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TitleStructural insights into the bi-specific cross-over dual variable antibody architecture by cryo-EM.
Journal, issue, pagesSci Rep, Vol. 13, Issue 1, Page 8694, Year 2023
Publish dateMay 29, 2023
AuthorsDavid Fernandez-Martinez / Mark D Tully / Gordon Leonard / Magali Mathieu / Eaazhisai Kandiah /
PubMed AbstractMulti-specific antibodies (msAbs) are being developed as next generation antibody-based therapeutics. Knowledge of the three-dimensional structures, in the full antibody context, of their fragment ...Multi-specific antibodies (msAbs) are being developed as next generation antibody-based therapeutics. Knowledge of the three-dimensional structures, in the full antibody context, of their fragment antigen-binding (Fab) moieties with or without bound antigens is key to elucidating their therapeutic efficiency and stability. However, the flexibility of msAbs, a feature essential for their multi specificity, has hindered efforts in this direction. Cross-Over Dual Variable immunoglobulin (CODV) is a promising bispecific antibody format, designed to simultaneously target the interleukins IL4 and IL13. In this work we present the biophysical and structural characterisation of a CODV:IL13 complex in the full antibody context, using cryo-electron microscopy at an overall resolution of 4.2 Å. Unlike the 1:2 stoichiometry previously observed for CODV:IL4, CODV:IL13 shows a 1:1 stoichiometry. As well as providing details of the IL13-CODV binding interface, including the residues involved in the epitope-paratope region, the structure of CODV:IL13 also validates the use of labelling antibody as a new strategy for the single particle cryo-EM study of msAbs in complex with one, or more, antigens. This strategy reduced the inherent flexibility of the IL13 binding domain of CODV without inducing either structural changes at the epitope level or steric hindrance between the IL4 and IL13 binding regions of CODV. The work presented here thus also contributes to the development of methodology for the structural study of msAbs, a promising platform for cancer immunotherapy.
External linksSci Rep / PubMed:37248285 / PubMed Central
MethodsEM (single particle)
Resolution3.97 Å
Structure data

16113

8blq

EMDB-16113, PDB-8blq:
Cryo-EM structure of the CODV-IL13-RefAb triple complex
Method: EM (single particle) / Resolution: 3.97 Å

Source
  • homo sapiens (human)
KeywordsIMMUNE SYSTEM / Cytosolic / bispecific / Fab / therapeutical

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