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Capillary circular dichroism

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Waldron, Daniel E., Marrington, Rachel, Grant, Marcus C., Hicks, Matthew R. and Rodger, Alison (2010) Capillary circular dichroism. Chirality, Vol.22 (No.1E Sp. Iss. SI). E136-E141. doi:10.1002/chir.20878 ISSN 0899-0042.

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Official URL: http://dx.doi.org/10.1002/chir.20878

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Abstract

Circular dichroism (CD) has become an increasingly important tool in the study of biological molecules as it enables structural information to be obtained non-destructively on solution-phase samples. However, sample requirements for CD are often seen as being too high with protein backbone measurements in standard cuvettes typically requiring similar to 100-300 mu L of 0.1 mg/ml protein. To address this issue, we have designed a new form of CD sample holder, which reduces the sample requirements of the technique by two orders of magnitude, with a sample requirement of less than 3 mu l. This sample saving has been achieved through the use of extruded quartz capillaries, the sample being held within the internal diameter of the quartz capillary through capillary action. The extruded quartz capillaries exhibit remarkably little birefringence, although still transmitting high energy UV circularly polarized light. The optics associated with capillaries were investigated. A configuration has been adopted with the light beam of the spectrophotometer being focused in front of the front face of the capillary using a biconvex lens and advantage being taken of the additional focusing effect of the capillary itself. The focusing is vital to the low wavelength performance of the cell, where we have acquired reliable data down to 180 nm using a Jasco J-815 spectrophotometer. The system performance was validated with Na[Co(EDDS)]center dot H2O (EDDS = N,N-ethylenediaminedisuccinic acid), concanavalin A, lysozyme, and progesterone. Chirality 22:E136-E141, 2010. (c) 2010 Wiley-Liss, Inc.

Item Type: Journal Article
Subjects: Q Science > QD Chemistry
Q Science > QP Physiology
Divisions: Faculty of Science, Engineering and Medicine > Science > Chemistry
Library of Congress Subject Headings (LCSH): Circular dichroism
Journal or Publication Title: Chirality
Publisher: John Wiley & Sons, Inc.
ISSN: 0899-0042
Official Date: 2010
Dates:
DateEvent
2010Published
Volume: Vol.22
Number: No.1E Sp. Iss. SI
Number of Pages: 6
Page Range: E136-E141
DOI: 10.1002/chir.20878
Status: Peer Reviewed
Publication Status: Published
Access rights to Published version: Restricted or Subscription Access
Funder: Engineering and Physical Sciences Research Council (EPSRC)
Grant number: EP/D075750/1 (EPSRC)

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