Waldron, Daniel E. (2011) Instrument design in UV polarised spectroscopy. PhD thesis, University of Warwick.

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Abstract

Biological macromolecules are becoming increasingly important as a target for
pharmaceutical research. Structural examination of this class of compounds is
vital, both in terms of understanding the three-dimensional structures these
molecules form as well as for quality control of bio-pharmaceutical products.
UV polarised spectroscopy, such as circular dichroism (CD) is a powerful tool
for this form of analysis, rapidly providing the user with structural information
on the sample. The ease of data analysis combined with its non-destructive
nature make UV polarised spectroscopy an ideal tool for this purpose.
However, limitations in the current instrumentation, especially in terms of
sample handling have placed significant barriers in the way of fully realising the
potential of these techniques.
To address the weaknesses of the currently available UV polarised spectroscopy
accessories, new devices have been designed and tested to increase the utility of
such techniques. A low volume capillary sample holder has been developed
which significantly reduces the sample requirements for circular dichroism
without loss of signal quality. This advance has been coupled with an HPLC
auto-sampler to create a device that can process 96-well and in some cases 384-
well plates. This device opens up a host of new applications for polarised UV
spectroscopy, including refolding screening as well as freeing up user time.
Additionally, a high precision demountable micro scale cuvette has been
developed that reproducibly assembles to the same path length every time. The
percentage error of path-length for this new cuvette is of the same order as
currently commercially available 1 mm cuvettes. This new accessory should
prove highly advantageous for the bio-pharmaceutical industry, as it allows
accurate path-length CD analysis for high concentration samples.
Additionally, the first confirmed linear dichroism (LD) spectrum of a bacterium
and flagella are reported, opening up the potential for real time UV
spectroscopic analysis of living bacteria.
By using the work contained within this thesis, it should be possible to construct
a new form of high path-length accuracy, low volume, multiple-sample UV
polarised spectroscopy accessory that could be used for both CD and LD
analysis.

Item Type:	Thesis or Dissertation (PhD)
Subjects:	Q Science > QD Chemistry
Library of Congress Subject Headings (LCSH):	Ultraviolet spectroscopy -- Instruments, Ultraviolet spectroscopy -- Methodology
Official Date:	July 2011
Dates:	Date Event July 2011 Submitted
Institution:	University of Warwick
Theses Department:	Department of Chemistry
Thesis Type:	PhD
Publication Status:	Unpublished
Supervisor(s)/Advisor:	Rodger, Alison ; Dafforn, Tim
Sponsors:	Engineering and Physical Sciences Research Council (EPSRC)
Extent:	195 leaves : illustrations, charts
Language:	eng
URI:	http://wrap.warwick.ac.uk/id/eprint/51549

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