دورية أكاديمية
Acoustic Focusing of Protein Crystals for In-Line Monitoring and Up-Concentration during Serial Crystallography.
| العنوان: | Acoustic Focusing of Protein Crystals for In-Line Monitoring and Up-Concentration during Serial Crystallography. |
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| المؤلفون: | Hammarström B; Department of Applied Physics, KTH Royal Institute of Technology, S-106 91 Stockholm, Sweden., Lane TJ; Center for Free-Electron Laser Science, Deutsches Elektronen-Synchrotron DESY, Notkestrasse 85, 22607 Hamburg, Germany., Batili H; Department of Applied Physics, KTH Royal Institute of Technology, S-106 91 Stockholm, Sweden., Sierra R; Linac Coherent Light Source, SLAC National Accelerator Laboratory, Menlo Park, California 94025, United States., Wiklund M; Department of Applied Physics, KTH Royal Institute of Technology, S-106 91 Stockholm, Sweden., Sellberg JA; Department of Applied Physics, KTH Royal Institute of Technology, S-106 91 Stockholm, Sweden. |
| المصدر: | Analytical chemistry [Anal Chem] 2022 Sep 20; Vol. 94 (37), pp. 12645-12656. Date of Electronic Publication: 2022 Sep 02. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: American Chemical Society Country of Publication: United States NLM ID: 0370536 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1520-6882 (Electronic) Linking ISSN: 00032700 NLM ISO Abbreviation: Anal Chem Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Washington, American Chemical Society. |
| مواضيع طبية MeSH: | Muramidase* , Synchrotrons*, Acoustics ; Crystallography ; Crystallography, X-Ray ; Proteins/chemistry ; Silicon Dioxide |
| مستخلص: | Serial femtosecond crystallography (SFX) has become one of the standard techniques at X-ray free-electron lasers (XFELs) to obtain high-resolution structural information from microcrystals of proteins. Nevertheless, reliable sample delivery is still often limiting data collection, as microcrystals can clog both field- and flow-focusing nozzles despite in-line filters. In this study, we developed acoustic 2D focusing of protein microcrystals in capillaries that enables real-time online characterization of crystal size and shape in the sample delivery line after the in-line filter. We used a piezoelectric actuator to create a standing wave perpendicular to the crystal flow, which focused lysozyme microcrystals into a single line inside a silica capillary so that they can be imaged using a high-speed camera. We characterized the acoustic contrast factor, focus size, and the coaxial flow lines and developed a splitting union that enables up-concentration to at least a factor of five. The focus size, flow rates, and geometry may enable an upper limit of up-concentration as high as 200 fold. The novel feedback and concentration control could be implemented for serial crystallography at synchrotrons with minor modifications. It will also aid the development of improved sample delivery systems that will increase SFX data collection rates at XFELs, with potential applications to many proteins that can only be purified and crystallized in small amounts. |
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| المشرفين على المادة: | 0 (Proteins) 7631-86-9 (Silicon Dioxide) EC 3.2.1.17 (Muramidase) |
| تواريخ الأحداث: | Date Created: 20220902 Date Completed: 20220922 Latest Revision: 20221017 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC9494305 |
| DOI: | 10.1021/acs.analchem.2c01701 |
| PMID: | 36054318 |
| قاعدة البيانات: | MEDLINE |
Find this issue from the American Chemical Society | تدمد: | 1520-6882 |
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| DOI: | 10.1021/acs.analchem.2c01701 |