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Rearranging the domain order of a diabody-based IgG-like bispecific antibody enhances its antitumor activity and improves its degradation resistance and pharmacokinetics
- Source :
- mAbs
- Publication Year :
- 2014
- Publisher :
- Informa UK Limited, 2014.
-
Abstract
- One approach to creating more beneficial therapeutic antibodies is to develop bispecific antibodies (bsAbs), particularly IgG-like formats with tetravalency, which may provide several advantages such as multivalent binding to each target antigen. Although the effects of configuration and antibody-fragment type on the function of IgG-like bsAbs have been studied, there have been only a few detailed studies of the influence of the variable fragment domain order. Here, we prepared four types of hEx3-scDb-Fc, IgG-like bsAbs, built from a single-chain hEx3-Db (humanized bispecific diabody [bsDb] that targets epidermal growth factor receptor and CD3), to investigate the influence of domain order and fusion manner on the function of a bsDb with an Fc fusion format. Higher cytotoxicities were observed with hEx3-scDb-Fcs with a variable light domain (VL)-variable heavy domain (VH) order (hEx3-scDb-Fc-LHs) compared with a VH-VL order, indicating that differences in the Fc fusion manner do not affect bsDb activity. In addition, flow cytometry suggested that the higher cytotoxicities of hEx3-scDb-Fc-LH may be attributable to structural superiority in cross-linking. Interestingly, enhanced degradation resistance and prolonged in vivo half-life were also observed with hEx3-scDb-Fc-LH. hEx3-scDb-Fc-LH and its IgG2 variant exhibited intense in vivo antitumor effects, suggesting that Fc-mediated effector functions are dispensable for effective anti-tumor activities, which may cause fewer side effects. Our results show that merely rearranging the domain order of IgG-like bsAbs can enhance not only their antitumor activity, but also their degradation resistance and in vivo half-life, and that hEx3-scDb-Fc-LHs are potent candidates for next-generation therapeutic antibodies.
- Subjects :
- CD3 Complex
Cell Survival
CD3
medicine.medical_treatment
Immunology
bispecific diabody
Antineoplastic Agents
Mice, SCID
Domain (software engineering)
Interferon-gamma
Antigen
Cancer immunotherapy
Cell Line, Tumor
Neoplasms
Antibodies, Bispecific
medicine
IgG-like bispecific antibody
Animals
Humans
Immunology and Allergy
Epidermal growth factor receptor
Cell Proliferation
Fusion
cancer immunotherapy
Binding Sites
antibody engineering
Dose-Response Relationship, Drug
biology
effective domain order
Chemistry
Xenograft Model Antitumor Assays
Molecular biology
Tumor Burden
ErbB Receptors
Area Under Curve
Immunoglobulin G
MCF-7 Cells
biology.protein
Biophysics
Female
Antibody
epidermal growth factor receptor
Function (biology)
Reports
Protein Binding
Subjects
Details
- ISSN :
- 19420870 and 19420862
- Volume :
- 6
- Database :
- OpenAIRE
- Journal :
- mAbs
- Accession number :
- edsair.doi.dedup.....75fcc54ee76c4befdde6124d04cac27f