To determine whether the antibody is inactivated, the core is to see if it can effectively bind to the antigen or exert a neutralizing effect. The commonly used methods are neutralization test and ELISA detection, one directly measuring the function and the other quantitatively assessing the binding ability.

1、 Neutralization test: measuring functional activity

Virus neutralization test: After mixing antibodies and viruses to infect cells, if the cell survival rate is high, it indicates that the antibodies have neutralized the virus and the activity is normal.

Toxicity neutralization test: For example, in the anti-O test, the antibody and toxin are mixed and red blood cells are added. If the red blood cells do not dissolve, it indicates that the antibody has neutralized the toxin and there is no problem with its activity.

2、 ELISA detection: quantitative binding ability

Principle: The antibody is adsorbed on a solid carrier, and after adding the antibody to be tested, it is labeled with an enzyme for secondary antibody color development. The degree of color development is related to the amount of antibody.

Steps:

Antigen encapsulation: Fixing antigens in a microplate.

Add the antibody to be tested: bind to the antigen.

Add enzyme-linked secondary antibody: bind to the antibody.

Color reaction: Adding substrate, the degree of color development reflects the amount of antibody.

Result analysis: Measure the absorbance value, compare it with the standard curve, and determine the antibody potency and activity.

3、 Other auxiliary methods

Immunohistochemistry (IHC): Assessing specificity by examining the binding of antibodies and tissue antigens.

Cross reactivity testing: Verify whether antibodies bind to non target antigens to avoid false positives.

4、 Precautions

Experimental conditions: Temperature, pH value, etc. should be strictly controlled to avoid affecting the results.

Antibody preservation: Avoid repeated freezing and thawing to prevent loss of activity.