Format of bispecific antibodies (BsAbs)-Diabody

SOCAIL MEDIA

Diabody is a noncovalent dimer of single-chain Fv (scFv) fragment that consists of the heavy chain variable (VH) and light chain variable (VL) regions connected by a small peptide linker. Another form of diabody is single-chain (Fv)2 in which two scFv fragments are covalently linked to each other. bispecific bivalent dimers are produced by using two different chains with the same orientation, the first containing the VH of Antibody 1 and the VL of Antibody 2, and the second containing the VH of Antibody 2 and the VL of Antibody 1 (Fig. 1). Varying the linker length by only one amino acid may shift the equilibrium completely to another oligomeric state with the lowest complexity being most favorable. Diabodies are one of the smallest recombinant BsAbs and the distance between the two antigen-binding sites is only 6.5 nm on average, which is less than half the distance in IgG. This compact size contribute to rapid pharmacokinetics, low immunogenicity and high tumor penetration.

Formats of bispecific antibodies (BsAbs)

Many formats have been developed for BsAb generation as listed in the following table.

Format	Description	Example BsAb	Trademark	Company
tandem VHH	Tandem VHH fragment-based BsAb	N/A
tandem scFv	Tandem ScFv fragment-based BsAb	AMG330	BiTE^TM	Amgen
Dual-affinity re-targeting antibody	Tandem domain-exchanged Fv (can also be used to fuse with Fc domain to create whole Abs)	Flotetuzumab	DART^TM	Macrogenics
Diabody	dimer of single-chain Fv (scFv) fragment	vixtimotamab	ReSTORE^TM	Amphivena Therapeutics
(scFv)2-Fab	a Fab domain and two scFv domains bind	A-337	ITab^TM	Generon/EVIVE Biotech
Rat–mouse hybrid IgG	Full-size IgG-like half antibodies from two different species	Catumaxomab	Triomab^TM	Trion Pharma
Hetero heavy chain, Common light chain	Hetero heavy chain, Common light chain	Emicizumab	ART-Ig^TM	Genentech/ Chugai/Roche
Controlled Fab arm exchange	Recombin the parental half antibodies	JNJ-64007957	Duobody^TM	Genmab/ Janssen
Hetero H, forced HL IgG1	KIH technology for heterodimerization of 2 distinct H chains, replacing the native disulfide bond in one of the CH1-CL interfaces with an engineered disulfide bond to enhance the cognate of H and L paring	MEDI5752	DuetMab^TM	MedImmune/ AstraZeneca
cH IgG1	Identical heavy chains; 2 different light chains: one kappa (κ) and one lambda (λ)	NI-1701	κλ body^TM	Novimmune SA
Hetero H, CrossMab	KIH technology; domain crossover of immunoglobulin domains in the Fab region	Vanucizumab	CrossMab^TM	Roche
scFv-Fab IgG	Fab-Fc; ScFv-Fc	Vibecotamab; M802	Xmab^TM (the engineered Fc to enhance the generation of heterodimeric Fc); YBODY^TM	Xencor/Amgen; YZYBio
VH1-VH2-CH1-Fc1(G1) x VL2-VL1-CL-Fc2(G1)	2 binding motif in one half antibody	SAR440234	CODV-Ig^TM	Sanofi
VL1-CL1-VH2-CH2-Fc x VH1-CH1 x VL2-CL2	2 binding motif in one half antibody	EMB-01	FIT-Ig^TM	EPIMAB BIOTHERAPEUTICS
VH-1-TCR Cα x VL-1-TCR Cβ; VH-2-CH-2-Fc x VL-2-CL-2	KIH technology; TCR Cα/Cβ is used to substitute the CH1 and CL domain in one arm		WuXibody^TM	WuXi Biologics
C-terminal linker of Fc	Link the other molecules at the C-terminal of Fc	APVO442	ADAPTIR-FLEX^TM	Aptevo Therapeutics
Fc antigen binding site	2 natural binding sites; 2 additional binding sites in the Fc loop	FS118	mAb²	F-star Therapeutics