Monoclonal antibodies against biotinylable AP-tag / Avitag
Chemokine receptors undergo internalization and desensitization in response to ligand activation. Internalized receptors are either preferentially directed towards recycling pathways (e.g. CCR5) or sorted for proteasomal degradation (e.g. CXCR4). Scientists at the Georg-August-University Göttingen developed a method monoclonal antibody against a biotinylable peptid (Epitope-Tag) called AP-tag or Avi Tag, GLNDIFEAQKIEWHE. The 15-residue peptide served as a substrate mimic for biotin ligase (BirA), which usually recognizes the much larger protein domain. Anti-AP antibodies are useful tools in the analysis / trafficking of AP-tag fusion proteins, e.g. quantification of receptor-internalization from the cell surface into the cell. Binding of EF10 antibodies to the target protein is not affected by biotinylation of the acceptor sequence. Using these antibodies an alternative method was developed to analyse receptor internalization and recycling which allows a more detailed analysis of receptor trafficking compared to classical antibody-based detection methods. This biotin-based detection system may be generally applicable to the analysis of transmembrane protein trafficking.
Double immunoflourescence (anti-AP/streptavidin) of CXCR4-/CCR5-expressing cells during ligand-induced internalization (30’) and recycling (60’).
Liebick M, Schläger C, Oppermann M (2016) Analysis of Chemokine Receptor Trafficking by Site-Specific Biotinylation. PLOSOne 11(6):e0157502.
We are looking for companies, who are interested in licensing these antibodies for selling them to industrial and scientific institutions or for developing advanced diagnostic tests and therapeutic solutions.
Dipl.-Biol. Christiane Bolli