دورية أكاديمية
أعرض في EDS

You get what you test for: The killing effect of phage lysins is highly dependent on buffer tonicity and ionic strength.

التفاصيل البيبلوغرافية
العنوان: You get what you test for: The killing effect of phage lysins is highly dependent on buffer tonicity and ionic strength.
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.)
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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
الوصف
تدمد:1751-7915
DOI:10.1111/1751-7915.14513