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ELISA Note

ELISA (Enzyme-Linked Immunosorbent Assay) is a biochemical technique used to detect and quantify proteins, antibodies, hormones, and antigens through antigen-antibody interactions on a solid surface. There are various types of ELISA, including direct, indirect, sandwich, and competitive ELISA, each with specific applications and methodologies. This technique is widely used in disease diagnosis, hormone level detection, drug screening, food allergen detection, and vaccine development due to its high sensitivity, specificity, and suitability for large-scale testing.

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10 views2 pages

ELISA Note

ELISA (Enzyme-Linked Immunosorbent Assay) is a biochemical technique used to detect and quantify proteins, antibodies, hormones, and antigens through antigen-antibody interactions on a solid surface. There are various types of ELISA, including direct, indirect, sandwich, and competitive ELISA, each with specific applications and methodologies. This technique is widely used in disease diagnosis, hormone level detection, drug screening, food allergen detection, and vaccine development due to its high sensitivity, specificity, and suitability for large-scale testing.

Uploaded by

catooa45
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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ELISA (Enzyme-Linked Immunosorbent Assay)

Definition:

ELISA is a biochemical technique used to detect and quantify substances such as proteins,

antibodies, hormones, and antigens using enzyme-linked antibodies.

Principle:

ELISA is based on antigen-antibody interaction. A specific antibody or antigen is bound to a solid

surface (usually a microplate), and detection is achieved using an enzyme-linked antibody and a

color-producing substrate.

Types of ELISA:

1. Direct ELISA: Antigen is immobilized; enzyme-linked primary antibody binds directly to it.

2. Indirect ELISA: Uses a secondary antibody for detection; more sensitive than direct ELISA.

3. Sandwich ELISA: Captures antigen between two antibodies; highly specific.

4. Competitive ELISA: Signal intensity is inversely proportional to antigen concentration.

Steps (Sandwich ELISA Example):

1. Coating: Capture antibody is attached to the microplate well.

2. Blocking: Non-specific binding sites are blocked.

3. Sample Addition: Antigen binds to the capture antibody.

4. Detection Antibody Addition: Binds to another epitope of the antigen.

5. Enzyme-linked Secondary Antibody: Binds to the detection antibody.

6. Substrate Addition: Enzyme reacts with substrate to produce a detectable signal (color change).

7. Result Measurement: Intensity is measured using a spectrophotometer.

Applications:

- Diagnosis of diseases (e.g., HIV, hepatitis, COVID-19)


- Hormone level detection (e.g., insulin, hCG)

- Drug screening

- Food allergen detection

- Vaccine development

Advantages:

- High sensitivity and specificity

- Quantitative results

- Suitable for large-scale testing

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