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Detergent Purification of Membrane Proteins01:18

Detergent Purification of Membrane Proteins

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Detergents are used to purify the integral proteins of the membrane. The hydrophobic portion of the detergent can replace membrane phospholipids while solubilizing the membrane proteins. When detergent monomers reach a specific concentration in a solution called critical micelle concentration (CMC), they form micelles. Above CMC, the concentration of the detergent monomers remains in equilibrium with the micelle. The number of detergent monomers present in the CMC varies for each detergent, and...
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Development of a Raman Model for Continuous Monitoring of Detergent Viral Inactivation.

Kurtis Denny1, Maria Znidarsic1, Hossein Hamedi1,2

  • 1Biogen, Morrisville, North Carolina, USA.

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Process analytical technology (PAT) using Raman spectroscopy enables real-time monitoring of detergent concentration in viral inactivation (VI) steps. This continuous measurement ensures effective virus inactivation during monoclonal antibody production.

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Area of Science:

  • Biopharmaceutical manufacturing
  • Process Analytical Technology (PAT)
  • Spectroscopic analysis

Background:

  • Process analytical technology (PAT) is crucial for continuous manufacturing (CM) in mAb production.
  • PAT enables real-time monitoring of critical attributes, ensuring consistent product quality.
  • Replacing offline measurements with in-line PAT enhances process efficiency.

Purpose of the Study:

  • To develop a Raman-based model for online detergent concentration measurement.
  • To implement real-time monitoring in a continuous viral inactivation (VI) step.
  • To replace traditional offline analysis with a rapid, in-line method.

Main Methods:

  • A 785 nm Raman spectroscopy system was employed.
  • A Partial Least Squares (PLS) model was developed for calibration using spiked samples.
  • The Raman flow cell was integrated into a pilot-scale continuous VI process.

Main Results:

  • A Raman-based model achieved an estimated accuracy of 0.02% w/v for detergent concentration.
  • The system demonstrated real-time monitoring of detergent addition during pilot-scale processing.
  • The developed method provides nearly instantaneous concentration measurements.

Conclusions:

  • Raman spectroscopy offers a viable PAT tool for real-time detergent monitoring in VI steps.
  • This technology facilitates continuous processing by ensuring adequate detergent levels for viral inactivation.
  • The developed online measurement system enhances process control and product safety.