(320f) Preparation of a Novel Tetrapeptide Biomimetic Chromatographic Resin and Its High Antibody-Binding Capacity
AIChE Annual Meeting
2019
2019 AIChE Annual Meeting
Separations Division
Chromatography Techniques for Bioprocessing
Tuesday, November 12, 2019 - 2:10pm to 2:30pm
The
separation and purification of protein drug, especial antibody is a big challenge
in biopharmaceutical process. Biomimetic affinity chromatography with short
peptide ligands, as a promising bioseparation technique, has great potential to
protein separation and purification. In this work, we focused on the exploring the new biomimetic
chromatographic resin preparation and properties with new tetrapeptide as
ligand. A tetrapeptide library with critical residues of natural ligands (ProA,
ProG) to hIgG was constructed and a ligand (Ac-FYWR) with high LibDock
score was selected by molecular docking. Then, the selected ligand was
evaluated by static/dynamic adsorption. It was found that ligand Ac-FYWR has
the high binding capacity and selectivity for hIgG. The results showed the Qm of Ac-FYWR-4FF resin to hIgG was 110.7
mg/g resin and the Kd of the resin was 0.10 mg/mL at
pH 8.0, while the resin almost could not adsorb hIgG at pH 5.0. Moreover, at pH
8.0, Q10%
of Ac-FYWR-4FF resin was 26.7 mg/mL for
hIgG but just 1.0 mg/mL at pH 5.0, which presented high selectivity of the
screened resin by absorbing at pH 8.0 and eluting at pH 5.0. Finally,
the Ac-FYWR-4FF resin was applied to separate mAb from
CHO cell culture supernatant with the purity of 97.34% by only one-step
separation. The results show that the
screened tetrapeptide resin (Ac-FYWR-4FF resin) could be well used in the antibody
separation and purification, be help
to develop the biomimetic chromatographic and
give the new direction for the design of the peptide ligand with high
adsorption for target proteins.
separation and purification of protein drug, especial antibody is a big challenge
in biopharmaceutical process. Biomimetic affinity chromatography with short
peptide ligands, as a promising bioseparation technique, has great potential to
protein separation and purification. In this work, we focused on the exploring the new biomimetic
chromatographic resin preparation and properties with new tetrapeptide as
ligand. A tetrapeptide library with critical residues of natural ligands (ProA,
ProG) to hIgG was constructed and a ligand (Ac-FYWR) with high LibDock
score was selected by molecular docking. Then, the selected ligand was
evaluated by static/dynamic adsorption. It was found that ligand Ac-FYWR has
the high binding capacity and selectivity for hIgG. The results showed the Qm of Ac-FYWR-4FF resin to hIgG was 110.7
mg/g resin and the Kd of the resin was 0.10 mg/mL at
pH 8.0, while the resin almost could not adsorb hIgG at pH 5.0. Moreover, at pH
8.0, Q10%
of Ac-FYWR-4FF resin was 26.7 mg/mL for
hIgG but just 1.0 mg/mL at pH 5.0, which presented high selectivity of the
screened resin by absorbing at pH 8.0 and eluting at pH 5.0. Finally,
the Ac-FYWR-4FF resin was applied to separate mAb from
CHO cell culture supernatant with the purity of 97.34% by only one-step
separation. The results show that the
screened tetrapeptide resin (Ac-FYWR-4FF resin) could be well used in the antibody
separation and purification, be help
to develop the biomimetic chromatographic and
give the new direction for the design of the peptide ligand with high
adsorption for target proteins.
Keywords:
biomimetic affinity
chromatography; adsorption behavior;
Ac-FYWR-4FF resin; CHO cell culture supernatant;