Affinity purification of IgG monoclonal antibodies using the D-PAM synthetic ligand: chromatographic comparison with protein A and thermodynamic investigation of the D-PAM/IgG interaction

D'Agostino, Biagio; Bellofiore, Piero; Martino, Tiziana; Punzo, Carmine; Rivieccio, Vincenzo; Verdoliva, Antonio

doi:10.1016/j.jim.2008.01.014

Cited by 39 publications

(23 citation statements)

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“…Ehrlish and Bailon, and Krook et al also reported IgGbinding peptides discovered using the filamentous phage display technique (5,6). Recently, Verdoliva et al screened a synthetic peptide library and identified an IgG-binding cyclic dimeric peptide that recognized the lower hinge region of IgG (7,8). Of these earlier attempts, the Fc-III peptide with an intramolecular disulfide bond reported by DeLano et al in 2000 (9) is a potential candidate that can displace Protein A functions.…”

mentioning

confidence: 99%

Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions

Sakamoto

Ito

Hatanaka

et al. 2009

Journal of Biological Chemistry

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In therapeutic antibody preparation, acidic pH conditions are generally used for elution from Protein A affinity column of IgG or for its viral inactivation. Exposing IgG to low pH conditions induces conformational changes, leading to its functional damage or loss, although the mechanisms have not been fully elucidated. In this study using random peptide T7 phage display libraries, we isolated a unique and novel peptide motif that specifically recognized the non-native conformer (acid conformer) of human IgG that was generated by the low pH treatment, but not the native conformer. We examined the generation conditions and biochemical properties of acid conformer using the peptide motif as an affinity ligand. The acid conformer was easily generated at acidic pH (25°C). The peptides isolated here could contribute to the elucidation of the mechanisms of antibody dysfunction or aggregation during acid exposure as well as storage of human IgG.

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confidence: 99%

Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions

Sakamoto

Ito

Hatanaka

et al. 2009

Journal of Biological Chemistry

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“…However, purification of antigens with antibodies should choose low-affinity antibodies to avoid denaturation of most proteins. With respect to the cost and safety, prompted research activities have being focused on novel synthetic ligands (synthetic mimic ligands of proteins A and L) with improved properties such as lower cost, avoidance of the risk of contamination associated with natural ligands of human or animal origin to isolate antibodies (D'Agostino et al, 2008). What's more, affinity chromatography (one of the protein separation approaches) on immobilized antibodies with specific binding to proteins plays an important role in the proteomic (phosphoprotemics or glycoproteomics) and genomic field.…”

Section: Discussionmentioning

confidence: 99%

“…Monoclonal antibodies (MAbs) are useful for the treatment of a wide array of indications including autoimmune diseases, infectious diseases, cardiovascular diseases, transplant rejection and cancer. For instance, IgA, IgE, IgM and IgY increasingly find application in the cure and/or diagnosis of important diseases, especially for the IgG class, which plays most important role in the clinical application (D'Agostino et al, 2008). Meanwhile, extremely purified-pooled polyclonal IgG, intravenous IgG and specific antibodies (hyperimmune IgG) have become the basis for standard therapies in a number of malignancies (Verdoliva et al, 2002).…”

Section: Review Articlementioning

confidence: 99%

“…Protein A mimetic ligands such as a synthetic peptide (TG19318) or PAM and its inverse derivative (D-PAM), as well as ligand 22/8 were researched (Kabir, 2002;Verdoliva et al, 2002;D'Agostino et al, 2008). TG 19318, comprising four identical tripeptide chains linked to a central polylysine core, has been used to isolate polyclonal and MAbs of different classes (IgG, IgM, IgA and IgE) from different sources (serum, ascites and cell supernatants) and species.…”

Section: Purification Of Antibodies Using Synthetic Mimic Ligands Of mentioning

confidence: 99%

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Separation of antigens and antibodies by immunoaffinity chromatography

Sheng

Kong

2012

Pharmaceutical Biology

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“…Synthetic peptide based affinity chromatography processes have been successfully applied for the isolation of antibodies from human serum and cell culture media (D'Agostino et al, 2008;Fassina et al, 1996;Linhult et al, 2005;Menegatti et al, 2012). Small ligands such as peptides have potential advantages in chromatography processes because they can be more stable and less immunogenic than protein ligands, can be constructed with a wide variety of bio-specificity and their production cost is low (Fassina et al, 1996;Yang et al, 2005).…”

Section: Introductionmentioning

confidence: 99%

Application of peptide chromatography for the isolation of antibodies from bovine skim milk, acid whey and colostrum

Billakanti

Fee

Naik

et al. 2014

Food and Bioproducts Processing

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Protein A mimetic peptide ligands have several benefits over conventional Protein A/G ligands, namely that they are small in size, have low production costs, are stable over a wide range of pH values and can withstand cleaning by harsh sanitization agents such as sodium hydroxide. In this paper, a hexamer peptide (HWRGWV) affinity matrix was used for the isolation of bovine immunoglobulins from various dairy streams (skim milk, acid whey and colostrum). Bound immunoglobulins were recovered in elution buffer (0.2 M sodium acetate buffer, pH 4.0) fractions with a purity of >85% in a single step. The peptide resin has achieved a maximum equilibrium adsorption capacity of 23±0.58 mg.mL-1 of resin for bovine IgG and had a dynamic binding capacity of 11.8±0.03 mg.mL-1 at residence time of 2 min. These results suggest that the hexamer peptide chromatography could potentially be used for the selective purification of bovine immunoglobulins from dairy streams. This method has promise as an alternative to conventional Protein A/G chromatography for direct capture of immunoglobulins from streams containing relatively high immunoglobulin concentrations such as colostrum, transgenic or hyper-immune milk. In the abstract, equilibrium adsorption capacity should appear with the associated error (in the previous submission this error appear).Decision: Authors are accepted the above and reviewed as suggested (23±0.58). Comment2.Page 12 line 10 "However, for experiments with standard bIgG at 1 mg mL-1 and 9 mg mL-1 were overloaded" The idea to answer comment 8, reviewer 1 is there but the sentence should better introduced in the text.Decision: Authors are accepted the above and reviewed as suggested in the draft. "However, for experiments with standard bIgG at 1 mg.mL -1 and 9 mg.mL -1 were overloaded since column reached its dynamic capacity approximately at 5.5 mL for 1 mg.mL -1 bIgG and 1.4 mL for 9 mg.mL -1 bIgG. Due to this, % of IgG bound data presented in table 1.1 for these experiments may not reflect the actual numbers". Thank you.Yours sincerely, Jagan M Billakanti, PhD. Cover LetterRevision 2 Reviewer #1 Comment1. In the abstract, equilibrium adsorption capacity should appear with the associated error (in the previous submission this error appear).Decision: Authors are accepted the above and reviewed as suggested (23±0.58) Comment2. Page 12 line 10 "However, for experiments with standard bIgG at 1 mg mL-1 and 9 mg mL-1 were overloaded" The idea to answer comment 8, reviewer 1 is there but the sentence should better introduced in the text.Decision: Authors are accepted the above and reviewed as suggested in the draft."However, for experiments with standard bIgG at 1 mg.mL -1 and 9 mg.mL -1 were overloaded since column reached its dynamic capacity approximately at 5.5 mL for 1 mg.mL -1 bIgG and 1.4 mL for 9 mg.mL -1 bIgG. Due to this, % of IgG bound data presented in table 1.1 for these experiments may not reflect the actual numbers". Detailed Response to ReviewersHighlights  HWRGWV peptide resin showed an equ...

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Affinity purification of IgG monoclonal antibodies using the D-PAM synthetic ligand: chromatographic comparison with protein A and thermodynamic investigation of the D-PAM/IgG interaction

Cited by 39 publications

References 40 publications

Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions

Discovery and Characterization of a Peptide Motif That Specifically Recognizes a Non-native Conformation of Human IgG Induced by Acidic pH Conditions

Separation of antigens and antibodies by immunoaffinity chromatography

Application of peptide chromatography for the isolation of antibodies from bovine skim milk, acid whey and colostrum

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