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Enhancing Protein Crystal Nucleation Using In Situ Templating on Bioconjugate-Functionalized Nanoparticles and
Caroline McCue1, Henri-Louis Girard1, Kripa K Varanasi1
1Department of Mechanical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, Massachusetts 02139, United States.
ACS Applied Materials & Interfaces
|February 28, 2023
Summary
Functionalized nanoparticles accelerate protein crystallization by acting as templates, reducing required protein concentration and time for manufacturing applications.
Area of Science:
- Biotechnology
- Materials Science
- Protein Science
Background:
- Protein crystallization is a cost-effective purification method but hindered by slow nucleation and high protein concentration needs.
- Current methods face challenges in achieving efficient protein crystallization for large-scale manufacturing.
Purpose of the Study:
- To investigate the use of bioconjugate-functionalized nanoparticles for inducing rapid protein crystallization.
- To overcome barriers in protein crystallization for downstream protein purification and manufacturing.
Main Methods:
- Utilized microbatch crystallization setup to assess nanoparticle effects on crystallization conditions.
- Employed a machine learning-enabled emulsion crystallization setup to quantify nucleation parameters.
- Tested bioconjugate-functionalized nanoparticles with model proteins.
Main Results:
- Functionalized nanoparticles expanded the range of successful crystallization conditions.
- Bioconjugate-functionalized nanoparticles decreased protein crystal induction time by up to 7-fold.
- Nucleation rates increased up to 3-fold in the presence of functionalized nanoparticles compared to controls.
Conclusions:
- Bioconjugate-functionalized nanoparticles serve as effective in situ templates for protein crystallization.
- This approach significantly reduces protein concentration and time requirements for nucleation.
- Enables potential for improved protein manufacturing processes through enhanced crystallization.
