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Preliminary Study on Interactions between Functionalized Nanoparticles and Protein

Author: LiZhenWei
Tutor: SiZhiKun；ZuoBing
School: Shandong University
Course: Analytical Chemistry
Keywords: Magnetic nanoparticles The surface functionalization multi -walled carbon nanotubes The nanoparticles - protein interactions Combinatorial Chemistry Biological Screening
CLC: TB383.1
Type: Master's thesis
Year: 2009
Downloads: 281
Quote: 2
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Abstract


Nanomaterials three-dimensional structure, at least there is a dimension size in nanoscale materials, its unique physical chemical properties of attracting the many scholars research interest, including biomedical applications for the goals of the exploratory research is rapid formation of a new research fields. Magnetic nanoparticles (MNPs) and carbon nanotubes (CNT) is more and more extensive application of two important nanomaterials be improved by the introduction of the functionalized groups, water-soluble and biocompatible nanomaterials, thus laid the foundation for its application in the biomedical field. The protein is an extremely important class of biological macromolecules, and is the basis of the substance of life. Studies show that some nanomaterials into the organism to interact with a variety of proteins thereby affecting the protein's structure and function, triggering the physiological effects of the unknown, so to study the interaction of the nanoparticles with proteins nanomedicine and nanotoxicology crucial. The study is divided into the following two aspects: the first part is to analyze the interactions of MNPs with bovine serum albumin. MNPs size of 200nm and 50 nm, respectively, the surface COOH, of PEG-COOH, NH 2 , and PEG-NH 2 the four kinds yl group modification. Total adsorption amount MicroBCA determined MNPs protein, SDS-PAGE electrophoresis and MALDI-TOF-MS were isolated and identified the MNPs by adsorption of protein, and choose the binding capacity of bovine serum albumin (BSA) steady-state fluorescence spectra and circular dichroism analysis of its interaction with the MNPs. Comprehensive analysis of the experimental results the following conclusions: a certain quality of MNPs surface binding protein amount by their size and surface modification group, and 50 nm of MNPs binding proteins were significantly more than 200nmMNPs with small particle size of the same quality nanoparticles provide more surface area of ??the combined space; carboxy-modified MNPs adsorbed proteins were significantly more with the other three, it is related to the surface charge of the nanoparticles; The surface PEG modified MNPs adsorption amount of protein was significantly lower than the PEG-modified the MNPs described hydrophobic role in their interactions are essential; MNPs in the serum protein adsorption has selective; BSA and different MNPs combined structure of the BSA has no effect. The second part of the study functionalized carbon nanotubes (f-MWNTs) to explore the interaction between chymotrypsin (ChT), f-MWNTs identification and regulation of the active site of ChT. Initial screening by the steady-state fluorescence and activity analysis of f-MWNTs ChT autofluorescence and activity. F-MWNTs filter results selected two groups of representative further study, the use of liquid chromatography determination of the ChT adsorption amount of f-MWNTs, circular dichroism analysis of the structure of ChT impact, further analysis of the kinetics of enzymatic reaction enzyme inhibition mechanism. Comprehensive analysis of the experimental results the following conclusions; most f-MWNTs and ChT nonspecific adsorption. The enzymatic kinetics analysis verified 29 #, 30 #, 34 #, 35 # # 36 belong to the competitive inhibition of of ChT the activity suppression, can identify the active site of chymotrypsin. Analyze the structure-activity relationship to the surface modification containing N, N-dibutylamine group f-MWNTs can identify ChT the active site, has good targeting. In summary, this thesis from the protein level using a variety of analytical methods to study the biological effects of the magnetic nanoparticles and the functionalized nanotubes, the surface modification of the nanoparticles with the protein binding have selective. Protein Conformation determine its physiological function, by changing the size of the nanoparticles, the shape, the surface chemical groups may be adjusted with the protein binding capacity and the binding sites, thereby changing biological toxicity, improve biocompatibility, so that the nano particles in the biomedical on broader application.

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