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Full Screen- TITLE
- DEDICATION
- CERTIFICATE-1
- CERTIFICATE-2
- DECLARATION
- A word of gratitude
- CONTENTS
- ABBREVIATIONS
- 1. Introduction
- lntroduction
- Definition and Subdivision of Lectin
- Occurrence and Distribution
- Physicochemical Properties
- Composition
- Table No 1.1 Well-characterised glycoprotein lectins
- Size and Subunit Structure
- Metal Ion Requirements
- Table 1.2 Metal content and metal requirement for activity of lectins
- Biological Activities
- Cell Binding
- Cell Agglutination
- Table No 1.3 Interaction of lectins with different cell types
- Table 1.4 Factors al.fecting cell agglutination by lectins
- Stimulation of Plant cells
- Inhibition of Fungal Growth
- Protection Against Phytopathogens
- Typing of Bacteria
- Role in Nature
- Cytotoxicity of Lectins
- Nutritional Significance
- Applications
- Cell Separation
- Isolation of Glycosylated Nucleic acids
- Characterisation of Cell Surfaces
- Histochemical and other Clinical Applications of Lectins
- Possible Use of Plant Lectins in Biotechnology and Transgenic Plants.
- Aim and Objectives
- 2. Purification, Molecular Weight Determination, Metal Ion Characterization, etc of BML
- Materials
- Chemicals
- Instruments
- Red Blood Cells
- Trypsinised Erythrocytes
- Methods
- Preparation of Sephadex Gel (Sephadex G-200)
- Preparation on of anticoagulant- Acid citrate Dextrose
- Hemagglutination assay
- Hemagglutination Inhibition assay
- Determination of Temperature stability of Lectin
- Protein Estimation
- Lowrys Method
- Reagents
- Dialysis- activation of dialysis tubing
- Preparation of Alkaline copper
- Procedure
- Electrophoretic Methods
- Polyacrylamide Gel Electrophoresis
- Laemmli System-SDS Phosphate Discontinuous
- Stock Solutions
- Protocol
- Weber and Osborn system - SDS- Phospate Continuous
- Stock solutions
- Native PAGE-Tube Gel
- SDS-PAGE-Slab Gel
- Protocol
- Gel Staining and Destaining
- Determination of molecular weight
- Glycoprotein Staining
- PAS staining
- Purification of Butea Monosperma Lectin
- Ammonium Sulphate Fractionation
- Purification by Affinity Chromatography
- Activation of Guar gum
- Affinity Chromatography
- Purification by Gel Permeation Chromatography
- Purity Test
- Determination of molecular weight
- Fig.2.2 SDS PAGE photograph of BML
- Results
- Hemagglutination Tests
- Agglutination Inhibition Assay
- Determination of Thermal Stability of Hemagglutinating Activity Hemagglutination Titration
- Hemagglutination Titration
- Table 2.1 Agglutinatiaon of BML
- Metal Ion Characterization
- Preparation of Demetallized Lectin
- Atomic Absorption Spectroscopy (AAS)
- Determination of Magnesium
- Preparation of Strontium Chloride 15,000 ppm
- Determination of Manganese
- Determination of Temperature Stability
- 3. Fluorescence Studies of BML
- Introduction
- Quenching Studies
- Unfolding Studies
- Guanidine Hydrogen Chloride Induced Denaturation
- Fig 4.Fluorescence spectrum of GuHCl induced denaturation of BML
- Fig.4.1c Gradation of em, with Concentration of the denaturant
- Urea Induced Denaturation
- Fig.4.2a Fluorescence spectrum of Urea induced Denaturation of BML
- Quenching Studies
- Fig 4.3a Fluorescence spectrum of acrylamide quenching at 280nm
- Fig 4.3b Fluorescence spectrum of acrylamide quenching at 295nm
- Fig 4.4b Stern-Volmer Plot
- Results
- 4. Study of Antibacterial Property
- Materials and Methods
- Bacterial Strains
- Preparation of the Culture Media
- Preparation of Discs
- Preparation of Cotton Swabs
- Plating the medium
- Testing of Anti-microbial Activity
- Determination of Minimum Inhibitory Concentration
- Determination of the Influence of Heat on the Antibacterial Activity of Lectin
- Results
- Table 4.1 Inhibition of bacteria by BML
- Table No. 4.2. Minirnum inhibitory concentration (MIC) of the lectin towardsthe tested bacterial strains
- Table 4.3 Effect of heat on the antibacterial property of lectin
- 5. Cell Proliferation Studies
- Cell Proliferation Studies
- Table 5.1 Lectins with mitogenic effect on human T-lymphocytes
- Materials and Methods
- Human Lymphocytes
- Medium
- Preparation of Giemsa stain
- Preparation of Giemsa working Solution
- Preparation of Tissue Culture Medium (for 500 ml) Fixative Reagents
- Cell Proliferation Assay
- Data Evaluation
- Results
- 6. Cytotoxic and Tumor Regression Studies
- Introduction
- Materials and Methods
- Invitro cytotoxicity studies of Butea Monosperma Lectin on Daltons
- In vivo studies of Lectin
- Results
- Table 6.1. DLA cells viability at various concentrations of Butea monospermalectin
- Table 6.2: Change in body weight of mice bearing tumour induced by DLAcells treated with Butea monosperma Lectin.
- Table 6.3. Life span of mice bearing tumour induced by DLA cells treatedwith Butea monosperma Lectin.
- Table 6.4.Mean change in tumor size (in cm) in mice bearing solid tumorinduced by DLA cells treated with Butea monosperma Lectin.
- DISCUSSION
- 7. Summary
- BIBILIOGRAPHY