Improved biocompatibility of protein encapsulation in sol-gel materials

A.M. Macmillan, D. Panek, C.D. McGuinness, J.C. Pickup, D. Graham, W.E. Smith, D.J.S. Birch, J. Karolin

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

By using the fluorescent dye 6-propionyl-2-(N,N-dimethylamino) naphthalene (PRODAN) to monitor methanol generated during tetramethyl orthosilicate polymerization we have optimised the encapsulation of protein in silica sol-gel monoliths with respect to completion of hydrolysis and distillation in order to remove methanol such that protein can be added without denaturation. A minimum of 24 h at +4 °C was found to be required before hydrolysis is complete and 3-5 min of vacuum distillation at 50 °C and 300 mbar needed to remove methanol before the gel is formed. The biocompatibility of a tetramethyl orthosilicate sol-gel monolith was demonstrated by preserving the trimer protein allophycocyanin (APC) in its native form for up to 500 h. This obviates the previously essential requirement of covalently binding the trimer together in order to prevent dissociation into monomers and has enabled observation of native APC trimer in a sol-gel pore for the first time down to the single molecule level using combined fluorescence spectroscopy and confocal microscopy. The higher stability afforded by the protocol we describe could impact on the application of sol-gel materials to single-molecule studies of wider bearing such as protein folding and aggregation.
LanguageEnglish
Pages380-384
Number of pages4
JournalJournal of Sol-Gel Science and Technology
Volume49
Issue number3
DOIs
Publication statusPublished - Mar 2009

Fingerprint

biocompatibility
Biocompatibility
Encapsulation
Sol-gels
gels
proteins
Proteins
Methanol
trimers
Distillation
methyl alcohol
distillation
Hydrolysis
Bearings (structural)
hydrolysis
Protein folding
Denaturation
Molecules
Confocal microscopy
Fluorescence spectroscopy

Keywords

  • PRODAN
  • Fluorescence
  • Allophycocyanin
  • Silica
  • Sol-gel
  • single molecule
  • Porous-media
  • aggregation

Cite this

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title = "Improved biocompatibility of protein encapsulation in sol-gel materials",
abstract = "By using the fluorescent dye 6-propionyl-2-(N,N-dimethylamino) naphthalene (PRODAN) to monitor methanol generated during tetramethyl orthosilicate polymerization we have optimised the encapsulation of protein in silica sol-gel monoliths with respect to completion of hydrolysis and distillation in order to remove methanol such that protein can be added without denaturation. A minimum of 24 h at +4 °C was found to be required before hydrolysis is complete and 3-5 min of vacuum distillation at 50 °C and 300 mbar needed to remove methanol before the gel is formed. The biocompatibility of a tetramethyl orthosilicate sol-gel monolith was demonstrated by preserving the trimer protein allophycocyanin (APC) in its native form for up to 500 h. This obviates the previously essential requirement of covalently binding the trimer together in order to prevent dissociation into monomers and has enabled observation of native APC trimer in a sol-gel pore for the first time down to the single molecule level using combined fluorescence spectroscopy and confocal microscopy. The higher stability afforded by the protocol we describe could impact on the application of sol-gel materials to single-molecule studies of wider bearing such as protein folding and aggregation.",
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Improved biocompatibility of protein encapsulation in sol-gel materials. / Macmillan, A.M.; Panek, D.; McGuinness, C.D.; Pickup, J.C.; Graham, D.; Smith, W.E.; Birch, D.J.S.; Karolin, J.

In: Journal of Sol-Gel Science and Technology, Vol. 49, No. 3, 03.2009, p. 380-384.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Improved biocompatibility of protein encapsulation in sol-gel materials

AU - Macmillan, A.M.

AU - Panek, D.

AU - McGuinness, C.D.

AU - Pickup, J.C.

AU - Graham, D.

AU - Smith, W.E.

AU - Birch, D.J.S.

AU - Karolin, J.

PY - 2009/3

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N2 - By using the fluorescent dye 6-propionyl-2-(N,N-dimethylamino) naphthalene (PRODAN) to monitor methanol generated during tetramethyl orthosilicate polymerization we have optimised the encapsulation of protein in silica sol-gel monoliths with respect to completion of hydrolysis and distillation in order to remove methanol such that protein can be added without denaturation. A minimum of 24 h at +4 °C was found to be required before hydrolysis is complete and 3-5 min of vacuum distillation at 50 °C and 300 mbar needed to remove methanol before the gel is formed. The biocompatibility of a tetramethyl orthosilicate sol-gel monolith was demonstrated by preserving the trimer protein allophycocyanin (APC) in its native form for up to 500 h. This obviates the previously essential requirement of covalently binding the trimer together in order to prevent dissociation into monomers and has enabled observation of native APC trimer in a sol-gel pore for the first time down to the single molecule level using combined fluorescence spectroscopy and confocal microscopy. The higher stability afforded by the protocol we describe could impact on the application of sol-gel materials to single-molecule studies of wider bearing such as protein folding and aggregation.

AB - By using the fluorescent dye 6-propionyl-2-(N,N-dimethylamino) naphthalene (PRODAN) to monitor methanol generated during tetramethyl orthosilicate polymerization we have optimised the encapsulation of protein in silica sol-gel monoliths with respect to completion of hydrolysis and distillation in order to remove methanol such that protein can be added without denaturation. A minimum of 24 h at +4 °C was found to be required before hydrolysis is complete and 3-5 min of vacuum distillation at 50 °C and 300 mbar needed to remove methanol before the gel is formed. The biocompatibility of a tetramethyl orthosilicate sol-gel monolith was demonstrated by preserving the trimer protein allophycocyanin (APC) in its native form for up to 500 h. This obviates the previously essential requirement of covalently binding the trimer together in order to prevent dissociation into monomers and has enabled observation of native APC trimer in a sol-gel pore for the first time down to the single molecule level using combined fluorescence spectroscopy and confocal microscopy. The higher stability afforded by the protocol we describe could impact on the application of sol-gel materials to single-molecule studies of wider bearing such as protein folding and aggregation.

KW - PRODAN

KW - Fluorescence

KW - Allophycocyanin

KW - Silica

KW - Sol-gel

KW - single molecule

KW - Porous-media

KW - aggregation

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SN - 0928-0707

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