Biosensors and Nanotheranostics
Bionanotechnology, Drug Delivery, Therapeutics | online ISSN 3064-7789
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RESEARCH ARTICLE   (Open Access)
Contents Vol 2 (1)

Enhanced Lysozyme Crystallization Using Nano-Templates: Effects of Pore Size and Surface Functionalization

Klepetsanis Pavlos 1*

+ Author Affiliations

Biosensors and Nanotheranostics 2 (1) 1-6 https://doi.org/10.25163/biosensors.219839

Submitted: 12 May 2023 Revised: 01 July 2023  Published: 02 July 2023 

Abstract

Protein crystallization is a pivotal method in bio-separation, utilized across various fields including structural biology and industrial enzyme production. Recent research has explored nano-templates as a promising approach to enhance protein crystallization by serving as effective nucleants. This study investigates the synthesis and application of nano-templates with varying porosities and surface chemistries to optimize lysozyme crystallization. Nano-templates were synthesized via a sol-gel method using nitric acid and 2M HCl, resulting in average pore sizes of ~4 nm and ~8 nm, respectively. These templates were further functionalized with phenyl, chloro, and methyl groups to modulate their surface properties. Characterization techniques such as nitrogen adsorption-desorption, TEM, FTIR, and zeta potential measurements were employed to assess the templates' properties. Lysozyme crystallization experiments demonstrated that nano-templates significantly reduced the induction time, with phenyl-functionalized templates and those with smaller pore sizes proving most effective. The results indicate that nano-templates with enhanced hydrophobicity and optimal surface charge facilitate faster and more efficient protein nucleation. These findings highlight the potential of nano-templates in advancing protein crystallization techniques, suggesting that further optimization could enhance their utility in bio-separation applications and offer a cost-effective solution for protein purification.

Keywords: Protein crystallization, nano-templates, lysozyme, surface functionalization, pore size

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