Peptides and pseudopeptide ligands: a powerful toolbox for the affinity purification of current and next-generation biotherapeutics

Chu, Wenning; Prodromou, Raphael; Day, Kevin; Schneible, John D.; Bacon, Kaitlyn; Bowen, John; Kilgore, Ryan; Catella, Carly M.; Moore, Brandyn; Mabe, Matthew D.; Alashoor, Kawthar; Xu, Yiman; Xiao, Yuanxin; Menegatti, Stefano

doi:10.1016/j.chroma.2020.461632

Cited by 20 publications

(12 citation statements)

References 433 publications

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“…The future manufacturing of therapeutic mAb will rely on novel chromatographic adsorbents that enable continuous and more affordable processes, and have an improved ability to remove the high‐risk HCPs that have been identified as persistent in current bioprocesses (Jones et al, 2021). In this context, our group has developed a downstream toolbox of peptide‐based chromatographic adsorbents that purify therapeutic proteins either in bind‐and‐elute mode (Barozzi et al, 2020; Chu et al, 2021; Day et al, 2019; Kish et al, 2017, 2018; Menegatti et al, 2016; Prodromou et al, 2021) or flow‐through mode (Lavoie, Chu, et al, 2021; Lavoie et al, 2020). The latter, named LigaGuard™, operates by capturing CHO HCPs while allowing the mAb product to flow through unbound as the clarified cell culture harvest is continuously fed to the adsorbent.…”

Section: Resultsmentioning

confidence: 99%

Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents

Sripada

Chu

Williams

et al. 2022

Biotech & Bioengineering

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The growth of advanced analytics in manufacturing monoclonal antibodies (mAbs) has highlighted the challenges associated with the clearance of host cell proteins (HCPs). Of special concern is the removal of "persistent" HCPs, including immunogenic and mAb-degrading proteins, that co-elute from the Protein A resin and can escape the polishing steps. Responding to this challenge, we introduced an ensemble of peptide ligands that target the HCPs in Chinese hamster ovary (CHO) cell culture fluids and enable mAb purification via flow-through affinity chromatography. This study describes their integration into LigaGuard™, an affinity adsorbent featuring an equilibrium binding capacity of ~30 mg of HCPs per mL of resin as well as dynamic capacities up to 16 and 22 mg/ml at 1-and 2-min residence times, respectively. When evaluated against cell culture harvests with different mAb and HCP titers and properties, LigaGuard™ afforded high HCP clearance, with logarithmic removal values (LRVs) up to 1.5, and mAb yield above 90%. Proteomic analysis of the effluents confirmed the removal of high-risk HCPs, including cathepsins, histones, glutathione-S transferase, and lipoprotein lipases. Finally, combining LigaGuard™ for HCP removal with affinity adsorbents for product capture afforded a global mAb yield of 85%, and HCP and DNA LRVs > 4.

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Section: Resultsmentioning

confidence: 99%

Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents

Sripada

Chu

Williams

et al. 2022

Biotech & Bioengineering

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“…Here, we found that R-Pro9-3D remains totally intact under various digestive conditions and like Pro9-3D, exerts significant antibacterial activity against Gram-negative bacteria. This suggested that inversion and RI may have provided R-Pro9-3D with significant protection against proteolysis, as proteases are less likely to target peptide bonds containing D-amino acids [62]. Almost all ESKAPE pathogens, including A. baumannii, can form biofilms on biotic (e.g., skin, mucosa, and wounds) and abiotic (e.g., catheter) surfaces, resulting in drug resistance and persistent infections [45].…”

Section: Discussionmentioning

confidence: 99%

Antiseptic 9-Meric Peptide with Potency against Carbapenem-Resistant Acinetobacter baumannii Infection

Krishnan

Choi

Jang

et al. 2021

IJMS

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Carbapenem-resistant A. baumannii (CRAB) infection can cause acute host reactions that lead to high-fatality sepsis, making it important to develop new therapeutic options. Previously, we developed a short 9-meric peptide, Pro9-3D, with significant antibacterial and cytotoxic effects. In this study, we attempted to produce safer peptide antibiotics against CRAB by reversing the parent sequence to generate R-Pro9-3 and R-Pro9-3D. Among the tested peptides, R-Pro9-3D had the most rapid and effective antibacterial activity against Gram-negative bacteria, particularly clinical CRAB isolates. Analyses of antimicrobial mechanisms based on lipopolysaccharide (LPS)-neutralization, LPS binding, and membrane depolarization, as well as SEM ultrastructural investigations, revealed that R-Pro9-3D binds strongly to LPS and impairs the membrane integrity of CRAB by effectively permeabilizing its outer membrane. R-Pro9-3D was also less cytotoxic and had better proteolytic stability than Pro9-3D and killed biofilm forming CRAB. As an LPS-neutralizing peptide, R-Pro9-3D effectively reduced LPS-induced pro-inflammatory cytokine levels in RAW 264.7 cells. The antiseptic abilities of R-Pro9-3D were also investigated using a mouse model of CRAB-induced sepsis, which revealed that R-Pro9-3D reduced multiple organ damage and attenuated systemic infection by acting as an antibacterial and immunosuppressive agent. Thus, R-Pro9-3D displays potential as a novel antiseptic peptide for treating Gram-negative CRAB infections.

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“…In general, a high-affinity ligands’ binding result was obtained from a greater intermolecular force between the ligands, which shows good activity. [ 24 25 ] The smaller the value, the stronger the energy used to form a bond. [ 26 ] Based on the molecular docking results in Table 1 , the compounds with strong bonds are catechin = epicatechin > anthocyanin > theobromine, with binding affinity values of −8.0, −8.0, −7.8, and −6.1 kcal/mol, respectively.…”

Section: Discussionmentioning

confidence: 99%

Potential anti-alopecia constituents from Theobroma cacao: An in silico study

Kurnia¹,

Kelutur²,

Mustarichie³

2021

J Adv Pharm Technol Res

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A BSTRACT Tinea capitis is local alopecia caused by a dermatophyte infection of the scalp. Trichophyton rubrum produces the squalene epoxidase enzyme, which has a crucial role in prolonged dermatophyte infection, as well as in synthesizing fatty acids in this dermatophyte group. This study analyzes Trichophyton cacao compounds as anti-alopecia by inhibiting the squalene epoxidase enzyme formation, in silico . The structure of T. cacao compounds was prepared using the MolView Web application. The compound docked to squalene epoxidase using AutoDock Vina in PyRx 0.8, followed by PyMOL for visualization, and the Proteins Plus program to analyze the complexity. The binding affinity value of catechin, epicatechin (−8.0 kcal/mol), and anthocyanin (−7.8 kcal/mol) compounds was higher than the positive control (terbinafine, −6.7 kcal/mol). Pre-ADMET demonstrated that catechin and epicatechin had moderate Human Intestinal Absorption (66.71%), but anthocyanin was very good (100%). Caco-2 parameters for catechin and epicatechin were relatively low (<4 nm s − 1 ), while anthocyanin, theobromine, and terbinafine were within 4–70 nm s − 1 . Plasma protein binding shows catechin, epicatechin, and anthocyanin diffuse through the plasma membrane and interact with plasma proteins. The toxicity results for all compounds are mutagenic, and only terbinafine is carcinogenic. Based on the Lipinski's “Rule of Five,” compounds from T. Cacao can be given orally. Catechin and epicatechin compounds have the potential to act as anti-alopecia. These two compounds can diffuse and interact with plasma proteins so they are directly on the target when given orally.

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Peptides and pseudopeptide ligands: a powerful toolbox for the affinity purification of current and next-generation biotherapeutics

Cited by 20 publications

References 433 publications

Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents

Towards continuous mAb purification: Clearance of host cell proteins from CHO cell culture harvests via “flow‐through affinity chromatography” using peptide‐based adsorbents

Antiseptic 9-Meric Peptide with Potency against Carbapenem-Resistant Acinetobacter baumannii Infection

Potential anti-alopecia constituents from Theobroma cacao: An in silico study

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