Determining the specific target of an antibody is important for applying its immunological properties to develop or evaluate immunotherapies. We need various methods to know which surface antigens are the targets of the antibodies present in the sample. Antibody depletion assays can be a useful tool to study antibody specificity.
For antibody specificity studies, Creative Biolabs provides specific protocols for antibody depletion assays, including general procedures for researchers to follow and design their own experiments.
|Stages||Solutions and Reagents|
|Antigen Setting||Known antigens, dilution buffer, blocking buffer|
|Antibody Depletion||Magnetic beads, washing buffer, elution buffer|
The presence of various antibodies in varying abundance in a number of serum samples. Removing some of the interfering species by antibody depletion will help us to improve the detection of the desired antibodies.
Prepare serial dilutions of known antigens in 96-well plates using buffer solutions. Keep an equal volume in each well after dilution and set up an antigen-free control. Add the blocking solution and incubate with shaking.
Add an equal amount of serum sample and incubate overnight with shaking. The interfering antibody will bind to the specific antigen. Then add magnetic beads for incubation. An antigen-antibody-magnetic bead complex is formed.
Add a magnetic field to separate the complex from the unbound antibody. The unbound fraction recovered is the depleted serum sample.
Incubate the depleted serum sample with an antigen library containing multiple antigens. Again, separate the antigen-antibody complexes by magnetic beads. Then isolate the antigens to verify specificity.
Poor antibody specificity
To enhance the discovery of antibody specificity in serum, antibody depletion is an excellent method. We can enhance the discovery of antibody specificity by removing the interfering antibody species with magnetic selection. If you have specific requirements for the application of an antibody depletion assay, contact us for communication and custom protocols.
For research use only. Not intended for any clinical use.