Small-angle neutron scattering (SANS) has been used to extend the structural characterization of the MS2 phage by examining its physical characteristics in solution. Specifically, the contrast variation technique was employed to determine the molecular weight of the individual components of the MS2 virion (protein shell and genomic RNA) and the spatial relationship of the genomic RNA to its protein shell. A consequence of this work was to evaluate a novel particle counting instrument, the integrated virus detection system (IVDS) that, in combination with SANS, has the potential to provide rapid quantitative physical characterization of unidentified viruses and phage.

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Public reporting burden for this collection of information is estimated to average 1 hour per response, including the time for reviewing instructions, searching existing data sources, gathering and maintaining the data needed, and completing and reviewing this collection of information. SPONSORING I MONITORING AGENCY NAME(S) AND ADDRESS(ES)10. SPONSORIMONITOR'S ACRONYM(S) SPONSORIMONITOR'S REPORT NUMBER(S) DISTRIBUTION I AVAILABILITY STATEMENTApproved for public release; distribution is unlimited. SUPPLEMENTARY NOTES ABSTRACTThis study demonstrates the characterization of the sample matrix composition of the MS2 virus using ESI-MS and IVDS detection systems. The MS2 samples were grown and purified using various techniques, which showed ESI-MS samples responding differently from IVDS samples. The LC-MS of the specific biomarker of the MS2 bacteriophage from an infected Escherichia coli (E. coli) sample was characterized in the presence of E. coli proteins. The significant impact of the sample matrix was observed upon the identification of MS2 using a database search. Escherichia coli infected with MS2 showed a different score from other uninfected scores. Only purified MS2, using CsCl and analyzed by LS-MS, showed a positive match in the database search. However, the variation in the MS2 sample matrix had no effect on the characterization of MS2.

Small-angle scattering (SAS) techniques, like small-angle X-ray scattering (SAXS) and small-angle neutron scattering (SANS), were used to measure and thus to validate the accuracy of a novel technology for virus sizing and concentration determination. These studies demonstrate the utility of SAS techniques for use in quality assurance measurements and as novel technology for the physical characterization of viruses. Published by Elsevier Ltd.