Enzymes, Chromatography & Specimens
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What is meant by the term “metabolism”?
A sequence of enzyme catalysed chemical reactions in the cell - both anabolic and catabolic reactions
Anabolic and catabolic pathways are different. Briefly explain two of the differences between an anabolic pathway and a catabolic pathway.
Anabolic: Concerned with the processes involved in the assembly of complex organic molecules, requires energy and has divergent pathway Catabolic: Processes related to degradation of complex substances, with resulting generation of energy, conserves energy and has a convergent pathway
What is an enzyme?
Catalysts that regulate the chemistry of cells and organisms
What is an enzyme active site?
Part of enzyme involved in the catalytic reaction
When referring to an enzymatic reaction, what is meant by the term “substrate”?
The reactant in an enzyme-catalysed reaction (Substance acted upon by the enzyme)
When referring to an enzymatic reaction, what is meant by the term “product”?
What is produced after the enzyme and substrate have bonded togther
List and briefly explain four (4) variables that can effect enzyme activity.
Temperature: Each enzyme has an optimum temperature, and can only work within a certain range of temperatures pH: Each enzyme has an optimum pH, and can only work within a certain range of pH levels Substrate concentration: The more substrates that bind with available enzymes, the faster the reaction Enzyme concentration; The greater the number of enzymes that are able to bind with a substrate, the faster the reaction
A laboratory assay uses an enzymatic reaction as part of the assay procedure. Explain why the incorrect incubation time for the laboratory assay may effect the results of the assay
This could affect the temperature that the enzyme is working in, and the enzyme may not be able to work effectively
List 4 different types of biological specimens that may be used in a Biomedical Science or Pathology laboratory.
• Blood • Urine • Faeces • Solid tissue
Name the cellular components of blood.
RBC, WBC, platelets
Name 3 components of blood required for clotting.
• Ca2+ • Platelets • Clotting proteins e.g. fibrinogen
Is there a difference between serum and plasma? Explain your answer.
Plasma: liquid portion of blood Serum: liquid portion of blood without clotting components
What is an anticoagulant?
A substance that allows the plasma portion of blood to be separated from the rest
List one anticoagulant and explain its mode of action.
Heparin: - inhibits clotting proteins (eg thrombin)
Why are different anticoagulants used when collecting blood?
Different coagulants have different purposes and modes of action, e.g. sodium fluoride preserves glucose in blood and oxalate joins calcium ions together
Why is correct identification of a patient sample important?
So that the patient waiting for results does not get an incorrect diagnosis, treatment or intervention
List 2 reasons that a patient sample may be unsuitable for a laboratory assay, necessitating the recollection of a sample.
• If the patient is identified incorrectly - If the sample is damaged while being transported
Haemophilia is a disease studied in a Coagulation laboratory. What is Haemophilia?
A blood disorder where blood is not able to clot properly
Why might a protein need to be separated from a mixture of other proteins?
There may only be one protein that is of interest to be analysed
What types of molecules can be separated from a mixture using Chromatography?
DNA, RNA, other organic molecules
List three Chromatography principles that can be used to separate a molecule of interest from a mixture.
Ion Exchange: - Solid stationary phase and liquid mobile phase Size Exclusion: - Liquid stationary phase and liquid mobile phase Affinity: - Stationary ligand and liquid mobile phase
For each of the principles that you listed in answer to the question above, what property of the molecule of interest is being utilised (eg size, charge, etc)
Ion exchange: charge Size exclusion: size
What factors may affect the reliability of a laboratory test result?
Accuracy and precision
Explain the term Accuracy when referring to laboratory results. Use a hypothetical example in your explanation.
Closeness of measured value to true value