Refinement of analytical technologies for detection of biomolecules of importance to the dairy sector : a thesis presented in partial fulfilment of the requirements for the degree of Master of Philosophy in Sciences at Massey University, Palmerston North, New Zealand
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Date
2011
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Massey University
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Abstract
There is a continuous push on the dairy industry to enhance livestock productivity to
meet with the demands of ever increasing human population. This demand can be
achieved by developing rapid and early diagnostic aids to help curtail various
problems encountered in the livestock production. The first study of the present
thesis was focussed on standardizing initial steps towards development of Surface
Plasmon Resonance for progesterone and oestradiol 17- , both of which are critically
implicated in animal reproduction. For progesterone, the binding response of two
different length linkers, and the antigen-antibody binding response of two different
source monoclonal antibodies (P1922 from Sigma vs SE-7720-1430 from Serotec)
were evaluated. It was concluded that the long length linker had better binding
response than the short length linker. The antibody obtained from Serotec
(SE-7720-1430) had greater sensitivity but its binding response was inconsistent. On
the other hand, the sensitivity of the monoclonal antibody from Sigma (P1922) was
lower, although its binding response was consistent. For oestradiol 17- , antibody
procured from Bio-trend (BT70-1020-06) was tested and its binding response was
consistently low on all the test days. This study thus suggests that careful testing and
selection of antibodies to achieve desired antigen-antibody binding response is a
critical step towards development of SPR for progesterone and oestradiol 17- . The
second study was undertaken to refine the currently existing fluorescent techniques
to measure phytoporphyrin in the peripheral circulation of cows. Phytoporphyrin is
implicated in facial eczema (FE), which is a photosensitization disease of high
economical importance. This occurs due to disturbances in the chlorophyll
metabolism as a result of liver damage and bile duct occlusions caused by fungal
toxicity. The present study described new modified fluorescent methods to measure
phytoporphyrin in the serum of cows. Further, the absorption and emission spectra of
phytoporphyrin were compared with that of other chlorophyll metabolites and thus a
currently existing anomaly in the chemical structure of phytoporphyrin was rectified.
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Keywords
Surface plasmon resonance, Antigen-antibody reactions, Progesterone, Oestradiol, Steroids, Phytoporphyrin, Facial eczema, Dairy cattle