I'm sure that many of you will relate to that experience of having an antibody that is just of bad quality. It creates problems because of cross-reactivity or non specific interaction and it's just impossible to work with, but is what you have or there is no other option but to use it.
So you soldier on, and develop new tricks to work with a fundamentally problematic system.
At what point you say.. "ok, it's enough" and admit defeat?
what is the last card you play before folding?
If you can't find a good commercial antibody, you can always reach out to laboratories that are working in the antibody engineering field. They can generate a custom antibody for your specific purpose. For example, the Technische Universität Braunschweig has two working groups dedicated to Antibody Phage Display (https://www.tu-braunschweig.de/bbt/biotech). You can contact them for custom and specific antibodies tailored to your needs. Additionally, there are companies that specialize in custom antibody design, such as Yumab (https://www.yumab.com) and Abcalis (https://abcalis.com). Feel free to reach out to them if you are interested in custom antibodies. Best wishes, Stöpa
Thank you for the links, but im more looking to start a conversation about common challenges and experiences during research and the balance between ingenuity and practicality
Pre-adsorbing the antiserum or antibody to remove the cross-reactivity can make a big improvement if you have a material to work with that is the same as the antigen-containing material but lacks the antigen. An example would be a cell extract from the cells used to express the antigen heterologously, but without induction of antigen expression.
Adam B Shapiro that is quite interesting. so it is a type of purification or selection to eliminate the most problematic elements in the serum sample? what happens if you have a problematic monoclonal antibody?
Since a mAb should be a single pure immunoglobulin, if it is cross-reacting with other antigens than the one it is intended to react with, then it means the other antigens have a similar or identical epitope as the intended antigen.
One scenario is that cross-reacting but unrelated antigens interact more weakly with the mAb than the intended antigen, in which case you can improve the specificity by reducing the antibody concentration.
Another scenario is that the cross-reacting antigens are closely related to the antigen of interest and contain the same epitope. It might be worth identifying these other antigens. In some cases, they may be proteolytic fragments of the antigen (lower molecular weight), or products of alternative splicing or gene duplication, or of alternative post-translational modifications. Different levels of glycosylation can result in multiple bands of the same antigen on a Western blot, for example.
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