Conformational Entropy as a Potential Liability of Computationally Designed Antibodies

In silico antibody discovery is emerging as a viable alternative to traditional in vivo and in vitro approaches. Many challenges, however, remain open to enabling the properties of designed antibodies to match those produced by the immune system. A major question concerns the structural features of computer-designed complementarity determining regions (CDRs), including the role of conformational entropy in determining the stability and binding affinity of the designed antibodies. To address this problem, we used enhanced-sampling molecular dynamics simulations to compare the free energy landscapes of single-domain antibodies (sdAbs) designed using structure-based (DesAb-HSA-D3) and sequence-based approaches (DesAbO), with that of a nanobody derived from llama immunization (Nb10). Our results indicate that the CDR3 of DesAbO is more conformationally heterogeneous than those of both DesAb-HSA-D3 and Nb10, and the CDR3 of DesAb-HSA-D3 is slightly more dynamic than that of Nb10, which is the original scaffold used for the design of DesAb-HSA-D3. These differences underline the challenges in the rational design of antibodies by revealing the presence of conformational substates likely to have different binding properties and to generate a high entropic cost upon binding.

Automated summary

The emerging field of in silico antibody discovery offers a promising alternative to traditional in vivo and in vitro approaches, but faces challenges in matching the properties of designed antibodies to those produced by the immune system. Structural features and conformational entropy play crucial roles in determining the stability and binding affinity of computer-designed antibodies. Enhanced-sampling molecular dynamics simulations reveal differences in conformational heterogeneity and dynamics of designed antibodies compared to a nanobody derived from llama immunization, highlighting the complexities in rational antibody design.