Design of protease-resistant peptide ligands for the purification of antibodies from human plasma

Menegatti, Stefano; Bobay, Benjamin G.; Ward, Kevin Lawrence; Islam, Tuhidul; Kish, William S.; Naik, Amith D.; Carbonell, Ruben G.

doi:10.1016/j.chroma.2016.03.087

Cited by 39 publications

(29 citation statements)

References 46 publications

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“…The synthesis procedure was performed according to a protocol adapted from Menegatti et al (2016) and is described in detail in prior work (Lavoie et al, 2019). Individual ligand candidates previously identified were synthesized and pooled as described in previous work (Lavoie et al, 2019).…”

Section: Toyopearl Peptide Resin Synthesis and Deprotectionmentioning

confidence: 99%

Targeted capture of Chinese hamster ovary host cell proteins: Peptide ligand binding by proteomic analysis

Lavoie

Fazio

Williams

et al. 2019

Biotech & Bioengineering

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The clearance of host cell proteins (HCPs) is of crucial importance in biomanufacturing, given their diversity in composition, structure, abundance, and occasional structural homology with the product. The current approach to HCP clearance in the manufacturing of monoclonal antibodies (mAbs) relies on product capture with Protein A followed by removal of residual HCPs in flow‐through mode using ion exchange or mixed‐mode chromatography. Recent studies have highlighted the presence of “problematic HCP” species, which are either difficult to remove (Group I), can degrade the mAb product (Group II), or trigger immunogenic reactions (Group III). To improve the clearance of these species, we developed a family of synthetic peptides that target HCPs and exhibit low binding to IgG product. In this study, these peptides were conjugated onto chromatographic resins and evaluated in terms of HCP clearance and mAb yield, using an industrial mAb‐producing CHO harvest as model supernatant. To gather detailed knowledge on the binding of individual HCPs, the unbound fractions were subjected to shotgun proteomic analysis by mass spectrometry. It was found that these peptide ligands exhibit superior HCP binding capability compared to those of the benchmark commercial resins commonly used in mAb purification. In addition, some peptide‐based resins resulted in much lower losses of product yield compared to these commercial supports. The proteomic analysis showed effective capture of many “problematic HCPs” by the peptide ligands, especially some that are weakly bound by commercial media. Collectively, these results indicate that these peptides show great promise toward the development of next‐generation adsorbents for safer and cost‐effective manufacturing of biologics.

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Section: Toyopearl Peptide Resin Synthesis and Deprotectionmentioning

confidence: 99%

Targeted capture of Chinese hamster ovary host cell proteins: Peptide ligand binding by proteomic analysis

Lavoie

Fazio

Williams

et al. 2019

Biotech & Bioengineering

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“…The purification of monoclonal IgG from industrial CHO cell culture supernatants was reported with a yield of ~96% and a purity of ~93%. Recently, the modification of the above described hexamers with unnatural amino acids was reported to gain resistance to proteases [57]. …”

Section: Immunoglobulin Binding Peptides and Peptidomimeticsmentioning

confidence: 99%

Fc-Binding Ligands of Immunoglobulin G: An Overview of High Affinity Proteins and Peptides

2016

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The rapidly increasing application of antibodies has inspired the development of several novel methods to isolate and target antibodies using smart biomaterials that mimic the binding of Fc-receptors to antibodies. The Fc-binding domain of antibodies is the primary binding site for e.g., effector proteins and secondary antibodies, whereas antigens bind to the Fab region. Protein A, G, and L, surface proteins expressed by pathogenic bacteria, are well known to bind immunoglobulin and have been widely exploited in antibody purification strategies. Several difficulties are encountered when bacterial proteins are used in antibody research and application. One of the major obstacles hampering the use of bacterial proteins is sample contamination with trace amounts of these proteins, which can invoke an immune response in the host. Many research groups actively develop synthetic ligands that are able to selectively and strongly bind to antibodies. Among the reported ligands, peptides that bind to the Fc-domain of antibodies are attractive tools in antibody research. Besides their use as high affinity ligands in antibody purification chromatography, Fc-binding peptides are applied e.g., to localize antibodies on nanomaterials and to increase the half-life of proteins in serum. In this review, recent developments of Fc-binding peptides are presented and their binding characteristics and diverse applications are discussed.

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“…However, examples in the literature pertaining to their development for downstream purification purposes are limited at present. Peptides however offer several advantages as affinity ligands due to their low cost, high shelf‐life, relative ease of chemical synthesis and purification (as opposed to biological expression and purification), and amenability to chemical modifications such as cyclization or the incorporation of non‐natural amino acids and moieties . Short peptides are also promising ligands for affinity chromatography since they are more stable than large proteins and antibodies under harsh elution and cleaning conditions.…”

Section: Introductionmentioning

confidence: 99%

“…For example, Kelley et al screened phage‐display libraries to identify peptides for the purification of recombinant B‐Domain Deleted Factor VIII . Menegatti et al and Naik et al have demonstrated the design of protease‐resistant, hexamer affinity peptides for monoclonal antibody (mAb) purification . Linear hexamer peptides developed in the earlier‐mentioned work were first identified by Yang et al via combinatorial screening and synthesis of a solid‐phase peptide library .…”

Section: Introductionmentioning

confidence: 99%

Design of peptide ligands for affinity purification of human growth hormone

Chandra

Timmick

Goodwine

et al. 2019

J of Chemical Tech & Biotech

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BACKGROUND: In this study, we have demonstrated the design, screening and selection of peptide ligands for the affinity capture of human growth hormone (hGH) from yeast cell cultures. RESULTS:Ligand design was carried out using multiple approaches based on primary sequence and structures of natural binding partners of hGH. Screening of potential affinity peptides was conducted using high throughput microarray platforms followed by assessment of in-solution binding to hGH using fluorescence polarization. Peptide leads were subsequently immobilized on chromatographic resins and the binding and desorption behavior was examined using batch adsorption studies. A lead candidate was examined in further details in column chromatography studies which indicated that while high purity was attained, further refinement was necessary for improved product recovery. Histidine scanning was employed to successfully improve the recovery of hGH from cell culture fluid while still maintaining high purity. Finally, proof-of-concept was demonstrated in the column format using complex feed stock where a product purity of 95% was attained at 80% yield. CONCLUSION: The approaches presented here can be translated to other biologics of interest for the rapid development of affinity based purification processes.

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Design of protease-resistant peptide ligands for the purification of antibodies from human plasma

Cited by 39 publications

References 46 publications

Targeted capture of Chinese hamster ovary host cell proteins: Peptide ligand binding by proteomic analysis

Targeted capture of Chinese hamster ovary host cell proteins: Peptide ligand binding by proteomic analysis

Fc-Binding Ligands of Immunoglobulin G: An Overview of High Affinity Proteins and Peptides

Design of peptide ligands for affinity purification of human growth hormone

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