution data: percentage of injected dose per gram of tissue (%ID/g SD) (large intestine) colitis mice with 64Cu-labeled FIB504, 6.49 2.25; control mice with 64Cu-labeled Sitagliptin phosphate monohydrate FIB504, 3.64 Sitagliptin phosphate monohydrate 1.12; colitis mice, 64Cu-labeled non-specific antibody 3.97 0.48 %ID/g ( 0.05 between groups). Conclusions The selective uptake of 64Cu-labeled FIB504 antibody in the gut of animals with colitis suggests that integrin 7 may be a promising target for radioimmunodetection of this disease, which would aid diagnosis, assessment and therapy guidance of this disease. tissue assay to measure radionuclide distribution in this acute model of colitis to investigate whether there was specific uptake in the inflamed gut. Materials and Methods General Chemicals and reagents were obtained from Sigma-Aldrich unless otherwise specified. EDC (1-ethyl-3-(3-(dimethylamino)propyl)carbodiimide hydrochloride) was purchased from Pierce (Rockford IL, USA). tissue biodistribution was carried out. Tissues were collected, weighed and radioactivity was assayed using a Packard Cobra II automated gamma counter (Meriden, CT). MicroPET and microCT images were registered manually using AMIDE software (A Medical Imaging Data Examiner, http://amide.sourceforge.net/index.html). Data from volumes of interest (VOIs) within relevant tissues were used to calculate biodistribution throughout the imaging study. Statistical analysis Statistical analysis was carried out using SPSS V 14.0 for Windows. The 48 h post-injection biodistribution data was analyzed for statistical significance by ANOVA, Sitagliptin phosphate monohydrate with analysis adjusted using the Bonferroni Ly6a correction. Differences were considered significant at the 5 % level ( 0.05). Results DSS model Including DSS in the drinking water resulted in body weight loss, as is usually shown for treated and control groups in Physique 1. Mean decrease in total body weight was 5.43 7.13 % for treated mice; body weight gain for non-treated mice was 5.9 0.75 % over the same period. In our experience, this pattern of initial increase and then decrease in body weight is usually common for mice treated with this level of DSS (2 % w/v) [12]. For all those DSS-treated mice feces became more liquid during the course of the study and hemorrhage from the bowel was observed at necropsy. Open in a separate window Physique 1 Bodyweight changes in mice treated with dextran sodium sulfate in order to experimentally induce colitisTotal body weights (means SD) of control (solid line) and DSS-treated mice (2 % w/v in water, dashed line). This pattern of increase and decrease is usually common. Immunoreactivity assay In order to determine whether Sitagliptin phosphate monohydrate the conjugation of the chelator reduced the binding capacity of the anti-7 integrin mAb (FIB504.64) to its target, we examined the binding of FIB504.64 to TK-1 model cells. Flow cytometry data (Physique 2) indicated that this immunoreactivity of the antibody was not impaired by the conjugation of the bifunctional chelator. Open in a separate window Physique 2 Conjugation of BiFunctional Chelator to FIB504.64 does not impair the binding of the antibodyA representative histogram confirming that this antigen binding of this anti-7 antibody was not impaired by conjugation with BFC NH2- 0.05) in the radioactivity in the blood between groups 1 (test group) and 3 (control group, non-specific antibody, DSS treated mice) at both 24 and 48 h with activity in group 1 lower at both time points (group 1; 12.93 2.12 percentage of injected dose per gram of tissue (%ID/g), 7.29 3.29 %ID/g: group 2; 20.63 3.69, 12.62 1.33: means SD at 24 Sitagliptin phosphate monohydrate and 48 hours, respectively). Open in a separate window Physique 3 Blood pharmacokinetics of radiolabeled antibodies in DSS induced colitis micePharmacokinetics (percentage.
