Wang et al. focused on improving the potency and specificity of mAbs targeting cancer- and infection-associated carbohydrates. Using structure-based rational design and directed evolution, variants of mAb735 – a modest-affinity, polysialic acid (polySia)-specific antibody – were generated (scFv and IgG formats) with significantly increased affinity (4- to 7-fold) compared to parental mAb735. Affinity-matured mAb735 IgG variants bound more avidly to polySia-positive tumor cell lines, and demonstrated increased functional potency and tumor cell killing, including ADCC and CDC, providing a framework for enhancing the promise of anti-glycan Abs.
Contributed by Katherine Turner
ABSTRACT: Monoclonal antibodies (mAbs) that specifically recognize cell surface glycans associated with cancer and infectious disease hold tremendous value for basic research and clinical applications. However, high-quality anti-glycan mAbs with sufficiently high affinity and specificity remain scarce, highlighting the need for strategies that enable optimization of antigen-binding properties. To this end, we engineered the affinity of a polysialic acid (polySia)-specific antibody called mAb735, which possesses only modest affinity. Using a combination of rational design and directed evolution, we isolated several affinity-matured IgG variants with _5- to 7-fold stronger affinity for polySia relative to mAb735. The higher affinity IgG variants opsonized polySia-positive cancer cells more avidly and triggered greater antibody-dependent cell-mediated cytotoxicity (ADCC) and complement-dependent cytotoxicity (CDC). Collectively, these results demonstrate the effective application of molecular evolution techniques to an important anti-glycan antibody, providing insights into its carbohydrate recognition and uncovering variants with greater therapeutic promise due to their enhanced affinity and potency.
