Bacterial Immunoglobulin–Binding Proteins

Contributors

Preface


1 Introduction to bacterial immunoglobulin-binding proteins

I. Introduction

II. The Distribution and Functional Reactivity of Bacterial IgG Fc-Binding Proteins

III. Bacterial IgG-Binding Proteins

IV. Second Generation Immunoglobulin-Binding Proteins

V. Summary

References


2 Detection and enhancement of expression of bacterial cell surface immunoglobulin-binding proteins

I. Introduction

II. Preparation of Bacteria

III. Standardization of Bacteria

IV. Assays for the Detection of Immunoglobulin-Binding Proteins

V. Direct Binding of Immunoglobulins to Bacteria

VI. Direct Binding Assay

VII. Absorption of IgG by Bacteria

VIII. Dot Blot Procedure

IX. Methods to Enhance Immunoglobulin-Binding Proteins

X. Mouse Passage

XI. Colony Blot Selection

XII. Storage of Strains

XIII. Conclusions

References


3 Extraction and monitoring of soluble immunoglobulin-binding proteins

I. Introduction

II. Extraction Procedures

III. Secreted IgG-Binding Proteins

IV. Summary

References


4 Isolation and functional characterization of bacterial immunoglobulin-binding proteins

I. Introduction

II. Selection of Immunoglobulin Source to Prepare Affinity Columns

III. Nature of the Sample to Be Purified

IV. Selection of Eluting Agent

V. Affinity Purification of a Type III Fc-Binding Protein Solubilized by Bacteriophage Lysis of the Group C Streptococcus 26RP66

VI. Characterization of Affinity-Purified Immunoglobulin-Binding Proteins

VII. Comparison of Functional Activities of Fc-Binding Proteins

VIII. Summary

References


5 Determination of protein-binding affinities among bacterial cell surface proteins

I. Introduction

II. Binding of Mammalian Proteins to Bacterial Surfaces: Screening Procedures

III. Solubilization of Immunoglobulin-Binding Bacterial Surface Proteins

IV. Determination of Immunoglobulin-Binding Protein in Low Concentrations

V. Analysis of the Binding Immunoglobulins and Other Host Proteins to Purified, Bacterial Surface Proteins

VI. Determination of the Affinity Constant for Binding between a Bacterial Cell Wall Protein and Its Ligand

References


6. Production of polyclonal antibodies to immunoglobulin-binding proteins

I. Introduction

II. Selection of the Species of Animal to Immunize

III. Preparation of Immunogen and Immunization

IV. Kinetics of Antibody Production

V. Detection of Antibody to Immunoglobulin-Binding Proteins

VI. Conclusion

References


7. Use of radiolabeled bacterial Fc-binding proteins as tracers for soluble antigens

I. Introduction

II. Competitive Inhibition Radioimmunoassay

III. Summary

References


8. Application of enzyme-labeled IgG-binding proteins in immunoassay

I. Introduction

II. Types of Assays

III. Preparation of Immunoglobulin Fc-Binding Protein-Enzyme Conjugate Tracers

IV. Development of an ELISA to Quantify Human IgA Using a Type III Fc-Binding Protein-Alkaline Phosphatase Conjugate as Tracer

V. Summary

References


9. Use of Fc-binding proteins to identify cell surface and secreted antigens associated with group B streptococci

I. Introduction

II. Group B Streptococcal Typing Nomenclature

III. Two-Stage Radioimmunoassay for Detection of Group B Streptococcal Type-Specific Antigens

IV. Adaptation of the Two-Stage RIA to a Dot Blot Assay

V. Adaptation of the Two-Stage RIA to an ELISA Typing Procedure

VI. Summary

References


10. Application of Fc-binding proteins for the detection of specific antibodies

I. Introduction

II. Development of an Assay for Antibodies to a Soluble Antigen

III. Detection of Antibodies to Tumor-Associated Antigens

IV. Detection of Rabbit IgM Antibodies to Sheep Erythrocytes

V. Summary

References


11. Use of fluorescent-conjugated bacterial immunoglobulin-binding proteins

I. Standard Methods

II. Conjugation Method Using GMBS

III. Comparison of Recombinant Protein G to Wild Type Protein G Isolated from Streptococcus Cell Membranes

References


12. Biotinylated IgG binding proteins—doubly versatile

I. Introduction

II. Biotinylation of IgG-Binding Protein

III. Immunohistochemical Staining Using Biotinylated IgG-Binding Proteins

References


13. Use of IgG-binding proteins in immunoelectronmicroscopy

I. Introduction

II. Preparation of Colloidal Gold

III. Coupling of Proteins to Colloidal Gold

IV. Use of Gold-Labeling for Localization of Immunoglobulin-Binding Sites and Antigen-Antibody Complexes in Bacteria

V. Double Labeling Techniques with Different Sizes of Colloidal Gold to Localize Two Different Antigens on Thin Sections

VI. Streptavidin/Avidin-Biotin Labeling for Detection of Immunoglobulin-Binding Proteins

VII. Replica Method with Plasma Polymerization Film by Glow Discharge for Three-Dimensional Demonstration of Colloidal Gold Particles

VIII. General Applications of Colloidal Gold Labeling

References


14. The use of bacterial Fc-binding proteins as probes for antigen-antibody complexes immobilized on nitrocellulose membranes

I. Dot Blot Assay

II. Colony and Plaque Blotting of Antigens Expressed by Bacteria

III. Western Blot Analysis

IV. Summary

References


15. Application of bacteria expressing immunoglobulin-binding proteins to immunoprecipitation reactions

I. Introduction

II. General Background

III. Preparation of Bacterial Immunosorbent Reagents

IV. Practical Applications Using Bacterial Immunosorbents

V. Summary

References


16. Use of bacteria expressing immunoglobulin-binding proteins in coagglutination assays

I. Introduction

II. Detection of Cell-Bound Antigens

III. Detection of Specific Antibody

IV. Procedure for Establishing a Coagglutination Assay to Measure a Polyvalent Soluble Antigen

V. Summary

References


17 Utilization of whole bacteria expressing IgG-binding proteins to detect cell surface antigens

I. Introduction

II. Reagents and Equipment

III. Preparation of Antibodies

IV. Preparation of Anti-Immunoglobulin-Coated Bacteria

V. Preparation of Hybridoma Antibody-Coated Bacteria

VI. Binding Assay

VII. Staining and Quantitation

VIII. Variations on the Theme

IX. Comments

References


18 Use of immobilized protein A to purify immunoglobulins

I. Introduction

II. Overview of Purification Procedure

III. Choice of Ligand

IV. Choice of Matrix

V. Immobilization Procedure

VI. Available Binding Capacity

VII. Column Preparation

VIII.Sample Preparation

IX. Sample Application

X. Elution Procedures

XI. Collection and Detection Methods

XII. Column Reequilibration, Reuse, and Storage

XIII. Limitations of Method

XIV. General Methods Using Protein A Sepharose CL-4B and Protein A Sepharose 4 Fast Flow for Mouse and Human IgG Purification

References


19. Purification and quantitation of monoclonal antibodies by affinity chromatography with immobilized protein A

I. Purification of IgGi Monoclonal Antibodies

II. Purification of IgM Monoclonal Antibodies

III. Quantitation of Monoclonal Antibodies

IV. Purification of Injectable-Grade Monoclonal Antibodies

V. Conclusions

References


20. Use of immobilized protein G to isolate IgG

I. Introduction

II. Determination of Optimal Conditions for Protein G Affinity Chromatography

III. Examples of Affinity Chromatography Using Protein G Agarose

IV. Use of Protein G Agarose to Make an Antigen-Binding Column

V. Summary

References


21. Bacterial immunoglobulin-binding proteins and complement

I. Introduction

II. Measurement of Functional Complement Activity

III. Detection of Classical Pathway Complement Activity

IV. Measurement of the Functional Activity of the Alternate Complement Pathway

V. Application of Functional Complement Titrations to Measurement of Activity of Immunoglobulin-Binding Proteins

VI. Measurement of the Generation of Complement Split Products

VII. Measurement of Complement Split Products Generated as a Consequence of Complement Activation Mediated by Bacterial Immunoglobulin-Binding Proteins

VIII. Studies of Binding of the First Component of Complement

IX. Antigenic Determination of Complement Activation

X. Analysis of Complexes Formed between Bacterial Immunoglobulin-Binding Proteins and IgG

XI. Summary

References


22. Activation and differentiation of human lymphocytes by bacterial Fc-binding proteins

I. Introduction

II. Lymphocyte Isolation and Purification

III. Assays for Lymphocyte Proliferation

IV. Assays of Lymphocyte Differentiation

V. Conclusion

References


23. Measurement of in vivo leucocyte chemotaxis mediated by Fc-binding proteins

I. Introduction

II. Air Sac Procedure

III. Use of Fc-Binding Proteins in the Air Sac Procedure

IV. Advantages and Limitations of the Air Sac Procedure

References


24. The cloning of streptococcal protein G genes

I. Colony Immunoassay

II. Streptococcal Clinical Isolates

III. Preparation of Streptococcal DNA

IV. Initial Gene Cloning

V. Cloning of Protein G Genes from Other Isolates

References


25. Bacterial immunoglobulin-binding proteins—future trends

I. Introduction

II. Role of Bacterial Immunoglobulin-Binding Proteins in Pathogenicity

III. Structure-Function Relationships of Bacterial Fc-Binding Proteins

IV. Applications Involving Immunoglobulin-Binding Proteins—Future Trends

V. Summary

References

Appendix

I. General Buffers

II. Iodination Buffers and Related Solutions

III. ELISA Buffers and Related Solutions

IV. Electrophoresis Buffers

V. Buffers for Use in Applications Involving Nitrocellulose

VI. General Buffers and Reagents

Index