BIOLOGICAL AMPLIFICATION SYSTEMS IN IMMUNOLOGY
CONTENTS 1 HISTORY OF IMMUNOLOGY1 1 MILESTONES IN IMMUNOLOGY 1 — 2 ARCHITECTURE OF IMMUNE SYSTEM 7— 2 1 INTRODUCTION 2 2 PRIMARY LYMPHOID ORGAN 2 3 THYMUS 2 3 1 SECTION OF THYMUS ENLARGED 2 3 2 THYMOSIN 2 3 3 THYMOPOIETIN 2 3 4 THYMIC FACTOR 2 3 5 THYMIC HUMORAL FACTOR 2 4 BONE MARROW 2 4 1 BURSA OF FABRICUS 2 5 T CELL MATURATION 2 5 1 MATURATION AND SELECTION OF T CELLS 2 6 BONEMARROW B CELL MATURATION 2 7 SECONDARY LYMPHOID ORGAN 2 8 THE SPLEEN 2 9 LYMPHNODES 2 9 1 LYMPH NODE 3 IMMUNE RESPONSE 18 — 3 1 T-LYMPHOCYTES 3 2 EFFECTOR RESPONSES IN HUMORAL BRANCHES 3 2 1 ANTIBODIES PERFORM THIS FUNCTION BY FOLLOWING MECHANISMS 3 2 2 PRIMARY AND SECONDARY RESPONSE 3 2 3 T CELL ACCESSORY MEMBRANE MOLECULES 3 2 4 CD4 AND CD8 RECEPTORS 3 2 5 CD4 3 2 6 CD8 3 2 6 1 PROPERTIES 3 2 7 SIGNAL TRANSDUCTION PROPERTIES 3 2 8 OTHER ACCESSORY MEMBRANE MOLECULES 3 2 9 T CELL ACTIVATION 3 2 10 EVIDENCE THAT TCR ABOVE RECOGNIZED PEPTIDE COMPLEXES 3 2 11 IMMEDIATE GENES 3 2 12 LATE GENES 3 2 13 TCR COATED SIGNAL PATHWAYS 3 2 14 EVIDENCE THAT TCR RECOGNIZED PEPTIDE MHC MOLECULES 3 2 15 COMPARISON OF PRIMARY AND SECONDARY IMMUNE RESPONSE 3 2 16 VIRAL NEUTRALIZATION BY HUMORAL ANTIBODY 3 2 17 POLIO OF TH CELLS IN HUMORAL RESPONSE 3 2 18 FORMATION OF TB CONJUGATE 3 2 19 CELL MEDIATED IMMUNE RESPONSE 3 2 20 FORMATION OF TB CONJUGATE 3 2 21 CONTACT DEPENDENT HELP MEDIATED BY CD40, CD40L INTERACTIONS 3 2 22 T CELL RECEPTOR COMPLEX 3 2 23 ITAM 3 2 23 1 ROLE 3 3 CELLULAR IMMUNITY 3 3 1 INTRODUCTION 3 3 2 ORIGIN OF LYMPHOCYTES 3 3 3 FUNCTION OF LYMPHOCYTES 4 IMMUNITY 30 — 4 1 INNATE IMMUNITY 4 1 1 INNATE NON SPECIFIC IMMUNITY 4 2 ACQUIRED IMMUNITY 4 3 INNATE OR NATURE IMMUNITY 4 4 RACIAL IMMUNITY 4 5 INDIVIDUAL IMMUNITY 4 5 1 AGE 4 5 2 HORMONAL INFLUENCE 4 5 3 NUTRITION 4 6 IMMUNITY 4 7 IMMUNE RESPONSE 4 8 INNATE IMMUNITY 4 8 1 SUMMARY OF NON SPECIFIC IMMUNITY 4 8 2 PHYSIOLOGICAL BARRIER 4 8 3 PHAGOCYTIC AND ENDOCYTIC BARRIERS 4 8 4 INFLAMMATORY BARRIERS 4 8 5 ANATOMY BARRIERS 4 8 6 PHYSIOLOGICAL BARRIER 4 8 7 ENDOCYTIC AND PHAGOCYTIC BARRIER 4 9 CONTAINING CLASS I AND CLASS II HLA MOLECULE 4 10 ANTIGEN PRESENTATION BY CLASS I AND CLASS II MAJOR HISTOCOMBATIBILITY COMPLEX PROTEINS 4 11 PEPTIDE BINDING BY CLASS I AND CLASS II MAJOR HISTOCOMBATIBILITY COMPLEX 5 EPITOPES 38 — 5 1 INTRODUCTION 5 2 EPITOPE CHARACTERISTICS 5 3 PROPERTIES OF OF B- CELL EPITOPE 5 4 B CELL EPITOPE CAN CONTAIN SEQUENTIAL OR NON SEQUENTIAL AMINO ACIDS 5 5 B CELL EPITOPES TEND TO BE LOCATED IN FLEXIBLE REGIONS OFTEN IMMUNOGEN AND DISPLAY SITE MOBILITY 5 6 COMPLEX PROTEINS CONTAIN MULTIPLE OVERLAPPING B CELL EPITOPE, SOME OF WHICH ARE IMMUNODOMINANCE 6 STRUCTURE AND FUNCTION OF SPECIFIC IMMUNOGLOBULINS 44 — 6 1 IGG 6 1 1 STRUCTURE 6 1 2 FUNCTION 6 2 IGA 6 2 1 SERUM IGA 6 2 2 SECRETORY IGA 6 2 3 STRUCTURE 6 2 4 BIOLOGICAL PROPERTIES OF IGA 6 3 IGM 6 3 1 BIOLOGICAL PROPERTIES OF IGM 6 4 IGD 6 5 IGE 6 6 ANTIGENIC DETERMINANTS ON IGS 6 6 1 ISOTYPIC DETERMINANTS 6 6 2 ALLOTYPIC DETERMINANTS 6 6 3 ISOTYPIC DETERMINANTS 6 6 4 DISEASE RELATED TO DEFICIENCY 6 6 5 COMPLEMENT DEFICIENCY 6 6 6 HEREDITARY ANGIO NEURO EDEMA 6 7 MODERN THEORY OF AB PRODUCTION 7 IMMUNO DIAGNOSTICS, ABO, RH, INCOMPATIBILITY, WIDAL TEST, 55 — ELISA, RIA TECHNIQUES AND THEIR APPLICATION 7 1 INTRODUCTION 7 2 ABO RH (D) INCOMBATIBILITY 7 3 WIDAL TEST 7 4 HEMOLYTIC TRANSFUSION REACTION 7 1 INTRAVASCULAR HEMOLYSIS OF RBC 7 2 NON HEMOCYTIC 7 3 NON DELAYED HEMOCYTIC 7 4 INFECTION DISEASES 7 5 IMMEDIATE HEMOCYTIC REACTION 7 6 EPIDEMOLOGY 7 7 SCIENCE AND SYMPTOMS 7 8 IMMUNOLOGY MAIN TITRATION 7 8 DELAYED REACTION 8 HYPERSENSITIVITY 59 — 8 1 TYPE I ANAPHYLACTIC REACTION 8 2 TYPE II CYTOLYTIC OR CYTOTOXIC REACTION 8 3 TYPE III CELL MEDIATED IMMUNITY 8 4 TYPE I 8 4 1 THE SENSITIZATION PHASE 8 4 2 ACTIVATION PHASE 8 4 3 EFFECTOR PHASE 8 4 4 SENSITIZATION PHASE 8 4 5 ACTIVATION PHASE 8 4 6 BY AN INFLUX OF CA2+ IONS 8 4 7 EFFECTOR PHASE 8 4 8 PREFORMED MEDICINE 8 4 9 SUBSTANCES SYNTHESIZED DENOVA 8 4 10 HISTAMINE 8 4 11 SEROTONIN 8 4 12 EOSINOPHILIC CHEMOTACTIC FACTOR OF ANAPHYLAXIS (ECF A) 8 4 13 SRS(A) 8 4 14 PLATELET ACTIVATION FACTOR 8 4 15 LOCALISED ANAPHYLAXIS (ATOPY) 8 5 ALLERGIC RHINITIS 8 6 ALLERGIC ASTHMA 8 7 INTRINSIC ASTHMA 8 8 EARLY RESPONSE 8 9 LATE RESPONSE 8 10 ATOPIC DERMATITIS 8 11 LATE PHASE REACTION 8 12 ANTIBODY MEDIATED CYTOTOXIC (TYPE II) HYPERSENSITIVITY 8 12 1 TRANSFUSION REACTION 8 12 2 HEMOLYTIC DISEASE OF THE NEW BORN 8 12 3 LOCALIZED TYPE III REACTION 8 12 4 GENERALIZED TYPE III REACTION 8 12 5 TDTH MEDIATED (TYPE IV) HYPERSENSITIVITY 9 COMPLEMENT 72 — 9 1 COMPLEMENT CASCADE 92 METABOLISM 9 2 1 SYNTHESIS 9 3 PROTEINS THAT REGULATE COMPLEMENT SYSTEM 9 4 GENE CLUSTURE 9 5 ALTERNATE PATHWAY 9 6 CLASSICAL PATHWAY 9 7 ALTERNATE PATHWAY (AMPLIFICATION OF LOOP) 9 7 1 REGULATION 9 8 BIOLOGICAL FUNCTION OF COMPLEMENT SYSTEM 9 8 1 CYTOLYSIS 9 8 2 OPSONISATION 9 8 3 VIRAL NEUTRALIZATION 9 8 4 INFLAMMATORY RESPONSE 9 8 5 CHAVARE OF IMMUNE COMPLEXES 9 9 LYMPHOID SYSTEM 9 9 1 PAYERS PATCH 10 AUTOIMMUNITY 81 — 10 1 INTRODUCTION10 2 CLASSIFICATION OF AUTO IMMUNE DISEASES 10 3 HEMOLYTIC AUTO IMMUNE DISEASES 10 3 1 AUTO IMMUNE HEMOLYTIC ANEMIAS 10 3 2 AUTO IMMUNE CHROMBO CYTOPENIA 10 3 3 AUTO IMMUNE LEUCOPENIA 10 3 4 LOCALIZED AUTOIMMUNE DISEASE (DISEASE MEDIATED BY CELLULAR DAMAGE) 10 3 5 ORGAN SPECIFIC AUTO IMMUNE DISEASE 10 3 6 HASHIMOTAS THYROIDITIS 10 3 7 AUTOIMMUNEANAEMIAS 10 3 8 PERINICIOUS ANEMIA 10 3 9 GOOD PASTEUR’S SYNDROME 10 3 10 INSULIN DEPENDENT DIABETES MELLITUS 10 3 11 DISEASES MEDIATED BY STIMULATING OR BLOCKING ABS 10 3 12 GRAVES DISEASES/MYASTHENIA GRAVIS 10 3 13 SYSTEMIC AUTO IMMUNE DISEASE 10 3 14 SYSTEMIC LUPUS ERYTHEMATOSUS 10 3 15 RHEUMATOID ARTHRITIS 10 3 16 MULTIPLE SCLEROSIS 10 3 17 TRANSITORY AUTOIMMUNE PROCESSES 11 TRANSPLANTATION 87 — 11 1 AUTOGRAFTS 11 2 ISOGRAFTS11 3 ALLOGRAFTS 11 4 XENOGRAFTS 11 5 TRANSPLANTATION AG 11 6 MECHANISM INVOLVED IN GRAFT REJECTION 11 7 SENSITIZATION STAGE 12 INTERFERON 90 — 12 1 CYTOKINES 12 2 WHY CYTOKINES ARE IMPORTANT 12 3 INTERFERON12 4 DEFINITION 12 5 ISSAC LINDEMANN EXPERIMENT 12 6 CHARACTERISTICS OF INTERFERON 12 7 TYPES OF INTERFERON 12 8 A INTERFERONS 12 9 ?-INTERFERONS 12 10 ? INTERFERONS 12 11 CLASS OF INTERFERONS 12 11 1 ?-INTERFERON 12 11 2 ?-INTERFERON 12 11 3 ?-INTERFERON 12 12 INTERFERONS AS HORMONES 12 13 INDUCTION OF INTERFERONS 12 13 1 PRODUCTION 12 13 1 1 CLASS A INDUCERS 12 13 1 2 CLASS B INDUCERS 12 13 2 TYPE II PRODUCTION 12 14 BIOLOGICAL PRODUCTION OF INTERFERONS 12 15 PRODUCTION OF INTERFERONS BY USING RDNA TECHNOLOGY 12 15 1 STEP I CDNA PREPARATIONS 12 15 2 STEP II LARGE SCALE PRODUCTION12 16 DISORDERS OF HUMAN INTERFERONS 13 CYTOKINES 96 — 13 1 PROPERTIES OF CYTOKINES 13 2 MONONUCLEAR PHAGOCYTES 13 2 1 DEVELOPMENT 13 2 2 AVIDITY 13 2 3 PHYTOHEMOAGGLUTINATION (PHA) 13 2 4 ADVANTAGES OF INACTIVATED VACCINES 13 2 5 ADVANTAGES OF LIVE VACCINES 14 TISSUE TYPING 101 — MOLECULAR MIMICRY 14 12 HEAT SHOCK PROTEINS 15 ADJUVANTS 108 — 16 ANTIBODY FORMING THEORY 110 — 16 1 SELECTIVE THEORY16 2 INSTRUCTIVE THEORY 16 3 CLONAL SELECTION THEORY 16 4 TEMPLATE OR DIRECTIVE OR INSTRUCTIVE THEORY 16 5 INSTRUCTIVE THEORY 16 6 POLYCLONAL ANTIBODIES 16 6 1 PRODUCTION 16 7 CELL AND CELL COLLABORATION 16 7 1 THREE TYPES OF CELLS ARE INVOLVED IN THE IMMUNE RESPONSE 16 7 2 T-B CELL CO-OPERATION 16 7 3 MECHANISM FOR T- B CELL CO-OPERATION 16 7 4 CARRIER EFFECT 17 ANTISERA, 115 — 17 1 FREUND’S ADJUVANT 17 2 RABBIT ANTIMOUSE IMMUNOGLOBULIN 17 2 1 MATERIALS AND EQUIPMENT 17 2 2 METHOD 17 3 AMMONIUM SULPHATE PRECIPITATION 17 3 1 METHODS 17 3 2 CALCULATION OF PROTEIN CONTENT 17 4 PURIFICATION OF ANTIBODIES 17 5 PREPARATION OF IMMUNOADSORBANT FOR ANTIBODY PURIFICATION 17 5 1 MATERIALS AND EQUIPMENTS 17 5 2 METHOD 17 5 3 USE OF IMMUNOADSORBANT FOR ANTIBODY PURIFICATION 17 5 3 1 MATERIALS AND EQUIPMENTS 17 5 3 2 METHOD 17 5 4 DISSOCIATION OF COMPLEX 17 5 5 CALCULATION OF RECOVERY FROM IMMUNOADSORBANT 17 5 6 ELUATES FROM IMMUNOADSORBENT TOTAL PROTEIN CONCENTRATION IN ELUATE IMMEDIATELY AFTER CONCENTRATION AND DIALYSIS 17 5 7 CALCULATION OF ANTIBODY CONTENT OF ELUATE 18 ANTIBODY-RECEPTOR STRUCTURE 120 — 18 1 INTRODUCTION 18 2 CELLULAR ASPECTS OF THE IMMUNE RESPONSE 18 3 RECEPTOR STRUCTURE – IMMUNOGLOBULIN 18 4 IMMUNOGLOBULIN GENE 18 5 THE T CELL RECEPTOR 18 6 MATURATION OF THE IMMUNE RESPONSE 18 7 THE USE OF ANTIBODIES IN MOLECULAR BIOLOGY 18 8 THE USE OF ANTIBODIES TO IDENTIFY POLYPEPTIDES 18 9 THE USE OF ANTIBODIES TO ASSAY MRNA 18 10 THE USE OF ANTIBODIES TO PURIFY MRNA 18 11 DIRECT SCREENING OF CDNA COUNTS WITH ANTIBODIES 18 12 IMMUNOBLOTTING FOR THE SCREENING OF MONOCLONAL ANTIBODIES 18 13 PURIFICATION OF ANTIBODIES FOR IMMUNOPRECIPITATION 18 14 IMMUNOPRECIPITATION OF POLYPEPTIDES FROM CELL FREE TRANSLATION 18 15 IMMUNOPRECIPITATION OF POLYSOME 18 16 DERIVATION OF HYBRIDS SECRETING SPECIFIC ANTIBODIES 18 16 1 MYELOMAS 18 16 2 SOMATIC CELL FUSION 18 16 3 HAT SELECTION 18 16 4 SELECTIVITY OF SOMATIC CELL FUSION 18 17 PLANNING OF THE FUSION EXPERIMENT 18 17 1 DIARY OF SUCCESSFUL EXPERIMENT 18 17 2 ANTIGEN PREPARATION 18 17 3 PLANNING THE IMMUNIZATION AND EXPERIMENTS 18 17 4 DAY – 56 18 17 5 DAY - 44 18 18 CELL FUSION AND SELECTION HYBRIDS 18 18 1 DAY - O 18 18 2 DAY - 1 18 19 SCREENING AND SELECTION OF MONOCLONAL ANTIBODIES SECRETING LINE 18 19 1 CONCLUSION OF THE EXPERIMENT 19 DESIGNING VACCINES FOR ACTIVE IMMUNIZATION 146 — 19 1 WHOLE ORGANISM VACCINES 19 2 CLASSIFICATION OF COMMON VACCINES FOR HUMANS 19 2 1 VIRAL PARTICLES 19 2 2 ATTENUATED VIRAL AND BACTERIAL VACCINES 19 3 DISADVANTAGES 19 4 INACTIVATED VIRAL AND BACTERIAL VACCINES 19 5 TOXOID VACCINES 19 6 RECOMBINANT VECTOR VACCINES 19 7 DNA VACCINES 19 8 SYNTHETIC PEPTIDE VACCINES 19 9 MULTIVATION SULE UNIT VACCINES 19 10 PURIFIED MUSCULAR VACCINES 19 11 TRANSGENIC PLANTS FOR PRODUCTION OF ANTIBODIES 20 CANCER, CANCER GENES AND GROWTH FACTORS 54 — 20 1 BIOCHEMICAL IMPORTANCE 20 2 PHYSICAL,CHEMICAL AND BIOLOGICAL AGENTS 20 2 1 CHEMICAL 20 2 2 TABLE 20 2 3 STRUCTURE 20 2 4 ACTION 20 3 METABOLISM OF CARCINOGEN 20 4 COVALENT BONDING 20 5 DAMAGE TO DNA 20 6 MUTAGENS 20 7 INITIATION AND PROMOTION 20 8 DIAGRAMATIC REPRESENTATION OF THE STAGE OF INITIATION AND PROMOTION OF CHEMICAL CARCINOGENESIS IN SKIN 20 9 SOME DNA AND RNA VIRUSES ARE CARCINOGENIC 20 10 CHROMOSOMAL TRANSLOCATION 20 11 GENE AMPLIFICATION 20 12 ACTIVATOR OF ONCOGENES 20 13 POLYPEPTIDE GROWTH FACTOR 20 14 MALIGNANCY OF TUMOR CELLS 20 15 METASTASIS, THE MOST DANGEROUS PROPERTY OF TUMOR CELLS 21 ACQUIRED IMMUNO DEFICIENCY SYNDROME 163 — 21 3 GENETIC VARIABILITY OF HIV –1 21 4 IMMUNOLOGICAL LESIONS IN AIDS 21 5 T CELL ABNORMALITIES 21 6 B CELL ABNORMALITIES 21 7 NK- CELL ABNORMALITIES 21 8 MACROPHAGE ABNORMALITIES 21 9 HIV IN OTHER CELL 21 10 AIDS THE DISEASE 21 11 SECONDARY INFECTIONS 21 12 NEUROLOGIC LESION 21 13 KAPOSI’S SARCOMA 21 14 GENETIC FACTORS 21 15 DIAGNOSIS OF HIV INFECTION 21 16 TREATMENT OF HIV INFECTION 21 17 AN AIDS VACCINE 21 18 AIDS IN ANIMALS 21 19 OTHER HUMAN VIRUSES AFFECTING THE HUMAN SYSTEM 21 20 AIDS PREVENTION AND ITS SOCIAL IMPLICATIONS 21 21 PREVENTION OF AIDS AN INDIAN PERSPECTIVE 21 22 AN OVERVIEW OF WORLD AIDS 21 23 AIDS IN INDIA 21 24 MOBILIZING YOUNG PEOPLE OF AIDS CONTROL 21 25 COMPAIGN OBJECTIVES 21 26 INCULCATING AWARENESS 21 27 CURRENT DEVELOPMENTS IN AIDS RESEARCH 21 28 BIOTECHNOLOGY POINT OF VIEW INTRODUCTION 21 29 PENETRATION AND DESTRUCTION OF CD4 T CELLS BY HIV 21 30 HYPOTHESIS FOR ETIOLOGY OF AIDS 21 31 SUGGESTION FOR FUTURE RESEARCH ON ETIOLOGY AND THERAPY 22 IMMUNOLOGICAL AND IMMUNOCHEMICAL TECHNIQUES 185 — 22 1 OUCHTERLONY DOUBLE IMMUNODIFFUSION TECHNIQUE 22 1 1 MATERIALS REQUIRED 22 1 2 PROCEDURE 22 2 OPTIMAL PROPORTIONAL ZONE OF EQUIVALENCE OF ANTIGEN- ANTIBODY REACTION 22 3 OUCHTERLONY DOUBLE IMMUNODIFFUSION TECHNIQUE-IDENTITY, PARTIAL IDENTITY AND NON IDENTITY TESTS 22 3 1 MATERIALS REQUIRED 22 3 2 PROCEDURE 22 4 QUANTITATIVE IMMUNODIFFUSION TECHNIQUE 22 4 1 PREER’S DOUBLE IMMUNODIFFUSION 22 4 1 1 PRINCIPLE 22 4 1 2 MATERIALS REQUIRED 22 4 1 3 PROCEDURE 22 4 2 OUDIN’S SINGLE IMMUNODIFFUSION 22 4 2 1 PRINCIPLE 22 4 2 2 MATERIALS REQUIRED 22 4 2 3 PROCEDURE 22 4 3 MANCINI’S SINGLE RADIAL IMMUNODIFFUSION 22 4 3 1 PRINCIPLE 22 4 3 2 MATERIALS REQUIRED 22 4 3 3 PROCEDURE 22 4 4 DETERMINATION OF THE DIFFUSION OF THE COEFFICIENT OF AN ANTIGEN 22 4 4 1 MATERIALS REQUIRED 22 4 4 2 PROCEDURE 22 4 5 RECORDING AND PRESERVATION OF THE GEL PLATES 22 4 5 1 MATERIALS REQUIRED 22 4 5 2 PROCEDURE 22 4 6 IMMUNOELECTROPHORESIS (GRABER AND WILLIAMS QUALITATIVE ANALYSIS) 22 4 6 1 PRINCIPLE 22 4 6 2 MATERIALS REQUIRED 22 4 6 3 PROCEDURE 22 4 7 COUNTER IMMUNOELECTROPHORESIS 22 4 7 1 MATERIALS REQUIRED 22 4 7 2 PROCEDURE 22 4 8 ROCKET IMMUNOELECTROPHORESIS 22 4 8 1 MATERIALS REQUIRED 22 4 8 2 PROCEDURE 22 4 9 TWO DIMENSIONAL CROSSED IMMUNO ELECTROPHORESIS 22 4 9 1 MATERIALS REQUIRED 22 4 9 2 PROCEDURE 22 5 APPENDIX 22 5 1 PRECOATING OF SLIDE 22 5 2 SANDWICH ASSEMBLY 22 5 3 PHOSPHATE BUFFER SALINE-AZIDE SOLUTION (PBS AZIDE) 22 5 4 1% AGAR 22 5 5 COOMASSIE BRILLIANT BLUE STAINING SOLUTION 22 5 6 DESTAINING SOLUTION 22 5 7 BARBITAL BUFFER 22 5 8 TRIS-GLYCINE BARBITAL BUFFER 22 5 8 1 BUFFER A 22 5 8 2 BUFFER B 23 STAPHYLOCOCCUS AUREUS PROTEIN – A PURIFICATION 198 — 23 1 TABLE 1 PROTEIN ‘A’ BINDING SPECIFICITY 23 2 EQUIPMENT 23 3 PREPARATION OF S AUREUS STOCK SOLUTION FOR IMMUNOADSORPTION 23 3 1 REAGENTS 23 3 2 PROCEDURE 23 3 3 TECHNICAL COMMENTS 23 4 DOUBLE DIFFUSION IN TWO DIMENSIONS 23 4 1 MATERIAL AND EQUIPMENT 23 4 2 METHOD 23 5 SUGGESTED ANTIBODY AND ANTIGEN PATTERNS 23 6 ANTISERUM 23 7 INTERPRETATION OF RESULT 24 AGGLUTINATION REACTION 202 — 24 APPLICATION OF AGGLUTINATION REACTION 24 1 SLIDE AGGLUTINATION TEST 24 2 TUBE AGGLUTINATION TEST 24 3 WIDAL TEST 24 4 TEST PROCEDURE 24 4 1 PROCEDURE 24 5 WELFLIX REACTION 24 6 HEME AGGLUTINATION TEST 24 6 1 BLOOD GROUPING CROSS MATCHING 24 7 COLD AGGLUTINATION TEST 24 8 PAUL BUNNEL TEST 24 9 ROSE WALAR’S TEST 24 9 1 TEST 24 10 MODIFIED ROSE WALAR’S TEST 24 11 COOMB’S TEST 24 11 1 PRINCIPLE 24 11 2 DIRECT COOMBS TEST 24 11 3 INDIRECT COOMBS TEST 24 12 COATED PARTICLE AGGLUTINATION TEST 24 13 LATEX AGGLUTINATION TEST 24 13 1 LATEX AGGLUTINATION TEST FOR RHEUMATOID FEVER 24 13 2 LATEX AGGLUTINATION TEST FOR ROTA VIRUS 24 14 MISCELLANEOUS AGGLUTINATION TEST 24 15 VIRUS AGGLUTINATION MONKEYS RBC’S 24 16 HAEMAGGLUTINATION INHIBITION TEST FOR INFLUENZA 25 PURIFICATION OF IG BY ION EXCHANGE CHROMATOGRAPHY 211 — 25 1 CHROMATOGRAPHY 25 1 1 PRINCIPLE 25 1 2 MATERIALS 25 1 3 PREPARATION OF COLUMN 25 1 4 SEPARATION OF IGS 25 2 IMMUNOFLORESCENT ASSAY 25 3 INDIRECT FLUORESCENT ANTIBODY TEST 25 4 COMPARISON OF TECHNIQUES INVOLVED IN DIRECT AND INDIRECT FLORESCENT ANTIBODY TEST 25 4 1 APPLICATIONS 26 RADIO IMMUNOASSAY AND RELATED TECHNIQUES 215 — 26 1 INTRODUCTION 26 2 RADIO IMMUNOASSAY 26 2 1 TECHNICAL REQUIREMENTS FOR RADIO IMMUNOASSAY 26 2 2 MONOCLONAL ANTIBODIES 26 2 3 LABELLED ANTIGEN 26 2 4 THE ANTIBODY 26 2 5 RADIO IODINATION PROCEDURES 26 2 6 ASSESSMENT OF THE QUALITY OF RADIO LABELS 26 2 7 STORAGE 26 2 8 SAFETY 26 2 9 SEPARATION OF FREE AND BOUND ANTIGENS 26 2 10 EFFICIENCY 26 2 11 PRACTICABILITY 26 2 12 SEPARATION METHODS 26 2 13 STANDARDS 26 2 14 ASSAY OPTIMIZATION 26 2 15 ASSAY CONDITION 26 2 16 LABELED ANTIGEN CONCENTRATION 26 2 17 SEPARATION SYSTEM 26 2 18 ANTIBODY CONCENTRATION (TITRE) 26 2 19 STANDARD CURVE 26 2 20 VALIDATION OF RADIO IMMUNOASSAY 26 2 21 ASSESSMENT OF SENSITIVITY ( DETECTION LIMIT ) 26 2 22 ASSESSMENT OF SPECIFICITY 26 2 23 STANDARD CURVE 26 2 24 PRECISION PROFILE 26 2 25 ASSESSMENT OF ACCURACY ( BIAS) 26 2 26 ASSESSMENT OF PRECISION 26 2 27 ASSESSMENT OF LINEARITY ( PARALLELISM) 26 2 28 OTHER ASPECTS 26 2 29 COUNTING AND CURVE FITTING PROCEDURES 26 2 30 QUALITY ASSESSMENT 26 2 31 INTERNAL QUALITY CONTROL 26 2 32 EXTERNAL QUALITY ASSESSMENT 26 2 33 AUTOMATION 26 2 34 FREE HORMONE MEASUREMENT 26 2 35 INDIRECT METHODS 26 2 36 DIRECT METHODS 26 2 37 TECHNIQUES RELATED TO RADIO IMMUNO ASSAY 26 2 38 LABELED ANTIBODY IMMUNOASSAY 26 3 ENZYME IMMUNO ASSAY 26 4 FLURO IMMUNOASSAY 26 5 LUMINESCENT IMMUNO ASSAY 26 5 1 CONCLUSION 26 6 RADIO IMMUNO ASSAY 26 6 1 TABLE COMPOUNDS QUANTITATED BY RADIO IMMUNO ASSAY 26 6 2 CONCEPTUAL OPERATION OF THE RADIO IMMUNOASSAY 26 6 3 125 I LABELING PROCEDURES 26 6 4 STANDARDISATION OF RADIO IMMUNOASSAY 27 ELISA (ENZYME LINKED IMMUNO SORBANT ASSAY) 237 — 27 1 EIA27 2 ELISA VS EMIT27 3 EMIT27 4 EMIT OUTLINE 27 5 ELISA 27 5 1 COMPETITIVE ( INHIBITION) METHOD FOR ANTIGEN 27 5 2 DOUBLE AB SANDWICH (DIRECT) METHOD FOR ANTIGEN 27 5 3 INDIRECT METHOD FOR ANTIBODY 27 5 4 CONJUGATION 27 6 ABEIA (AVIDIN BIOTIN EIA) 27 6 1 COMPETITIVE DIRECT ELISA FOR ZEALENONE (MYCOTOXIN) 27 7 IMMUNOBLOT 27 7 1 BLOTTING COMPRISES OF TWO STEPS 27 8 CHARACTERIZATION OF ANTIGENS BY WESTERN BLOTTING (PAMELA LAME MORICARTY) 27 8 1 INTRODUCTION 27 8 2 MATERIALS AND METHODS 27 8 2 1 STEP-1 SDS-PAGE 27 8 2 2 EQUIPMENT AND MATERIALS 27 8 3 STEP 2 BLOT 27 8 3 1 EQUIPMENT AND MATERIALS 27 8 3 2 REAGENTS 27 8 3 3 PROCEDURE 27 8 3 4 STEP-3 ANTIBODY PROBE TREATMENT 27 8 3 4 1 EQUIPMENT AND MATERIALS 27 8 3 4 2 REAGENTS 27 8 3 4 3 PROCEDURE 27 8 4 APPENDICES 27 8 4 1 RECRYSTALLIZATION OF ACRYLAMIDE 27 8 4 2 PROCEDURE 27 8 4 3 POWER SUPPLY 27 8 4 4 COMPONENTS 27 8 4 5 SALT FRACTIONATION OF SERUM 27 8 4 6 IMMUNIZATION 27 8 5 AFFINITY PURIFICATION 27 9 DETECTING SPECIFIC AB IN PATIENT’S SERUM BY ELISA 27 10 WESTERN BLOT ANALYSIS 27 2 PCR (POLYMERASE CHAIN REACTION) 28 NEW VISTAS FOR MONOCLONAL ANTIBODIES 254 —