SURFACE IMPRINTED SILICA PARTICLES FOR CONCANAVALIN A PURIFICATION

Abstract

ABSTRACT

SURFACE IMPRINTED SILICA PARTICLES FOR CONCANAVALIN A PURIFICATION

Galiya RAZYM Master of Science, Department of Bioengineering Supervisor: Prof. Dr. Handan Yavuz Alagöz January 2018, 65 pages

Concanavalin A (Con A) is a representative of plant proteins group known as lectins. Many of lectin proteins have useful characteristic for studies of cell division and cell surfaces. It generally employed in affinity chromatography for characterization and purification of cell structures and carbohydrate containing molecules. In this research, a new adsorbent for specific separation of Con A were prepared by applying core-shell surface imprinting method. Silica microspheres were employed as a core material to prepare specific surface imprinted polymer contrary to Con A. Silica surface were stimulated by acidic treatment and modified with 3-methacryloyloxypropyl trimethoxysilane (MPTMS). N-methacryloyl-L-histidine methyl ester was used as a functional monomer to imprint Con A molecules on the surface of silica particles. The core-shell silica particles were characterized using Zeta Size analysis, SEM equipment and FTIR spectroscopy. The effect of parameters such as pH, concentration of Con A and temperature on the adsorption of Con A were tested and calculated. Progress of adsorption of Con A was checked by measuring it in different period of time. 1.0 M NaCl was used as removal agent. To compare the effectivness of MIPs, Non-Imprinted Polymers (NIP) were also prepared and tested the same way as MIP. Additionally we used lysozyme and hemoglobin molecules instead of Con A to check the specificity of cavities on imprinted silica particles. The selectivity of the imprinted silica particles was also investigated by isolating Con A from Jack Bean. The purity of the Con A was tested by sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). Finally Con A imprinted silica particles were tested for reusability by exploiting it in 10 subsequent cycles.

Keywords: Concanavalin A, lectins, molecularly imprinted polymers, silica particles.