Fig. 4: Purification and characterization of RV3 preF lead candidates. | Nature Communications

Fig. 4: Purification and characterization of RV3 preF lead candidates.

From: Universal paramyxovirus vaccine design by stabilizing regions involved in structural transformation of the fusion protein

Fig. 4

A Analytical SEC trace of wildtype (gray) and stabilized OnlyEcto2P (purple) in supernatant. B SEC-MALS trace of purified OnlyEcto2P. C Melting temperature as determined by DSF of purified OnlyEcto2P of (B). The first derivative of the fluorescence signal is plotted. BLI using kinetic Octet with PIA174 immobilized to anti-human Fc sensors (not shown), followed subsequently by capture of purified preF variants (0−900 s phase) and association of sdAb’s 4C06 (D) and 4C03 (E) (900−1800 s phase). F SEC-MALS trace of purified OnlyEcto. G Melting temperature as determined of OnlyEcto as in (C). H Analytical SEC trace of purified OnlyEcto after storage at either 4 °C (left panel) or 37 °C (right panel) for up to 24 weeks. I Percentage trimer recovery of (H). Shown is the average of two replicates. J Expression and melting temperature of OnlyEcto and back-substitutions as measured by analytical SEC and DSF on crude cell supernatant.

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