Agglutination, Complement, Neutralization, and Inhibition - 1st Edition - ISBN: 9780127544045, 9781483220598

Agglutination, Complement, Neutralization, and Inhibition

1st Edition

Methods in Immunology and Immunochemistry, Vol. 4

Editors: Curtis A. Williams Merrill W. Chase
eBook ISBN: 9781483220598
Imprint: Academic Press
Published Date: 28th November 1977
Page Count: 488
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Methods in Immunology and Immunochemistry, Volume IV: Agglutination, Complement, Neutralization, and Inhibition provides information pertinent to direct and indirect agglutination reactions. This book covers a variety of topics, including complement-fixation procedures, isolation of complement components, hemolytic intermediates, complement-related proteins, and neutralization reactions.

Organized into three chapters, this volume begins with an overview of test-tube agglutinations that are preferred for blood grouping with saline agglutinins that require more than a few minutes for agglutination. This text then describes blood group antibodies that agglutinate red blood cells suspended in saline. Other chapters consider the classical pathway of complement utilization. This book discusses as well the complexity of events leading to hemolysis of erythrocytes by complement. The final chapter deals with the ability of antitoxin to neutralize diphtheria toxin and explains the quantitative relationships between antigen and antibody.

This book is a valuable resource for immunologists, scientists, and research workers.

Table of Contents

Contributors to Volume IV


Contents of Other Volumes

Chapter l6. Agglutination and Fiocculation

A. Direct Hemagglutination

Slide Agglutination

Tube Agglutination: Saline Agglutinins

Quantitative Cell-Counting Method

Tube Agglutination: Incomplete Antibodies and Cold Agglutinins

Enzyme-Treated Erythrocytes


B. Indirect (Passive) Hemagglutination

Direct Passive-Polysaccharide Adsorption on Untreated Erythrocytes

Hemagglutination with Tannic Acid-Treated Erythrocytes

Antigen-Erythrocyte Conjugates Prepared with Bisdiazobenzidine

Preparation of Stable Formalinized Antigen-Erythrocyte Conjugates

Special Methods of Chemical Bonding for Preparing Antigen-Erythrocyte Conjugates

Chromic Chloride, Carbodiimide Coupling, Difluorodinitrobenzene

Specific Coupling to Erythrocyte Antigenic Sites

Antiglobulin Reactions with Erythrocytes

Direct, Indirect Reactions

Inhibition Tests

Antiglobulin Consumption Test

The Red Cell-Linked Antigen Test

Mixed Agglutination and Erythrocytes

The Mixed Agglutination Reaction

The Mixed Antiglobulin Reaction

The Mixed Hemadsorption Test

C. Inhibition of Hemagglutination

Inhibition by Antibodies and Antibody-like Reagents: Semiquantitative Tube Tests

Quantitative Inhibition (Cell-Counting Method)

D. Bacterial Agglutination

Preparation of Bacteria as Antigens

Special Antigens


Agglutination Tests

Tube Tests

Cover Slip Tests

Macroscopic Slide Tests

Enumeration of Bacterial Colonies in Liquid Media

Antigens Inhibiting Agglutination

Interpretation and Applications

E. Immobilization of Motile Bacteria by Anti-Flagellar Antibody

F. Agglutination of Spermatozoa

General Considerations

The Sperm Antigens

Preparation of Heteroand Autospermagglutinins

Semen Examination

Spermagglutination: Macroscale



Comparisons of the Various Spermagglutination Methods

G. Agglutination with Antigen on Inert Particles

1. Latex Fixation Techniques

2. The Bentonite Flocculation Test

Chapter 17. Complement

A. Introduction

The Classical Pathway of Complement Utilization

Alternative Pathway for C3 Activation

Principles of Quantitation of Complement Components

Complement Fixation Reactions

Topical Presentation of Complement Components

Key to Buffers for the Complement System

B. Immune Hemolysis and Complement Fixation

1. Diluents and Reagents

2. General Procedures

3. Complement Fixation Tests

4. Passive Immune Hemolysis

C. Preparation of Hemolytic Intermediates

1. General Considerations

2. Hemolytic Intermediates from Guinea Pig Complement

3. Measurement of Hemolytic Intermediates of Guinea Pig Complement

4. Hemolytic Intermediates from Human Complement

D. Isolation and Assay of Complement Components

1. Introduction

2. Complement Components from Human Serum

3. Complement Components from Guinea Pig Serum

E. Isolation and Assay of Complement-Related Proteins

1. Introduction

2. Isolation and Assay of Human Serum C1-Inhibitor (C1-INH)

3. Human C3b-Inactivator (C3b-In, or Conglutinogen-Activating Factor, KAF)

4. Isolation and Assay of Human Properdin

5. Isolation and Assay of Properdin Factor D

6. Purification of Properdin Factor B (C3PA, GBG, C3-Proactivator). Preparations of Human Properdin Factor B

7. Isolation of Cobra Venom Factor (CoF, CVF)

8. Formation and Isolation of Anaphylatoxin C3a

9. Anaphylatoxin C5a (Human, Guinea Pig)

F. Antisera to Complement Proteins

1. Introduction

2. Antisera to Guinea Pig Components

3. Rabbit Antisera to Human Complement Components

4. Detection of Complement in Tissues by Fluorochrome-Coupled Anti-C3

Chapter 18. Neutralization Reactions

A. Toxin-Antitoxin Systems

1. Introduction

2. Methods for Toxin and Antitoxin Assay in Vitro

3. Neutralization of Toxin in Vivo

B. Enzyme-Antibody Interactions

1. Characterization of the Enzyme-Antibody Interaction

2. Mechanism of Modification of Enzyme Activity by Antibody Introduction

3. Immunoenzymological Comparisons

C. Virus Neutralization

1. Immunological Reactions of Bacteriophages

2. Antibody Reactions with Chemically Modified Bacteriophages

Appendix I World Health Organization Report on Nomenclature of Complement

Nomenclature of Complement

Components of Complement

First Component (C1)

Intermediate Complexes

Complement Components Showing Altered Activity

Fragments and Chains

Complement Factors Not Directly Involved in the Haemolytic Sequence


Appendix II The International System of Units: SI Nomenclature


Basic Units and Multiples

Supplementary Units

Derived SI Units with Special Names

Examples of Other Derived SI Units

Units to Be Allowed in Conjunction with SI

Examples of Units Contrary to SI, with their Equivalents

Author Index

Subject Index


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© Academic Press 1977
Academic Press
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About the Editor

Curtis A. Williams

Merrill W. Chase

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