Antibody Purification, Conjugation and Modification
Purification - From Raw Antiserum to Purified Antibody
In case you have sourced or produced raw antiserum and are in need of a simple purification step prior to further processing, DiaMed, Inc. can help. We can purify your raw material to different degrees of purity based on your needs. In general, you can choose between:
- Purification by delipidation and precipitation with multiple dialysis steps.
- Purification by delipidation, precipitation and ion exchange chromatography with multiple dialysis steps.
- The antigen1 is bound to a solid phase column material and the antibodies are eluted from the column using optimized buffer systems.
1Note: The antigen is usually supplied by the client.
For all purification methods mentioned above, the final concentration of the product can be adjusted to your needs.
Also, the buffer system for the final product can be adjusted to your specifications. Common buffers used are TRIS and PBS.
Additional purification steps can be performed upon request (e.g. Protein A, Protein G, Protein L etc.).
Conjugation - Add Enzymes, Fluorophores or other Molecules.
Through the process of (bio-)conjugation, antibodies can be labeled with fluorophores, enzymes or other molecules, as required by our clients. DiaMed, Inc. can perform small scale to bulk (> 10 L) labeling of antibodies.
For example, some of the most requested conjugates are FITC or AP. Other fluorophores, enzymes or molecules can be used for conjugation upon request2.
For conjugation of antibodies with fluorophores, a prior purification step (e.g. γ-fractionation) is strongly advised. For enzyme conjugation, a prior affinity purification is recommended.
2Note: It is important to note, that the use of some fluorophores, enzymes or other molecules may be restricted by patents. Therefore, it may be necessary to obtain a license from the licensor (proprietor of the patent) prior to performing the work.
Modification - Customize Your Antibody
Do you have a specific application in mind that necessitates the modification of your antibody, such as enzymatic digestion with or without partial reduction to produce F(ab')2-, Fab'- or Fab-fragments? DiaMed, Inc. can help to modify antibodies to your needs3. It is strongly recommended to purify antibodies prior to modification.
Most antibody fragments can also be conjugated to fluorophores, enzymes or other small molecules.
3Note: Not all antibody subtypes from all organisms are suitable for this type of modification.