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Colloid and Polymer Science

, Volume 293, Issue 12, pp 3517–3526 | Cite as

pH-responsive double hydrophilic protein-polymer hybrids and their self-assembly in aqueous solution

  • Naipu HeEmail author
  • Zhenwu Lu
  • Weigang Zhao
Original Contribution

Abstract

Double hydrophilic protein-polymer hybrids were successfully prepared by “grafting-to” method. Sodium polyacrylate (PANa) with an activated end group was prepared using acrylic acid as initial monomer by atom transfer radical polymerization (ATRP) in the water/DMF mixture at pH 7.8. Then, the activated end group of PANa directly covalently binds to the primary amino groups of the lysine of bovine serum albumin (BSA) in phosphate buffer solution (PBS) at pH 7.4. BSA-PANa hybrids were characterized by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and matrix-associated laser desorption/ionization time-of-flight mass spectra (MALDI-ToF MS). UV-vis spectrometer, transmission electron microscopy (TEM), and dynamic light scatter (DLS) were used to study their self-assembly in PBS at different pH. The resulting BSA-PANa hybrids were pH-responsive. They were found to form spherical structures with narrow distribution in PBS at pH 7.4. In PBS at pH 2.5, fewer spherical particles with broad distribution were visible. Meanwhile, spherical particles had a very thin wall with thickness of 10 nm.

Graphical Abstract

The table of contents (ToC)

The resulting BSA-PANa hybrids were found to be pH-responsive. The self-assembly aggregates were found to form spherical structures with a diameter of approximately 100 nm phosphate buffer solution (PBS) at pH 7.4. In PBS at pH 2.5, the diameters of spherical particles varied between 80 and 200 nm. Meanwhile, spherical particles had a very thin wall with thickness of 10 nm.

Keywords

pH-responsive polymer Double hydrophilic block copolymer Protein-polymer hybrids Self-assembly 

Abbreviations

PPHs

Protein-polymer hybrids

AA

Acrylic acid

PAA

Poly(acrylic acid)

PANa

Sodium polyacrylate

ATRP

Atom transfer radical polymerization

BSA

Bovine serum albumin

SDS-PAGE

Sodium dodecyl sulfate polyacrylamide gel electrophoresis

MALDI-ToF MS

Matrix-associated laser desorption/ionization time-of-flight mass spectra

TEM

Transmission electron microscopy

PBS

Phosphate buffer solution

DLS

Dynamic light scatter

Notes

Acknowledgments

We gratefully acknowledge the National Natural Science Foundation of China (21164003) and Gansu Province Natural Science Foundation (1308RJZA137) for financial support.

References

  1. 1.
    Maynard HD (2013) Proteins in a pill. Nat Chem 5:557–558CrossRefGoogle Scholar
  2. 2.
    Klok H-A (2009) Peptide/Protein-synthetic polymer conjugates: Quo Vadis. Macromolecules 42:7990–8000CrossRefGoogle Scholar
  3. 3.
    Witus LS, Francis MB (2011) Using synthetically modified proteins to make new materials. Acc Chem Res 44:774–783CrossRefGoogle Scholar
  4. 4.
    Börner HG (2009) Strategies exploiting functions and self-assembly properties of bioconjugates for polymer and materials sciences. Prog Polym Sci 34:811–851CrossRefGoogle Scholar
  5. 5.
    Velonia K, Rowan AE, Nolte RJM (2002) Lipase polystyrene giant amphiphiles. J Am Chem Soc 124:4224–4225CrossRefGoogle Scholar
  6. 6.
    Boerakker MJ, Botterhuis NE, Bomans PHH, Frederik PM, Meijer EM, Nolte RJM, Sommerdijk NAJM (2006) Aggregation behavior of giant amphiphiles prepared by cofactor reconstitution. Chem Eur J 12:6071–6080CrossRefGoogle Scholar
  7. 7.
    Reynhout IC, Cornelissen JJLM, Nolte RJM (2009) Synthesis of polymer-biohybrids: from small to giant surfactants. Acc Chem Res 42:681–692CrossRefGoogle Scholar
  8. 8.
    Thomas CS, Glassman MJ, Olsen BD (2011) Solid-state nanostructured materials from self-assembly of a globular protein-polymer diblock copolymer. ACS Nano 5:5697–5707CrossRefGoogle Scholar
  9. 9.
    Lam CN, Kim M, Thomas CS, Chang D, Sanoja GE, Okwara CU, Olsen BD (2014) The nature of protein inter-actions governing globular protein-polymer block copolymer self-assembly. Biomacromolecules 15:1248–1258CrossRefGoogle Scholar
  10. 10.
    Thomas CS, Xu L, Olsen BD (2013) Effect of small molecule osmolytes on the self-assembly and functionality of globular protein-polymer diblock copolymers. Biomacromolecules 14:3064–3072CrossRefGoogle Scholar
  11. 11.
    Chang D, Lam CN, Tang S, Olsen BD (2014) Effect of polymer chemistry on globular protein-polymer block copolymer self-assembly. Polym Chem 5:4884–4895CrossRefGoogle Scholar
  12. 12.
    Lavigueur C, Garc JG, Hendriks L, Hoogenboom R, Cornelissen JJLM, Nolte RJM (2011) Thermoresponsive giant biohybrid amphiphiles. Polym Chem 2:333–340CrossRefGoogle Scholar
  13. 13.
    Matsumoto NM, Prabhakaran P, Rome LH, Maynard HD (2013) Smart Vaults: thermally-responsive protein nanocapsules. ACS Nano 7:867–874CrossRefGoogle Scholar
  14. 14.
    He N, He Y, Wang R, Song P, Zhou Y (2010) Protein-polymer conjugates. Prog Chem 22:2388–2396Google Scholar
  15. 15.
    He N, Wang R (2012) Self-assembly of protein with polymer. Prog Chem 24:94–100Google Scholar
  16. 16.
    He N, Lu S, Zhao W, Du X, Huang S, Wang R (2014) Fabrication of the self-assembly systems based on protein molecules. Prog Chem 26:303–309Google Scholar
  17. 17.
    Velonia K (2010) Protein-polymer amphiphilic chimeras: recent advances and future challenges. Polym Chem 1:944–952CrossRefGoogle Scholar
  18. 18.
    He W, Jiang H, Zhang L, Cheng Z, Zhu X (2013) Atom transfer radical polymerization of hydrophilic monomers and its applications. Polym Chem 4:2919–2938CrossRefGoogle Scholar
  19. 19.
    Colombani O, Ruppel M, Schubert F, Zettl H, Pergushov DV, Müller AHE (2007) Synthesis of poly(n-butyl acrylate)-block-poly(acrylic acid) diblock copolymers by ATRP and their micellization in water. Macromolecules 40:4338–4350CrossRefGoogle Scholar
  20. 20.
    Shedge A, Colombani O, Nicolai T, Chassenieux C (2014) Charge dependent dynamics of transient networks and hydrogels formed by self-assembled pH-sensitive triblock copolyelectrolytes. Macromolecules 47:2439–2444CrossRefGoogle Scholar
  21. 21.
    Ye P, Dong H, Zhong M, Matyjaszewski K (2011) Synthesis of binary polymer brushes via two-step reverse atom transfer radical polymerization. Macromolecules 44:2253–2260CrossRefGoogle Scholar
  22. 22.
    Cheng G, Bolker A, Zhang M, Krausch G, Müller AHE (2001) Amphiphilic cylindrical core-shell brushes via a “grafting from” process using ATRP. Macromolecules 34:6883–6888CrossRefGoogle Scholar
  23. 23.
    Delcroix MF, Huet GL, Conard T, Demoustier-Champagne S, Du Prez FE, Landoulsi J, Dupont-Gillain CC (2013) Design of mixed PEO/PAA brushes with switchable properties toward protein adsorption. Biomacromolecules 14:215–225CrossRefGoogle Scholar
  24. 24.
    Zhang W, Yuan J, Weiss S, Ye X, Li C, Müller AHE (2011) Telechelic hybrid poly(acrylic acid)s containing polyhedral oligomeric silsesquioxane (POSS) and their self-assembly in water. Macromolecules 44:6891–6898CrossRefGoogle Scholar
  25. 25.
    Chu C-C, Tsai Y-J, Hsiao L-C, Wang L (2011) Controlled self-aggregation of C60-anchored multiarmed polyacrylic acids and their cytotoxicity evaluation. Macromolecules 44:7056–7061CrossRefGoogle Scholar
  26. 26.
    Ashford EJ, Naldi V, O’Dell R, Billingham NC, Armes SP (1999) First example of the atom transfer radical polymerisation of an acidic monomer: direct synthesis of methacrylic acid copolymers in aqueous media. Chem Commun (14):1285–1286. doi: https://doi.org/10.1039/A903773J
  27. 27.
    Sumerlin BS (2012) Proteins as initiators of controlled radical polymerization: grafting from via ATRP and RAFT. ACS Macro Lett 1:141–145CrossRefGoogle Scholar
  28. 28.
    Le Droumaguet B, Nicolas J (2010) Recent advances in the design of bioconjugates from controlled/living radical polymerization. Polym Chem 1:563–598CrossRefGoogle Scholar
  29. 29.
    Averick S, Simakova A, Park S, Konkolewicz D, Magenau AJD, Mehl RA, Matyjaszewski K (2012) ATRP under biologically relevant conditions: grafting from a protein. ACS Macro Lett 1:6–10CrossRefGoogle Scholar
  30. 30.
    Osborne V L, Jones DM, Huck WTS (2002) Controlled growth of triblock polyelectrolyte brushes. Chem Commun (17):1838–1839. doi: https://doi.org/10.1039/B204737C
  31. 31.
    Dong R, Krishnan S, Baird BA, Lindau M, Ober CK (2007) Patterned biofunctional poly(acrylic acid) brushes on silicon surfaces. Biomacromolecules 8:3082–3092CrossRefGoogle Scholar
  32. 32.
    Lele BS, Murata H, Matyjaszewski K, Russell AJ (2005) Synthesis of uniform protein-polymer conjugates. Biomacromolecules 6:3380–3387CrossRefGoogle Scholar
  33. 33.
    Abuchowski A, van Es T, Palczuk NC, Davis FF (1997) Alteration of immunological properties of bovine serum albumin by covalent attachment of polyethylene glycol. J Biol Chem 252:3578–3581Google Scholar
  34. 34.
    Abuchowski A, van Es T, Palczuk NC, Davis FF (1997) Effect of covalent attachment of polyethylene glycol on immunogenicity and circulating life of bovine liver catalase. J Biol Chem 252:3582–3586Google Scholar
  35. 35.
    Bontempo D, Heredia KL, Fish BA, Maynard HD (2004) Cysteine-reactive polymers synthesized by atom transfer radical polymerization for conjugation to proteins. J Am Chem Soc 126:15372–15373CrossRefGoogle Scholar
  36. 36.
    Lecolley F, Tao L, Mantovani G, Durkin I, Lautru S, Haddleton DM (2004) A new approach to bioconjugates for proteins and peptides (“pegylation”) utilising living radical polymerization. Chem Commun (18):2026–2027. doi: https://doi.org/10.1039/B407712A
  37. 37.
    Bontempo D, Maynard HD (2005) Streptavidin as a macroinitiator for polymerization: in situ protein-polymer conjugate formation. J Am Chem Soc 127:6508–6509CrossRefGoogle Scholar
  38. 38.
    Simakova A, Averick SE, Konkolewicz D, Matyjaszewski K (2012) Aqueous ARGET ATRP. Macromolecules 45:6371–6379CrossRefGoogle Scholar
  39. 39.
    He N, Wang R, He Y, Dang X (2012) Fabrication, structure and surface charges of albumin-chitosan hybrids. Sci China Chem 55:1788–1795CrossRefGoogle Scholar
  40. 40.
    Le Droumaguet B, Mantovani G, Haddletonb DM, Velonia K (2007) Formation of giant amphiphiles by post-functionalization of hydrophilic protein-polymer conjugates. J Mater Chem 17:1916–1922CrossRefGoogle Scholar
  41. 41.
    Le Droumaguet B, Velonia K (2008) In situ ATRP-mediated hierarchical formation of giant amphiphile Bionanoreactors. Angew Chem Int Ed 47:6263–6266CrossRefGoogle Scholar
  42. 42.
    Dirks AJ, van Berkel SS, Hatzakis NS, Opsteen JA, van Delft FL, Cornelissen JJLM, Rowan AE, van Hest JCM, Rutjes FPJT, Nolte RJM (2005) Preparation of biohybrid amphiphiles via the copper catalysed Huisgen [3 + 2] dipolar cycloaddition reaction. Chem Commun (33):4172–4174. doi: https://doi.org/10.1039/B508428H
  43. 43.
    Liu Z, Dong C, Wang X, Wang H, Li W, Tan J, Chang J (2014) Self-assembled biodegradable protein-polymer vesicle as a tumor-targeted nanocarrier. ACS Appl Mater Interfaces 6:2393–2400CrossRefGoogle Scholar
  44. 44.
    Reynhout IC, Cornelissen JJLM, Nolte RJM (2007) Self-assembled architectures from biohybrid triblock copolymers. J Am Chem Soc 129:2327–2332CrossRefGoogle Scholar
  45. 45.
    Kadir MA, Lee C, Han HS, Kim B-S, Ha E-J, Jeong J, Song JK, Lee S-G, And SSA, Paik H (2013) In situ formation of polymer–protein hybrid spherical aggregates from (nitrilotriacetic acid)-endfunctionalized polystyrenes and His-tagged proteins. Polym Chem 4:2286–2292CrossRefGoogle Scholar

Copyright information

© Springer-Verlag Berlin Heidelberg 2015

Authors and Affiliations

  1. 1.College of Chemical and Biological EngineeringLanzhou Jiaotong UniversityLanzhouChina

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