The process of antibody production, whether hybridoma or recombinant protein production in mammalian cells or bacteria, necessitates antibody isolation from other proteins present in the sample (cell lysate, medium, ascites fluid, sera, etc.).
Isolating the antibody of interest requires applying the proper strategy, depending on the class, species, and status (tagged or untagged) of the antibody; its intrinsic chemistry; and the required purity level.
We’ve prepared this guide to help you take the best route toward antibody purification.
Antibody Purification Affinities
For monoclonal antibodies, purification affinities tend to be confined to Protein A, Protein G, and Protein L chromatography usage.
The binding capacities, however, depend on the antibody species and classes. Beads complexed with Protein A and G tend to be the most commonly used antibodies that are utilized for the capture step of human IgG.
However, they aren’t very efficient and aren’t adapted to isolate human IgM, IgE, IgD, and IgA. Therefore, it’s best to opt for other strategies.
Strategies for Antibody Purification
Conventional approaches to antibody purification involve the use of protein L, G, and A resins.
They can be relatively easy to use and are rather inexpensive. However, in some instances, approaching antibody purification in a different way may yield better results.
Certain species or classes of antibodies and antibody fragments are challenging to capture. In such cases, performing an ion exchange purification is a useful strategy. However, it isn’t very efficient because several purification steps must be followed.
Here are some effective solutions that’ll help you control your budget and resources for experiments better.
- NHS Activated Beads: Using N-hydroxysuccinimide activated beads allows the ligand to be coupled to a preactivated medium through its primary amine group. One of its primary benefits is that it allows the capture of an antibody-based solely on its affinity for the target.
- Multimodal Beads:Multimodal beads are a great option for more complicated scenarios, such as other strategies prove futile, when the number of purification steps needs to be reduced, and when protein stability becomes an issue.
As a reputable manufacturer of high-quality biological products for research in the United States, our team at Pel-Freez Biologicals is committed to providing its clients with the solutions they need for a rapidly advancing fields of research. We hold an Animal Welfare Certification, as well as an ISO 9001:2015 certification.
Visit our website to purchase animal serum, plasma, complement, and ancillary products. We offer gamma globulins found in plasma function, human gamma globulin, goat gamma globulin, IgG/IgM/antibody depleted complement, adult rabbit complement, and more.