Labeling yield (decay corrected), radiochemical purity and stability of ⁶⁸Ga-labeled Affibody molecules.

*<p>To evaluate stability, conjugates were incubated with 500-fold excess of EDTA for 1 h and then analyzed using ITLC. Control samples were incubated in PBS. The experiments were performed in duplicates. The data present an average conjugate-associated radioactivity and maximum error.</p

Tumor-to-organ ration data 2h after injection for NODAGA-Z_HER2:S1, NOTA-Z_HER2:S1 and DOTA-Z_HER2:S1 in mice bearing SKOV-3 xenografts.

<p>Data are presented as an average and standard deviation for four mice. Data concerning bone from one mice injected with ⁶⁸Ga-NOTA-ZHER2:S1 and bone and muscle form one mice injected with ⁶⁸Ga-NODAGA-ZHER2:S1were excluded due to low counts.</p>a<p>significant difference (p<0.05) between ⁶⁸Ga-DOTA-Z_HER2:S1 and ¹¹¹In-DOTA-Z_HER2:S1.</p>b<p>significant difference (p<0.05) between ⁶⁸Ga-NOTA-Z_HER2:S1 and ¹¹¹In-NOTA-Z_HER2:S1.</p>c<p>significant difference (p<0.05) between ⁶⁸Ga-NODAGA-Z_HER2:S1 and ¹¹¹In-NODAGA-Z_HER2:S1.</p>d<p>significant difference (p<0.05) between ⁶⁸Ga-DOTA-Z_HER2:S1 and ⁶⁸Ga-NOTA-Z_HER2:S1.</p>e<p>significant difference (p<0.05) between ⁶⁸Ga-NOTA-Z_HER2:S1 and ⁶⁸Ga-NODAGA-Z_HER2:S1.</p>f<p>significant difference (p<0.05) between ⁶⁸Ga-NODAGA-Z_HER2:S1 and ⁶⁸Ga-DOTA-Z_HER2:S1.</p

Comparative biodistribution of NODAGA-Z_HER2:S1, NOTA-Z_HER2:S1 and DOTA-Z_HER2:S1 labelled with gallium-68 and indium-111 after intravenous injection in female BALB/C nu/nu mice bearing SKOV-3 xenografts.

<p>Data are presented as an average % IA/g and standard deviation for four mice. Data concerning bone from one mice injected with ⁶⁸Ga-NOTA-Z_HER2:S1 and bone and muscle form one mice injected with ⁶⁸Ga-NODAGA-Z_HER2:S1were excluded due to low counts.</p>a<p>significant difference (p<0.05) between ⁶⁸Ga-DOTA-Z_HER2:S1 and ¹¹¹In-DOTA-Z_HER2:S1.</p>b<p>significant difference (p<0.05) between ⁶⁸Ga-NOTA-Z_HER2:S1 and ¹¹¹In-NOTA-Z_HER2:S1.</p>c<p>significant difference (p<0.05) between ⁶⁸Ga-NODAGA-Z_HER2:S1 and ¹¹¹In-NODAGA-Z_HER2:S1.</p>d<p>significant difference (p<0.05) between ⁶⁸Ga-DOTA-Z_HER2:S1 and ⁶⁸Ga-NOTA-Z_HER2:S1.</p>e<p>significant difference (p<0.05) between ⁶⁸Ga-NOTA-Z_HER2:S1 and ⁶⁸Ga-NODAGA-Z_HER2:S1.</p>f<p>significant difference (p<0.05) between ⁶⁸Ga-NODAGA-Z_HER2:S1 and ⁶⁸Ga-DOTA-Z_HER2:S1.</p>*<p>Data for gastrointestinal (GI) tract and carcass are presented as %IA per whole organ.</p

Biophysical characteristics of the Affibody conjugates [22].

<p>Biophysical characteristics of the Affibody conjugates <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0070028#pone.0070028-Malmberg2" target="_blank">[22]</a>.</p

Specificity of targeting of SKOV-3 xenografts in BALB/C nu/nu mice using ⁶⁸Ga-DOTA-Z_HER2:S1, ⁶⁸Ga-NOTA-Z_HER2:S1 and ⁶⁸Ga-NODAGA-Z_HER2:S1 at 1h after injection.

<p>The total injected radiolabeled peptide dose per mouse was 3 µg (0.42 nmol). The blocked group was subcutaneously pre-injected with an excess amount (1000 µg, 150 nmol) of non-labelled Z_HER2∶342 to saturate binding sites of HER2. Results are expressed as percentage of injected activity per gram of tissue (%IA/g) and presented as mean values for four mice and standard deviations. Uptake of radioactivity was significant lower (p<0.0005) for pre-saturated SKOV-3 tumors.</p

MicroPET/CT imaging of HER2 expression in SKOV3 xenografts in BALB/C nu/nu mice using ⁶⁸Ga-NODAGA-Z_HER2:S1.

<p>The image was acquired 2 h after administration of the tracer. Arrows point at tumor (T) and kidneys (K).</p

Cellular processing of ⁶⁸Ga-NODAGA-Z_HER2:S1, ⁶⁸Ga-NOTA-Z_HER2:S1 and ⁶⁸Ga-DOTA-Z_HER2:S1 by HER2-expressing cells SKOV-3 in vitro.

<p>Cells were incubated with labeled compound at 37°C. Cell bound activity is normalized to the maximum uptake. Data are presented as mean values for three cell dishes and standard deviations. Error bars might be smaller than the symbols.</p

Radiolabeled HRG homes to the perivascular area and to inflammatory cells.

<p>Micro-autoradiography of ¹²⁵I-mHRG in the spleen and T241 fibrosarcomas (Tumor) tissue at 15 min post injection of radiolabeled HRG. A. Panels show tumor and spleen tissues from mice injected with PBS (left) or ¹²⁵I-mHRG (middle and right). Immunohistochemical staining with CD31 antibodies show endothelial cells colocalized with ¹²⁵I-mHRG in the middle panels (arrows). Panels to the right show the retention of ¹²⁵I-mHRG alone. B. Panels as above but immunohistochemical staining with CD45 antibodies to identify leukocytes. Arrowheads in the middle panel indicate colocalization of CD45-positive leukocytes and ¹²⁵I-mHRG. Panels to the right show the retention of ¹²⁵I-mHRG alone.</p

Biodistribution of ¹²⁵I-mHRG in normal and tumor-bearing C57Bl/6 mice.

<p>Uptake is expressed as % ID/g and presented as an average value from 4 animals ± S.D.</p><p>“Naive” and “Tumor”, indicate healthy mice and mice challenged with subcutaneous T241 fibrosarcomes, respectively.</p>§<p>) Data for intestines with content and carcass are presented as %ID per whole sample.</p><p>*) Significant (<i>p</i><0.05) difference between normal and tumor bearing mice at this time point.</p>#<p>) Significant (<i>p</i><0.05) difference between hHRG and mHRG at this time point.</p>‡<p>) Significant (<i>p</i><0.05) difference between tumor and muscle at this time point.</p><p>Biodistribution of ¹²⁵I-mHRG in normal and tumor-bearing C57Bl/6 mice.</p

Schematic outline of HRG's interaction with its receptor on mononuclear phagocytes.

<p>HRG is shown produced in the liver, distributed in the circulation bound to mononuclear phagocytes which home to sites of inflammation. Binding of HRG to the HRG receptor (HRGR) leads internalization, degradation and thereby regulation of HRG turnover.</p