Particle assisted protein crystal nucleation as a protein purification platform for pharmaceutical manufacturing

Slow protein crystal nucleation is a major barrier to using crystallization as a separation and purification strategy in protein drug manufacturing. This work investigates the use of functionalized nanoparticles in solution to act as in situ templates for initiating crystal nucleation. We used lysozyme, a protein with well-characterized crystallization conditions, to evaluate the nucleation rates of crystals grown on the functionalized particles. The nucleation rates were measured using an emulsion based technique that involves imaging protein-containing drops at regular intervals for several hours and then using an image recognition algorithm to count the number of crystals that formed in the drops. On a microfluidic chip, a supersaturated solution of lysozyme is mixed with a stream containing precipitants and functionalized nanoparticles. The functional groups used include bio-conjugates, which bind covalently with biomolecules, and are commonly used in diagnostic imaging and targeted drug delivery. The nucleation kinetics of lysozyme were compared in cases with and without added nanoparticles, both bare and functionalized. This work suggests that nanoparticles functionalized with bio-conjugates such as n-hydroxysuccinimide and maleimide may enhance crystal nucleation.

Name thesis:degree_name

Master

Department dc:contributor.department

Massachusetts Institute of Technology. Department of Mechanical Engineering

Grantor dc:publisher

Massachusetts Institute of Technology

Year dc:date.issued

2019

Author dc:creator

• McCue, Caroline.

Advisor dc:contributor.advisor

• Kripa K. Varanasi.

• Mechanical Engineering.

Statement dc:rights

• MIT theses may be protected by copyright. Please reuse MIT thesis content according to the MIT Libraries Permissions Policy, which is available through the URL provided.

Licence dc:rights.uri

• http://dspace.mit.edu/handle/1721.1/7582

Language dc:language.iso

eng

Handle dc:identifier.uri

https://hdl.handle.net/1721.1/128336

OAI identifier oai:identifier

oai:dspace.mit.edu:1721.1/128336