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Bio-Modification of Non-Biological Surfaces with Function-Spacer-Lipid Constructs by Methods Including Bioprinting

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Bio-Modification of Non-Biological Surfaces with Function-Spacer-Lipid Constructs by Methods Including Bioprinting Bio-Modification of Non-Biological Surfaces with Function-Spacer-Lipid Constructs by Methods Including Bioprinting Katie Barr Thesis submitted in fulfilment of the degree of Doctorate of Philosophy Auckland University of Technology Auckland New Zealand 2013 Table of Contents Table of Contents ......................................................................................................................... i List of figures .............................................................................................................................. iii List of tables................................................................................................................................ iv Attestation of Authorship .......................................................................................................... vi Acknowledgements ................................................................................................................... vii Abstract ..................................................................................................................................... viii Abbreviations .............................................................................................................................. x Chapter 1 Introduction ............................................................................................................. 1 1.1 Bio-modification of non-biological surfaces with biological molecules ...................... 1 1.1.1 Immobilisation principles ..................................................................................... 1 1.1.2 Deposition techniques ......................................................................................... 23 1.2 KODE™ technology ................................................................................................... 35 1.3 Aim ............................................................................................................................. 39 Chapter 2 Methods and Results ............................................................................................. 40 2.1 Methods of delivery .................................................................................................... 40 2.1.1 Detection method ................................................................................................ 40 2.1.2 Painting ............................................................................................................... 42 2.1.3 Soaking/contact ................................................................................................... 43 2.1.4 Inkjet printing ...................................................................................................... 44 2.2 Non-biological surfaces .............................................................................................. 49 2.2.1 Cellulose membranes .......................................................................................... 50 2.2.2 Paper ................................................................................................................... 54 2.2.3 Silica ................................................................................................................... 58 2.2.4 Polymers ............................................................................................................. 61 2.2.5 Natural fibres ...................................................................................................... 66 2.2.6 Glass .................................................................................................................... 68 2.2.7 Metals .................................................................................................................. 70 2.2.8 Non-planar surfaces ............................................................................................ 73 2.2.9 Summary table of non-biological surfaces tested with FSLs .............................. 78 2.3 Testing limitations of FSL immobilisation ................................................................. 81 2.3.1 F (functional head group) versus FSL ................................................................. 81 2.3.2 Lipid tail variations ............................................................................................. 83 2.3.3 Detergent/solvent washing tests .......................................................................... 87 2.3.4 Detergents/solvents in printing solution .............................................................. 89 2.3.5 Serum washing tests ............................................................................................ 90 2.3.6 Drying versus not drying tests ............................................................................ 91 2.3.7 Attachment time .................................................................................................. 94 i 2.3.8 Stability tests ....................................................................................................... 94 2.3.9 Resolution tests ................................................................................................... 97 2.4 Inkjet printed FSL constructs for bioassays ................................................................ 99 2.4.1 Different formats ................................................................................................. 99 2.4.2 Mapping monoclonal reagents with inkjet printed FSL constructs ................... 103 2.4.3 Using printed peptide FSL constructs as disease markers ................................ 107 2.4.4 Attaching recombinant proteins to printed FSLs for immunoassays ................ 112 2.5 Cell attachment ......................................................................................................... 116 2.5.1 Establishing FSL attachment to bacteria ........................................................... 116 2.5.2 Mammalian cells ............................................................................................... 121 Chapter 3 Discussion ............................................................................................................. 135 References ................................................................................................................................ 147 ii List of figures Figure 1. Schematic representation of the main different methods of immobilisation. ..................................................2 Figure 2. Examples of covalent coupling chemistries for immobilising biomolecules ...................................................6 Figure 3. Examples of FSL constructs. ......................................................................................................................... 36 Figure 4: Schematic diagram of FSL-Atri .................................................................................................................... 37 Figure 5. Schematic diagram of FSL-biotin .................................................................................................................. 37 Figure 6. Examples of painted FSL-A. ......................................................................................................................... 43 Figure 7. Image of microbeads soaked in FSL-biotin. .................................................................................................. 43 Figure 8. The inkjet printer used for printing biological molecules. ............................................................................. 44 Figure 9. Example of printed FSL solution containing a blue dye for visualisation. .................................................... 45 Figure 10. An example of FSL-biotin applied to a surface by inkjet printing. ............................................................. 45 Figure 11. Thermal inkjet printer. Anisaldehyde stained TLC and TLC immunoassay. .............................................. 47 Figure 12. Piezoelectric inkjet printer. Anisaldehyde stained TLC and TLC immunoassay ........................................ 48 Figure 13. An example of stripy staining and an example of satellite droplets ............................................................. 49 Figure 14. FSL-Atri and FSL-biotin painted onto the right hand side of fibre glass treads.. ....................................... 78 Figure 15. TLC of FSL-Atri and FSS-Atri printing solutions and fresh solutions ........................................................ 86 Figure 16. TLC of FSL-Btri and FSL-B monoacyl printing solutions and fresh solutions ........................................... 86 Figure 17. 2mm thick acrylic plate with laser cut holes to make reaction wells ......................................................... 100 Figure 18. Examples of different printing patterns.. ................................................................................................... 101 Figure 19. An example of a printed FSL enzyme immunoassay (pFSL-EIA).. ......................................................... 104 Figure 20. Comparison of kodecytes serological results and pFSL-EIA results against monoclonal and polyclonal ABO reagents .......................................................................................................................... 106 Figure 21. Examples of FSL-Chagas inkjet printed well tested against positive and negative sera ........................... 109 Figure 22. Design of printed reaction well with 2 FSL constructs at 3 concentrations
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