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Disulfide-stabilized diabody antiCD19/antiCD3 exceeds its parental antibody in tumor-targeting activity

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Abstract

Background

A diabody is a bispecific antibody that is capable of recruiting a polyclonal T cell to antibody target-expressing tumor cells. However, the two chains of diabodies tend to dissociate because they are integrated non-covalently. Therefore, it is necessary to remodel the diabody to increase its stability in order to enhance the antitumor activity.

Methods

We constructed an antiCD3×antiCD19 diabody with one binding site for the T cell antigen receptor (TCRCD3) and the other for the B cell-specific antigen (CD19) by recombinant gene engineering technology. Cysteine residues were introduced into the V domains of the anti-CD3 segment. The stability and cytotoxicity of the two diabodies were compared in vitro and vivo.

Results

The disulfide-stabilized (ds) diabodies produced by Escherichia coli were secreted with high yields in a fully active form without a decrease in affinity. Compared with the parental diabody, the disulfide-stabilized (ds) diabody proved more stable in vitro and in vivo without reducing binding affinity. Both were able to effectively eliminate human lymphoma Raji cells by redirecting T lymphocytes in vitro and in vivo, but the ds diabody was more effective in inhibiting the growth of xenografts transplanted in BALB/C nude mice.

Conclusion

The antiCD3×antiCD19 ds diabody is more suitable for a controlled polyclonal T cell therapy of human CD19-positive B cell malignancies than its parental diabody.

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Acknowledgments

This research was supported by grants from the Natural Science Foundation of China (Grant Nos. 30873091 and 30971291), the Natural Science Foundation of Tianjin (Grant No. 05YFGZGX02800), the National Science and Technology Major Project (Grant No. 2009ZX09103-720).

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Authors and Affiliations

  1. State key Laboratory of Experimental Hematology, Institute of Hematology and Hospital of Blood Diseases, Chinese Academy of Medical Sciences and Peking Union Medical College, Nanjing Road 288, Tianjin, 300020, China

    Li Wei, Fan DongMei, Yang Ming, Yan Yan, Jiang Linlin, Yan Cihui, Li Shuangjing, Wang Min, Wang Jianxiang & Xiong DongSheng

  2. Tianjin Medical University Cancer Institute and Hospital, Huan-Hu-Xi Road, Ti-Yuan-Bei, He Xi District, Tianjin, 300060, People’s Republic of China

    Li Wei

  3. Department of Pharmacology, Shanxi Medical University, No.56 Xinjian Nanlu, Taiyuan, Shanxi, 030001, China

    Shi RuiZan

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  1. Li Wei
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  2. Fan DongMei
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  3. Yang Ming
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  5. Yan Yan
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  8. Li Shuangjing
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  9. Wang Min
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Corresponding authors

Correspondence to Yang Ming, Wang Jianxiang or Xiong DongSheng.

Additional information

Li Wei and Fan DongMei contributed equally to this work.

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Wei, L., DongMei, F., Ming, Y. et al. Disulfide-stabilized diabody antiCD19/antiCD3 exceeds its parental antibody in tumor-targeting activity. Cell Oncol. 35, 423–434 (2012). https://doi.org/10.1007/s13402-012-0101-9

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