The Design, Preparation and Application of Novel Boronate Affinity Materials for Recognition of Cis-diol Biomolecules

Author:Wang Shuang Shou

Supervisor:liu zhen


Degree Year:2015





Cis-diol containing biomolecules,which have important significance in life-analysis science,are not only the important target analytes in-omics analysis but also the important disease markers,such as CEA,PSA and AFP and so on.Moreover,abnormal expression of glycosylation is uaually the hallmark of canceration of cells or tissues.Therefore,specific recognition and separation of cis-diol containing biomolecules have important practical significance.However,all existing methods for specific recognition and separation of cis-diol containing biomolecules have many deficiencies,such as poor storagy stability,hard to purification and preparation,exorbitant price,low reuse efficiency,and harsh operated condition for antibody,aptamer,and lectin,although them have excellent specific recognition property;hydrazine chemical method,hydrophilic interaction chromatography and ion exchange chromatography have poor selectivity and easy to suffer interference;in contrast,boronate affinity chromatography own many advanties,such as straitforward operation,fine compatibility with MS and good anti-interference performance,but it is still a universal method.Therefore,exploitation and application of materials which not only can overcome these defects but also with perfect specific recognition and separation properties will be of great significance.At recent years,application of molecular imprinting and fluorescence resonance energy transfer(FRET)in life-analysis has gradually be knowen.Considering the excellent specific recognition property of imprinting cavities produced by molecular imprinting method and the selective recognition of FRET achieved via competitive effect,both of them can be the new elements for exploitation of novel boronate affinity materials for specific recognition of cis-diol biomolecules.At first,to surmount the poor selectivity of existing methods and materials for recognition of cis-diol containing biomolecules,a new method,namely boronate affinity-based controllable oriented surface imprinting,which combining boronate affinity effect with molecular imprinting,was provided in this article for preparation of boronate affinity-based imprinted materials,and for the first time presented the unique affinity-tunable duol-mode binding mechanism.Specific recognition and separation of glycoprotein was achieved successfully in this way.Comparison with existing imprinting methods,this method exhibits many advantages,including excellent specificity,high imprinting efficiency,tunable affinity and strong university,and so on.This method not only provided the possibility for keeping the specificity while enables the binding strength to be adjusted,but also presented new sight on the role of affinity-determining factors in the process of molecular imprinting.The new understanding of the binding mechanism in molecular imprinting is favorable not only for the exploitation of new imprinting polymers but also for the design of other biomimetic materials.Secondly,combining boronate affinity effect,molecular imprinting and fluorescence detection,a new monosaccharide-targeted boronate affinity-based imprinted material,namely monosaccharide imprinted fluorescence nanoparticles,was developed for specific targeting and imaging of Cancer cells.Fluorescent imaging of human hepatoma cells(HegG-2)over normal hepatic cells(L-02)and mammary cancer cells(MCF-7)over normal mammary epithelial cells(MCF-10A)by these NPs was demonstrated.In addition,the differential expression of fucose and mannose on the surface of HepG-2 and MCF-7 cells was demonstrated by fluorescence imaging.As the imprinting approach employed herein is generally applicable,monosaccharide-imprinted NPs can find more promising appplications in cancer diagnostics and therapy,also open up new horizon in bioimaging,disease diagnosis and therapy.Finally,based on the difference of binding force between boronic acid and different cis-diol containing biomolecules,combining boronate affinity and FRET effect,a new FRET system was developed for selective recognition of cis-diol containing biomolecules.This system was consisted of polyAPBA NPs(energy donor)and AMP-GO(energy accepter).Boronate affinity played a key role in this system and provided nice selectivity for recognition of cis-diol containing biomolecules,therefore,polyAPBA NPs-AMP-GO FRET system was a universal sensing platform for cis-diol containing biomolecules.Because of the important clinical or biological significance of many cis-diol containing biomolecules(such as TRF,AFP and CEA,and so on),this new FRET system presented in this article has great potential application value,such as biosensing,clinical diagnosis and so on.