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Low Volume in Vitro Diagnostic Proton NMR Spectroscopy of Human Blood Plasma for Lipoprotein and Metabolite Analysis: Application to SARS-CoV-2 Biomarkers #MMPMID33491459
Lodge S; Nitschke P; Loo RL; Kimhofer T; Bong SH; Richards T; Begum S; Spraul M; Schaefer H; Lindon JC; Holmes E; Nicholson JK
J Proteome Res 2021[Feb]; 20 (2): 1415-1423 PMID33491459show ga
The utility of low sample volume in vitro diagnostic (IVDr) proton nuclear magnetic resonance ((1)H NMR) spectroscopic experiments on blood plasma for information recovery from limited availability or high value samples was exemplified using plasma from patients with SARS-CoV-2 infection and normal controls. (1)H NMR spectra were obtained using solvent-suppressed 1D, spin-echo (CPMG), and 2-dimensional J-resolved (JRES) spectroscopy using both 3 mm outer diameter SampleJet NMR tubes (100 muL plasma) and 5 mm SampleJet NMR tubes (300 muL plasma) under in vitro diagnostic conditions. We noted near identical diagnostic models in both standard and low volume IVDr lipoprotein analysis (measuring 112 lipoprotein parameters) with a comparison of the two tubes yielding R(2) values ranging between 0.82 and 0.99 for the 40 paired lipoprotein parameters samples. Lipoprotein measurements for the 3 mm tubes were achieved without time penalty over the 5 mm tubes as defined by biomarker recovery for SARS-CoV-2. Overall, biomarker pattern recovery for the lipoproteins was extremely similar, but there were some small positive offsets in the linear equations for several variables due to small shimming artifacts, but there was minimal degradation of the biological information. For the standard untargeted 1D, CPMG, and JRES NMR experiments on the same samples, the reduced signal-to-noise was more constraining and required greater scanning times to achieve similar differential diagnostic performance (15 min per sample per experiment for 3 mm 1D and CPMG, compared to 4 min for the 5 mm tubes). We conclude that the 3 mm IVDr method is fit-for-purpose for quantitative lipoprotein measurements, allowing the preparation of smaller volumes for high value or limited volume samples that is common in clinical studies. If there are no analytical time constraints, the lower volume experiments are equally informative for untargeted profiling.
|Adult[MESH]
|Aged[MESH]
|Biomarkers/blood/metabolism[MESH]
|COVID-19/blood/*diagnosis/virology[MESH]
|Female[MESH]
|Humans[MESH]
|Lipoproteins/blood/*metabolism[MESH]
|Male[MESH]
|Metabolomics/*methods[MESH]
|Middle Aged[MESH]
|Protein Interaction Maps[MESH]
|Proteomics/*methods[MESH]
|Proton Magnetic Resonance Spectroscopy/*methods[MESH]
J+Proteome+Res 2021 ; 20 (2): 1415-1423
