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Questions and Answers
What is the principle behind direct ELISA?
What is the principle behind direct ELISA?
- Competition between target and labeled antigens
- Using two antibodies specific to different epitopes
- Using an unlabeled primary antibody followed by a labeled secondary antibody
- Binding of an enzyme-linked antibody to an immobilized antigen (correct)
What is a significant advantage of indirect ELISA?
What is a significant advantage of indirect ELISA?
- Fewer steps compared to direct ELISA
- Increased sensitivity due to signal amplification (correct)
- Limited sensitivity
- Higher complexity in procedure
Which ELISA format is known for quantifying small antigens?
Which ELISA format is known for quantifying small antigens?
- Sandwich ELISA
- Indirect ELISA
- Competitive ELISA (correct)
- Direct ELISA
What limitation does direct ELISA have compared to other ELISA formats?
What limitation does direct ELISA have compared to other ELISA formats?
Which of the following statements is true about sandwich ELISA?
Which of the following statements is true about sandwich ELISA?
What is a common application of indirect ELISA?
What is a common application of indirect ELISA?
What is a key limitation of competitive ELISA?
What is a key limitation of competitive ELISA?
Which ELISA type is suitable for the detection of low-abundance antigens?
Which ELISA type is suitable for the detection of low-abundance antigens?
In what scenario would you prefer to use an indirect ELISA over a direct ELISA?
In what scenario would you prefer to use an indirect ELISA over a direct ELISA?
What is a disadvantage of sandwich ELISA compared to direct ELISA?
What is a disadvantage of sandwich ELISA compared to direct ELISA?
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Study Notes
Direct ELISA
- Detects antigens directly using enzyme-linked antibodies
- Used for:
- Identifying viral antigens in patient samples
- Advantages:
- Fewer steps, simplifies the process
- Less non-specific binding occurs
- Limitations:
- Lower sensitivity compared to other ELISA formats
Indirect ELISA
- Detects antibodies using two antibodies:
- Unlabeled primary antibody binds to the antigen
- Labeled secondary antibody binds to the primary antibody
- Used for:
- Detecting host antibodies against viruses
- Advantages:
- More sensitive due to signal amplification
- Can be used for various antibody classes
- Limitations:
- Potential for increased non-specific binding
- Requires careful optimization of assay conditions
Sandwich ELISA
- Recognizes antigens using two antibodies:
- Capture antibody immobilizes the antigen on the plate
- Detection antibody binds to a different epitope on the target antigen and is enzyme-linked
- Used for:
- Detecting viral antigens in samples, with high specificity and sensitivity
- Advantages:
- Very specific and sensitive, ideal for low-abundance antigens
- Limitations:
- Needs two high-affinity antibodies, which makes the assay more complex and costly
Competitive ELISA
- Measures antigens by their competition with a labelled version for binding to a specific antibody
- Used for:
- Quantifying small antigens
- Measuring antibody levels
- Advantages:
- Suitable for detecting small molecules
- Provides precise quantification
- Limitations:
- May be more difficult to interpret
- Requires careful standardization
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