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A side-by-side biophysical comparison of five single-domain antibody scaffolds used for synthetic display libraries - PubMed

a day ago
  • #biophysical comparison
  • #synthetic libraries
  • #single-domain antibodies
  • Systematic comparison of five single-domain antibody (sdAb) scaffolds used in synthetic display libraries using sequence analysis, biophysical assays, and AlphaFold-based structure prediction.
  • Evaluated key parameters include humanness, expression yield, hydrophobicity, monomericity, stability, and display efficiency of sdAb scaffolds.
  • Camelid sdAbs showed superior biophysical traits but the lowest humanness, while human or humanized VH-based sdAbs had high human sequence homology but biophysical limitations.
  • Mutations for improving stability or humanization were observed to compromise other biophysical properties in the studied sdAb scaffolds.
  • Findings underscore the complexity of multi-parameter optimization in sdAb development, highlighting the value of synthetic display libraries for therapeutic antibody discovery.