Noncovalent PEGylation, An Innovative Subchapter in the Field of Protein Modification

Reichert, Christian; Borchard, Gerrit

doi:10.1002/jps.24692

Cited by 25 publications

(18 citation statements)

References 31 publications

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“…Compared to the SS-βCD branches, PEGylated HER branches (SS-PEG-SS-HER) were long, bulky, hydrophilic, and flexible. In an aqueous environment, SS-PEG-SS-HER chains surround the small SS-βCD blocks on the surface of CNPs to improve the solubility of DL-CNPs and the protection of DOX in βCD from premature release in blood vessels [41][42][43]. When the DL-CNPs are delivered to the tumor, they are degraded in response to the high concentration of GSH, and DOX in βCD is released with exposure to an acidic environment.…”

Section: Loading and External Stimuli-trigger Release Of Doxmentioning

confidence: 99%

Dual pH- and GSH-Responsive Degradable PEGylated Graphene Quantum Dot-Based Nanoparticles for Enhanced HER2-Positive Breast Cancer Therapy

Van

Hong

et al. 2020

Nanomaterials

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Dual stimuli-responsive degradable carbon-based nanoparticles (DS-CNPs) conjugated with Herceptin (HER) and polyethylene glycol (PEG) have been designed for the treatment of HER2-positive breast cancer. Each component has been linked through disulfide linkages that are sensitive to glutathione in a cancer microenvironment. β-cyclodextrin (β-CD) on the surface of DS-CNPs formed an inclusion complex (DL-CNPs) with doxorubicin (DOX) at a high loading capacity of 5.3 ± 0.4%. In response to a high level of glutathione (GSH) and low pH in a tumor environment, DL-CNPs were rapidly degraded and released DOX in a controlled manner via disruption of host–guest inclusion. These novel DL-CNPs exhibited high cellular uptake with low toxicity, which induced the efficient inhibition of antitumor activity both in vitro and in vivo. Cell viability, confocal laser scanning microscopy, and animal studies indicate that DL-CNPs are a great platform with a synergistically enhanced antitumor effect from the dual delivery of HER and DOX in DL-CNPs.

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Section: Loading and External Stimuli-trigger Release Of Doxmentioning

confidence: 99%

Dual pH- and GSH-Responsive Degradable PEGylated Graphene Quantum Dot-Based Nanoparticles for Enhanced HER2-Positive Breast Cancer Therapy

Van

Hong

et al. 2020

Nanomaterials

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Section: Introductionmentioning

confidence: 99%

“…Moreover, due to the flexible PEG chain around the protein, it wraps around the molecule masking sites or epitopes recognized by the immune cells, receptors or proteolytic enzymes, thus increasing its half-life and lowering the immunotoxicity. 5 Several therapeutic PEGylated proteins have recently been employed clinically such as those for interferon-α, granulocyte colony-stimulating factor, urate oxidase and a monoclonal antibody against tumor necrosis factor-α. [6][7][8][9] We have previously succeeded in conjugating a PEG polymer to the therapeutic enzyme glucarpidase (carboxypeptidase G2 [CPG2]) known to detoxify methotrexate (MTX) in an overdose and to activate the benzoic mustard inactive prodrugs to highly cytotoxic drugs, and we have generated a PEGylated biobetter glucarpidase variant (CPG2).…”

Section: Introductionmentioning

confidence: 99%

Production of Long-Acting CNGRC–CPG2 Fusion Proteins: New Derivatives to Overcome Drug Immunogenicity of Ligand-Directed Enzyme Prodrug Therapy for Targeted Cancer Treatment

Al-Mansoori

Qahtani

Elsinga

et al. 2021

Technol Cancer Res Treat

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Objectives: Aminopeptidase N (APN) is an enzyme highly expressed in metastatic cancers and could be used in targeted cancer therapy. Our previous work showed the successful construction of CNGRC–carboxypeptidase G2 (CPG2) and CNGRC–CPG2–CNGRC fusion proteins. Our conjugates and prodrugs were effective in targeting high APN-expressing cancer cells. In the present study, we aim to produce long-acting fusion proteins to overcome 2 of the main drawbacks of antibody-directed enzyme prodrug therapy. Methods: N-terminal and N-, C-terminal fusion CPG2, CNGRC–CPG2, and CNGRC–CPG2–CNGRC, respectively, were PEGylated using polyethylene glycol (PEG) maleimide (40K). We examined the effect of PEGylation on the therapeutic efficacy of the new products. The resulting PEGylated fusion proteins were tested for their stability, ex vivo immunotoxicity, binding capacity to their target on high HT1080, and low A549 APN-expressing cells. The catalytic activity of the resulting PEGylated fusion CPG2 proteins was investigated. Pro-drug “ZD2767P” cytotoxic effect in association with PEG CPG2–CNGRC fusion proteins on cancer cells was studied. Results: Our work demonstrated that the properties of the PEGylated single-fused proteins were significantly improved over that of un-PEGylated fused CPG2, and its kinetic activity and APN-binding affinity were not negatively affected by the PEGylation. Significantly, The PEGylated single-fused CPG2 had lower immunogenicity than the un-PEGylated CPG2. Our results, however, were different in the case of the PEGylated double-fused CPG2. Although its stability in human serum under physiological conditions was not significantly affected, the kinetic activity and its binding affinity to their cellular marker (APN) were substantially reduced. When the study was performed with high and low APN-expressing cancer cell lines, using the prodrug ZD2767p, the PEGylated fusion CPG2 demonstrated cancer cell killing effects. Conclusion: We have successfully produced PEGylated-CNGRC–CPG2, which is bioactive and with lower immunogenicity in ligand-directed enzyme prodrug therapy for cancer treatment.

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“…Non-covalent PEGylation is an innovative approach in which a chemical reaction between protein and PEG is avoided (Reichert and Borchard, 2016). It is based on the mechanisms of hydrophobic interactions (Mueller et al, 2011a;Mueller et al, 2011b;Mueller et al, 2012), ionic interactions (Khondee et al, 2011), protein polyelectrolyte complex (Kurinomaru and Shiraki, 2015;Kurinomaru et al, 2017), or chelation (Mero et al, 2011).…”

Section: Non-covalent Pegylationmentioning

confidence: 99%

“…It is based on the mechanisms of hydrophobic interactions (Mueller et al, 2011a;Mueller et al, 2011b;Mueller et al, 2012), ionic interactions (Khondee et al, 2011), protein polyelectrolyte complex (Kurinomaru and Shiraki, 2015;Kurinomaru et al, 2017), or chelation (Mero et al, 2011). The main advantage of this technique is that it eliminates a potential loss of product due to additional purification processes (Reichert and Borchard, 2016). However, the release of the protein during storage is an important shortcoming for this approach (Santos et al, 2018).…”

Section: Non-covalent Pegylationmentioning

confidence: 99%

From Synthesis to Characterization of Site-Selective PEGylated Proteins

et al. 2019

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Covalent attachment of therapeutic proteins to polyethylene glycol (PEG) is widely used for the improvement of its pharmacokinetic and pharmacological properties, as well as the reduction in reactogenicity and related side effects. This technique named PEGylation has been successfully employed in several approved drugs to treat various diseases, even cancer. Some methods have been developed to obtain PEGylated proteins, both in multiple protein sites or in a selected amino acid residue. This review focuses mainly on traditional and novel examples of chemical and enzymatic methods for site-selective PEGylation, emphasizing in N-terminal PEGylation, that make it possible to obtain products with a high degree of homogeneity and preserve bioactivity. In addition, the main assay methods that can be applied for the characterization of PEGylated molecules in complex biological samples are also summarized in this paper.

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Noncovalent PEGylation, An Innovative Subchapter in the Field of Protein Modification

Cited by 25 publications

References 31 publications

Dual pH- and GSH-Responsive Degradable PEGylated Graphene Quantum Dot-Based Nanoparticles for Enhanced HER2-Positive Breast Cancer Therapy

Dual pH- and GSH-Responsive Degradable PEGylated Graphene Quantum Dot-Based Nanoparticles for Enhanced HER2-Positive Breast Cancer Therapy

Production of Long-Acting CNGRC–CPG2 Fusion Proteins: New Derivatives to Overcome Drug Immunogenicity of Ligand-Directed Enzyme Prodrug Therapy for Targeted Cancer Treatment

From Synthesis to Characterization of Site-Selective PEGylated Proteins

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