5a700db578551d14cb9a92e21df19966 pharmaceutics_a2020v12n11p1004.pdf 89ca45b3e9c7b78c96093a4aa33fa36e8fa3de22 pharmaceutics_a2020v12n11p1004.pdf c5a8df4aa7ce6461f9d66237a9050599031ebd493244f9313a5de8c9f65db5ec pharmaceutics_a2020v12n11p1004.pdf Title: Fluorescent Dye Labeling Changes the Biodistribution of Tumor-Targeted Nanoparticles Subject: Fluorescent dye labeling is a common strategy to analyze the fate of administered nanoparticles in living organisms. However, to which extent the labeling processes can alter the original nanoparticle biodistribution has been so far neglected. In this work, two widely used fluorescent dye molecules, namely, ATTO488 (ATTO) and Sulfo-Cy5 (S-Cy5), have been covalently attached to a well-characterized CXCR4-targeted self-assembling protein nanoparticle (known as T22-GFP-H6). The biodistribution of labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles has been then compared to that of the non-labeled nanoparticle in different CXCR4+ tumor mouse models. We observed that while parental T22-GFP-H6 nanoparticles accumulated mostly and specifically in CXCR4+ tumor cells, labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles showed a dramatic change in the biodistribution pattern, accumulating in non-target organs such as liver or kidney while reducing tumor targeting capacity. Therefore, the use of such labeling molecules should be avoided in target and non-target tissue uptake studies during the design and development of targeted nanoscale drug delivery systems, since their effect over the fate of the nanomaterial can lead to considerable miss-interpretations of the actual nanoparticle biodistribution. Keywords: protein nanomaterials; functional materials; self-assembling nanoparticles; fluorescent labeling; biodistribution; targeting Author: Patricia Álamo, Victor Pallarès, María Virtudes Céspedes, Aïda Falgàs, Julieta M. Sanchez, Naroa Serna, Laura Sánchez-García, Eric Voltà-Duràn, Gordon A. Morris, Alejandro Sánchez-Chardi, Isolda Casanova, Ramón Mangues, Esther Vazquez, Antonio Villaverde and Ugutz Unzueta Creator: LaTeX with hyperref package Producer: pdfTeX-1.40.18 CreationDate: Mon Oct 26 14:10:24 2020 CET ModDate: Mon Oct 26 14:52:30 2020 CET Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2696920 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- EZRNSZ+Pxsy Type 1 Builtin yes yes no 10 0 HKBJPQ+VnURWPalladioL Type 1 Custom yes yes no 14 0 WKRZDD+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 IJLDYL+URWPalladioL-Bold Type 1 Custom yes yes no 24 0 WQTNKU+URWPalladioL-Roma Type 1 Custom yes yes no 28 0 FATHTC+Rpxr Type 1 Builtin yes yes no 32 0 CCUJRL+VnURWPalladioL-Bold Type 1 Custom yes yes no 36 0 QTLELQ+Pxbsy Type 1 Builtin yes yes no 40 0 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2019-04-17) Date: 2020-11-18 07:09:33 CET RepresentationInformation: pharmaceutics_a2020v12n11p1004.pdf ReportingModule: PDF-hul, Rel. 1.12.1 (2019-04-17) LastModified: 2020-11-17 10:34:45 CET Size: 2696920 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 527 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@13c27452 Item: Title: Materials and Methods Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@262b2c86 Children: Item: Title: Protein Production, Purification and Characterization Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@371a67ec Item: Title: Fluorescent Dye Conjugation Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@5ed828d Item: Title: Morphometric Characterization Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@50d0686 Item: Title: Optical Properties of Nanoparticles and Free Dyes Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@7a3d45bd Item: Title: Selection of Cell Lines for the Generation of In Vitro and In Vivo Models Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1e7c7811 Item: Title: Cell Culture, Protein Internalization and Competition Assays Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77ec78b9 Item: Title: CXCR4 Receptor Binding Affinity Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@1a3869f4 Item: Title: Biodistribution in Subcutaneous DLBCL Mouse Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@a38d7a3 Item: Title: Biodistribution in Subcutaneous Colorectal Cancer Mouse Model Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@77f99a05 Item: Title: Ex Vivo Biodistribution Fluorescent Image Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@63440df3 Item: Title: Statistical Analysis Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@3aeaafa6 Item: Title: Results Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@76a3e297 Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4d3167f4 Item: Title: Conclusions Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@ed9d034 Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6121c9d6 Info: Title: Fluorescent Dye Labeling Changes the Biodistribution of Tumor-Targeted Nanoparticles Author: Patricia Álamo, Victor Pallarès, María Virtudes Céspedes, Aïda Falgàs, Julieta M. Sanchez, Naroa Serna, Laura Sánchez-García, Eric Voltà-Duràn, Gordon A. Morris, Alejandro Sánchez-Chardi, Isolda Casanova, Ramón Mangues, Esther Vazquez, Antonio Villaverde and Ugutz Unzueta Subject: Fluorescent dye labeling is a common strategy to analyze the fate of administered nanoparticles in living organisms. However, to which extent the labeling processes can alter the original nanoparticle biodistribution has been so far neglected. In this work, two widely used fluorescent dye molecules, namely, ATTO488 (ATTO) and Sulfo-Cy5 (S-Cy5), have been covalently attached to a well-characterized CXCR4-targeted self-assembling protein nanoparticle (known as T22-GFP-H6). The biodistribution of labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles has been then compared to that of the non-labeled nanoparticle in different CXCR4+ tumor mouse models. We observed that while parental T22-GFP-H6 nanoparticles accumulated mostly and specifically in CXCR4+ tumor cells, labeled T22-GFP-H6-ATTO and T22-GFP-H6-S-Cy5 nanoparticles showed a dramatic change in the biodistribution pattern, accumulating in non-target organs such as liver or kidney while reducing tumor targeting capacity. Therefore, the use of such labeling molecules should be avoided in target and non-target tissue uptake studies during the design and development of targeted nanoscale drug delivery systems, since their effect over the fate of the nanomaterial can lead to considerable miss-interpretations of the actual nanoparticle biodistribution. 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