Modular, bioorthogonal strategy for the controlled loading of cargo into a protein nanocage

Lise Schoonen, Selma Eising, Mark B. van Eldijk, Jaleesa Bresseleers, Margo Van Der Pijl, Roeland J.M. Nolte, Kimberly M. Bonger, Jan C.M. van Hest

Research output: Contribution to journalArticleAcademicpeer-review

3 Citations (Scopus)

Abstract

Virus capsids, i.e., viruses devoid of their genetic material, are suitable nanocarriers for biomedical applications such as drug delivery and diagnostic imaging. For this purpose, the reliable encapsulation of cargo in such a protein nanocage is crucial, which can be accomplished by the covalent attachment of the compounds of interest to the protein domains positioned at the interior of the cage. This approach is particularly valid for the capsid proteins of the cowpea chlorotic mottle virus (CCMV), which have their N-termini located at the inside of the capsid structure. Here, we examined several site-selective modification methods for covalent attachment and encapsulation of cargo at the N-terminus of the CCMV protein. Initially, we explored approaches to introduce an N-terminal azide functionality, which would allow the subsequent bioorthogonal modification with a strained alkyne to attach the desired cargo. As these methods showed compatibility issues with the CCMV capsid proteins, a strategy based on 2-pyridinecarboxaldehydes for site-specific N-terminal protein modification was employed. This method allowed the successful modification of the proteins, and was applied for the introduction of a bioorthogonal vinylboronic acid moiety. In a subsequent reaction, the proteins could be modified further with a fluorophore using the tetrazine ligation. The application of capsid assembly conditions on the functionalized proteins led to successful particle formation, showing the potential of this covalent encapsulation strategy.

LanguageEnglish
Pages1186-1193
Number of pages8
JournalBioconjugate Chemistry
Volume29
Issue number4
DOIs
StatePublished - 18 Apr 2018

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Bromovirus
Viruses
Proteins
Capsid
Encapsulation
Capsid Proteins
Alkynes
Azides
Diagnostic Imaging
Fluorophores
Ligation
Drug delivery
Acids
Imaging techniques
Pharmaceutical Preparations
Genes

Cite this

Schoonen, Lise ; Eising, Selma ; van Eldijk, Mark B. ; Bresseleers, Jaleesa ; Van Der Pijl, Margo ; Nolte, Roeland J.M. ; Bonger, Kimberly M. ; van Hest, Jan C.M./ Modular, bioorthogonal strategy for the controlled loading of cargo into a protein nanocage. In: Bioconjugate Chemistry. 2018 ; Vol. 29, No. 4. pp. 1186-1193
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Schoonen, L, Eising, S, van Eldijk, MB, Bresseleers, J, Van Der Pijl, M, Nolte, RJM, Bonger, KM & van Hest, JCM 2018, 'Modular, bioorthogonal strategy for the controlled loading of cargo into a protein nanocage' Bioconjugate Chemistry, vol. 29, no. 4, pp. 1186-1193. DOI: 10.1021/acs.bioconjchem.7b00815

Modular, bioorthogonal strategy for the controlled loading of cargo into a protein nanocage. / Schoonen, Lise; Eising, Selma; van Eldijk, Mark B.; Bresseleers, Jaleesa; Van Der Pijl, Margo; Nolte, Roeland J.M.; Bonger, Kimberly M.; van Hest, Jan C.M.

In: Bioconjugate Chemistry, Vol. 29, No. 4, 18.04.2018, p. 1186-1193.

Research output: Contribution to journalArticleAcademicpeer-review

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Schoonen L, Eising S, van Eldijk MB, Bresseleers J, Van Der Pijl M, Nolte RJM et al. Modular, bioorthogonal strategy for the controlled loading of cargo into a protein nanocage. Bioconjugate Chemistry. 2018 Apr 18;29(4):1186-1193. Available from, DOI: 10.1021/acs.bioconjchem.7b00815