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. 2022 Apr 25;25(5):104294.
doi: 10.1016/j.isci.2022.104294. eCollection 2022 May 20.

An SI-traceable reference material for virus-like particles

Andrea Briones  1   2 , Gustavo Martos  2 , Magali Bedu  2 , Tiphaine Choteau  2 , Ralf D Josephs  2 , Robert I Wielgosz  2 , Maxim G Ryadnov  1   3
Affiliations

An SI-traceable reference material for virus-like particles

Andrea Briones et al. iScience. .

Abstract

A reference material for virus-like particles traceable to the International System of Units (Système International d'Unités - the SI) is reported. The material addresses the need for developing reference standards to benchmark virus-like gene delivery systems and help harmonize measurement approaches for characterization and testing. The material is a major component of synthetic polypeptide virus-like particles produced by the state-of-the-art synthetic and analytical chemistry methods used to generate gene delivery systems. The purity profile of the material is evaluated to the highest metrological order demonstrating traceability to the SI. The material adds to the emerging toolkit of reference standards for quantitative biology.

Keywords: Analytical chemistry; Bioengineering; Biomaterials; Materials science; Nanotechnology.

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Conflict of interest statement

The authors declare no competing interests.

Figures

None
Graphical abstract
Figure 1
Figure 1
Virus-like particle material and its major component: C3-triskelion peptide (A) A molecular model of the C3-triskelion peptide (blue) in combination with its assembly partner peptide (red) (PDB entry 4DMD rendered by Macromodel Schrödinger). Each subunit of the triskelion corresponds to the altitude of an equilateral triangle. The peptide adopts a 3-fold symmetry (C3) matching that of triangular faces in an icosahedron, assembling into a capsid. (B) A molecular model of T = 4 icosahedron assembled from the C3-triskelions. For clarity, only one 5-fold axis is shown in color. (C) A molecular model of one arm of the triskelion (blue), its corresponding sequence, and the chemical structure (box). In the sequence, lysine residues are highlighted in blue, leucines and isoleucines are in orange.
Figure 2
Figure 2
The purity of the C3+ material established by amino acid analysis (A) Purity values for peptide in C3+ material calculated from the contributions of leucine and isoleucine residues, with the averaged value of 636 ± 45 mg/g. Error bars are expanded uncertainties (k = 2). (B) Relative uncertainty contributions to the final assigned purity value of the C3+ material, where u(Wp) is the standard uncertainty of the peptide mass fraction, u(IntPre) is an uncertainty due to the variance in the hydrolysis and the MS signal between injections, u(IDMS) is the combined uncertainty of the standard amino acid solution mass fraction and weighing, my is the mass of labeled amino acid solution in sample, myc is the mass of labeled amino acid solution in the calibrant, mz is the mass of standard amino acid solution in calibrant, Wz is the mass fraction of the standard amino acid solution, mx is the mass of peptide solution in sample.
Figure 3
Figure 3
Quantitative analysis of the C3+ material by qNMR (A) 2D homonuclear COSY (top) and TOCSY (bottom) correlations for the C3+ material. Cross signals highlighted in blue and green indicate proton-proton interactions within the same lysine spin systems. (B) 1H NMR spectrum of the peptide containing BTMSB-F4 as internal standard. Partially assigned regions are indicated with letters (see Table 3) and are integrated to display the relative proton stoichiometry. (C) Comparison of results of the mass balance method and AAA & qNMR. Where, the value obtained for the C3+ peptide in the mass balance method was of 669 ± 15 mg/g (k = 2) and the averaged value obtained for the measurements with the AAA & qNMR was 638 ± 24 mg/g (k = 2). Error bars are expanded uncertainties (k = 2).
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