Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct
Recently, the development of T-cell engager cancer therapeutics, consisting of anticancer and anti-T-cell antibody parts to engage the T-cell to the cancer site, has gained interest. Anti-CD3 antibodies are predominantly used to achieve specific binding to T-cells for the T-cell engager construction...
Saved in:
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2025-06-01
|
| Series: | Biochemistry and Biophysics Reports |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2405580825001025 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1849687258494402560 |
|---|---|
| author | Takuya Asanuma Peiwei Ding Yuki Kato Takeshi Nakanishi Ryutaro Asano Koki Makabe |
| author_facet | Takuya Asanuma Peiwei Ding Yuki Kato Takeshi Nakanishi Ryutaro Asano Koki Makabe |
| author_sort | Takuya Asanuma |
| collection | DOAJ |
| description | Recently, the development of T-cell engager cancer therapeutics, consisting of anticancer and anti-T-cell antibody parts to engage the T-cell to the cancer site, has gained interest. Anti-CD3 antibodies are predominantly used to achieve specific binding to T-cells for the T-cell engager construction. Various kinds of anti-CD3 IgG clones have been developed and engineered, but the available anti-CD3 VHH, a single variable domain of a dromedary heavy-chain antibody, clones are limited in number. Thus, the assessment of the available anti-CD3 VHHs is important for therapeutic applications. Here, we demonstrated the expression and characterization of an anti-CD3 VHH clone, 117G03, and evaluated this T-cell engager with an anti-EFGR VHH in a bispecific format to assess the developability of this anti-CD3 VHH clone. 117G03 was expressed as a monomer in the soluble fraction and demonstrated decent thermal stability with binding activity against a CD3-positive cell line. The T-cell engager construct was prepared using the refolding method and exhibited enhanced cytotoxic activity against the epidermal growth factor receptor (EGFR)-positive cell line mediated by activated T-cells. These results indicate that 117G03 can be utilized for T-cell engager applications. |
| format | Article |
| id | doaj-art-81896830f9a0443fa0bc2fa651553550 |
| institution | DOAJ |
| issn | 2405-5808 |
| language | English |
| publishDate | 2025-06-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Biochemistry and Biophysics Reports |
| spelling | doaj-art-81896830f9a0443fa0bc2fa6515535502025-08-20T03:22:22ZengElsevierBiochemistry and Biophysics Reports2405-58082025-06-014210201510.1016/j.bbrep.2025.102015Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug constructTakuya Asanuma0Peiwei Ding1Yuki Kato2Takeshi Nakanishi3Ryutaro Asano4Koki Makabe5Graduate School of Science and Engineering, Yamagata University, 4-3-16 Jyonan, Yonezawa, Yamagata, 992-8510, JapanGraduate School of Science and Engineering, Yamagata University, 4-3-16 Jyonan, Yonezawa, Yamagata, 992-8510, JapanDepartment of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, JapanDepartment of Chemistry and Bioengineering, Division of Science and Engineering for Materials, Chemistry and Biology, Graduate School of Engineering, Osaka Metropolitan University, Sugimoto 3-3-138, Sumiyoshi-ku, Osaka, 558-8585, JapanDepartment of Biotechnology and Life Science, Graduate School of Engineering, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo, 184-8588, JapanGraduate School of Science and Engineering, Yamagata University, 4-3-16 Jyonan, Yonezawa, Yamagata, 992-8510, Japan; Corresponding author.Recently, the development of T-cell engager cancer therapeutics, consisting of anticancer and anti-T-cell antibody parts to engage the T-cell to the cancer site, has gained interest. Anti-CD3 antibodies are predominantly used to achieve specific binding to T-cells for the T-cell engager construction. Various kinds of anti-CD3 IgG clones have been developed and engineered, but the available anti-CD3 VHH, a single variable domain of a dromedary heavy-chain antibody, clones are limited in number. Thus, the assessment of the available anti-CD3 VHHs is important for therapeutic applications. Here, we demonstrated the expression and characterization of an anti-CD3 VHH clone, 117G03, and evaluated this T-cell engager with an anti-EFGR VHH in a bispecific format to assess the developability of this anti-CD3 VHH clone. 117G03 was expressed as a monomer in the soluble fraction and demonstrated decent thermal stability with binding activity against a CD3-positive cell line. The T-cell engager construct was prepared using the refolding method and exhibited enhanced cytotoxic activity against the epidermal growth factor receptor (EGFR)-positive cell line mediated by activated T-cells. These results indicate that 117G03 can be utilized for T-cell engager applications.http://www.sciencedirect.com/science/article/pii/S2405580825001025 |
| spellingShingle | Takuya Asanuma Peiwei Ding Yuki Kato Takeshi Nakanishi Ryutaro Asano Koki Makabe Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct Biochemistry and Biophysics Reports |
| title | Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct |
| title_full | Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct |
| title_fullStr | Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct |
| title_full_unstemmed | Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct |
| title_short | Evaluation of an anti-CD3 VHH and construction of an anti-CD3/anti-EGFR bispecific tandem VHH as a cancer cell targeting drug construct |
| title_sort | evaluation of an anti cd3 vhh and construction of an anti cd3 anti egfr bispecific tandem vhh as a cancer cell targeting drug construct |
| url | http://www.sciencedirect.com/science/article/pii/S2405580825001025 |
| work_keys_str_mv | AT takuyaasanuma evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct AT peiweiding evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct AT yukikato evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct AT takeshinakanishi evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct AT ryutaroasano evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct AT kokimakabe evaluationofananticd3vhhandconstructionofananticd3antiegfrbispecifictandemvhhasacancercelltargetingdrugconstruct |