The Use of Hydrogen Peroxide for Closing Disulfide Bridges in Peptides

Sidorova, M. V.; Молокоедов, А. С.; Aa, Az'muko; Ev, Kudriavtseva; Krause, Eberhard; Mv, Ovchinnikov; ZhD, Bespalova

doi:10.1023/b:rubi.0000023093.05123.31

Cited by 25 publications

(13 citation statements)

References 7 publications

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“…For disulfide bond conjugation, after dispersion of 0.1 g of the VCM@Fe 3 O 4 /HNT–MPS or OFX@Fe 3 O 4 /HNT–MPS particles in 3.0 mL of cold deionized water (4 °C), H 2 O 2 (30 wt%, 0.2 mL) and Cys-containing peptide chain (3.0 mL, 0.5 M, in ethanol) were added to the mixture drop-by-drop using two separate syringes, during the shaking for 1.5 h. 56 After completion of the process, the obtained VCM@Fe 3 O 4 /HNT–C(WR) 3 and OFX@Fe 3 O 4 /HNT–C(WR) 3 nanocargoes were separated via centrifugation (4000 rpm, 15 min), and washed for two times with deionized water, via vortex mixing and centrifugation. Finally, the particles were redispersed in the deionized water, transferred to the glass Petri dishes, and dried by freeze drier, for 48 h.…”

Section: Methodsmentioning

confidence: 99%

Enhanced antimicrobial treatment by a clay-based drug nanocarrier conjugated to a guanidine-rich cell penetrating peptide

Khodabakhshi

Baghersad

2021

RSC Adv.

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Section: Methodsmentioning

confidence: 99%

Enhanced antimicrobial treatment by a clay-based drug nanocarrier conjugated to a guanidine-rich cell penetrating peptide

Khodabakhshi

Baghersad

2021

RSC Adv.

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“…17 Hydrogen peroxide has been used in the formation of disulfide bonds in multiple synthetic peptides of various lengths and structures. 18 The process is controlled under mild conditions without noticeable side reactions with Trp, Tyr and Met residues. In the biopharmaceutical industry, hydrogen peroxide is frequently used in forced degradation studies to determine possible biologics degradation pathways.…”

Section: Introductionmentioning

confidence: 99%

Using hydrogen peroxide to prevent antibody disulfide bond reduction during manufacturing process

Cheng

Huang

Borwankar

et al. 2018

mAbs

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During large-scale monoclonal antibody manufacturing, disulfide bond reduction of antibodies, which results in generation of low molecule weight species, is occasionally observed. When this happens, the drug substance does not meet specifications. Many investigations have been conducted across the biopharmaceutical industry to identify the root causes, and multiple strategies have been proposed to mitigate the problem. The reduction is correlated with the release of cellular reducing components and depletion of dissolved oxygen before, during, and after harvest. Consequently, these factors can lead to disulfide reduction over long-duration storage at room temperature prior to Protein A chromatography. Several strategies have been developed to minimize antibody reduction, including chemical inhibition of reducing components, maintaining aeration before and after harvest, and chilling clarified harvest during holding. Here, we explore the use of hydrogen peroxide in clarified harvest bulk or cell culture fluid as a strategy to prevent disulfide reduction. A lab-scale study was performed to demonstrate the effectiveness of hydrogen peroxide in preventing antibody reduction using multiple IgG molecules. Studies were done to define the optimal concentration of hydrogen peroxide needed to avoid unnecessary oxidization of the antibody products. We show that adding a controlled amount of hydrogen peroxide does not change product quality attributes of the protein. Since hydrogen peroxide is soluble in aqueous solutions and decomposes into water and oxygen, there is no additional burden involved in removing it during the downstream purification steps. Due to its ease of use and minimal product impact, we demonstrate that hydrogen peroxide treatment is a powerful, simple tool to quench reducing potential by simply mixing it with harvested cell culture fluid.

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“…After that, the resin of peptide was removed by the cleavage mixture with trifluoroacetic acid (TFA), water, thioanisole (TIS), and 1, 2-ethanedithiol (EDT) (94/2/2/2; V / V / V ). Hydrogen peroxide (H 2 O 2 ) was executed to oxidise linear peptides as described previously [ 18 ]. Disulphide bridges were formed in the presence of 0.5% ( w / v ) H 2 O 2 for 0.5 h at room temperature before lyophilisation.…”

Section: Methodsmentioning

confidence: 99%

Modification and Targeted Design of N-Terminal Truncates Derived from Brevinin with Improved Therapeutic Efficacy

Chen

et al. 2020

Biology

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Antimicrobial peptides (AMPs) are a class of molecules that play an essential role in innate immune regulation. The Brevinin-1 family are AMPs that show strong pharmacological and antimicrobial potential. A novel peptide, B1A, was designed based on the primary structure of brevinin-1PLb and brevinin-1PLc. Subsequently, a synthesised replicate was subjected to a series of bioassays and was found to display antimicrobial activity. However, it also displayed high levels of haemolysis in a horse red blood cell haemolytic assay, suggesting potential toxicity. Therefore, we rationally designed a number of B1A analogues with aim of retaining antimicrobial activity, lowering toxicity, and to explore the structure–activity relationship of its N-terminus. B1A and its analogues still retained the “Rana Box” and the FLP-motif, which is a feature of this subfamily. However, the introduction of Lys and Trp residues into the peptide sequences revealed that antimicrobial activity of these analogues remained unchanged once the hydrophobicity and the charge reached the threshold. Hence, the idea that the hydrophobicity saturation in different situations is related to antimicrobial activity can be understood via the structure–activity relationship. Meanwhile, it could also be the starting point for the generation of peptides with specific antimicrobial activity.

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The Use of Hydrogen Peroxide for Closing Disulfide Bridges in Peptides

Cited by 25 publications

References 7 publications

Enhanced antimicrobial treatment by a clay-based drug nanocarrier conjugated to a guanidine-rich cell penetrating peptide

Enhanced antimicrobial treatment by a clay-based drug nanocarrier conjugated to a guanidine-rich cell penetrating peptide

Using hydrogen peroxide to prevent antibody disulfide bond reduction during manufacturing process

Modification and Targeted Design of N-Terminal Truncates Derived from Brevinin with Improved Therapeutic Efficacy

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