A megaporous material harbouring a peptide ligand for affinity IgG purification

Abstract

Common limitations of Protein A affinity chromatography include high adsorbent costs, ligand instability and possible ligand leakage. In this study, a short peptide with affinity for IgG was synthesized chemically and subsequently immobilized on a megaporous support. The support was prepared utilising the cryogel technique while the peptide-ligand was covalently immobilised via thiol-epoxy click chemistry. The cryogel support was chemically grafted to increase the number of reaction sites. This adsorbent was designated as "MP-Pep". Adsorption isotherms were employed to evaluate protein binding capacity. A maximum static binding capacity within the range of 30-60 mg/mL was observed for hIgG. This parameter compares well with other commercial and non-commercial adsorbents, as reported in the literature. As a control material, a Protein A grafted megaporous cryogel was synthesized. Dynamic binding capacity values were obtained by breakthrough analysis. The peptide cryogel showed a dynamic capacity value 9.0 mg/mL in comparison to 9.7 mg/mL in the case of the Protein A based adsorbent. The ratio of dynamic binding capacity to static binding capacity was 20%, indicating suboptimal product capture. However, the advantage of MP-Pep lies in its cost-effective preparations while maintaining a reasonable binding capacity for the targeted product. The presence of cooperative effects during protein binding could also represent an advantage during the processing of a feedstock containing a product in high concentration.

In vitro/in vivo degradation analysis of trastuzumab by combining specific capture on HER2 mimotope peptide modified material and LC-QTOF-MS

“…S4). The binding capacity at 10% breakthrough was found to be 11.3 mg/mL, which is comparable to that previously reported for a Fc-specific peptide based polymer towards hIgG (9.0 mg/mL) [42]. The density (4.0 μmol/mL or 11 mg/mL) and affinity (K d = 2.6 μM) of HH24 in the new material are lower than the peptide ligand density (12.3 μmol/mL or 25 mg/mL) and affinity (K d = 345 nM) of the KH19 immobilized membrane towards trastuzumab.…”

Supramolecular Mimotope Peptide Nanofibers Promote Antibody–Ligand Polyvalent and Instantaneous Recognition for Biopharmaceutical Analysis

“…Eventually, standing for 1 min was selected as the optimal preparation method of mimotope peptide nanofibers. Under optimal conditions, the mimotope peptide nanofibers exhibit instantaneous capture and high enrichment yields, overcoming the drawbacks of low capture efficiency observed with previously reported affinity materials − and showing potential to be advantageous in practical applications. Moreover, the remaining nanofibers after enrichment could be recycled back into an aqueous solution and reused for additional enrichment cycles (up to three times).…”

Affibody-Binding Ligands

“…The development of an affinity-based capture technology targeting the constant regions of affibodies in α-helix 3 and α-helix 1 holds great promise toward streamlining the manufacturing of affibodies and reducing their cost. Synthetic peptides are ideal scaffolds to develop cost-effective ligands with excellent biorecognition ability and high biochemical stability [30][31][32][33][34][35].…”