Well-defined protein-polymer conjugates via in situ RAFT polymerization

Boyer, C., Bulmus, V., Liu, J., Davis, T. P., Stenzel, M. H., & Barner-Kowollik, C. (2007) Well-defined protein-polymer conjugates via in situ RAFT polymerization. Journal of the American Chemical Society, 129(22).

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Abstract

Biotechnology, biomedicine, and nanotechnology applications would benefit from methods generating well-defined, monodisperse protein-polymer conjugates, avoiding time-consuming and difficult purification steps. Herein, we report the in situ synthesis of protein-polymer conjugates via reversible addition-fragmentation chain transfer polymerization (RAFT) as an efficient method to generate well-defined, homogeneous protein-polymer conjugates in one step, eliminating major postpolymerization purification steps. A water soluble RAFT agent was conjugated to a model protein, bovine serum albumin (BSA), via its free thiol group at Cys-34 residue. The conjugation of the RAFT agent to BSA was confirmed by UV-visible spectroscopy, matrix-assisted laser desorption ionization - time of flight (MALDI-TOF), and 1H NMR. BSA-macroRAFT agent was then used to control the polymerization of two different water soluble monomers, N-isopropylacrylamide (NIPAAm) and hydroxyethyl acrylate (HEA), in aqueous medium at 25 °C. The growth of the polymer chains from BSA-macroRAFT agent was characterized by size exclusion chromatography (SEC), 1H NMR, MALDI-TOF, and polyacrylamide gel electrophoresis (PAGE) analyses. The controlled character of the RAFT polymerizations was confirmed by the linear evolution of molecular weight with monomer conversion. The SEC analyses showed no detectable free, nonconjugated polymer formation during the in situ polymerization. The efficiency of BSA-macroRAFT agent to generate BSA-polymer conjugates was found to be ca. 1 by deconvolution of the SEC traces of the polymerization mixtures. The structural integrity and the conformation-related esterase activity of BSA were found to be unaffected by the polymerization conditions and the conjugation of the polymer chain. BSA-poly(NIPAAm) conjugates showed hybrid temperature-dependent phase separation and aggregation behavior. The lower critical solution temperature values of the conjugates were found to increase with the decrease in molecular weight of poly(NIPAAm) block conjugated to BSA. © 2007 American Chemical Society.

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ID Code: 99144
Item Type: Journal Article
Refereed: Yes
Additional Information: Cited By :253
Export Date: 5 September 2016
CODEN: JACSA
Correspondence Address: Bulmus, V.; Centre for Advanced Macromolecular Design (CAMD), School of Chemical Sciences and Engineering, University of New South Wales, Sydney, NSW 2052, Australia; email: camd@unsw.edu.au
Chemicals/CAS: poly(n isopropylacrylamide), 25189-55-3; Acrylamides; Acrylates; N-isopropylacrylamide, 2210-25-5; Polymers; Proteins; Serum Albumin, Bovine
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Keywords: Hydroxyethyl acrylate (HEA), Laser desorption ionization, Protein-polymer conjugates, Reversible addition-fragmentation chain transfer polymerization (RAFT), Molecular weight, Phase separation, Polymerization, Proteins, Size exclusion chromatography, Synthesis (chemical), Ultraviolet spectroscopy, Conjugated polymers, bovine serum albumin, hydroxyethyl acetic acid, monomer, poly(n isopropylacrylamide), unclassified drug, article, matrix assisted laser desorption ionization time of flight mass spectrometry, polyacrylamide gel electrophoresis, protein polymerization, protein structure, proton nuclear magnetic resonance, reversible addition fragmentation chain transfer polymerization, Acrylamides, Acrylates, Electrophoresis, Polyacrylamide Gel, Nuclear Magnetic Resonance, Biomolecular, Polymers, Serum Albumin, Bovine, Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization, Temperature
DOI: 10.1021/ja070956a
ISSN: 00027863
Divisions: Current > Schools > School of Chemistry, Physics & Mechanical Engineering
Current > Institutes > Institute for Future Environments
Current > QUT Faculties and Divisions > Science & Engineering Faculty
Deposited On: 22 Sep 2016 04:50
Last Modified: 06 Oct 2016 04:17

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