PDBsum entry 5m3v

Immune system

PDB id: 5m3v

Contents

Protein chains

208 a.a.

176 a.a.

Ligands

NAG-NAG-BMA-MAN-NAG-GAL-MAN-NAG-FUL ×2

Waters ×493

PDB id:

5m3v

Name:

Immune system

Title:

Beat fc

Structure:

Ig gamma-1 chain c region,ig gamma-3 chain c region. Chain: a. Synonym: hdc,heavy chain disease protein, beat fc (a). Engineered: yes. Mutation: yes. Ig gamma-1 chain c region. Chain: b. Synonym: beat fc (b). Engineered: yes.

Source:

Homo sapiens. Human. Organism_taxid: 9606. Gene: ighg1, ighg3. Expressed in: homo sapiens. Expression_system_taxid: 9606. Gene: ighg1. Expression_system_taxid: 9606

Resolution:

1.97Å R-factor: 0.213 R-free: 0.279

Authors:

D.Skegro,C.Stutz,F.Bourquin,S.Blein

Key ref:

D.Skegro et al. (2017). Immunoglobulin domain interface exchange as a platform technology for the generation of Fc heterodimers and bispecific antibodies. J Biol Chem, 292, 9745-9759. PubMed id: 28450393

Date:

17-Oct-16 Release date: 10-May-17

Protein chain

P01857  (IGHG1_HUMAN) -  Immunoglobulin heavy constant gamma 1 from Homo sapiens

Seq: 399 a.a.

Struc: 208 a.a.*

Protein chain

P01860  (IGHG3_HUMAN) -  Immunoglobulin heavy constant gamma 3 from Homo sapiens

Seq: 446 a.a.

Struc: 208 a.a.*

Protein chain

P01857  (IGHG1_HUMAN) -  Immunoglobulin heavy constant gamma 1 from Homo sapiens

Seq: 399 a.a.

Struc: 176 a.a.*

* PDB and UniProt seqs differ at 44 residue positions (black crosses)

J Biol Chem 292:9745-9759 (2017)

PubMed id: 28450393

Immunoglobulin domain interface exchange as a platform technology for the generation of Fc heterodimers and bispecific antibodies.

D.Skegro, C.Stutz, R.Ollier, E.Svensson, P.Wassmann, F.Bourquin, T.Monney, S.Gn, S.Blein.

ABSTRACT

Bispecific antibodies (bsAbs) are of significant importance to the development of novel antibody-based therapies, and heavy chain (Hc) heterodimers represent a major class of bispecific drug candidates. Current technologies for the generation of Hc heterodimers are suboptimal and often suffer from contamination by homodimers posing purification challenges. Here, we introduce a new technology based on biomimicry wherein the protein-protein interfaces of two different immunoglobulin (Ig) constant domain pairs are exchanged in part or fully to design new heterodimeric domains. The method can be applied across Igs to design Fc heterodimers and bsAbs. We investigated interfaces from human IgA CH3, IgD CH3, IgG1 CH3, IgM CH4, T-cell receptor (TCR) α/β, and TCR γ/δ constant domain pairs, and we found that they successfully drive human IgG1 CH3 or IgM CH4 heterodimerization to levels similar to or above those of reference methods. A comprehensive interface exchange between the TCR α/β constant domain pair and the IgG1 CH3 homodimer was evidenced by X-ray crystallography and used to engineer examples of bsAbs for cancer therapy. Parental antibody pairs were rapidly reformatted into scalable bsAbs that were free of homodimer traces by combining interface exchange, asymmetric Protein A binding, and the scFv × Fab format. In summary, we successfully built several new CH3- or CH4-based heterodimers that may prove useful for designing new bsAb-based therapeutics, and we anticipate that our approach could be broadly implemented across the Ig constant domain family. To our knowledge, CH4-based heterodimers have not been previously reported.