Probing the limits of Q-tag bioconjugation of antibodies

Site-selective labelling of antibodies (Abs) can circumvent problems from heterogeneity of conventional conjugation. Here, we evaluate the industrially-applied chemoenzymatic ‘Q-tag’ strategy based on transglutaminase-mediated (TGase) amide-bond formation in the generation of (89)Zr-radiolabelled antibody conjugates. We show that, despite previously suggested high regioselectivity of TGases, in the anti-Her2 Ab Herceptin™ more precise native MS indicates only 70–80% functionalization at the target site (Q298(H)), in competition with modification at other sites, such as Q3(H) critically close to the CDR1 region.