tetano
Editor, Senior Moderator
J Am Soc Mass Spectrom. 2019 Oct 8. doi: 10.1007/s13361-019-02318-z. [Epub ahead of print] [h=1]Using LC-MS to Identify Clipping in Self-Assembled Nanoparticles During Vaccine Development.[/h]
Schneck NA[SUP]1[/SUP], Ivleva VB[SUP]1[/SUP], Rosales-Zavala E[SUP]1[/SUP], Wang X[SUP]1[/SUP], Gollapudi D[SUP]1[/SUP], Cooper JW[SUP]1[/SUP], Lei QP[SUP]2[/SUP].
[h=3]Author information[/h] 1 Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9 West Watkins Mill Rd., Gaithersburg, MD, 20878, USA. 2 Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9 West Watkins Mill Rd., Gaithersburg, MD, 20878, USA. paula.lei@nih.gov.
[h=3]Abstract[/h] A hemagglutinin stabilized stem nanoparticle (HA-SS-np) that is designed to provide broad protection against influenza is being developed as a potential vaccine. During an early formulation screening study, reducing gel (rCGE) analysis indicated product degradation in a few candidate buffers at the first-week accelerated stability point, whereas no change was shown in the size exclusion chromatography (SEC) measurement. A LC-MS workflow was therefore applied to investigate the integrity of this large HA-SS-np vaccine molecule (≈ 1 MDa). Application of LC-MS was critical to rationalize the conflicting results from the rCGE and SEC assays and led to the discovery that (1) an unexpected sequence clipping in the HA-SS-np subunits occurred, explaining the atypical reducing gel profile, and (2) an undisrupted disulfide bond held the two fragments together, explaining the unchanged SEC profile. This analytical case study led to a formulation buffer redesign, which mitigated the issue.
[h=4]KEYWORDS:[/h] Characterization; Clipping; Influenza; LC-MS; Nanoparticles; Vaccine
PMID: 31595432 DOI: 10.1007/s13361-019-02318-z
Schneck NA[SUP]1[/SUP], Ivleva VB[SUP]1[/SUP], Rosales-Zavala E[SUP]1[/SUP], Wang X[SUP]1[/SUP], Gollapudi D[SUP]1[/SUP], Cooper JW[SUP]1[/SUP], Lei QP[SUP]2[/SUP].
[h=3]Author information[/h] 1 Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9 West Watkins Mill Rd., Gaithersburg, MD, 20878, USA. 2 Vaccine Production Program, Vaccine Research Center, National Institute of Allergy and Infectious Diseases, National Institutes of Health, 9 West Watkins Mill Rd., Gaithersburg, MD, 20878, USA. paula.lei@nih.gov.
[h=3]Abstract[/h] A hemagglutinin stabilized stem nanoparticle (HA-SS-np) that is designed to provide broad protection against influenza is being developed as a potential vaccine. During an early formulation screening study, reducing gel (rCGE) analysis indicated product degradation in a few candidate buffers at the first-week accelerated stability point, whereas no change was shown in the size exclusion chromatography (SEC) measurement. A LC-MS workflow was therefore applied to investigate the integrity of this large HA-SS-np vaccine molecule (≈ 1 MDa). Application of LC-MS was critical to rationalize the conflicting results from the rCGE and SEC assays and led to the discovery that (1) an unexpected sequence clipping in the HA-SS-np subunits occurred, explaining the atypical reducing gel profile, and (2) an undisrupted disulfide bond held the two fragments together, explaining the unchanged SEC profile. This analytical case study led to a formulation buffer redesign, which mitigated the issue.
[h=4]KEYWORDS:[/h] Characterization; Clipping; Influenza; LC-MS; Nanoparticles; Vaccine
PMID: 31595432 DOI: 10.1007/s13361-019-02318-z