Protein conjugation with amphiphilic block copolymers for enhanced cellular delivery

Xiang Yi, Elena Batrakova, William A. Banks, Serguei V Vinogradov, Alexander V. Kabanov

Research output: Contribution to journalArticle

39 Citations (Scopus)

Abstract

Modification of a model protein, horseradish peroxidase (HRP), with amphiphilic block copolymer poly(ethylene oxide)-b-poly(propylene oxide)-b-poly(ethylene oxide) (Pluronic), was previously shown to enhance the transport of this protein across the blood-brain barrier in vivo and brain microvessel endothelial cells in vitro. This work develops procedures for synthesis and characterization of HRP with Pluronic copolymers, having different lengths of poly(ethylene oxide) (PEO) and poly(propylene oxide) (PPO) blocks. Four monoamine Pluronic derivatives (L81, P85, L121, P123) were synthesized and successfully conjugated to a model protein, HRP, via biodegradable or nondegradable linkers (dithiobis(succinimidyl propionate) (DSP), dimethyl 3,3′-dithiobispropionimidate (DTBP), and disuccinimidyl propionate (DSS)). The conjugation was confirmed by HRP amino group titration, matrix-assisted laser desorption/ionization-time of flight spectroscopy, and cation-exchange chromatography. HRP conjugates containing an average of one to two Pluronic moieties and retaining in most cases over 70% of the activity were synthesized. Increased cellular uptake of these conjugates was demonstrated using the Mardin-Derby canine kidney cell line and primary bovine brain microvessel endothelial cells. The optimal modifications included Pluronic L81 and P85. These copolymers have shorter PPO chains compared to Pluronic P123 and L121, which were less efficient. There was little if any dependence of the uptake on the length of the hydrophilic PEO block for the optimal modifications. The proposed modifications may be used to increase cellular uptake of other proteins.

Original languageEnglish (US)
Pages (from-to)1071-1077
Number of pages7
JournalBioconjugate Chemistry
Volume19
Issue number5
DOIs
StatePublished - May 1 2008

Fingerprint

Poloxamer
Horseradish Peroxidase
Polyethylene oxides
Block copolymers
Ethylene Oxide
Proteins
Polypropylene oxides
Microvessels
Endothelial cells
Endothelial Cells
Brain
Copolymers
Propionates
Blood-Brain Barrier
Cations
Canidae
Chromatography
Spectrum Analysis
Titration
Carrier Proteins

ASJC Scopus subject areas

  • Chemistry(all)
  • Organic Chemistry
  • Clinical Biochemistry
  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Yi, X., Batrakova, E., Banks, W. A., Vinogradov, S. V., & Kabanov, A. V. (2008). Protein conjugation with amphiphilic block copolymers for enhanced cellular delivery. Bioconjugate Chemistry, 19(5), 1071-1077. https://doi.org/10.1021/bc700443k

Protein conjugation with amphiphilic block copolymers for enhanced cellular delivery. / Yi, Xiang; Batrakova, Elena; Banks, William A.; Vinogradov, Serguei V; Kabanov, Alexander V.

In: Bioconjugate Chemistry, Vol. 19, No. 5, 01.05.2008, p. 1071-1077.

Research output: Contribution to journalArticle

Yi, X, Batrakova, E, Banks, WA, Vinogradov, SV & Kabanov, AV 2008, 'Protein conjugation with amphiphilic block copolymers for enhanced cellular delivery', Bioconjugate Chemistry, vol. 19, no. 5, pp. 1071-1077. https://doi.org/10.1021/bc700443k
Yi, Xiang ; Batrakova, Elena ; Banks, William A. ; Vinogradov, Serguei V ; Kabanov, Alexander V. / Protein conjugation with amphiphilic block copolymers for enhanced cellular delivery. In: Bioconjugate Chemistry. 2008 ; Vol. 19, No. 5. pp. 1071-1077.
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