a0f7dd23cef42878f97bc83fd9a60af7 pmc_35161907.pdf 8f9f1c73885363b956a032ac54a8bdf716f628fb pmc_35161907.pdf 1655b0df27f6abbf28e92127733d81e9dda8c147bc03cd5576e5a728dad46f0a pmc_35161907.pdf Title: Patterns in Dried Droplets to Detect Unfolded BSA Subject: The morphological analysis of patterns in dried droplets has allowed the generation of efficient techniques for the detection of molecules of medical interest. However, the effectiveness of this method to reveal the coexistence of macromolecules of the same species, but different conformational states, is still unknown. To address this problem, we present an experimental study on pattern formation in dried droplets of bovine serum albumin (BSA), in folded and unfolded conformational states, in saline solution (NaCl). Folded proteins produce a well-defined coffee ring and crystal patterns all over the dry droplet. Depending on the NaCl concentration, the crystals can be small, large, elongated, entangled, or dense. Optical microscopy reveals that the relative concentration of unfolded proteins determines the morphological characteristics of deposits. At a low relative concentration of unfolded proteins (above 2%), small amorphous aggregates emerge in the deposits, while at high concentrations (above 16%), the ``eye-like pattern'', a large aggregate surrounded by a uniform coating, is produced. The radial intensity profile, the mean pixel intensity, and the entropy make it possible to characterize the patterns in dried droplets. We prove that it is possible to achieve 100% accuracy in identifying 4% of unfolded BSA contained in a protein solution. Keywords: patterns; dried droplets; folded proteins; unfolded proteins Author: Yojana J. P. Carreón, Mary Luz Gómez-López, O. Díaz-Hernández, Pamela Vazquez-Vergara, R. E. Moctezuma, José M. Saniger and Jorge González-Gutiérrez Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Mon Feb 7 11:56:15 2022 CET ModDate: Mon Feb 7 12:16:53 2022 CET Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 14 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 85205877 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- EZZFBA+URWPalladioL-Bold Type 1 Custom yes yes no 10 0 PHZRCW+URWPalladioL-Roma Type 1 Custom yes yes no 15 0 OZUIMD+URWPalladioL-Ital Type 1 Custom yes yes no 19 0 MCBCCI+SourceSansRoman-Regular Type 1C Custom yes yes no 27 0 MCBCCM+SourceSansRoman.613wght Type 1C Custom yes yes no 31 0 MCBCCO+SourceSansRoman-Bold Type 1C Custom yes yes no 35 0 RVSEMT+TimesNewRomanPS-BoldItalicMT Type 1C WinAnsi yes yes no 47 0 TJDGTQ+PazoMath-Italic Type 1 Builtin yes yes no 72 0 EIMZQL+PazoMath Type 1 Builtin yes yes no 76 0 CZKGAS+CMR10 Type 1 Builtin yes yes no 80 0 OJYQXC+CMSY10 Type 1 Builtin yes yes no 84 0 OVNEXW+EURM10 Type 1 Builtin yes yes no 88 0 UYVLXV+CMEX10 Type 1 Builtin yes yes no 95 0 RLYTRW+CMMI10 Type 1 Builtin yes yes no 108 0 Jhove (Rel. 1.28.0, 2023-05-18) Date: 2024-05-07 04:29:41 CEST RepresentationInformation: pmc_35161907.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2022-02-27 18:42:00 CET Size: 85205877 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 344 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Protein Solutions Preparation Destination: subsection.2.1 Item: Title: Droplet Evaporation Destination: subsection.2.2 Item: Title: Image Acquisition Destination: subsection.2.3 Item: Title: Image Analysis Destination: subsection.2.4 Item: Title: Results Destination: section.3 Children: Item: Title: Patterns in Dried Droplets of Proteins Destination: subsection.3.1 Item: Title: Accuracy to Identify Unfolded Proteins. Destination: subsection.3.2 Item: Title: Pattern Formation in Dried Droplets of Folded and Unfolded Proteins Destination: subsection.3.3 Item: Title: Discussion Destination: section.4 Item: Title: References Destination: section.4 Info: Title: Patterns in Dried Droplets to Detect Unfolded BSA Author: Yojana J. P. Carreón, Mary Luz Gómez-López, O. Díaz-Hernández, Pamela Vazquez-Vergara, R. E. Moctezuma, José M. Saniger and Jorge González-Gutiérrez Subject: The morphological analysis of patterns in dried droplets has allowed the generation of efficient techniques for the detection of molecules of medical interest. However, the effectiveness of this method to reveal the coexistence of macromolecules of the same species, but different conformational states, is still unknown. To address this problem, we present an experimental study on pattern formation in dried droplets of bovine serum albumin (BSA), in folded and unfolded conformational states, in saline solution (NaCl). Folded proteins produce a well-defined coffee ring and crystal patterns all over the dry droplet. Depending on the NaCl concentration, the crystals can be small, large, elongated, entangled, or dense. Optical microscopy reveals that the relative concentration of unfolded proteins determines the morphological characteristics of deposits. At a low relative concentration of unfolded proteins (above 2%), small amorphous aggregates emerge in the deposits, while at high concentrations (above 16%), the ``eye-like pattern'', a large aggregate surrounded by a uniform coating, is produced. The radial intensity profile, the mean pixel intensity, and the entropy make it possible to characterize the patterns in dried droplets. We prove that it is possible to achieve 100% accuracy in identifying 4% of unfolded BSA contained in a protein solution. 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