Polyclonal Antibody Purification Techniques Study Guide
Learn about the essential techniques for purifying polyclonal antibodies, crucial for various applications in immunoanalysis and research. This guide covers: * **Ammonium Sulphate Precipitation**: A bulk purification method to concentrate antibodies. * **Immunoaffinity Column Chromatography**: Advanced methods for high-purity isolation. * **Antigen-Based Purification**: Utilizes the specific binding between antibodies and immobilized antigens. * **Protein A-Based Purification**:
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Introduction to Immunoanalysis and Antibody Principles
Introduction to Immunoanalysis and Antibodies
An introduction to immunoanalysis using antibodies as specific binding reagents to understand antigen-antibody interactions and immunoglobulin structure.
- Immunoanalysis
- Immunoassays
- Antibodies
- Antigens
- Immunoglobulin G (IgG)
- IgA
- IgD
- IgM
- Epitope
- Paratope
- Hapten
- Plasma cells
- Light polypeptide chains
- Heavy polypeptide chains
- Continuous epitopes
- Discontinuous conformational epitopes
Principles and Applications of Immunoassays
This content covers basic principles of ELISA and diagnostic tests across clinical, food, environmental, forensic, and pharmaceutical applications.
- Enzyme-linked immunosorbent assay (ELISA)
- Lateral flow assays
- Clinical diagnostics
- Food & environmental testing
- Pharmaceutical testing
- Forensics
- Multiplex plates
- Biochips
- Microneedle biosampling device
- Antigen-antibody interaction
Characteristics and Production of Monoclonal Antibodies
Characteristics and Production of Monoclonal Antibodies
This content describes the characteristics, therapeutic suffixes, and production processes of monoclonal antibodies including hybridoma technology and oncology applications.
- Monoclonal antibodies (mAbs)
- Hybridomas
- Murine, Chimeric, Humanized, and Human suffixes
- Naked, Conjugated, and Bispecific mAbs
- Rituximab, Trastuzumab, and Ofatumumab
- Immunization and isolation of splenocytes
- Myeloma cell preparation
- Cell fusion using polyethylene glycol (PEG)
- Clone screening and functional characterization
Characteristics and Production of Monoclonal Antibodies
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Comparing Monoclonal and Polyclonal Antibodies
Comparison of monoclonal antibodies and polyclonal antibodies regarding production cost, time, specificity, and market growth.
- Monoclonal antibodies (mAbs)
- Polyclonal antibodies (PAbs)
- Epitope binding
- Batch-to-batch reproducibility
- High specificity
- Cross-reactivity
- Hybridized clone
- Naked mAb
- Conjugated mAb
- Bispecific mAb
Enzyme Labels and Endpoints in Colourimetry
Enzyme Labels and Colourimetry
- Horseradish Peroxidase (HRP)
- Alkaline Phosphatase (AP)
- Enzyme-conjugated antibody
- Chromogenic substrates
- Specific activity
- Catalytic turnover
- Sensitivity
- Conjugation
- Oxidoreductase
- Hydrolysis of phosphate esters
- Non-Competitive Assay
- Competitive Assay
FluoroPhore Labels and Fluorescent Detection
Fluorophores and Fluorescent Immunoassay Detection
Understanding absorption and emission spectroscopy, the Stokes shift, and the application of fluorophores and lanthanide probes in fluorescent immunoassays.
- Absorption spectroscopy
- Emission spectroscopy
- Direct and indirect fluorescent immunoassays
- Fluorescein isothiocyanate (FITC)
- Rhodamine probes
- Time resolved fluorescent immunoassays
- Fluorescence decay
- Signal-to-noise ratio
- DELFIA automatic immunoassay system
Introduction to Immunoanalysis and Polyclonal Antibody Production
Polyclonal Antibody Purification Techniques
Methods for polyclonal antibody purification including ammonium sulphate precipitation and immunoaffinity column chromatography using antigen or Protein A.
- Polyclonal Antibody Purification
- Ammonium sulphate precipitation
- Immunoaffinity column chromatography
- Cyanogen bromide activated Sepharose
- Protein A
- IgG Fc region
- FAb or F(ab)2 region
- Antigen-antibody dissociation
- Primary and secondary immune response
Polyclonal Antibody Purification Techniques
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