Date of Award

1998

Document Type

Master Thesis

Degree Name

Masters of Science (Research)

Department

Department of Biological Sciences

First Advisor

Dr. John O'Mullane

Abstract

α-Amylase is a heterogeneous polypeptide, primarily found in the Salivary and Pancreatic glands, giving rise to two isoenzymes, namely Salivary- and Pancreatic-type Amylases. Total α-Amylase activity measurements are routinely carried out in hospital laboratories to aid in the diagnosis of diseases of the pancreas and in the investigation of pancreatic function in patients with severe abdominal pains. With the development and wide availability of modem chromogenic oligosaccharide substrates specific for Amylase, assays have become more sensitive, rapid and easier to use.

Differential Amylase Isoenzyme assays ware developed to distinguish isoenzyme levels. Several methods were developed for isoenzyme determination. Of these. Selective Inhibition was found to be rapid, selective and sensitive. Two selective inhibitors were tested, W.G.I. and DOG3. The inhibition from DOG3 lacked the specificity required for a selective assay, however, the W.G.I. inhibited the Salivary and Pancreatic isoenzymes by 88% and 13% respectively. Upon incorporation with a chromogenic oligosaccharide such as EPS or CNPG3, an assay was developed which could be used in the clinical field for rapid differentiated analysis of the two isoenzymes of Amylases.

Comparative trials were carried out using a commercially available kit incorporating monoclonal antibodies against S.Amylase. However, the cost of reagents was a significant drawback.

In conclusion, both the EPS and CNPG3 Assay’s incorporating the W.G.I., provide conclusive isoenzyme analysis in addition to being effective, sensitive and rapid techniques. In addition these assays have the appropriate accuracy and precision.

Comments

Submitted to the National Council for Educational Awards. August 1998

Access Level

info:eu-repo/semantics/openAccess

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