Cyclic peptide ligand with high binding capacity for affinity purification of immunoglobulin G

Kang, Hyo Jin; Choe, Weonu; Kim, Jung Min; Lee, Young-Mi; Kim, B. Moon; Chung, Sang J.

doi:10.1016/j.chroma.2016.09.007

Cited by 23 publications

(13 citation statements)

References 29 publications

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“…As can be seen from Table 4 the wash steps increased HCP LRV considerably, with values of 2.07 and 2.15 for Tris pH 9 and 1 M NaCl respectively. This level of HCP reduction is the highest among the synthetic adsorbents reported in literature [23,24,60,61].…”

Section: Resultsmentioning

confidence: 94%

“…Initially, the display of the peptide ligands on the surface of agarose-based WorkBeads resins (WB) was optimized using spacer arms, resulting in values of static and dynamic binding capacity (DBC) of about 85 mg/mL and 65 mg/mL, respectively. These values are unprecedented in the literature on synthetic affinity ligands [23,24,60,61]. The optimization of the peptide sequence and the chromatographic conditions enabled a noticeable increase of HCP clearance, from 1.04 obtained with HWRGWVC-TREN-WB, to 2.15 obtained with Ac-HWCitGWVC-TREN-WB coupled with a pre-elution wash with 1 M NaCl in PBS.…”

Section: Discussionmentioning

confidence: 89%

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Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Islam

Naik

Hashimoto

et al. 2019

IJMS

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This work presents the use of peptide ligand HWRGWV and its cognate sequences to develop affinity adsorbents that compete with Protein A in terms of binding capacity and quality of the eluted product. First, the peptide ligand was conjugated to crosslinked agarose resins (WorkBeads) at different densities and using different spacer arms. The optimization of ligand density and display resulted in values of static and dynamic binding capacity of 85 mg/mL and 65 mg/mL, respectively. A selected peptide-WorkBeads adsorbent was utilized for purifying Mabs from Chinese Hamster Ovary (CHO) cell culture supernatants. The peptide-WorkBeads adsorbent was found able to withstand sanitization with strong alkaline solutions (0.5 M NaOH). The purity of the eluted product was consistently higher than 95%, with logarithmic removal value (LRV) of 1.5 for host cell proteins (HCPs) and 4.0 for DNA. HCP clearance was significantly improved by adding a post-load washing step with either 0.1 M Tris HCl pH 9 or 1 M NaCl. The cognate peptide of HWRGWV, constructed by replacing arginine (R) with citrulline, further increased the HCP LRV to 2.15. The peptide-based adsorbent also showed a remarkable performance in terms of removal of Mab aggregates; unlike Protein A, in fact, HWRGWV was found to bind only monomeric IgG. Collectively, these results demonstrate the potential of peptide-based adsorbents as alternative to Protein A for the purification of therapeutic antibodies.

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Section: Resultsmentioning

confidence: 94%

Section: Discussionmentioning

confidence: 89%

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Islam

Naik

Hashimoto

et al. 2019

IJMS

View full text Add to dashboard Cite

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“…The potential of Fc-III was demonstrated with the development of a re-useable (up to 60 runs) affinity purification column that gives high purity Ab (95%) [49]. Immobilization of Fc-III on sepharose had no significant effect on the binding affinity of Fc-III towards IgG and allowed elution of the bound Ab at pH comparable to Protein G affinity columns (pH = 3.2).…”

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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“…One such cyclic peptide, the FC‐III, is a 13 amino acid cyclic peptide binder of the human immunoglobulin G (IgG) Fc domain with K d =16 n m (Blue β‐sheet in Figure ) . Previously, it has been employed as an affinity ligand in column purification of IgGs and as a half‐life‐increasing tag when expressed in fusion with a protein of interest (POI) . Very recently, methods to covalently link Fc binding peptides to the IgG through either a photo‐crosslinker or NHS‐coupling were demonstrated .…”

Section: Figurementioning

confidence: 99%

Peptide‐Directed DNA‐Templated Protein Labelling for The Assembly of a Pseudo‐IgM

et al. 2019

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The development of methods for conjugation of DNA to proteins is of high relevance for the integration of protein function and DNA structures. Here, we demonstrate that protein‐binding peptides can direct a DNA‐templated reaction, selectively furnishing DNA–protein conjugates with one DNA label. Quantitative conversion of oligonucleotides is achieved at low stoichiometries and the reaction can be performed in complex biological matrixes, such as cell lysates. Further, we have used a star‐like pentameric DNA nanostructure to assemble five DNA–Rituximab conjugates, made by our reported method, into a pseudo‐IgM antibody structure that was subsequently characterized by negative‐stain transmission electron microscopy (nsTEM) analysis.

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Cyclic peptide ligand with high binding capacity for affinity purification of immunoglobulin G

Cited by 23 publications

References 29 publications

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

Optimization of Sequence, Display, and Mode of Operation of IgG-Binding Peptide Ligands to Develop Robust, High-Capacity Affinity Adsorbents That Afford High IgG Product Quality

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

Peptide‐Directed DNA‐Templated Protein Labelling for The Assembly of a Pseudo‐IgM

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