This work was supported by grants from the National Research Foundation of Korea, funded by the government of the Republic of Korea, (Grant nos. 2012M2B2B1055245 and 2012M2A2A6011335) and by the RI-Biomics Center of Korea Atomic Energy Research Institute.

זהירות: צורת החמצון של יוד רדיואקטיבי היא תנודתי למדי ויש לטפל עם מגינים להוביל נאותים בקבוקוני עופרת. כל צעדי radiochemical צריכות להתבצע במנדף פחם-מסונן מאוורר היטב, ואת הפרוצדורות צריכות להיות פיקוח על ידי זיהוי התקני רדיואקטיביות. 1. הכנת הכימיקלים והמחסנית הפוך שלב לסינתזה של 125 I-שכותרתו יזיד <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> הכנת ריאגנטים בתמיסה <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> ממסי 1 מ&quot;ג של המבשר יזיד (2) ב 150 μl אתנול אבסולוטי (איור 1). הערה: הליך סינטטי מפורט המבשר יזיד (2) דווחה בעיתון הקודם 22. </li><li style=";text-align:right;direction:rtl"> ממיסים T chloramine 1 מ&quot;ג ב 20 μl של חיץ מלוחים פוספט 1x (pH = 7.4). </li><li style=";text-align:right;direction:rtl"> ממיסים metabisulfite נתרן 2 מ&quot;ג ב 20 μl H 2 O. </li></ol></li><li style=";text-align:right;direction:rtl"> preparation של המחסנית <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לשטוף את המחסנית tC18 עם אתנול אבסולוטי 10 מ&quot;ל ואחריו 10 מ&quot;ל H 2 O. אין לייבש את המטריצה ​​של מחסנית עם אוויר. </li></ol></li></ol> 2. Radiosynthesis של 125 הקבוצה התותבת תזיד שכותרתו לי <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> תגובת Radioiodination של המבשר <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> מוסיפים את פתרון מבשר אזיד (1 מ&quot;ג ב 150 μl של אתנול אבסולוטי) וחומצה אצטית (10 μl) לצינור 1.5 מ&quot;ל microcentrifuge. </li><li style=";text-align:right;direction:rtl"> להוסיף 150 MBq של [125 I] נאה ב 0.1 M NaOH (50 μl) לתערובת התגובה. </li><li style=";text-align:right;direction:rtl"> הוספת פתרון T chloramine (1 מ&quot;ג ב 20 μl של חיץ מלוחים פוספט 1x) ולסגור את תערובת התגובה המכיל microcentrifuge צינור. </li><li style=";text-align:right;direction:rtl"> דגירה תערובת התגובה בטמפרטורת החדר למשך 15 דקות עד השלמת התגובה radioiodination. </li><li style=";text-align:right;direction:rtl"> הוספת פתרון metabisulfite נתרן (2 מ&quot;ג ב 20 μl H 2 O) לתערובת התגובהכדי להרוות את התגובה radioiodination. </li><li style=";text-align:right;direction:rtl"> לסגת 0.2 μl של המוצר הגולמי ואז לדלל אותו עם 100 μl של פתרון (H 2 O / CH 3 CN, 1: 1) עבור ניתוח HPLC. הערה: עבור כל הניסויים HPLC, השתמש 0.1% חומצה פורמית המכיל H 2 O (ממס) ו -0.1% אצטוניטריל המכילים חומצה פורמית (B ממס) כמו eluents. </li><li style=";text-align:right;direction:rtl"> נתח את המוצר הגולמי מדולל באמצעות הפוכה פאזיים אנליטיים רדיו-HPLC (C18 הפוכה שלב בטור; קצב הזרימה: 1 מ&quot;ל / דקה; שיפוע eluent: 20% B ממס עבור 0-2 דקות, 20-80% ב ממס עבור 2-22 דקות, B ממס 80-100% עבור 22-23 דקות, ולאחר 100% B ממס עבור 23-28 דקות; זמן השמירה: 16.4 דק &#39;) (איור 2). </li></ol></li><li style=";text-align:right;direction:rtl"> טיהור של המוצר הגולמי עם HPLC preparative הערה: ספק מספיק שריון עופרת סביב חלקי HPLC כגון מזרק, עמודה, גלאי, בקבוקונים אוספים, ואת המכל שבו הקולחים נאספו. <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לסגת הדואר תערובת תגובה כולו לתוך בקבוקון HPLC. יש לשטוף את צינור התגובה עם אצטוניטריל (0.5 מ&quot;ל) ולהוסיף השטיפה לתוך אותו בקבוקון ההזרקה. לדלל את הפתרון שנאספו עם H 2 O (1 מ&quot;ל). </li><li style=";text-align:right;direction:rtl"> להזריק את המוצר הגולמי על רדיו-HPLC preparative (C18 הפוכה שלב בטור; קצב הזרימה: 10 מ&quot;ל / דקה; שיפוע eluent: 20% B ממס עבור 0-2 דקות, 20-80% ב ממס עבור 2-22 דקות, 80-100% B ממס עבור 22-23 דקות, ולאחר 100% B ממס עבור 23-28 דקות). </li><li style=";text-align:right;direction:rtl"> אספו את שיא רדיואקטיבי המייצג 125 יזיד שכותרתו אני (1) (R t בתנאי HPLC אלה הוא 17.8-18.8 דקות) במבחנת זכוכית (איור 2). </li><li style=";text-align:right;direction:rtl"> מדוד את תשואת radiochemical של השבר באמצעות כיל רדיואקטיביות מינון על פי הפרוטוקול של היצרן. </li><li style=";text-align:right;direction:rtl"> להזריק את המוצר המטוהר תתפס-HPLC רדיו אנליטיים באמצעות באותם התנאים HPLC לקביעת טוהר radiochemical של המוצר. </li></ol></li><li style=";text-align:right;direction:rtl"> מיצוי שלב מוצק של המוצר <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לדלל את שבריר המכיל את המוצר הרצוי (1) עם 40 מ&quot;ל טהור H 2 O. </li><li style=";text-align:right;direction:rtl"> מוסיף את הפתרון המדולל לתוך מחסנית tC18 preconditioned. </li><li style=";text-align:right;direction:rtl"> לשטוף את המחסנית עם H מ&quot;ל 15 נוספים 2 O. </li><li style=";text-align:right;direction:rtl"> Elute המוצר (1) לכוד בתוך מחסנית עם 2 מ&quot;ל אצטון לתוך בקבוקון זכוכית 10 מ&quot;ל כי הוא מוגן על ידי מגן עופרת. מדוד את הרדיואקטיביות של מוצר eluted באמצעות כיל רדיואקטיביות מינון על פי הפרוטוקול של היצרן. הערה: sulfoxide דימתיל (DMSO) או אתנול אבסולוטי יכול לשמש גם עבור elution של המוצר מהמחסנית. כ 5-10% של רדיואקטיביות בדרך כלל מקלות על המחסנית, המוצר radiolabeled הנותרים לא ניתן eluted מלא באמצעות כמויות גדולות של ממיס אורגני. </li><li style=";text-align:right;direction:rtl"> לאדות את אצטון עם זרם של חנקן או גז ארגון. </li><li style=";text-align:right;direction:rtl"> ממיסים את מחדשsidue עם DMSO (100-200 μl) עבור השלב radiolabeling הבא. </li></ol></li></ol> סינתזת 3. DBCO-group-מצומדות חלקיקי זהב <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> שינוי פני שטח של זהב 13 ננומטר בגודל חלקיקים עם פוליאתילן גליקול קבוצה-DBCO המכיל <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כן חלקיקי זהב נתרן מיוצב-ציטראט (3) (= גודל הממוצע 13 ננומטר) על פי דו&quot;ח קודם 24. </li><li style=";text-align:right;direction:rtl"> להוסיף בתמיסה מימית של 20 Tween (1 מ&quot;מ, 1.5 מ&quot;ל) אל חלקיקי זהב מיוצב ציטרט (10 ננומטר, 15 מ&quot;ל). לנער את הפתרון עבור 20 דקות על שייקר מסלולית. </li><li style=";text-align:right;direction:rtl"> להוסיף בתמיסה מימית של תיאול פוליאתילן גליקול DBCO המכילים-קבוצה (משקל מולקולרי ממוצע = 5000, 100 מיקרומטר, 1.5 מ&quot;ל). לנער את פתרון 2 שעות על שייקר מסלולית. </li></ol></li><li style=";text-align:right;direction:rtl"> טיהור של חלקיקי זהב DBCO-שונה-group <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לטהר את חלקיקי זהב DBCO-שונה-קבוצה (4) &lt;/ Strong&gt; על ידי צנטריפוגה רצוף (11,400 XG, 15 דק &#39;x 3). </li><li style=";text-align:right;direction:rtl"> למזוג supernatant ולהוסיף מים טהורים עבור resuspension של כדורי ננו-חלקיקים מזהב. </li></ol></li></ol> 4. Radiolabeling של ננו-חלקיקי זהב DBCO-שונה-קבוצה דרך התגובה לחץ חופשי הנחושת <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> סינתזה של 125 ננו-חלקיקי זהב שכותרתו לי באמצעות 125 I-שכותרתו יזיד (1) <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> כן פתרון מרוכז של חלקיקי זהב DBCO-שונה-קבוצה באמצעות צנטריפוגה (11,400 XG, 15 דק &#39;), ולהתאים את הריכוז של חלקיקי זהב עד 2 מיקרומטר. </li><li style=";text-align:right;direction:rtl"> להוסיף 4.1 MBq של אזיד 125 שכותרתו אני (1) ב DMSO (5 μl) כדי השעיה של חלקיקי זהב (4) (2 מיקרומטר, 50 μl). </li><li style=";text-align:right;direction:rtl"> דגירת תערובת התגובה וכתוצאה מכך ב -40 מעלות צלזיוס במשך 60 דקות. </li><li style=";text-align:right;direction:rtl"> משיכת דוגמא מתמיסה בסיסית (0.2 μl) מהמוצר הגולמי ולהחיל אותו על שיתוף סיליקהכרומטוגרפיה בשכבה דקה ated (TLC) צלחת. </li><li style=";text-align:right;direction:rtl"> לפתח את צלחת TLC באמצעות אתיל יצטט כשלב נייד. </li><li style=";text-align:right;direction:rtl"> מניחים את צלחת TLC על סורק רדיו-TLC ולהפעיל את הסורק כדי לפקח על התגובה radiolabeling (איור 3) על פי פרוטוקול של היצרן. </li></ol></li><li style=";text-align:right;direction:rtl"> טיהור של המוצר הגולמי <ol style=";text-align:right;direction:rtl"><li style=";text-align:right;direction:rtl"> לטהר את תערובת התגובה המכיל את חלקיקי זהב 125 שכותרתו לי (4) על ידי צנטריפוגה (11,400 XG, 15 דק &#39;). </li><li style=";text-align:right;direction:rtl"> למזוג supernatant ולהוסיף מים טהורים עבור resuspension של כדורי ננו-חלקיקים מזהב. </li><li style=";text-align:right;direction:rtl"> משיכת דוגמא מתמיסה בסיסית (0.2 μl) מהמוצר המטוהר ולהחיל אותו על צלחת TLC מצופה סיליקה. </li><li style=";text-align:right;direction:rtl"> לפתח את צלחת TLC באמצעות אתיל יצטט כשלב הנייד. </li><li style=";text-align:right;direction:rtl"> מניחים את צלחת TLC על סורק רדיו-TLC ולהפעיל את הסורק כדי לקבוע את התשואה radiochemical וטוהר radiochemical של 125 gol שכותרתו ליחלקיקי ד (4) (איור 3) על פי הפרוטוקול של היצרן. </li></ol></li></ol>

<ol>
	<li>Jewett, J. C., Bertozzi, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cu-free+Click+Cycloaddition+Reactions+in+Chemical+Biology.">Cu-free Click Cycloaddition Reactions in Chemical Biology.</a> Chem. Soc. Rev. 39, 1272-1279 (2010).</li><li>Debets, M. F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioconjugation+with+Strained+Alkenes+and+Alkyne.">Bioconjugation with Strained Alkenes and Alkyne.</a> Acc. Chem. Res. 44, 805-815 (2011).</li><li>Sletten, E. M., Bertozzi, C. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=From+Mechanism+to+Mouse:+A+Tale+of+Two+Bioorthogonal+Reactions.">From Mechanism to Mouse: A Tale of Two Bioorthogonal Reactions.</a> Acc. Chem. Res. 44, 666-676 (2011).</li><li>Koo, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioorthogonal+Cu-Free+Click+Chemistry+in+vivo+for+Tumor-Targeted+Delivery+of+Nanoparticles.">Bioorthogonal Cu-Free Click Chemistry in vivo for Tumor-Targeted Delivery of Nanoparticles.</a> Angew. Chem. Int. Ed. 51, 11836-11840 (2012).</li><li>Chang, P. V., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper-Free+Click+Chemistry+in+Living+Animals.">Copper-Free Click Chemistry in Living Animals.</a> Proc. Natl. Acad. Sci. U. S. A. 107, 1821-1826 (2010).</li><li>Bostic, H. E., Smith, M. D., Poloukhtine, A. A., Popik, V. V., Best, M. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+Labeling+and+Immobilization+via+copper-free+Click+Chemistry.">Membrane Labeling and Immobilization via copper-free Click Chemistry.</a> Chem. Commun. 48, 1431-1433 (2012).</li><li>Someya, T., Ando, A., Kimoto, M., Hirao, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Site-Specific+Labeling+of+RNA+by+Combining+Genetic+Alphabet+Expansion+Transcription+and+Copper-Free+Click+Chemistry.">Site-Specific Labeling of RNA by Combining Genetic Alphabet Expansion Transcription and Copper-Free Click Chemistry.</a> Nucl. Acids Res. 43, 6665-6676 (2015).</li><li>Lee, S. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mesoporous+Silica+Nanoparticle+Pretargeting+for+PET+Imaging+Based+on+a+Rapid+Bioorthogonal+Reaction+in+a+Living+Body.">Mesoporous Silica Nanoparticle Pretargeting for PET Imaging Based on a Rapid Bioorthogonal Reaction in a Living Body.</a> Angew. Chem. Int. Ed. 52, 10549-10552 (2013).</li><li>Sachin, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=F-18+Labeling+Protocol+of+Peptides+Based+on+Chemically+Orthogonal+Strain-Promoted+Cycloaddition+under+Physiologically+Friendly+Reaction+Conditions.">F-18 Labeling Protocol of Peptides Based on Chemically Orthogonal Strain-Promoted Cycloaddition under Physiologically Friendly Reaction Conditions.</a> Bioconjugate Chem. 23, 1680-1686 (2012).</li><li>Evans, H. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper-Free+Click+-+A+Promising+Tool+for+Pre-targeted+PET+Imaging.">Copper-Free Click - A Promising Tool for Pre-targeted PET Imaging.</a> Chem. Commun. 48, 991-993 (2012).</li><li>Campbell-Verduyn, L. S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strain-Promoted+Copper-Free+&amp;#34;Click&amp;#34;+Chemistry+for+18F+Radiolabeling+of+Bombesin.">Strain-Promoted Copper-Free &amp;#34;Click&amp;#34; Chemistry for 18F Radiolabeling of Bombesin.</a> Angew. Chem. Int. Ed. 50, 11117-11120 (2011).</li><li>Arumugam, S., Chin, J., Schirrmacher, R., Popik, V. V., Kostikov, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=18F]Azadibenzocyclooctyne+([18F]ADIBO):+A+Biocompatible+Radioactive+Labeling+Synthon+for+Peptides+using+Catalyst+Free+[3+2]+Cycloaddition.">18F]Azadibenzocyclooctyne ([18F]ADIBO): A Biocompatible Radioactive Labeling Synthon for Peptides using Catalyst Free [3+2] Cycloaddition.</a> Bioorg. Med. Chem. Lett. 21, 6987-6991 (2011).</li><li>Bouvet, V., Wuest, M., Wuest, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Copper-Free+Click+Chemistry+with+the+Short-Lived+Positron+Emitter+Fluorine-18.">Copper-Free Click Chemistry with the Short-Lived Positron Emitter Fluorine-18.</a> Org. Biomol. Chem. 9, 7393-7399 (2011).</li><li>Satpati, D., Bauer, N., Hausner, S. H., Sutcliffe, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+of+[64Cu]DOTA-ADIBON3-Ala-PEG28-A20FMDV2+via+Copper-Free+Click+Chemistry+for+PET+Imaging+of+Integrin+&#945;v&#946;6.">Synthesis of [64Cu]DOTA-ADIBON3-Ala-PEG28-A20FMDV2 via Copper-Free Click Chemistry for PET Imaging of Integrin &#945;v&#946;6.</a> J. Radioanal. Nucl. Chem. 302, 765-771 (2014).</li><li>Lee, D. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Facile+Method+To+Radiolabel+Glycol+Chitosan+Nanoparticles+with+64Cu+via+Copper-Free+Click+Chemistry+for+MicroPET+Imaging.">Facile Method To Radiolabel Glycol Chitosan Nanoparticles with 64Cu via Copper-Free Click Chemistry for MicroPET Imaging.</a> Mol. Pharmaceutics. 10, 2190-2198 (2013).</li><li>Zeng, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=64Cu+Core-Labeled+Nanoparticles+with+High+Specific+Activity+via+Metal-Free+Click+Chemistry.">64Cu Core-Labeled Nanoparticles with High Specific Activity via Metal-Free Click Chemistry.</a> ACS Nano. 6, 5209-5219 (2012).</li><li>Jeon, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiosynthesis+and+in+vivo+Evaluation+of+[125I]2-4(iodophenethyl)-2-Methylmalonic+Acid+as+a+Potential+Radiotracer+for+Detection+of+Apoptosis.">Radiosynthesis and in vivo Evaluation of [125I]2-4(iodophenethyl)-2-Methylmalonic Acid as a Potential Radiotracer for Detection of Apoptosis.</a> J. Radioanal. Nucl. Chem. 308, 23-29 (2016).</li><li>Adam, M. J., Wilbur, D. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiohalogens+for+Imaging+and+Therapy.">Radiohalogens for Imaging and Therapy.</a> Chem. Soc. Rev. 34, 153-163 (2005).</li><li>Jeon, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Radiosynthesis+of+123I-Labeld+Hesperetin+for+Biodistribution+Study+of+Orally+Administered+Hesperetin.">Radiosynthesis of 123I-Labeld Hesperetin for Biodistribution Study of Orally Administered Hesperetin.</a> J. Radioanal. Nucl. Chem. 306, 437-443 (2015).</li><li>Kil, K. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+[123I]IPEB+and+[123I]IMPEB+as+SPECT+Radioligands+for+Metabotropic+Glutamate+Receptor+Subtype.">Development of [123I]IPEB and [123I]IMPEB as SPECT Radioligands for Metabotropic Glutamate Receptor Subtype.</a> ACS Med. Chem. Lett. 5, 652-656 (2014).</li><li>Chen, M. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Utility+of+I-123+Pretherapy+Scan+in+I-131+Radioiodine+Therapy+for+Thyroid+Cancer.">The Utility of I-123 Pretherapy Scan in I-131 Radioiodine Therapy for Thyroid Cancer.</a> Thyroid. 22, 304-309 (2012).</li><li>Jeon, J., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Efficient+Method+for+Iodine+Radioisotope+Labeling+of+Cyclooctyne-Containing+Molecules+using+Strain-Promoted+Copper-Free+Click+Reaction.">Efficient Method for Iodine Radioisotope Labeling of Cyclooctyne-Containing Molecules using Strain-Promoted Copper-Free Click Reaction.</a> Bioorg. Med. Chem. 23, 3303-3308 (2015).</li><li>Choi, M. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Synthesis+and+Evaluation+of+an+125I-Labeled+Azide+Prosthetic+Group+for+Efficient+and+Bioorthogonal+Radiolabeling+of+Cyclooctyne-Group+Containing+Molecules+using+Copper-Free+Click+Reaction.">Synthesis and Evaluation of an 125I-Labeled Azide Prosthetic Group for Efficient and Bioorthogonal Radiolabeling of Cyclooctyne-Group Containing Molecules using Copper-Free Click Reaction.</a> Bioorg. Med. Chem. Lett. 26, 875-878 (2016).</li><li>Kim, Y. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+Targeting+and+Imaging+Using+Cyclic+RGD-PEGylated+Gold+Nanoparticle+Probes+with+Directly+Conjugated+Iodine-125.">Tumor Targeting and Imaging Using Cyclic RGD-PEGylated Gold Nanoparticle Probes with Directly Conjugated Iodine-125.</a> Small. 7, 2052-2060 (2011).</li></ol>

The authors have nothing to disclose.

באופן כללי, התשואה radiochemical הנצפה של אזיד מטוהרים 125 שכותרתו-I (1) היה 75 ± 10% (n = 8). Radiolabeling הושג עם 50-150 MBq של רדיואקטיביות, ותוצאות radiochemical הן די עקביות. אם [125 I] Nai (t 1/2 = 59.4 ד) כי עבר התפרקות רדיואקטיבית במשך יותר מחודש שמשה תגובת radioiodination, תשואת ra...

The strain-promoted copper-free click reaction between azides and cyclooctynes has been extensively applied to the efficient bioorthogonal labeling of a wide range of biomolecules, nanomaterials, and living subjects1-7. Due to the excellent site-specificity and rapid reaction rate of this conjugation reaction, it has also been used to synthesize radiolabeled tracers. A few 18F-labeled azide or DBCO prosthetic groups have been prepared for in vitro labeling of various cancers targeting peptides and antibodies, as well as for in vivo pre-targeted imaging of tumors8-13. In addition to these examples, the same conjugation reaction was applied to the metal-radioisotope-labeling of nanomaterials for positron emission tomography (PET) imaging studies14-16.
For several decades, radioactive iodines have been used for biomedical research and clinical trials through PET imaging (124I), single-photon emission computed tomography (SPECT) imaging (123I, 125I), and thyroid cancer treatment (131I)17-21. Therefore, an efficient method for radioactive iodine labeling is fundamentally important for various investigations, including molecular imaging studies, analysis of organ distribution of biomolecules, biomarker identification, and drug development. A copper-free click reaction strategy could be used in radioactive iodine labeling. However, this application has not been investigated as extensively as 18F-labeled biomolecules22-23. Here, we will provide a step-by-step protocol for the synthesis of an 125I-labeled azide for radiolabeling of DBCO-group-derived molecules. The procedures in the present report will include radioiodination of the stannylated precursor, purification steps with HPLC, and solid phase extraction. We also demonstrate efficient radiolabeling of DBCO-group-modified 13-nm-sized gold nanoparticles using the 125I-labeled azide. The detailed protocol in this report will help synthetic chemists understand a new radiolabeling methodology for the synthesis of radiolabeled products.

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		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Chloramine T trihydrate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">402869</td>
			<td style="width:167px;"></td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:255px;">[125I]NaI in aq. NaOH</td>
			<td style="width:101px;">Perkin-Elmer</td>
			<td style="width:160px;">NEZ033A010MC</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Sodium metabisulfite&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">S9000</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Formic acid</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">251364</td>
			<td></td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;">Sep-Pak tC18 plus cartridge</td>
			<td style="width:101px;">Waters</td>
			<td>WAT036800</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Dimethyl sulfoxide&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">D2650</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Acetone</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">650501</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Ethanol</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">459844</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Gold(III) chloride trihydrate</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">520918</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Tween 20&#160;</td>
			<td style="width:101px;">Sigma</td>
			<td style="width:160px;">P1379</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">DBCO PEG SH (MW 5000)</td>
			<td style="width:101px;">NANOCS</td>
			<td style="width:160px;">PG2-DBTH-5k</td>
			<td></td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;width:255px;">TLC silica gel 60 F254</td>
			<td style="width:101px;">Merck</td>
			<td></td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Analytical HPLC</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">1290 Infinity</td>
			<td>Model number</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Preparative HPLC</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">1260 Infinity</td>
			<td>Model number</td>
		</tr>
		<tr height="25">
			<td height="25" style="height:25px;">Analytical C18 reverse-phase column</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">Zorbax Eclipse XDB-C18</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Preparative C18 reverse-phase column</td>
			<td style="width:101px;">Agilent</td>
			<td style="width:160px;">PrepHT XDB-C18</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:255px;">Radio TLC scanner</td>
			<td style="width:101px;">Bioscan</td>
			<td style="width:160px;">AR-2000</td>
			<td>Model number</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:255px;">Radioisotope dose calibrator</td>
			<td style="width:101px;">Capintec, Inc</td>
			<td style="width:160px;">CRC -25R dose calibrator</td>
			<td>Model number</td>
		</tr>
	</tbody>
</table>

an optimized protocol for the efficient radiolabeling of gold nanoparticles by using a 125i-labeled azide prosthetic group

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient radiolabeling of DBCO-group-functionalized gold nanoparticles using a copper-free click reaction. Radioiodination of the stannylated precursor (2) was carried out by using [125I]NaI and chloramine T as an oxidant at room temperature for 15 min. After HPLC purification of the crude product, the purified 125I-labeled azide (1) was obtained with high radiochemical yield (75 &plusmn; 10%, n = 8) and excellent radiochemical purity (&gt;99%). For the synthesis of radiolabeled 13-nm-sized gold nanoparticles, the DBCO-functionalized gold nanoparticles (3) were prepared by using a thiolated polyethylene glycol polymer. A copper-free click reaction between 1 and 3 gave the 125I-labeled gold nanoparticles (4) with more than 95% of radiochemical yield as determined by radio-thin-layer chromatography (radio-TLC). These results clearly indicate that the present radiolabeling method using a strain-promoted copper-free click reaction will be useful for the efficient and convenient radiolabeling of DBCO-group-containing nanomaterials.

תגובת radioiodination של מבשר stannylated (2) בוצעה באמצעות 150 MBq של [125 I] נאה, חומצה אצטית, ו chloramine T בטמפרטורת חדר למשך 15 דקות כדי לספק את המוצר רדיואקטיבי (1). לאחר HPLC טיהור preparative של התערובת הגולמית, המוצר הרצוי הושג עם 75 ± 10% (n = 8) של תשואת radioche...

Watch this Scientific Journal Video about An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group at JoVE.com

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

נוהל מפורט לסינתזה של אזיד 125 I-שכותרתו ואת radiolabeling של dibenzocyclooctyne (DBCO) -group מצומדות, 13 ננומטר בגודל ננו-חלקיקי זהב באמצעות תגובת לחץ ללא נחושת מתואר.

פרוטוקול אופטימיזציה עבור Radiolabeling היעיל של חלקיקי זהב על ידי שימוש<sup&gt; 125</sup&gt; שכותרתו לי אזיד תותבת קבוצה

Here, we demonstrate a detailed protocol for the radiosynthesis of a 125I-labeled azide prosthetic group and its application to the efficient ...

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Chemistry

An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

8.1K Views.  Korea Atomic Energy Research Institute.  נוהל מפורט לסינתזה של אזיד 125 I-שכותרתו ואת radiolabeling של dibenzocyclooctyne (DBCO) -group מצומדות, 13 ננומטר בגודל ננו-חלקיקי זהב באמצעות תגובת לחץ ללא נחושת מתואר. 

Video: An Optimized Protocol for the Efficient Radiolabeling of Gold Nanoparticles by Using a 125I-labeled Azide Prosthetic Group

Stabilizing Hepatocellular Phenotype Using Optimized Synthetic Surfaces

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing and maintaining primary and pluripotent stem cell derived hepatocytes. While biological matrices are useful, they permit short term culture of hepatocytes, limiting their widespread application. We have attempted to overcome the limitations using a synthetic polymer coating. Polymers represent one of the broadest classes of biomaterials and possess a wide range of mechanical, physical and chemical properties, which can be fine-tuned for purpose. Importantly, such materials can be scaled to quality assured standards and display batch-to-batch consistency. This is essential if cells are to be expanded for high through-put screening in the pharmaceutical testing industry or for cellular based therapy. Polyurethanes (PUs) are one group of materials that have shown promise in cell culture. Our recent progress in optimizing a polyurethane coated surface, for long-term culture of human hepatocytes displaying stable phenotype, is presented and discussed.

This article will focus on developing polymer coated surfaces for long-term, stable culture of stem cell derived human hepatocytes.

ייצוב Hepatocellular הפנוטיפ באמצעות משטחים סינטטיים אופטימליים

Currently, one of the major limitations in cell biology is maintaining differentiated cell phenotype. Biological matrices are commonly used for culturing ...

A Simple Method for the Size Controlled Synthesis of Stable Oligomeric Clusters of Gold Nanoparticles under Ambient Conditions

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like oligomeric clusters of these yellow nanoparticles of narrow size distribution are synthesized under ambient conditions via two methods. The delay-time method controls the number of subunits in the oligoclusters by varying the time between the addition of HAuCl4 to alkaline solution and the subsequent addition of reducing agent, NaSCN. The yellow oligoclusters produced range in size from ~3 to ~25 nm. This size range can be further extended by an add-on method utilizing hydroxylated gold chloride (Na+[Au(OH4-x)Clx]-) to auto-catalytically increase the number of subunits in the as-synthesized oligocluster nanoparticles, providing a total range of 3 nm to 70 nm. The crude oligocluster preparations display narrow size distributions and do not require further fractionation for most purposes. The oligoclusters formed can be concentrated &#62;300 fold without aggregation and the crude reaction mixtures remain stable for weeks without further processing. Because these oligomeric clusters can be concentrated before derivatization they allow expensive derivatizing agents to be used economically. In addition, we present two models by which predictions of particle size can be made with great accuracy.

We describe a simple method for producing highly stable oligomeric clusters of gold nanoparticles via the reduction of chloroauric acid (HAuCl4) with sodium thiocyanate (NaSCN). The oligoclusters have a narrow size distribution and can be produced with a wide range of sizes and surface coats.

שיטה פשוטה עבור סינתזת מבוקרת הגודל של אשכולות Oligomeric היציבים של זהב חלקיקים בתנאי סביבה

Reducing dilute aqueous HAuCl4 with sodium thiocyanate (NaSCN) under alkaline conditions produces 2 to 3 nm diameter nanoparticles. Stable grape-like ...

A Protocol for the Production of Gliadin-cyanoacrylate Nanoparticles for Hydrophilic Coating

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray coating. These nanoparticles are produced by the polymerization reaction of alkyl cyanoacrylate on the surface of cereal protein (gliadin) molecules. Alkyl cyanoacrylate is a monomer that instantly polymerizes at RT when it is applied to the surface of materials. Its polymerization reaction is initiated by the trace amounts of weakly basic or nucleophilic species on the surface, including moisture. Once polymerized, the polymerized alkyl cyanoacrylates show a strong affinity with the object materials because nitrile groups are in the backbone of poly (alkyl cyanoacrylate). Proteins also work as initiator for this polymerization because they contain amine groups that can initiate the polymerization of cyanoacrylate. If aggregated protein is used as an initiator, protein aggregate is surrounded by the hydrophobic poly(alkyl cyanoacrylate) chains after the polymerization reaction of alkyl cyanoacrylate. By controlling the experimental condition, particles in the nanometer range are produced. The produced nanoparticles readily adsorb to the surface of most materials including glass, metals, plastics, wood, leather, and fabrics. When the surface of a material is sprayed with the produced nanoparticle suspension and rinsed with water, the micellar structure of nanoparticle changes its conformation, and the hydrophilic proteins are exposed to the air. As a result, the nanoparticle-coated surface changes to hydrophilic.

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic. The produced nanoparticle is an assembly of gliadin-cyanoacrylate diblock copolymers. Spray coating with the produced nanoparticle changes the surface of target material to a hydrophilic surface.

פרוטוקול עבור ההפקה של חלקיקי גליאדין-cyanoacrylate לציפוי הידרופיליות

This article presents a protocol for the production of protein-based nanoparticles that changes the hydrophobic surface to hydrophilic by a simple spray ...

A Simple and Efficient Protocol for the Catalytic Insertion Polymerization of Functional Norbornenes

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic nature of the monomer is conserved. The resulting polymer, polynorbornene, has a very high glass transition temperature, Tg, and interesting optical and electrical properties. However, the polymerization of functional norbornenes by this mechanism is complicated by the fact that the endo substituted norbornene monomer has, in general, a very low reactivity. Furthermore, the separation of the endo substituted monomer from the exo monomer is a tedious task. Here, we present a simple protocol for the polymerization of substituted norbornenes (endo:exo ca. 80:20) bearing either a carboxylic acid or a pendant double bond. The process does not require that both isomers be separated, and proceeds with low catalyst loadings (0.01 to 0.02 mol%). The polymer bearing pendant double bonds can be further transformed in high yield, to afford a polymer bearing pendant epoxy groups. These simple procedures can be applied to prepare polynorbornenes with a variety of functional groups, such as esters, alcohols, imides, double bonds, carboxylic acids, bromo-alkyls, aldehydes and anhydrides.

We describe the catalytic insertion polymerization of 5-norbornene-2-carboxylic acid and 5-vinyl-2-norbornene to form functional polymers with a very high glass transition temperature.

פרוטוקול פשוט ויעיל עבור פילמור קטליטי קלטי Norbornenes פונקציונלית

Norbornene can be polymerized by a variety of mechanisms, including insertion polymerization whereby the double bond is polymerized and the bicyclic ...

A Protocol for Safe Lithiation Reactions Using Organolithium Reagents

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents warrants proper technique, thorough training, and proper personal protective equipment. To aid in the training of researchers using organolithium reagents, a thorough, step-by-step protocol for the safe and effective use of tert-butyllithium on an inert gas line or within a glovebox is described. As a model reaction, preparation of lithium tert-butyl amide by the reaction of tert-butyl amine with one equivalent of tert-butyl lithium is presented.

The safe and proper use of organolithium reagents is described.

פרוטוקול תגובות בטוח Lithiation באמצעות ריאגנטים Organolithium

Organolithium reagents are powerful tools in the synthetic chemist&#39;s toolbox. However, the extreme pyrophoric nature of the most reactive reagents ...

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity and convenient colorimetric readout. However, high densities of nanoparticles are typically needed for detection. The available synthesis-based enhancement protocols are either limited to gold and silver nanoparticles or rely on precise enzymatic control and optimization. Here, we present a protocol to enhance the colorimetric readout of gold, silver, silica, and iron oxide nanoprobes. It was observed that the colorimetric signal can be improved by up to a 10000-fold factor. The basis for such signal enhancement strategies is the chemical reduction of Au3+ to Au0. There are several chemical reactions that enable the reduction of Au3+ to Au0. In the protocol, Good&#39;s buffers and H2O2 are used and it is possible to favor the deposition of Au0 onto the surface of existing nanoprobes, in detriment of the formation of new gold nanoparticles. The protocol consists of the incubation of the microarray with a solution consisting of chloroauric acid and H2O2 in 2-(N-morpholino)ethanesulfonic acid pH 6 buffer following the nanoprobe-based detection assay. The enhancement solution can be applied to paper and glass-based sensors. Moreover, it can be used in commercially available immunoassays as demonstrated by the application of the method to a commercial allergen microarray. The signal development requires less than 5 min of incubation with the enhancement solution and the readout can be assessed by naked eye or low-end image acquisition devices such as a table-top scanner or a digital camera.

Rapid Nanoprobe Signal Enhancement by In Situ Gold Nanoparticle Synthesis

In this work, a protocol for signal enhancement of nanoprobe-based biosensing is presented. The protocol is based on the reduction of chloroauric acid onto the surface of existing nanoprobes that consist of gold, silver, silica or iron-oxide nanoparticles.

שיפור האות Nanoprobe מהירה על ידי באתרו בתוך Nanoparticle זהב סינתזה

The use of nanoprobes such as gold, silver, silica or iron-oxide nanoparticles as detection reagents in bioanalytical assays can enable high sensitivity ...

The MPLEx Protocol for Multi-omic Analyses of Soil Samples

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and characterize microbial communities in environmental and biological systems. These measurements are even enabling enhanced analyses of complex soil microbial communities, which are the most complex microbial systems known to date. Multi-omic analyses, however, do have sample preparation challenges, since separate extractions are typically needed for each omic study, thereby greatly amplifying the preparation time and amount of sample required. To address this limitation, a 3-in-1 method for the simultaneous extraction of metabolites, proteins, and lipids (MPLEx) from the same soil sample was created by adapting a solvent-based approach. This MPLEx protocol has proven to be both simple and robust for many sample types, even when utilized for limited quantities of complex soil samples. The MPLEx method also greatly enabled the rapid multi-omic measurements needed to gain a better understanding of the members of each microbial community, while evaluating the changes taking place upon biological and environmental perturbations.

A protocol is presented for simultaneously extracting metabolites, proteins, and lipids from a single soil sample, allowing reduced sample preparation times and enabling multi-omic mass spectrometry analyses of samples with limited quantities.

פרוטוקול MPLEx עבור ניתוחים מולטי-omic של דגימות אדמה

Mass spectrometry (MS)-based integrated metaproteomic, metabolomic, and lipidomic (multi-omic) studies are transforming our ability to understand and ...

Luminophore Formation in Various Conformations of Bovine Serum Albumin by Binding of Gold(III)

The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (&#955;em = 640 nm) of BSA-Au(III) complexes. The method adjusts the pH to show that the emergence of the red fluorescence is correlated with the pH-induced equilibrium transitions of the BSA conformations. Red fluorescent BSA-Au(III) complexes can only be formed with an adjustment of pH at or above 9.7, which corresponds to the &#34;A-form&#34; conformation of BSA. The protocol to adjust the BSA to Au molar ratio and to monitor the time-course of the process of Au(III) binding is described. The minimum number of Au(III) per BSA, to produce the red fluorescence, is less than seven. We describe the protocol in steps to illustrate the presence of multiple Au(III) binding sites in BSA. First, by adding copper (Cu(II)) or nickel (Ni(II)) cations followed by Au(III), this method reveals a binding site for Au(III) that is not the red fluorophore. Second, by modifying BSA by thiol capping agents, another nonfluorophore-forming Au(III) binding site is revealed. Third, changing the BSA conformation by cleaving and capping of the disulfide bonds, the possible Au(III) binding site(s) are illustrated. The protocol described, to control the BSA conformations and Au(III) binding, can be generally applied to study the interactions of other proteins and metal cations.

Luminophore Formation in Various Conformations of Bovine Serum Albumin by Binding of GoldIII

The protocols for studying the binding of gold cations (Au(III)) to various conformations of bovine serum albumin (BSA) as well as for characterizing the conformational dependent unique BSA-Au fluorescence are presented.

Luminophore-צורה הייצורים החיים השונים של אלבומין שור על-ידי איגוד של Gold(III)

The purpose of the presented protocols is to study the process of Au(III) binding to BSA, yielding conformation change-induced red fluorescence (&#955;em ...

Synthesis and Characterization of Amphiphilic Gold Nanoparticles

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of their interactions with cell membranes, lipid bilayers, and viruses. The hydrophilic ligands make these particles colloidally stable in aqueous solutions and the combination with hydrophobic ligands creates an amphiphilic particle that can be loaded with hydrophobic drugs, fuse with the lipid membranes, and resist nonspecific protein adsorption. Many of these properties depend on nanoparticle size and the composition of the ligand shell. It is, therefore, crucial to have a reproducible synthetic method and reliable characterization techniques that allow the determination of nanoparticle properties and the ligand shell composition. Here, a one-phase chemical reduction, followed by a thorough purification to synthesize these nanoparticles with diameters below 5 nm, is presented. The ratio between the two ligands on the surface of the nanoparticle can be tuned through their stoichiometric ratio used during synthesis. We demonstrate how various routine techniques, such as transmission electron microscopy (TEM), nuclear magnetic resonance (NMR), thermogravimetric analysis (TGA), and ultraviolet-visible (UV-Vis) spectrometry, are combined to comprehensively characterize the physicochemical parameters of the nanoparticles.

Amphiphilic gold nanoparticles can be used in many biological applications. A protocol to synthesize gold nanoparticles coated by a binary mixture of ligands and a detailed characterization of these particles is presented.

סינתזה ואפיון חלקיקי זהב אמפיפילי

Gold nanoparticles covered with a mixture of 1-octanethiol (OT) and 11-mercapto-1-undecane sulfonic acid (MUS) have been extensively studied because of ...

Synthesis of Aptamer-PEI-g-PEG Modified Gold Nanoparticles Loaded with Doxorubicin for Targeted Drug Delivery

Due to drug resistance and toxicity in healthy cells, use of doxorubicin (DOX) has been limited in clinical cancer therapy. This protocol describes the designing of poly(ethylenimine) grafted with polyethylene glycol (PEI-g-PEG) copolymer functionalized gold nanoparticles (AuNPs) with loaded aptamer (AS1411) and DOX through amide reactions. AS1411 is specifically bonded with targeted nucleolin receptors on cancer cells so that DOX targets cancer cells instead of healthy cells. First, PEG is carboxylated, then grafted to branched PEI to obtain a PEI-g-PEG copolymer, which is confirmed by 1H NMR analysis. Next, PEI-g-PEG copolymer coated gold nanoparticles (PEI-g-PEG@AuNPs) are synthesized, and DOX and AS1411 are covalently bonded to AuNPs gradually via amide reactions. The diameter of the prepared AS1411-g-DOX-g-PEI-g-PEG@AuNPs is ~39.9 nm, with a zeta potential of -29.3 mV, indicating that the nanoparticles are stable in water and cell medium. Cell cytotoxicity assays show that the newly designed DOX loaded AuNPs are able to kill cancer cells (A549). This synthesis demonstrates the delicate arrangement of PEI-g-PEG copolymers, aptamers, and DOX on AuNPs that are achieved by sequential amide reactions. Such aptamer-PEI-g-PEG functionalized AuNPs provide a promising platform for targeted drug delivery in cancer therapy.

In this protocol, doxorubicin-loaded AS1411-g-PEI-g-PEG modified gold nanoparticles are synthesized via three-step amide reactions. Then, doxorubicin is loaded and delivered to target cancer cells for cancer therapy.

סינתזה של Aptamer-PEI-g-PEG שונה חלקיקי זהב טעון עם וקסורוביצ'ין עבור משלוח תרופות ממוקדות

Due to drug resistance and toxicity in healthy cells, use of doxorubicin (DOX) has been limited in clinical cancer therapy. This protocol describes the ...