Surface functionalization of Feraheme (FH, 1) with azide or alkyne groups and radiolabeling functionalized NPs by HIR.

<p>a) Using FH (<b>1</b>)<b>,</b> the Azide-FH (<b>4</b>) and Alkyne-FH (<b>5</b>) were synthesized. Portions of Azide-FH <b>(4)</b> and Alkyne-FH (<b>5</b>) were then radiolabeled by HIR, yielding ⁸⁹Zr-Azide-FH (^<b>89</b><b>Zr-4</b>) and ⁸⁹Zr-Alkyne-FH (^<b>89</b><b>Zr-5</b>). To determine reactive azide or reactive alkynes, NPs were reacted with the appropriate click reactive Cy5.5 fluorochromes, with Cy5.5s shown as the yellow stars of <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172722#pone.0172722.g001" target="_blank">Fig 1</a>. After removal of the unreacted Cy5.5s (DBCO-Cy5.5, <b>6</b> or Azide-Cy5.5, <b>7</b>), the number of Cy5.5’s per NP was determined from absorption spectra examples of which are shown in Fig 2b–2e. Controls for covalent binding were a reaction of FH (<b>1</b>) and DBCO-Cy5.5 (<b>6</b>) and a reaction of Azide-FH <b>(4)</b> and DBCO-Cy5.5 (<b>6</b>) preoccupied with DBCO-NH₂. Values in parentheses are the numbers of reactive groups per NP with values summarized in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172722#pone.0172722.t001" target="_blank">Table 1</a>.</p

Outline of heat induced radiolabeling (HIR) and click chemistry surface functionalization used to obtain multimodal, targeted NPs.

<p>Our previous HIR of FH NPs (top) yielded non-surface functionalized, radioactive NPs (red core). The alkyne and azide functionalized FH intermediates were synthesized (Azide-FH, <b>4</b> and Alkyne-FH, <b>5</b>) and labeled by HIR reaction, to yield ⁸⁹Zr-Azide-FH (^<b>89</b><b>Zr-4</b>) and ⁸⁹Zr-Alkyne-FH (^<b>89</b><b>Zr-5</b>). Imaging detection modalities for the NPs are in bold. NPs targeted to folate receptors (⁸⁹Zr-Folate-FH, ^<b>89</b><b>Zr-11</b>), integrins (RGD-FH, <b>14</b>) or NPs with protamines (⁸⁹Zr-Cy5.5-Protamine-FH, ^<b>89</b><b>Zr-16</b>) were then synthesized. Detailed synthetic schemes are given in Figs <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172722#pone.0172722.g002" target="_blank">2</a>–<a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0172722#pone.0172722.g004" target="_blank">4</a>.</p

Summary of HIR NP’s and Surface Functionalization Reactions.

<p>Summary of HIR NP’s and Surface Functionalization Reactions.</p

Additional file 1 of Radiosynthesis automation, non-human primate biodistribution and dosimetry of K+ channel tracer [11C]3MeO4AP

Additional file 1: Supplementary Information

Parametric images of V_T and ΔΨ_T.

<p>Each panel of three x two images shows short axis, vertical and horizontal slices. Images of a representative control pig are shown in the left panel. Images of a representative scar pig are shown in the right panel. The top row of each panel depicts the TPP⁺ volume of distribution and bottom row the membrane potential.</p

Dependence of <i>V</i>_<i>T</i> on mitochondrial membrane potential and fractional ECS computed with Eq 3.

<p><i>ΔΨ</i>_<i>T</i> is more negative inside membranes. Black lines indicate variation (systematic error) in <i>ΔΨ</i>_<i>T</i> range at constant volume of distribution due to neglecting fractional ECS volume.</p

Volume of distribution model for¹⁸F-TPP⁺ in a PET image voxel.

<p>The outer black line represents the voxel boundary. C_p, C_inter, C_cyto, and C_mito represent the concentrations of the plasma, interstitial space, cytosol, and mitochondria respectively. The arrows represent ¹⁸F-TPP⁺ transport between the different compartments. <i>f</i>_ECS represents the voxel volume fraction occupied by ECS and <i>f</i>_mito represents the cellular volume fraction occupied by mitochondria.</p

Measured values of mitochondrial membrane potential (milliVolts).

<p>Measured values of mitochondrial membrane potential (milliVolts).</p

Typical ¹⁸F-TPP⁺ SUV image, integrated from 60–120 minutes after IV bolus injection.

<p>SUV is highest in liver, followed by LV myocardium, with lower activity in the visible in bone marrow.</p

Comparison of <i>ΔΨ</i>_<i>T</i> in 17 "bull’s eye" segments Results shown for five control (blue squares) and three pigs with injury to the LAD territory (red circles).

<p>Comparison of <i>ΔΨ</i>_<i>T</i> in 17 "bull’s eye" segments Results shown for five control (blue squares) and three pigs with injury to the LAD territory (red circles).</p