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URN etd-0721109-155941 Statistics This thesis had been viewed 3128 times. Download 1854 times. Author Yi-chen Chen Author's Email Address No Public. Department Bioengineering Year 2008 Semester 2 Degree Master Type of Document Master's Thesis Language zh-TW.Big5 Chinese Page Count 153 Title Preparation of high affinity molecularly imprinted
polymers selective for denatured creatine kinase and
their application to the extraction of native and
Keyword creatine kinase denatured protein molecularly imprinted polymer sodium dodecyl sulfate sodium dodecyl sulfate molecularly imprinted polymer denatured protein creatine kinase Abstract The inter-relationship of protein structure and function is of interest to
many researchers; however, formal academic study of denatured proteins,
arising e.g. from genetic mutations, has until now been limited. Of critical
significance are the degenerative processes that lead to the conversion of the
α-helix to the β-sheet, and the generation of amyloid plaques in for example:
Alzheimer’s disease, Parkinson’s disease and degenerative conditions
associated with prions.
Here a simple synthetic approach based on micro- contact imprinting
has been used to form imprints of the secondary structure of creatine kinase
(CK), denatured by treatment with SDS. The imprinted materials, formed with MMA and PEG400DMA, in a volume ratio of 5:95, have been
demonstrated to be able to separate denatured CK from its native from.
Using NaOH/trypsin, a template extraction-efficiency, i.e. denatured CK
removal from the film, of 90% was obtained. Subsequent re-binding of the
template, i.e. denatured CK, was typically 70%, (template solution 3.5 ×10 -7
mol, MIP surface area 1.69 cm2), with an imprinting efficiency of 9.7.
Evaluation of the imprinted polymers in non-competitive re-binding
experiments with native and denatured proteins, showed re-binding of 68.3%,
95.3%, 92.3% and 62.4% to the native forms of CK, IgG, HSA and myoglobin respectively; while the respective binding to denatured forms of
IgG, HSA and myoglobin was 94.1%, 92.7%, 71.7%. In a competitive binary
system, using the denatured forms of: IgG, HSA and myoglobin; the
selectivity was 98.7%, 96.8% and 63.8%, respectively.
The method used has been shown to be a successful approach for the
formation of high-affinity materials able to separate denatured CK from its
native form and from other denatured proteins. Such materials may find
future applications in practical sensing devices.
Advisor Committee Chung-yih Wang - advisor
Tse-chuan Chou - advisor
Kuo-chuan Ho - co-chair
Files Date of Defense 2009-06-26 Date of Submission 2009-07-21