Protein Stabilization and Formulation Techniques Quiz
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Questions and Answers

Which amino acid residues are the principle sites of deamidation in alkaline pH for hGH?

  • Asn-149 and Gln-13 (correct)
  • Asn-9 and Gln-13
  • Asn-9 and Asn-149
  • Asn-9 and Gln-13
  • Which amino acid residue is the major oxidative site in hGH?

  • Tyrosine
  • Histidine
  • Tryptophan
  • Methionine (correct)
  • How many Zn+2 binding sites are present in the primary structure of hGH?

  • 3
  • 1
  • 4
  • 2 (correct)
  • Where does deamidated hGH occur inside the body?

    <p>Pituitary gland in the brain</p> Signup and view all the answers

    What can counteract hGH aggregation due to hydrophobic interactions when unfolded?

    <p>Zn binding sites and surfactants like polysorbate-20</p> Signup and view all the answers

    How do sugar and glycine stabilize proteins in the liquid state?

    <p>By interacting with water</p> Signup and view all the answers

    What technique traps proteins and stabilizes the formulation through glass stabilization?

    <p>Amorphous sugar states</p> Signup and view all the answers

    How should reconstituted hGH be stored to prevent concentration-induced aggregation?

    <p>Refrigerated</p> Signup and view all the answers

    What does freeze-drying involve to produce proteins in a dry form?

    <p>Creating a vacuum, freezing, and evaporating water</p> Signup and view all the answers

    What can impact pH due to lower solubility at lower temperatures?

    <p>Phosphate buffers</p> Signup and view all the answers

    What can affect the stability of molecules during freezing?

    <p>Phosphate buffers</p> Signup and view all the answers

    What can cause protein degradation during freezing and drying?

    <p>Low temperature and dehydration</p> Signup and view all the answers

    What is spray drying used for?

    <p>Producing inhalable insulin powder</p> Signup and view all the answers

    What factors can be controlled in spray drying to affect the drying rate?

    <p>Inlet temperature, drying gas humidity, and aspirator air flow</p> Signup and view all the answers

    What increases the dry capacity of gas in spray drying, affecting liquid evaporation and drying rate?

    <p>High temperature and low humidity</p> Signup and view all the answers

    What is the likely outcome to the residual water content in the powder using a drying gas with high temperature and low humidity at a high aspirator flow?

    <p>High</p> Signup and view all the answers

    What is the purpose of using a freezing medium like liquid nitrogen in spray freeze drying?

    <p>To turn the liquid into frozen droplets for subsequent drying under vacuum</p> Signup and view all the answers

    Which chromatographic method is suitable for detecting protein aggregation?

    <p>Size Exclusion HPLC</p> Signup and view all the answers

    What information does Far UV CD (190-240nm) provide in circular dichroism (CD) spectroscopy?

    <p>Information about amide bonds and secondary structures</p> Signup and view all the answers

    What does Fourier transform infrared (FTIR) spectroscopy focus on in the Amide 1 region (1600-170 cm^-1)?

    <p>Secondary structures</p> Signup and view all the answers

    What is the purpose of SDS-page in electrophoresis?

    <p>To separate molecules according to size</p> Signup and view all the answers

    What does Isoelectric focusing gel separate molecules according to?

    <p>Charge</p> Signup and view all the answers

    Why may bioactivity assays exhibit variability?

    <p>Structural changes might not always affect bioactivity</p> Signup and view all the answers

    What is the likely outcome of decreasing the liquid feed rate in spray drying?

    <p>Faster drying</p> Signup and view all the answers

    What is the effect of increasing the aspirator flow on cyclone efficiency for powder collection?

    <p>Increases efficiency</p> Signup and view all the answers

    What is the likely outcome of increasing the spray gas flow (atomizing pressure) in spray drying?

    <p>Finer powder</p> Signup and view all the answers

    How does the solid concentration in the liquid affect the dry particles after evaporation of the liquid?

    <p>Higher solid concentration leads to bigger dry particles</p> Signup and view all the answers

    What is the likely outcome to the residual water content in the powder using a drying gas with high temperature and low humidity at a high aspirator flow?

    <p>Low</p> Signup and view all the answers

    What does increasing aspirator flow lead to in terms of cyclone efficiency for powder collection?

    <p>Increase efficiency</p> Signup and view all the answers

    What effect does decreasing liquid feed rate have on the drying process?

    <p>Faster drying</p> Signup and view all the answers

    What is the likely outcome of increasing spray gas flow (atomizing pressure) on the fineness of the powder?

    <p>Finer powder</p> Signup and view all the answers

    What effect does a higher solid concentration in the liquid have on the size of dry particles after evaporation?

    <p>Larger particles</p> Signup and view all the answers

    What is the purpose of spray freeze drying?

    <p>To produce dry particles with void space</p> Signup and view all the answers

    What does Size Exclusion HPLC detect in proteins?

    <p>Aggregation</p> Signup and view all the answers

    What does Circular Dichroism (CD) spectroscopy reveal about proteins?

    <p>Amide bonds and secondary structures</p> Signup and view all the answers

    What does Fourier transform infrared (FTIR) spectroscopy focus on in proteins?

    <p>Secondary structures</p> Signup and view all the answers

    What does SDS-page electrophoresis separate molecules according to?

    <p>Size</p> Signup and view all the answers

    What does isoelectric focusing gel electrophoresis separate molecules according to?

    <p>Charge</p> Signup and view all the answers

    Why might bioactivity assays exhibit variability?

    <p>Bioactivity of protein may not be ordered</p> Signup and view all the answers

    Study Notes

    Protein Stabilization and Formulation Techniques

    • Proteins, including hGH, can aggregate due to hydrophobic interactions when unfolded.
    • Zn binding sites and surfactants like polysorbate-20 can counteract hGH aggregation.
    • Sugar and glycine stabilize proteins in liquid state by interacting with water.
    • Amorphous sugar states can trap proteins, stabilizing the formulation through glass stabilization.
    • Reconstituted hGH should be refrigerated to prevent concentration-induced aggregation.
    • Freeze-drying involves creating a vacuum, freezing, and evaporating water to produce proteins in a dry form.
    • Phosphate buffers can impact pH due to lower solubility at lower temperatures.
    • Phosphate buffers can affect the stability of molecules during freezing.
    • Low temperature and dehydration during freezing and drying can cause protein degradation.
    • Spray drying is a one-step process to turn liquid into powder, used for producing inhalable insulin powder.
    • Controlling factors for spray drying include inlet temperature, drying gas humidity, and aspirator air flow.
    • High temperature and low humidity increase the dry capacity of gas, increasing liquid evaporation and drying rate.

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    Description

    Test your knowledge of protein stabilization and formulation techniques with this quiz. Learn about methods to prevent protein aggregation, the role of surfactants and sugars in stabilizing proteins, and the processes of freeze-drying and spray drying. Understand the impact of temperature, humidity, and buffers on protein stability and formulation.

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