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Surface Immobilization of Synthetic Proteins Via Plasma Polymer Interlayers

Published online by Cambridge University Press:  10 February 2011

Hans J Griesser*
Affiliation:
CSIRO Molecular Science, Clayton Laboratory, Private Bag 10. Clayton South, Victoria 3169, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
Keith M McLean
Affiliation:
CSIRO Molecular Science, Clayton Laboratory, Private Bag 10. Clayton South, Victoria 3169, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
Gerrit J Beumer
Affiliation:
CSIRO Molecular Science, Clayton Laboratory, Private Bag 10. Clayton South, Victoria 3169, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
Xiaoyi Gong
Affiliation:
CSIRO Molecular Science, Clayton Laboratory, Private Bag 10. Clayton South, Victoria 3169, Australia.
Peter Kingshot
Affiliation:
CSIRO Molecular Science, Clayton Laboratory, Private Bag 10. Clayton South, Victoria 3169, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
Graham Johnson
Affiliation:
CSIRO Molecular Science, North Ryde Laboratory, PO Box 184, North Ryde, New South Wales, 2113, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
John G Steele
Affiliation:
CSIRO Molecular Science, North Ryde Laboratory, PO Box 184, North Ryde, New South Wales, 2113, Australia. Co-operative Research Centre for Eye Research and Technology, University of New South Wales, Sydney 2052, Australia.
*
*corresponding author; e-mail: h.griesser@molsci.csiro.au
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Abstract

Coatings of biologically active molecules on synthetic ”bulk“materials are of much interest for biomedical applications since they can in principle elicit specific, predictable. controlled responses of the host environment to an implanted device. However, issues such as shelf life. storage conditions, biological safety, and enzymatic attack in the biological environment must be considered; synthetic proteins may offer advantages. In this study we investigated the covalent immobilization onto polymeric materials of synthetic proteins which possess some properties that mimic those of the natural protein collagen, particularly the ability to form triple helical structures, and thus may provide similar bio-responses while avoiding enzymatic degradation. In order to perform immobilization of these collagen-like molecules (CLMs) under mild reaction conditions, the bulk materials are first equipped with suitable surface groups using rf plasma methods. Plasma polymer interlayers offer advantages as versatile reactive platforms for the immobilization of proteins and other biologically active molecules. Application of a thin plasma polymer coating from an aldehyde monomer is particularly suitable as it enables direct immobilization of CLMs by reaction with their terminal amine groups, using reductive amination chemistry. An alternative route is via plasma polymer layers that contain carboxylic acid groups and using carbodiimnide chemistry. A third route makes use of alkylamme plasma polymer interlayers, which are less process sensitive than aldehyde and acid plasma coatings. A layer of poly-carboxylic acid compounds such as carboxylic acid terminated PAMAM-starburst dendrimers or carboxymethylated dextran is then attached by carbodiimide chemistry onto the amine plasma layer. Amine-terminated CLMs can then be immobilized onto the poly-carboxylic acid layer. Surface analytical methods have been used to characterize the immobilization steps and to assess the surface coverage. Initial cell attachment and growth assays indicate that the biological performance of the CLMs depends on their amino acid sequence.

Type
Research Article
Copyright
Copyright © Materials Research Society 1999

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