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A new biosynthetic tracer for the inline measurement of virus retention in membrane processes: part II. Biochemical and physicochemical characterizations of the new tracer
Authors:Soussan L  Guigui C  Alfenore S  Mathe S  Cabassud C
Institution:Université de Toulouse, INSA, LISBP, 135 Avenue de Rangueil, F-31077 Toulouse, France;INRA, UMR792 Ingénierie des Systèmes Biologiques et des Procédés, F-31400 Toulouse, France;CNRS, UMR5504, F-31400 Toulouse, France
Abstract:In a previous work, a reproducible procedure to produce a new biosynthetic tracer was developed. This new tracer is an MS2 bacteriophage with enzymatic probes grafted on its surface, which can induce enzymatic activity of the tracer. In this paper, the biochemical and physicochemical characteristics of this new tracer are determined. A protocol was developed to determine the specific enzymatic activity kcatTRACER of the tracer, which was found to be 2.93 ± 0.78 × 104 min−1 on average. Physicochemical characterizations of this new tracer showed that it is representative of viruses and may thus be used as a virus surrogate to assess the virus retention of membrane systems inline. Notably, the mean diameter and molecular weight of the tracer were found to be respectively 64.1 ± 0.3 nm and 12140 ± 3654 kDa, which are within the size and molecular weight ranges of pathogenic viruses carried by water. The tracer surface was also studied and revealed the considerable porosity of the grafted probe layer, with a mean porosity of 88%, which could explain why the zeta potential of the tracers (−14.34 ± 1.66 mV) was nearly the same as that of the native MS2 phages. Finally, a comparison between filtration of the reference microorganism used for membrane performance assessment (the MS2 phage) and the tracer suspensions showed the same filtration behaviour.
Keywords:Abbreviations: HPLC&ndash  SEC  high performance liquid chromatography&ndash  size exclusion chromatography  HRP  horseradish peroxidase  Ti  the peak or the fraction of tracers at the ith HPLC&ndash  SEC purification step  Si  the peak or the fraction of free probes at the ith HPLC&ndash  SEC purification step  〈 〉  the mean values of the terms concerned
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