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Elisa 1

ELISA (Enzyme-Linked Immunosorbent Assay) is a technique used to detect and quantify antigens or antibodies, with four main types: Direct, Indirect, Sandwich, and Competitive ELISA, each having specific principles, advantages, and uses. Direct ELISA is fast but less sensitive, Indirect ELISA is more sensitive but has cross-reactivity risks, Sandwich ELISA is highly specific but requires two antibodies, and Competitive ELISA is suitable for small antigens but involves complex data interpretation. Applications include virus testing, food allergen detection, and tracking disease outbreaks.

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8 views6 pages

Elisa 1

ELISA (Enzyme-Linked Immunosorbent Assay) is a technique used to detect and quantify antigens or antibodies, with four main types: Direct, Indirect, Sandwich, and Competitive ELISA, each having specific principles, advantages, and uses. Direct ELISA is fast but less sensitive, Indirect ELISA is more sensitive but has cross-reactivity risks, Sandwich ELISA is highly specific but requires two antibodies, and Competitive ELISA is suitable for small antigens but involves complex data interpretation. Applications include virus testing, food allergen detection, and tracking disease outbreaks.

Uploaded by

gogoibhoirab78
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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ELISA

Types of ELISA

ELISA (Enzyme-Linked Immunosorbent Assay) is a widely used immunoassay technique that


detects and quantifies antigens or antibodies. Depending on the experimental need, different
ELISA formats are used. The four main types are:

1. Direct ELISA

Principle:

The primary antibody is directly conjugated with an enzyme.

Antigen is immobilized on the plate, and the enzyme-linked antibody binds to it.

Steps:

1. Coat the plate with antigen.

2. Add enzyme-linked primary antibody.

3. Wash to remove unbound antibodies.

4. Add substrate → color develops.

Advantages:

Fast (fewer steps).

No cross-reactivity from secondary antibodies.


Disadvantages:

Less sensitive (no signal amplification).

Requires labeled primary antibodies for every target.

Uses:

Detecting small molecules or peptides.

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2. Indirect ELISA

Principle:

Uses two antibodies:

Primary antibody (unlabeled, binds antigen).

Secondary antibody (enzyme-labeled, binds primary antibody).

Steps:

1. Coat plate with antigen.

2. Add primary antibody.

3. Add enzyme-linked secondary antibody.

4. Wash → add substrate → measure color.

Advantages:
More sensitive (signal amplification via secondary antibody).

Flexible (same secondary antibody can detect different primary antibodies).

Disadvantages:

Risk of cross-reactivity.

More steps than direct ELISA.

Uses:

Antibody detection (e.g., HIV, autoimmune diseases).

3. Sandwich ELISA

Principle:

Uses two antibodies (capture & detection) to "sandwich" the antigen.

Highly specific and sensitive.

Steps:

1. Coat plate with capture antibody.

2. Add sample (antigen binds to capture antibody).

3. Add enzyme-linked detection antibody.

4. Wash → add substrate → measure color.


Advantages:

High specificity (two antibodies bind to the antigen).

Works well for complex samples (e.g., serum, cell lysates).

Disadvantages:

Requires two antibodies that bind to different epitopes.

More expensive.

Uses:

Measuring cytokines, hormones (e.g., insulin, TSH).

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4. Competitive ELISA

Principle:

Measures antigen concentration based on competition between sample antigen and a reference
antigen.

Used when antigen is small or only one antibody is available.

Steps:

1. Coat plate with antigen (or antibody).

2. Add sample (unknown antigen) + enzyme-linked antigen (or antibody).

3. Both compete for binding sites.

4. Wash → add substrate → measure color (inversely proportional to antigen concentration).


Advantages:

Works with small antigens (e.g., drugs, hormones).

Less affected by sample impurities.

Disadvantages:

Complex data interpretation.

Uses:

Drug testing (e.g., THC, opioids).

Measuring hormone levels (e.g., testosterone, cortisol).

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Exam Tips

✔ Sandwich ELISA is best for large antigens (e.g., proteins).


✔ Competitive ELISA is used for small molecules (e.g., drugs).
✔ Indirect ELISA is more sensitive than direct ELISA.

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APPLICATION

1. It can be used to detect the presence of antigen or antibody in a sample.

2. It can be used to determine serum antibody concentrations in a virus test

3. It can be used in food industry to detect potential food allergens.

4. It can be used in disease outbreaks to track the spreading of diseases HIV, bird flu, common,
colds, cholera, STDs, etc.

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