Correct option is D
Ans. (d) Sandwich ELISA with monoclonal capture and detection antibodies against different epitopes of human IL-17
Sandwich ELISA is the most accurate approach for quantifying serum levels of IL-17 in a clinical setting. In this method, a monoclonal capture antibody is used to specifically bind to the IL-17, and a different monoclonal detection antibody is used to bind to another epitope on the same protein. This method minimizes the chances of cross-reactivity and ensures specificity and sensitivity. Using antibodies against different epitopes of IL-17 reduces the potential for interference that could occur if the same epitope were targeted for both capture and detection, leading to a more precise and accurate quantification of IL-17 levels in serum samples.
Information Booster
Sandwich ELISA: This method involves the use of two antibodies: one to capture the analyte (IL-17) and the other for detection. The use of monoclonal antibodies against different epitopes ensures that the detected signal is specific to the target and that any interference due to antibody competition is minimized. This results in a highly specific and reproducible method for quantifying IL-17 in clinical samples.
Monoclonal antibodies against different epitopes: When different epitopes of a protein are targeted by monoclonal antibodies, the risk of the antibody not binding due to conformational changes or steric hindrance is reduced. This provides a more reliable and accurate measurement in a clinical context.
Additional Information
Option (a): Sandwich ELISA, with monoclonal capture and detection antibodies against the same epitope of human IL-17 is less accurate than using antibodies against different epitopes. When both antibodies target the same epitope, there is a risk of competition for binding, reducing the assay's sensitivity and specificity.
Option (b): Fractionation of the serum sample on SDS-PAGE followed by Western blotting with polyclonal anti-human IL-17 antibody is more complex and time-consuming than ELISA. While Western blotting can detect specific proteins, it is not suitable for accurate quantification in a clinical setting due to its semiquantitative nature and the need for additional processing steps.
Option (c): Direct ELISA by coating the plate with patient serum and detection with polyclonal anti-human IL-17 antibody is less specific than Sandwich ELISA. The polyclonal antibody may bind to multiple forms of IL-17, leading to cross-reactivity, and it may not provide the same level of specificity or reproducibility as the Sandwich ELISA method with monoclonal antibodies.
