tetano
Editor, Senior Moderator
Vaccine. 2017 May 2. pii: S0264-410X(17)30558-3. doi: 10.1016/j.vaccine.2017.04.066. [Epub ahead of print]
[h=1]Nile Red fluorescence spectrum decomposition enables rapid screening of large protein aggregates in complex biopharmaceutical formulations like influenza vaccines.[/h] Sahin Z[SUP]1[/SUP], Akkoc S[SUP]1[/SUP], Neeleman R[SUP]2[/SUP], Haines J[SUP]2[/SUP], Kayser V[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] The extensive presence of large (high molecular weight) protein aggregates in biopharmaceutical formulations is a concern for formulation stability and possibly safety. Tests to screen large aggregate content in such bioformulations are therefore needed for rapid and reliable quality control in industrial settings. Herein, non-commercial seasonal influenza split-virus vaccine samples, produced using various strains and extracted from selected industrial processing steps, were used as model complex bioformulations. Orthogonal characterization through transmission electron microscopy, UV-Vis absorption spectroscopy, fluorescence emission spectroscopy, high-performance liquid chromatography and single-radial immunodiffusion revealed that large, amorphous protein aggregates are formed after virus splitting and their presence is linked mainly, albeit not only, to surfactant (Triton X-100) content in a sample. Importantly, the presence of large virus aggregates in purified whole virus samples and large protein aggregates in vaccine samples was found to correlate with broadening/shouldering in Nile Red fluorescence spectra. Accordingly, decomposition of Nile Red spectra into components allowed the development of a novel, rapid, reliable and user-friendly test with high-throughput potential for screening large aggregate content in influenza split-virus vaccines. The test can be adapted for screening other complex biopharmaceutical formulations, provided relevant controls are done for informed decomposition of fluorescence spectra into their components.
Copyright ? 2017. Published by Elsevier Ltd.
[h=4]KEYWORDS:[/h] Biopharmaceutical formulation stability; Influenza split-virus vaccine; Nile Red fluorescence; Protein aggregate screening; Triton X-100
PMID: 28476626 DOI: 10.1016/j.vaccine.2017.04.066
[h=1]Nile Red fluorescence spectrum decomposition enables rapid screening of large protein aggregates in complex biopharmaceutical formulations like influenza vaccines.[/h] Sahin Z[SUP]1[/SUP], Akkoc S[SUP]1[/SUP], Neeleman R[SUP]2[/SUP], Haines J[SUP]2[/SUP], Kayser V[SUP]3[/SUP].
[h=3]Author information[/h]
[h=3]Abstract[/h] The extensive presence of large (high molecular weight) protein aggregates in biopharmaceutical formulations is a concern for formulation stability and possibly safety. Tests to screen large aggregate content in such bioformulations are therefore needed for rapid and reliable quality control in industrial settings. Herein, non-commercial seasonal influenza split-virus vaccine samples, produced using various strains and extracted from selected industrial processing steps, were used as model complex bioformulations. Orthogonal characterization through transmission electron microscopy, UV-Vis absorption spectroscopy, fluorescence emission spectroscopy, high-performance liquid chromatography and single-radial immunodiffusion revealed that large, amorphous protein aggregates are formed after virus splitting and their presence is linked mainly, albeit not only, to surfactant (Triton X-100) content in a sample. Importantly, the presence of large virus aggregates in purified whole virus samples and large protein aggregates in vaccine samples was found to correlate with broadening/shouldering in Nile Red fluorescence spectra. Accordingly, decomposition of Nile Red spectra into components allowed the development of a novel, rapid, reliable and user-friendly test with high-throughput potential for screening large aggregate content in influenza split-virus vaccines. The test can be adapted for screening other complex biopharmaceutical formulations, provided relevant controls are done for informed decomposition of fluorescence spectra into their components.
Copyright ? 2017. Published by Elsevier Ltd.
[h=4]KEYWORDS:[/h] Biopharmaceutical formulation stability; Influenza split-virus vaccine; Nile Red fluorescence; Protein aggregate screening; Triton X-100
PMID: 28476626 DOI: 10.1016/j.vaccine.2017.04.066