Enzymatic approaches to new protein conjugates

Grigoletto, Antonella; Maso, Katia; Pasut, Gianfranco

doi:10.1016/b978-0-444-64081-9.00013-9

Cited by 4 publications

(8 citation statements)

References 87 publications

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“…For conjugation to terminal Gal, recombinant GalOx was used to oxidize Gal at position C6, again generating an aldehyde and allowing specific conjugation with biotin-PEG 3 -ONH 2 label. 27–29 Oxidation/ligation proceeded efficiently, but significant side oxidation was observed (Figure 1B). In an alternative approach, we used recombinant human ST6Gal sialyltransferase 1 to derivatize the terminal Gal with an azido-sialic acid (AzSia) residue, which allows subsequent conjugation with alkyne-containing molecules in a click reaction.…”

Section: Resultsmentioning

confidence: 99%

“…64,72 More recently, methodologies harnessing the power of specific enzymes like Sortase, Transglutaminase or Formylglycine-Generating enzyme have expanded the toolbox for natural amino acid-directed conjugation. 29,70,[73][74][75][76][77] These chemoenzymatic strategies allow more control over label:protein ratio and site-specificity as they target specific AA stretches or so-called 'tags'. On the other hand, their very dependency on tags also confines their use, as the protein of interest must tolerate the introduction of a specific consensus sequence without repercussions on protein structure and activity.…”

Section: Discussionmentioning

confidence: 99%

Section: Exploring the Glycan-based Conjugation Landscapementioning

confidence: 99%

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GlyConnect: a glycan-based conjugation extension of the GlycoDelete technology

Thooft

Santens

et al. 2021

Preprint

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Recently, our lab developed GlycoDelete, a technology suite that allows a radical simplification of eukaryotic N-glycosylation. The technology allows to produce glycoproteins that carry single GlcNAc, LacNAc, or LacNAc-Sia type glycans on their N-linked glycosylation sequons. GlycoDelete-type N-glycans are uniquely suited for glycan-based conjugation purposes, as these provide a short, homogeneous and hydrophilic link to the protein backbone. Targeting GlycoDelete-glycans allows for highly site-specific conjugation at sites in the protein which are normally occupied by bulky glycans, thus ensuring minimal interference with protein structure and function. The current manuscript describes the evaluation and optimization of both chemical and chemo-enzymatic conjugation of molecules onto the GlycoDelete-type glycans of a limited set of benchmark proteins

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

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GlyConnect: a glycan-based conjugation extension of the GlycoDelete technology

Thooft

Santens

et al. 2021

Preprint

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“…At the beginning, the chemistry of conjugation was very simple; it was based on the coupling of an electrophilic group in a polymer (i.e., an activated carboxylic of carbonate group) with lysine's nucleophilic ε‐amino group, the most represented and reactive nucleophile in a protein. Following its initial positive results, conjugation chemistry has continued to experience advances with the development of new selective methods of conjugation, some based on the exploitation of enzymes as tools to couple polymers and proteins (Grigoletto & Maso, 2020) or on genetic methods (e.g., thiol conjugation on an inserted cysteine; Basu et al, 2006 or genetic code expansion with Amber codon technique; Wals & Ovaa, 2014), as shown in Figure 5. Proteins have also witnessed new developments; at the beginning they were mainly non‐human enzymes, then recombinant proteins, such as human recombinant proteins expressed in Escherichia coli with or without genetic modification were developed, and now conjugates are prepared with proteins containing a non‐natural amino acid for selective conjugation.…”

Section: Polymer Conjugation For Biotech Drugsmentioning

confidence: 99%

“…Significant advances have been made in conjugation chemistry in connection to: new conjugation sites (Balan et al, 2007; Giorgi, Agusti, & de Lederkremer, 2020; Katre, 2020; Picken, Awwad, Zloh, Khalili, & Brocchini, 2020), the development of enzymatic coupling methods (Grigoletto & Maso, 2020; Grigoletto, Mero, Maso, & Pasut, 2017), and a new protein expression system that makes it possible to incorporate unnatural amino acids into the protein sequence with unique reactivities towards the desired polymer (Axup et al, 2012; Nairn, Grabstein, Shanebeck, Li, & Wang, 2020).…”

Section: Polymer Conjugation For Biotech Drugsmentioning

confidence: 99%

The evolution of polymer conjugation and drug targeting for the delivery of proteins and bioactive molecules

Grigoletto

Tedeschini

Canato

et al. 2020

WIREs Nanomed Nanobiotechnol

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Polymer conjugation can be considered one of the leading approaches within the vast field of nanotechnology‐based drug delivery systems. In fact, such technology can be exploited for delivering an active molecule, such as a small drug, a protein, or genetic material, or it can be applied to other drug delivery systems as a strategy to improve their in vivo behavior or pharmacokinetic activities such as prolonging the half‐life of a drug, conferring stealth properties, providing external stimuli responsiveness, and so on. If on the one hand, polymer conjugation with biotech drug is considered the linchpin of the protein delivery field boasting several products in clinical use, on the other, despite dedicated research, conjugation with low molecular weight drugs has not yet achieved the milestone of the first clinical approval. Some of the primary reasons for this debacle are the difficulties connected to achieving selective targeting to diseased tissue, organs, or cells, which is the main goal not only of polymer conjugation but of all delivery systems of small drugs. In light of the need to achieve better drug targeting, researchers are striving to identify more sophisticated, biocompatible delivery approaches and to open new horizons for drug targeting methodologies leading to successful clinical applications. This article is categorized under: Therapeutic Approaches and Drug Discovery > Emerging Technologies Therapeutic Approaches and Drug Discovery > Nanomedicine for Oncologic Disease Toxicology and Regulatory Issues in Nanomedicine > Regulatory and Policy Issues in Nanomedicine

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Lipoic Acid Ligase A‐Mediated Ligation: Mechanism, Applications, and Emerging Innovations in Bioconjugation

Yamazaki,

Matsuda

2023

ChemistrySelect

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This review outlines the latest findings on bioconjugation using lipoic acid ligase A (LplA), a key enzyme that allows the introduction of new functional groups into biomolecules. LplA functions via a characteristic mechanism of covalently binding orthologous lipoic acid derivatives in vivo to a specific 13 amino acid sequence called the LplA acceptor peptide (LAP). This unique functionality has enabled a wide range of applications, including cell‐surface modification, protein immobilization, fluorescent labeling of antibodies. Recently, the modification of proteins without LAP sequences has been demonstrated, suggesting their potential for future biopharmaceutical production. Although several review articles on enzyme ligation have been published, research on LplA remains limited. This review attempts to fill this gap by introducing LplA based on its mechanism, applications, and recent research reports.

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Enzymatic approaches to new protein conjugates

Cited by 4 publications

References 87 publications

GlyConnect: a glycan-based conjugation extension of the GlycoDelete technology

GlyConnect: a glycan-based conjugation extension of the GlycoDelete technology

The evolution of polymer conjugation and drug targeting for the delivery of proteins and bioactive molecules

Lipoic Acid Ligase A‐Mediated Ligation: Mechanism, Applications, and Emerging Innovations in Bioconjugation

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