Many Routes to an Antibody Heavy-Chain CDR3: Necessary, Yet Insufficient, for Specific Binding

• Published in: Frontiers in immunology 2018-03, Vol.9, p.395-395
• Main Authors: D'Angelo, Sara, Ferrara, Fortunato, Naranjo, Leslie, Erasmus, M Frank, Hraber, Peter, Bradbury, Andrew R M
• Format: Article
• Language: English
• Subjects: BASIC BIOLOGICAL SCIENCES; binding specificity; Biological Science; heavy-chain complementarity-determining region 3; Immunology; inverse PCR; rearrangement; single-chain Fv display
• ISSN: 1664-3224; 1664-3224
• DOI: 10.3389/fimmu.2018.00395

Because of its great potential for diversity, the immunoglobulin heavy-chain complementarity-determining region 3 (HCDR3) is taken as an antibody molecule's most important component in conferring binding activity and specificity. For this reason, HCDR3s have been used as unique identifiers to investigate adaptive immune responses and to characterize selection outputs where display systems were employed. Here, we show that many different HCDR3s can be identified within a target-specific antibody population after selection. For each identified HCDR3, a number of different antibodies bearing differences elsewhere can be found. In such populations, all antibodies with the same HCDR3 recognize the target, albeit at different affinities. In contrast, within populations, the majority of antibodies with the same HCDR3 sequence do not bind the target. In one HCDR3 examined in depth, all target-specific antibodies were derived from the same VDJ rearrangement, while non-binding antibodies with the same HCDR3 were derived from many different V and D gene rearrangements. Careful examination of previously published datasets reveals that HCDR3s shared between, and within, different individuals can also originate from rearrangements of different V and D genes, with up to 26 different rearrangements yielding the same identical HCDR3 sequence. On the basis of these observations, we conclude that the same HCDR3 can be generated by many different rearrangements, but that specific target binding is an outcome of unique rearrangements and VL pairing: the HCDR3 is necessary, albeit insufficient, for specific antibody binding.