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    ELISA (Enzyme-Linked Immunosorbent Assay) is a test that uses antibodies and enzyme-linked substrates to detect the presence of antigens or antibodies in a sample. It works based on the antibody-antigen lock and key principle. In ELISA, antigens are immobilized on a plate and antibodies in a sample are detected through enzyme-conjugated secondary antibodies. This produces a color change showing the amount of antibodies present. ELISA has many advantages like speed, sensitivity, specificity, and ability to test many samples at once. It is widely used to detect viruses, hormones, infections, allergens, toxins and more.

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    SummerThe Privy Council vs CCJ Debate

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    ELISA (Enzyme-Linked Immunosorbent Assay) is a test that uses antibodies and enzyme-linked substrates to detect the presence of antigens or antibodies in a sample. It works based on the antibody-antigen lock and key principle. In ELISA, antigens are immobilized on a plate and antibodies in a sample are detected through enzyme-conjugated secondary antibodies. This produces a color change showing the amount of antibodies present. ELISA has many advantages like speed, sensitivity, specificity, and ability to test many samples at once. It is widely used to detect viruses, hormones, infections, allergens, toxins and more.

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    © All Rights Reserved
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    11 views11 pages

    SummerThe Privy Council Vs CCJ Debate

    Uploaded by

    Lorenzini Grant
    ELISA (Enzyme-Linked Immunosorbent Assay) is a test that uses antibodies and enzyme-linked substrates to detect the presence of antigens or antibodies in a sample. It works based on the antibody-antigen lock and key principle. In ELISA, antigens are immobilized on a plate and antibodies in a sample are detected through enzyme-conjugated secondary antibodies. This produces a color change showing the amount of antibodies present. ELISA has many advantages like speed, sensitivity, specificity, and ability to test many samples at once. It is widely used to detect viruses, hormones, infections, allergens, toxins and more.

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    © All Rights Reserved
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    of 11

    Enzyme Linked Immunosorbent Assay

    (ELISA)
    Applied Food Safety Education Laboratory Course III
    NFS 492/592
    Hi-Yung Hsueh, Extension Graduate Assistant
    Joan Hegerfeld-Baker, Extension Food Safety Specialist
    July, 2011

    Introduction of ELISA
    Introduced by Engvall and Pearlmann in 1971.
    Main points covered in Lecture
    Basic principles of ELISA
    Advantages of using ELISA
    Uses of ELISA widely used

    Basic Principles of ELISA


    Based on Basic Immunology Response
    Lock and Key Concept
    Antigen (key): substance when introduced into the body produces
    antibodies
    Antibody (lock): protein in the body that is used by immune system to
    identify and neutralize foreign targets (referred to as antigens)
    Key fits into the lock

    Enzyme conjugate substrates


    Enzyme that converts colorless substrates to a colored product
    Bound to the antibody that is part of the antibody-antigen complex
    Or
    Bound to a secondary antibody that binds with the antibody-antigen
    complex

    Antibody-Antigen Complex

    Conducting an ELISA test


    Measuring amount of Antibodies in solution:
    Antigens (Ag) fixed to a solid surface
    (immobilized)
    96-wells plastic plate

    Antibodies (Ab) in solution to be tested


    i.e. urine or serum

    Enzyme-conjugated Anti-immunoglobulin
    Antibody against the antibodies being tested for.
    Enzyme linked (conjugated) to the Ab

    Substrate binds to enzyme and produces color


    Color intensity proportional to bound enzyme-Ab

    Conducting ELISA (cont.)


    Quantitative & Qualitative Measurements
    Four common ELISA tests based on the
    binding structure between the Antibody and
    Antigen.
    1.
    2.
    3.
    4.

    Direct-ELISA
    Indirect-ELISA
    Sandwich-ELISA
    Competition-ELISA

    Common ELISA tests


    Direct-ELISA
    Enzyme conjugated Ab is directly bound to the Ag

    Indirect-ELISA

    Common ELISA tests (contd.)


    Sandwich-ELISA

    Competition-ELISA
    Competition between two antibodies for one antigen, or
    two antigens for one antibody.

    Advantages of ELISA

    Fast90 samples tested in 2-3 hr


    Sensitivity (up to 10 pg/mL)
    Specificity (sample with high concentration contaminants
    Many samples can be processed at once
    Small sample size required (10 L ~ 100L )
    Colorimetric results easily observed and measured
    (spectrophotometer)
    Test for presence of Ag or Ab
    Flexible usage for research design
    Easy to learn, simple procedure

    Hundreds of ELISA Kits Manufactured

    Viral Contamination
    Hormone levels
    Infections
    Specific disease factors
    Drugs
    Allergens in food
    Residues in food
    Toxins in food
    Others

    End of part I

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