How PEGylation influences protein conformational stability

Lawrence, Paul B.; Price, Joshua L.

doi:10.1016/j.cbpa.2016.08.006

Cited by 97 publications

(89 citation statements)

References 58 publications

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“…In this work, a β-sheet conformation was observed for DDAAAAAA and DDVVVVVV peptides, which was not affected after conjugation with PEG. Actually, the effect of PEGylation on conformational stability of proteins or peptides is not well clarified 26 . In some studies, no effect was observed on the secondary structure of epta-or octapeptides after PEGylation 27 .…”

Section: Resultsmentioning

confidence: 99%

PEGylation affects the self-assembling behaviour of amphiphilic octapeptides

Perinelli

Campana

Singh

et al. 2019

International Journal of Pharmaceutics

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Surfactant-like peptides are a class of amphiphilic macromolecules, which are able to self-assemble in water forming different supramolecular structures. Among them, octapeptides composed of six hydrophobic and two hydrophilic residues have attracted interest since they have a length similar to those of natural phospholipids. Supramolecular structures of different amphiphilic octapeptides have been widely reported, but no study has been performed aimed at investigating the effect of PEGylation on their self-assembling behaviour. The aim of the present work was to synthesize and characterise the self-assembling behaviour of PEGylated alanine-or valine based amphiphilic octapeptides (mPEG1.9kDa-DDAAAAAA and mPEG1.9kDa-DDVVVVVV) in comparison to the non-PEGylated ones (DDAAAAAA and DDVVVVVV). The self-aggregation process in ultrapure water was investigated by fluorescence spectroscopy, small angle neutron scattering (SANS), dynamic light scattering (DLS), while the secondary structure was assessed by circular dichroism. PEGylation markedly affects the self-assembling behaviour of these amphiphilic octapeptides in terms of both critical aggregation concentration (CAC) and shape of the formed supramolecular aggregates. Indeed, PEGylation increases CAC and prevents the self-aggregation into fibrillary supramolecular aggregates (as observed for non-PEGylated peptides), by promoting the formation of micelle-like structures (as demonstrated for valine-based octapeptide). 2 On the other side, the secondary structure of peptides seems not to be affected by PEGylation. Overall, these results suggest that self-assembling behaviour of amphiphilic octapeptides can be modified by PEGylation, with a great potential impact for the future applications of these nanomaterials.

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Section: Resultsmentioning

confidence: 99%

PEGylation affects the self-assembling behaviour of amphiphilic octapeptides

Perinelli

Campana

Singh

et al. 2019

International Journal of Pharmaceutics

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“…due to proteolytic degradation 26 . In this study, therefore, we adopted a different approach to produce biobetter glucarpidases, which should both reduce the potential for antibodies production and also protect the protein against endogenous proteases.…”

Section: Discussionmentioning

confidence: 99%

Production of “biobetter” glucarpidase variants to improve drug detoxification and antibody directed enzyme prodrug therapy for cancer treatment

Al-Qahtani

Al-Mansoori

Bashraheel

et al. 2019

European Journal of Pharmaceutical Sciences

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“…In a Fab, both VH/VL and CH/CL interactions contribute to the functional stability of the antibody fragment and potentially to the in vivo toxin neutralization capacity as suggested previously [66]. In addition, one can consider site-specific PEGylation of scFvs for enhancing the pharmacokinetic properties, the conformational stability, protection from proteolysis and immunogenicity [67]. Altogether, these findings represent a proof of concept on humanized mouse antibodies specific to animal toxins, encouraging the optimized production of molecules for in vivo assessment.…”

Section: Discussionmentioning

confidence: 99%

Loxoscelism: Advances and Challenges in the Design of Antibody Fragments with Therapeutic Potential

Karim-Silva

Becker-Finco

Jiacomini

et al. 2020

Toxins

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Envenoming due to Loxosceles spider bites still remains a neglected disease of particular medical concern in the Americas. To date, there is no consensus for the treatment of envenomed patients, yet horse polyclonal antivenoms are usually infused to patients with identified severe medical conditions. It is widely known that venom proteins in the 30-35 kDa range with sphingomyelinase D (SMasesD) activity, reproduce most of the toxic effects observed in loxoscelism. Hence, we believe that monoclonal antibody fragments targeting such toxins might pose an alternative safe and effective treatment. In the present study, starting from the monoclonal antibody LimAb7, previously shown to target SMasesD from the venom of L. intermedia and neutralize its dermonecrotic activity, we designed humanized antibody V-domains, then produced and purified as recombinant single-chain antibody fragments (scFvs). These molecules were characterized in terms of humanness, structural stability, antigen-binding activity, and venom-neutralizing potential. Throughout this process, we identified some blocking points that can impact the Abs antigen-binding activity and neutralizing capacity. In silico analysis of the antigen/antibody amino acid interactions also contributed to a better understanding of the antibody's neutralization mechanism and led to reformatting the humanized antibody fragment which, ultimately, recovered the functional characteristics for efficient in vitro venom neutralization. Key Contribution:The humanization of a mouse antibody V-domains able to neutralize Loxosceles intermedia venom paves the way for a new generation of anti-venom therapy. This study highlights the importance of understanding the antibody's mechanism of neutralization for appropriate design of innovative antidotes. Likewise, preserving favorable physico-chemical properties and antigen-binding activity is a challenge yet to be further addressed when considering pre-clinical trials.

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How PEGylation influences protein conformational stability

Cited by 97 publications

References 58 publications

PEGylation affects the self-assembling behaviour of amphiphilic octapeptides

PEGylation affects the self-assembling behaviour of amphiphilic octapeptides

Production of “biobetter” glucarpidase variants to improve drug detoxification and antibody directed enzyme prodrug therapy for cancer treatment

Loxoscelism: Advances and Challenges in the Design of Antibody Fragments with Therapeutic Potential

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