Development of a thermometric sensor for fructosyl valine and fructose using molecularly imprinted polymers as a recognition element [Elektronische Ressource] / von Rajagopal Rajkumar

Rajagopal Rajkumar

Sujets

• Biologie

Informations

• Publié par: universitat_potsdam
• Publié le: 01 janvier 2007
• Nombre de lectures: 22
• Langue: English
• Poids de l'ouvrage: 3 Mo

Extrait

Institut für Biochemie und Biologie
Professur für Analytische Biochemie

Development of a Thermometric Sensor for Fructosyl Valine and Fructose
using Molecularly Imprinted Polymers as a Recognition Element











Dissertation
zur Erlangung des akademischen Grades
"doctor rerum naturalium"
(Dr. rer. nat.)
in der Wissenschaftsdisziplin Analytische Biochemie










eingereicht an der
Mathematisch-Naturwissenschaftlichen Fakultät
der Universität Potsdam



von
Rajagopal Rajkumar





Potsdam, November 2007















































Elektronisch veröffentlicht auf dem
Publikationsserver der Universität Potsdam:
http://opus.kobv.de/ubp/volltexte/2008/1727/
urn:nbn:de:kobv:517-opus-17272
[http://nbn-resolving.de/urn:nbn:de:kobv:517-opus-17272]

Acknowledgements


My very special gratitude is directed to my supervisor Prof. Dr. Frieder W. Scheller for giving me
this great opportunity to work in his internationally acclaimed group. Thank you for your
continuous support during hard times, encouragement and fruitful discussions.

I thank Dr. Martin Katterle who guided me through my work and for all the good advice and
creative suggestions concerning the project.

I would like to thank Dr. Axel Warsinke for helping me with the project, advice and being very
kind to me.

I am grateful to Prof. Dr. Helmuth Möhwald, Max Planck Institute of Colloids and Interfaces for
the physical chemist view of the project and valuable discussions and support.

I thank Prof. Günter Wulff, Heinrich-Heine University, Düsseldorf for his tips on making
imprints. Thanks to Dr. Kristian Lettau for helping me to make my first imprints and introducing
me to the thermistor.

My special thanks go to Prof. Ulla Wollenberger and Dr. Walter Stöcklein for the scientific
discussions. I like to acknowledge Prof. Frank Bier for the support from Fraunhofer IBMT.

I thank Ms. Angelika Lehmann, Ms. Andrea Kühn, Mr. Joachim Jacob at Analytical
Biochemistry, Uni-Potsdam for their friendly assistance in the lab. Further I would like to thank
Ms. Annemarie Martins, Ms. Irena Shikova, Ms. Regina Rothe, Ms. Annegret Praast and Dr.
Jürgen Hartmann at Max Planck Institute of Colloids and Interfaces for the technical help.

I am thankful to the Dr. Angelo Valleriani, the coordinator of IMPRS on biomimetic systems for
the administrative and financial support. Further financial support from BMBF (InnoRegio
03i1314 and BioHySys 0311993) is acknowledged.





I like to thank moritz for nice little chats and for the help with the translation. Thanks roman for
nice chats in the lab and for giving me company for long stays in the lab. I wish to thank roberto
for nice jokes, for teaching me Italian ☺ and for friendship. Further I thank noya, carsten, katrin,
merlin for a relaxed atmosphere in the lab and for the social activities. I also extend my thanks to
the people at IBMT, soeren, umporn oliver, kai, & erika for the help and for social activities.

Special thanks to birgit for organising a pleasant trip to wöbbelin. It was a great time i had with
your entire family. Thanks to Prof. Burkhard Micheel and Mrs Edith Micheel for the wonderful
pinnow trip and showing me the interesting aspects of life at the countryside in germany.

Many thanks to buddy (lakshmanan) ☺, raveendra, sajini, bala, priya, sangita and saravana for all
the fun I had in germany and who never let me feel alone, lifted my spirits up in the bad times and
gave me a feeling of home away from home.

Further I thank my T3-friends atul, arun, tijo, amit, mishra, paruthi & robert for the funny
evenings and parties. I wish to thank my friends sarath, sekhar, sridhar & ashoka for the nice
moments i shared with them in india.


I admire the support given to me from all my teachers from my school days in SRSI, friends at the
Anna University, Chennai and colleagues at CFTRI, Mysore who contributed one way or the
other in fulfilling this task.

And last but certainly not least, I would like to thank my dad rajagopal, mom kanagam and my
brother naveen for the moral support and love throughout these years.













i
Table of contents


I. Table of contents...................................................................................i
II. List of figures................................................iii
III. List of tables ...................................................v
IV. Abbreviations................................................vi
V. Thesis structure............................................viii

I. Table of contents
1 INTRODUCTION ................................................................................................. 1
1.1 Biosensors ................................................................................................................................................ 1
1.2 Recognition Elements ............................................................................................................................. 2
1.2.1 Enzymes............................................................................................................................................... 2
1.2.2 Microbial cells...................................................................................................................................... 3
1.2.3 Antibodies ............................................................................................................................................ 3
1.2.4 Receptors.............................................................................................................................................. 4
1.2.5 Tissue materials and organelles............................................................................................................ 5
1.2.6 Nucleic acids........................................................................................................................................ 5
1.2.7 Molecularly imprinted polymers (MIPs).............................................................................................. 6
1.3 Transducer .............................................................................................................................................. 7
1.4 Molecular Imprinting ............................................................................................................................. 8
1.4.1 Background .......................................................................................................................................... 9
1.4.2 Molecular imprinting strategies and approaches.................................................................................10
1.4.3 MIP formats ........................................................................................................................................14
1.4.4 MIP sensors.........................................................................................................................................16
1.5 Enzyme Thermistor ...............................................................................................................................26
1.6 Boronic Acid for Saccharide Recognition............................................................................................28
1.7 Biological Significance of the Template used - Fructosyl Valine (Fru-Val).....................................29
2 OBJECTIVES.....................................................................................................32
3 MATERIALS AND METHODS...........................................................................34
3.1 Materials.................................................................................................................................................34
3.1.1 Chemicals and biochemicals ...............................................................................................................34
3.1.2 Instruments..........................................................................................................................................35
3.1.3 Buffers and reagents............................................................................................................................35
3.2 Methods ..................................................................................................................................................36
3.2.1 Synthesis of template, functional monomer and template-functional monomer complex ..................36
3.2.2 Prepara