دورية أكاديمية
You get what you test for: The killing effect of phage lysins is highly dependent on buffer tonicity and ionic strength.
العنوان: | You get what you test for: The killing effect of phage lysins is highly dependent on buffer tonicity and ionic strength. |
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المؤلفون: | Vázquez R; Department of Biotechnology, Ghent University, Ghent, Belgium.; Centro de Investigación Biomédica en Red de Enfermedades Respiratorias (CIBERES), Madrid, Spain., Gutiérrez D; Department of Biotechnology, Ghent University, Ghent, Belgium., Dezutter Z; Department of Biotechnology, Ghent University, Ghent, Belgium., Criel B; Department of Biotechnology, Ghent University, Ghent, Belgium., de Groote P; Department of Biotechnology, Ghent University, Ghent, Belgium., Briers Y; Department of Biotechnology, Ghent University, Ghent, Belgium. |
المصدر: | Microbial biotechnology [Microb Biotechnol] 2024 Jul; Vol. 17 (7), pp. e14513. |
نوع المنشور: | Journal Article |
اللغة: | English |
بيانات الدورية: | Publisher: Wiley-Blackwell Country of Publication: United States NLM ID: 101316335 Publication Model: Print Cited Medium: Internet ISSN: 1751-7915 (Electronic) Linking ISSN: 17517915 NLM ISO Abbreviation: Microb Biotechnol Subsets: MEDLINE |
أسماء مطبوعة: | Publication: Hoboken, NJ : Wiley-Blackwell Original Publication: Oxford : Blackwell, c2008- |
مواضيع طبية MeSH: | Bacteriophages*/chemistry , Bacteriophages*/physiology , Bacteriophages*/genetics , Acinetobacter baumannii*/drug effects , Acinetobacter baumannii*/virology, Osmolar Concentration ; Microbial Viability/drug effects ; Buffers ; Humans ; Viral Proteins/genetics ; Viral Proteins/metabolism ; Viral Proteins/chemistry ; Endopeptidases/metabolism ; Endopeptidases/chemistry |
مستخلص: | The phage lysin field has done nothing but grow in the last decades. As a result, many different research groups around the world are contributing to the field, often with certain methodological differences that pose a challenge to the interpretation and comparison of results. In this work, we present the case study of three Acinetobacter baumannii-targeting phage lysins (wild-type endolysin LysMK34 plus engineered lysins eLysMK34 and 1D10) plus one lysin with broad activity against Gram-positive bacteria (PlySs2) to provide exemplary evidence on the risks of generalization when using one of the most common lysin evaluation assays: the killing assay with resting cells. To that end, we performed killing assays with the aforementioned lysins using hypo-, iso- and hypertonic buffers plus human serum either as the reaction or the dilution medium in a systematic manner. Our findings stress the perils of creating hypotonic conditions or a hypotonic shock during a killing assay, suggesting that hypotonic buffers should be avoided as a test environment or as diluents before plating to avoid overestimation of the killing effect in the assayed conditions. As a conclusion, we suggest that the nature of both the incubation and the dilution buffers should be always clearly identified when reporting killing activity data, and that for experimental consistency the same incubation buffer should be used as a diluent for posterior serial dilution and plating unless explicitly required by the experimental design. In addition, the most appropriate buffer mimicking the final application must be chosen to obtain relevant results. (© 2024 The Author(s). Microbial Biotechnology published by John Wiley & Sons Ltd.) |
References: | Front Cell Infect Microbiol. 2021 Mar 02;11:637313. (PMID: 33738267) J Biol Chem. 2011 Sep 30;286(39):34391-403. (PMID: 21816821) Antimicrob Agents Chemother. 2013 Jun;57(6):2743-50. (PMID: 23571534) J Virol. 2021 Jun 24;95(14):e0032121. (PMID: 33883227) Appl Environ Microbiol. 2020 Sep 17;86(19):. (PMID: 32709718) Sci Adv. 2020 Jun 03;6(23):eaaz1136. (PMID: 32537492) mBio. 2014 Jul 01;5(4):e01379-14. (PMID: 24987094) J Clin Invest. 2020 Jul 1;130(7):3750-3760. (PMID: 32271718) Curr Opin Biotechnol. 2021 Apr;68:51-59. (PMID: 33126104) Viruses. 2018 May 29;10(6):. (PMID: 29844287) Appl Microbiol Biotechnol. 2011 Apr;90(2):529-39. (PMID: 21264466) Microb Biotechnol. 2024 Jul;17(7):e14513. (PMID: 38962879) Antimicrob Agents Chemother. 2016 May 23;60(6):3480-8. (PMID: 27021321) Front Microbiol. 2014 Oct 16;5:542. (PMID: 25360133) Lancet Infect Dis. 2020 Sep;20(9):e216-e230. (PMID: 32653070) J Med Microbiol. 2013 Oct;62(Pt 10):1506-1516. (PMID: 23813275) Viruses. 2018 Jun 06;10(6):. (PMID: 29882827) Antibiotics (Basel). 2020 Sep 19;9(9):. (PMID: 32961696) J Microbiol. 2022 Aug;60(8):859-866. (PMID: 35614377) Biophys J. 2013 Jun 18;104(12):2733-42. (PMID: 23790382) Front Microbiol. 2016 Sep 07;7:1402. (PMID: 27656173) Virol Sin. 2015 Feb;30(1):45-51. (PMID: 25680444) mBio. 2018 Jan 23;9(1):. (PMID: 29362234) Sci Rep. 2017 Nov 28;7(1):16506. (PMID: 29184097) PLoS One. 2017 Jan 3;12(1):e0169180. (PMID: 28046082) Crit Rev Food Sci Nutr. 2023;63(27):8919-8938. (PMID: 35400249) FEMS Microbiol Rev. 2002 Mar;26(1):49-71. (PMID: 12007642) Clin Infect Dis. 2024 Jun 14;78(6):1473-1481. (PMID: 38297916) Curr Opin Microbiol. 2018 Apr;42:62-70. (PMID: 29125939) Antimicrob Agents Chemother. 2014 Jun;58(6):3073-84. (PMID: 24637688) Clin Infect Dis. 2022 Aug 25;75(2):338-341. (PMID: 34894129) Front Pharmacol. 2024 May 20;15:1385261. (PMID: 38831886) Acta Crystallogr D Struct Biol. 2022 Apr 1;78(Pt 4):435-454. (PMID: 35362467) Appl Environ Microbiol. 2022 Jan 11;88(1):e0151521. (PMID: 34669452) Microb Biotechnol. 2021 Mar;14(2):403-418. (PMID: 32519416) |
معلومات مُعتمدة: | 01P10022 Bijzonder Onderzoeksfonds UGent; 1S38519N Fonds Wetenschappelijk Onderzoek; G066919N Fonds Wetenschappelijk Onderzoek |
المشرفين على المادة: | 0 (Buffers) EC 3.4.99.- (endolysin) 0 (Viral Proteins) EC 3.4.- (Endopeptidases) |
تواريخ الأحداث: | Date Created: 20240704 Date Completed: 20240704 Latest Revision: 20240706 |
رمز التحديث: | 20240706 |
مُعرف محوري في PubMed: | PMC11222872 |
DOI: | 10.1111/1751-7915.14513 |
PMID: | 38962879 |
قاعدة البيانات: | MEDLINE |
تدمد: | 1751-7915 |
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DOI: | 10.1111/1751-7915.14513 |