TOPIC: “ELISA”

PRESENTER: AREEJ TAHIR

COURSE INSTRUCTOR:
DATE:
 ABOUT ME

• Former General Secretary at IEEE-WIE SSUET

• Volunteer at HIWT
• Active member of Literary arts & culture Forum
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 STEPS OF PERFORMING ELISA
Coating
The antigen or antibody is adsorbed onto
the microplate wells to create a solid-phase
for subsequent binding reactions.
Detection
Enzyme-linked antibodies are added to
bind specifically to the target antigen or
antibody, facilitating the detection process.
Measurement
The signal intensity is quantified using a
spectrophotometer, providing data on the
concentration of the target antigen or
antibody in the sample.

1 3 5

2 4

Blocking Subrstrate addition

A blocking solution is added to cover A substrate solution is added, which reacts
unoccupied surfaces in the wells, with the enzyme linked to the detection
preventing non-specific binding and antibody to produce a measurable signal.
ensuring assay specificity.
TYPES OF ELISA

Direct ELISA:
1.Utilizes a labeled primary antibody to directly detect the
antigen.
2.Simple and straightforward.
3.Suitable for detecting large antigens.
4.Applications: Medical diagnostics, environmental
monitoring.
TYPES OF ELISA

Indirect ELISA:
1.Employs a secondary antibody labeled with an enzyme to
detect the primary antibody.
2.Amplifies the signal, increasing sensitivity.
3.Useful for detecting small antigen concentrations.
4.Applications: Antibody titer determination, autoimmune
disease detection.
TYPES OF ELISA

Sandwich ELISA:
1.Involves capturing the antigen between two antibodies
(capture and detection antibodies).
2.Highly specific and sensitive.
3.Ideal for detecting antigens with multiple epitopes.
4.Applications: Biomarker detection, protein quantification.
TYPES OF ELISA

Competitive ELISA:
1. Involves competition between the sample antigen and a labeled
antigen for binding to a limited amount of antibody.
2. Useful for detecting small antigens that might be difficult to
capture directly.
3. Provides accurate quantification of antigens.
4. Applications: Hormone levels measurement, drug testing, toxin
detection.
COMPARATIVE ANALYSIS
OF ELISA TYPES

DIRECT

INDIRECT

COMPETITIVE

SANDWICH
USES OF ELISA
• Disease Diagnosis: Detects pathogens (HIV, hepatitis, COVID-19) and autoimmune diseases.

• Hormone Level Measurement: Quantifies hormones like insulin and hCG.

• Allergen Detection: Identifies food and environmental allergens.

• Drug Testing: Monitors therapeutic drug levels and detects drugs of abuse.

• Vaccine Development: Assesses immune responses by measuring antibodies.

• Cancer Marker Detection: Identifies tumor markers for diagnosis and treatment monitoring.

• Quality Control in Food Industry: Ensures food safety by detecting contaminants.

• Research Applications: Studies protein interactions and biomarkers.

• Blood Screening: Screens blood donations for infectious agents.

COMPARISON ON THE BASIS OF APPLICATIONS
• Direct ELISA:
• Indirect ELISA:
Best for detecting large antigens,
Offers increased sensitivity, suitable for
used in medical diagnostics where
detecting low antigen concentrations in
straightforward detection is needed.
applications like autoimmune disease
detection.

• Competitive ELISA: • Sandwich ELISA:

Useful for detecting small molecules
like hormones, toxins, and drugs,
especially when the antigen is small
and has only one epitope, making it
ideal for applications in pharmacology
and environmental monitoring.
Highly specific and sensitive, ideal for
detecting biomarkers and quantifying
proteins accurately, making it valuable in
research and clinical settings for detailed
analyses.
THANK YOU