Related Experiment Video
Updated: Jul 1, 2026
Tracking Bispecific Antibody-Induced T Cell Trafficking Using Luciferase-Transduced Human T Cells
Published on: May 12, 2023
Preclinical Comparison of Two CD3xB7H4 Bispecific Antibodies for Solid Tumor Therapy Reveals CD3 Affinity-Dependent
Louise A Koopman1, Farshid Alemdehy1, Stefanie A H de Poot1
1Genmab (Netherlands) Utrecht Netherlands.
Abstract:
The immune checkpoint protein B7H4 is overexpressed in tumors compared to normal tissues, making it an attractive target for cancer immunotherapy. Here, we compare two B7H4-targeting CD3 bispecific antibodies (bsAbs) with different CD3 affinities across in vitro and in vivo characterization studies. In vitro, the CD3xB7H4 bsAb variant with higher CD3 affinity showed greater potency for tumor cell killing and cytokine secretion than the lower affinity variant. In nonclinical toxicology assessments in cynomolgus monkeys, the CD3xB7H4 bsAb variant with lower CD3 affinity induced less cytokine secretion and was tolerated at a higher maximum plasma concentration. Both CD3xB7H4 bsAbs demonstrated antitumor activity in vivo in an ovarian cancer patient-derived xenograft (PDX) mouse model and ex vivo in dissociated primary human ovarian tumor samples. These results demonstrate that increased CD3 affinity enhances cytotoxic potency but results in cytokine secretion at lower antibody concentrations in vitro and in vivo, highlighting an important design trade-off in CD3 bsAb development. The therapeutic potential of CD3xB7H4 bsAbs, particularly in relation to CD3 affinity, warrants further investigation in the clinic.
Insights
This study compares two B7H4-targeting bispecific antibodies (bsAbs) for cancer immunotherapy. Higher CD3 affinity enhances tumor cell killing but increases cytokine release, indicating a key design trade-off for bsAb development.
Area of Science:
- Immunology
- Oncology
- Biotechnology
Background:
- B7H4 is an immune checkpoint protein overexpressed in tumors.
- B7H4 is a promising target for cancer immunotherapy.
- Bispecific antibodies (bsAbs) engaging CD3 and tumor antigens offer therapeutic potential.
Purpose of the Study:
- To compare two B7H4-targeting CD3 bsAbs with differing CD3 affinities.
- To evaluate the impact of CD3 affinity on in vitro and in vivo efficacy and safety.
- To identify design trade-offs in bsAb development for cancer immunotherapy.
Main Methods:
- In vitro characterization of bsAb-mediated tumor cell killing and cytokine secretion.
- In vivo efficacy studies in ovarian cancer patient-derived xenograft (PDX) mouse models.
- Nonclinical toxicology assessments in cynomolgus monkeys.
- Ex vivo analysis of bsAb activity in human ovarian tumor samples.
Main Results:
- The higher CD3 affinity bsAb variant showed increased in vitro tumor cell killing and cytokine secretion.
- The lower CD3 affinity bsAb variant induced less cytokine secretion and was better tolerated in nonclinical toxicology studies.
- Both bsAb variants demonstrated antitumor activity in vivo and ex vivo.
- Increased CD3 affinity enhances cytotoxic potency but also cytokine release at lower concentrations.
Conclusions:
- CD3 affinity is a critical parameter in designing B7H4-targeting CD3 bsAbs.
- A balance between enhanced potency and manageable cytokine release is crucial for bsAb development.
- Further clinical investigation of CD3xB7H4 bsAbs, considering CD3 affinity, is warranted.
More Related Videos
07:25In Vitro Methods for Comparing Target Binding and CDC Induction Between Therapeutic Antibodies: Applications in Biosimilarity Analysis
Published on: May 4, 2017
15:04Potentiation of Anticancer Antibody Efficacy by Antineoplastic Drugs: Detection of Antibody-drug Synergism Using the Combination Index Equation
Published on: January 19, 2019