Cell Bio Lab Elisa

Nonspecific barriers (Innate)

Function without regard to specific pathogen.

• Physical barriers: Saliva, mucus, skin, lysozymes

• Immune Response: Fever, inflammation, antiviral interferons

• Can distinguish between own cells and foreign ones

Specific barriers (Adaptive)

Function for a specific pathogen

• Macrophages, B cells, T cells

Antibody and antigen relationship

Antibodies are produced to have highly specific regions that attach to a specific antigen which signals for its destruction.

Direct vs Indirect ELISA

• Direct: presence of certain antigen in patients blood

• Indirect: Presence of antibodies in patients blood (if antibody is present than there was an infection and an immune response to a certain antigen)

Indirect ELISA steps

• 1) pre-coat well with antigen

• 2) Block to prevent non specific binding

• 3) Wash

• 4) Add secondary antibody

• 5) Wash

• 6) Add substrate

Explaining Indirect ELISA results

• Positive: Specific antibody for certain antigen is produced which binds to form antigen-antibody complex. This allows for the secondary antibody to bind to that complex and have an enzyme that will produce a color change to indicate that the original antibody was able to bind

• Negative: Antibody is not present in patients blood and does not bind to antigen. When secondary antibody is added it will have no complex to bind to and therefore the enzyme cannot bind and no color change will occur