High Capacity Temperature-responsive Affinity Chromatography

发布时间：2023-08-03 17:53

　　

【文章页数】：150 页

【学位级别】：博士

【文章目录】：
ABSTRACT
Chapter 1 Literature Review
    1.1 Introduction
    1.2 Immunoglobulin G(IgG)
    1.3 General Overview of Analytical Method in HPLC for Separation
        1.3.1 Protein-A Affinity Chromatography
        1.3.2 Short peptide biomimetic affinity chromatography
        1.3.3 Mixed mode chromatography
        1.3.4 Ion exchange chromatography
        1.3.5 Hydrophobic interaction chromatography(HIC)
        1.3.6 Size exclusion chromatography(SEC)
    1.4 Dendrimers
        1.4.1 Structure and characteristics of dendrimers
        1.4.2 Synthesis of PAMAM dendrimer
        1.4.3 Significance of dendrimer
    1.5 Temperature-responsive polymer
    1.6 The research purpose,significance and main research content of topic
        1.6.1 Research background and significance of the topic
        1.6.2 Research Objectives
        1.6.3 The main contents of the project
        1.6.4 The special features and innovations of the project
    References
Chapter 2 Preparation and Characterization of Thermo-responsive Affinity Chromatography Stationary Phase with Linear Spacer Arms,and Its Application for Antibody Separation
    2.1 Introduction
    2.2.Experimental Section
        2.2.1 Reagents and materials
        2.2.2 Pretreatment of Reagent
        2.2.3 Experimental instruments
        2.2.4 Preparation of materials
        2.2.5 Material Characterization
        2.2.6 Packing of the temperature-responsive affinity chromatography column
        2.2.7 Chromatographic separation
        2.2.8 Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis(SDS–PAGE)
        2.2.9 Determination of static adsorption capacity of protein
        2.2.10 Pretreatment of Egg Yolk Sample
    2.3.Results and Discussion
        2.3.1.Preparation and Characterization of thermo-responsive affinity chromatography stationary phase
        2.3.2.Separation of BSA andγ-globulins via temperature-responsive affinity chromatography stationary phase(TRAC0)
        2.3.3.Effect of temperature on separation of BSA fromγ-globulins
        2.3.4.Adsorption Isotherm
        2.3.5.Separation of antibody by a non-temperature-sensitive HCIC chromatographic stationary phase(Si O2-MEP)and temperature-sensitive affinity chromatography stationary phase(TRAC0)with linear spacer arm
        2.3.6 Application of thermo-responsive stationary phase TRAC0 in separation of Ig G and Ig Y from real sample
    2.4.Conclusions
    References
Chapter 3 Preparation and Characterization of High Capacity Thermo-responsive Affinity Chromatography Stationary Phases with Different Generation of PAMAM Dendrimer as Spacer Arms
    3.1 Introduction
    3.2 Experimental Section
        3.2.1 Reagents and materials
        3.2.2 Experimental instruments
        3.2.3 Preparation of materials
        3.2.4 Characterization of temperature-responsive affinity chromatography materials
    3.3 Results and discussion
        3.3.1 Preparation of high capacity temperature-responsive affinity chromatography stationary phase
        3.3.2 Characterization of high capacity temperature-responsive affinity chromatography stationary phase
    3.4 Conclusions
    References
Chapter 4 Antibody Separation Via High Capacity Thermo-responsive Affinity Chromatography Stationary Phases with PAMAM Dendritic Spacer Arms
    4.1 Introduction
    4.2 Experimental Section
        4.2.1 Reagents and materials
        4.2.2 Experimental instrument
        4.2.3 Preparation of high capacity thermo-responsive stationary phases with thermos-sensitive dendritic polymer PAMAM-PNIPAM as spacer arms
        4.2.4 Packing of High Capacity Thermo-responsive affinity Chromatography Column
        4.2.5 Chromatographic separation
        4.2.6 Characterization of the protein structure
        4.2.7 Sodium dodecyl sulphate-polyacrylamide gel electrophoresis analysis(SDS–PAGE)
        4.2.8 Determination of static adsorption capacity of protein
    4.3 Results and discussion
        4.3.1 Preparation and characterization of high capacity thermo-responsive stationary phases
        4.3.2.Application of high capacity temperature-responsive affinity chromatography stationary phase for separation of antibody
        4.3.3 Effect of temperature on separation of antibodies by temperature-sensitive stationary phases
        4.3.4 Effect of temperature on stationary phase silica-PAMAM without the PNIPAAm and ligands MEP
        4.3.5 Separation and Adsorption Capacity of BSA andγ-globulins by TRAC stationary phases with different ligand number and spacer arms
        4.3.6 Application of thermo-responsive stationary phases in separation of Ig G and Ig Y from real sample
        4.3.7 Application of thermo-responsive stationary phase in separation of recombinant monoclonal antibody(inflixmab)and IgG
    4.4 Conclusions
    References
Chapter 5 Mechanism of Antibody Separation by High Capacity Temperature-responsive Affinity Chromatography Stationary Phases
    5.1 Introduction
    5.2 Experimental Section
        5.2.1 Reagents and materials
        5.2.2 Experimental instruments
        5.2.3 Preparation of thermo-responsive stationary phase with linear spacer arms PNIPAAm and dendritic PAMAM spacer arms
        5.2.4 Packing of the temperature-responsive affinity chromatography column
        5.2.5 Chromatographic separation
        5.2.6 Adsorption isotherm
    5.3 Results and discussion
        5.3.1 Preparation and characterization of thermo-responsive and high capacity thermo-responsive stationary phases
        5.3.2 Mechanism of antibody separation by temperature-responsive affinity chromatography
    5.4 Conclusion
    References
Conclusion
Scientific Achievement During Ph.D Degree
Acknowledgements



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