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Protein Aggregation and

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Nina Uchida

on 26 April 2017

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Transcript of Protein Aggregation and

Intensity fluctuates due to Brownian motion
Protein Aggregation and
Dynamic Light Scattering

Proteins as Biopharmaceuticals
Challenges in
Therapeutic Protein Development
Dynamic Light Scattering
DLS
In Conclusion
Protein aggregation and stability is an important aspect of biopharmaceutical development
As biopharma grows, novel methods of characterizing protein aggregation will be needed
DLS is a fast and relatively easy method of characterizing size distributions in a sample
Growth in Biopharmaceuticals
DLS correlates fluctuations of scattered light with particle size
Experiment
Bovine Serum Albumin (BSA) in phosphate buffered saline (PBS)
30 mg/mL (~3.8 volume%)
Heated in water bath at 55C for 20 min to 120 min
Analyzed for size distribution with DLS
Nina Uchida | CHBE 333 Tech Topic
Insulin
Factor VIII
[1]
[2]
[3]
30mg/mL BSA in PBS,
55°C for 40 min
Brownian Motion
DLS setup
Light scatters off particles
Smaller particles: faster Brownian motion
Translational diffusion coefficient
Autocorrelation function
r=kT/6πμD
Stokes-Einstein Equation:
[4]
[5]
[6]
[Courtesy Dr. Marc Taraban,
University of Maryland School of Pharmacy]
References
[1] “RCSB Protein Data Bank - RCSB PDB.” [Online]. Available: http://www.rcsb.org/pdb/home/home.do. [Accessed: 19-Apr-2017].
[2] “World Preview 2016, Outlook to 2022.” (Sep-2016). EvaluatePharma. [Online.] Available: http://info.evaluategroup.com/rs/607-YGS-364/images/wp16.pdf. [Accessed 19-Apr-2017].
[3] “protein ‘egg’-regation,” (11-Mar-2011). Bite-Sized Biology. [Online]. Available: http://bitesizedbiology.tumblr.com/post/3787483400/protein-egg-regation. [Accessed: 19-Apr-2017].
[4] Rutger Saly. (19-Nov-2009). Brownian Motion - nanoparticles in water. [YouTube Video]. https://youtu.be/cDcprgWiQEY. [Accessed: 19-Apr-2017].
[5] “Dynamic Light Scattering for particle sizing and measurement of particle size distribution.” [Online]. Available: http://www.lsinstruments.ch/technology/dynamic_light_scattering_dls/. [Accessed: 19-Apr-2017].
[6] “Dynamic Light Scattering (DLS).” Nano Research Facility. [Online]. Available: http://www.nano.wustl.edu/doc/Instrument%20Manuals%20and%20Protocols/DLS%20Final.pdf. [Accessed: 19-Apr-2017].
[7] T.J. Woehl, private communication, April 2017.
[8] M. Taraban, private communication, April 2017.

30mg/mL BSA in PBS,
60°C for 15 min
[7]
[7]
Full transcript