Advanced analytical techniques Flashcards
what is Spectrophotometry
measure how much a chemical substance absorbs light by measuring the intensity of light as a beam of light passes through sample solution.
What is Calorimetry
measuring the amount of heat released or absorbed during a chemical reaction. By knowing the change in heat, it can be determined whether or not a reaction is exothermic (releases heat) or endothermic (absorbs heat).
How does a Spectrofluorimetric assay determine the formation of a product?
The formation of product or reduction of reactant concentration is measured by attaching a moiety that fluoresce at defined wave lengths an example being Dibutyryl Fluorescein which is injected into the patients veins and due to the action of lipase becomes Flurescein which is fluorescent.
How would you use Spectrofluorimetric Bioluminescent enzyme Assay to measure antibody dependent cell-mediated cytotoxicity (ADCC)?
Assay quantitatively measures release of Glyceraldehyde- 3-Phosphate Dehydrogenase (GAPDH) which is released during apoptosis. Further in this reaction luciferase catalyses luciferin to release light which is then measured. (indirect measurement of GAPHD)
What is Manometer?
Records a reduction in conc/ and increase in gaseous release ( useful for measure enzyme activity if one of the component is in gaseous form)
Describe how Manometry technique would be carried out?
- The samples and reagents are placed in separate compartments and mixed at defined time period and the reaction can be followed as the reaction proceeds.
- Eg: Oxygen consumption is measured in glucose oxidase, Carbon-dioxide production
produced in reactions catalysed by decarboxylases
What is Enthalpimetry ?
Measures the enthalpy change during the course
of a reaction
What conditions would you need for an enthalpimetry measurement and why?
- Requires high sensitivity and a closed circuit with a net entropy of 0 so that minute changes can be recorded.
How would you use Radiochemical methods for an enzyme assay?
Radioactively labelled substrate is used to follow the enzymatic reaction
The substrate is then separated from the product using electrophoresis or chromatography and the radioactive fraction of the product or the substrate is used to estimate activity of the enzyme.
What is the function of a Lipase and how is it produced in an industrial scale?
Breaks down Fat and is taken from Animal pancreases
What is the function of Trypsin and how is it produced on an industrial scale
Trypsin helps break down protein within small intestine and is sourced through Ox bile within industry
What is the function of Urokinase and how is it produced within industry?
Urokinase (made in kidney) converts inactive plasminogen into active plasmin and is sourced from Cow urine and human plasma within industry.
What is a Lysozyme, where can it be found within the body and how is it produced within industry?
Lysozyme is an antimicrobial agent that acts by cleaving the peptidoglycan component of bacterial cell walls, which leads to cell death.
This can be found within bodily secretions such as tears, saliva, and milk, within industry this is sourced from transgenic plants, animals and microorganisms that can produce human lysozyme
What is Adenosine Deaminase function, where is it sourced and how can it be produced within industry?
eliminates deoxyadenosine, which is a product of DNA breakdown which is toxic to lymphocytes.
This is produced in all cells but higher levels in lymphocytes and can be sourced from Bovine intestine.
What is pepsin’s function, where is it sourced and how can it be produced within industry?
An enzyme that breaks down proteins in food during digestion within the stomach, this can be sourced from Hog pancreas.
What is Dornase α
Is an inhaled medication (synthetic deoxyribonuclease) that thins mucus. Reduce the number of lung infections and to improve lung function in patients with cystic fibrosis by breaking down extracellular DNA in the lungs which coats bacteria.
