Generating a Novel Bispecific Nanobody to Enhance Antitumor Activity

Ge, Qiuhan; Sun, Tze-Chien; Bian, Yanlin; Zhu, Jianwei

doi:10.1055/s-0040-1714138

Cited by 2 publications

(2 citation statements)

References 38 publications

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“…Based on cumulative research on cancer immunotherapy in our laboratory, we have established a symmetric research program from designing bispecific antibodies to comprehensively characterize a wide spectrum of tumours in vitro and in vivo [24][25][26][27][28]. Here, we report recent progress on the design and characterization of a novel bispecific nanobody targeting PD-L1 and CXCR4.…”

Section: Introductionmentioning

confidence: 99%

Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4

Hao

et al. 2022

BMC Cancer

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Background: Antibodies and derivative drugs targeting immune checkpoints have been approved for the treatment of several malignancies, but there are fewer responses in patients with pancreatic cancer. Here, we designed a nanobody molecule with bi-targeting on PD-L1 and CXCR4, as both targets are overexpressed in many cancer cells and play important roles in tumorigenesis. We characterized the biochemical and anti-tumour activities of the bispecific nanobodies in vitro and in vivo. Methods: A nanobody molecule was designed and constructed. The nanobody sequences targeting PD-L1 and CXCR4 were linked by the (G4S)3 flexible peptide to construct the anti-PD-L1/CXCR4 bispecific nanobody. The bispecific nanobody was expressed in E. coli cells and purified by affinity chromatography. The purified nanobody was biochemically characterized by mass spectrometry, Western blotting and flow cytometry to confirm the molecule and its association with both PD-L1 and CXCR4. The biological function of the nanobody and its anti-tumour effects were examined by an in vitro tumour cell-killing assay and in vivo tumour inhibition in mouse xenograft models. Results: A novel anti-PD-L1/CXCR4 bispecific nanobody was designed, constructed and characterized. The molecule specifically bound to two targets on the surface of human cancer cells and inhibited CXCL12-induced Jurkat cell migration. The bispecific nanobody increased the level of IFN-γ secreted by T-cell activation. The cytotoxicity of human peripheral blood mononuclear cells (hPBMCs) against pancreatic cancer cells was enhanced by the molecule in combination with IL-2. In a human pancreatic cancer xenograft model, the anti-PD-L1/CXCR4 nanobody markedly inhibited tumour growth and was superior to the combo-treatment by anti-PD-L1 nanobody and anti-CXCR4 nanobody or treatment with atezolizumab as a positive control. Immunofluorescence and immunohistochemical staining of xenograft tumours showed that the anti-tumour effects were associated with the inhibition of angiogenesis and the infiltration of immune cells. Conclusion: These results clearly revealed that the anti-PD-L1/CXCR4 bispecific nanobody exerted anti-tumour efficacy in vitro and inhibited tumour growth in vivo. This agent can be further developed as a therapeutic reagent to treat human pancreatic cancer by simultaneously blocking two critical targets.

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Section: Introductionmentioning

confidence: 99%

Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4

Hao

et al. 2022

BMC Cancer

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“…12,[16][17][18] Besides, we have established a number of relevant technology platforms including immunotoxin and bispecific nanobody to facilitate the study on enhancing immunoactivity through bispecific drug conjugates. [19][20][21] We established a BsADC construction method based on BAPTS, which could conjugate about three payloads into the BsAbs. In this study, we further constructed a PRLR Â HER2-targeting BsADC conjugating MMAE as payloads, and characterized its affinity, internalization as well as cytotoxicity in vitro.…”

Section: Introductionmentioning

confidence: 99%

Generating a Bispecific Antibody Drug Conjugate Targeting PRLR and HER2 with Improving the Internalization

Zong

Zhang

Zhu

2022

Pharmaceutical Fronts

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Antibody drug conjugate (ADC) therapy has become one of the most promising approaches in cancer immunotherapy. The bispecific targeting could improve the specificity, affinity, and internalization of the ADC molecules. Prolactin preceptor (PRLR) and HER2 have crosstalk signaling in breast cancer, and PRLR undergoes a rapid internalization compared with HER2. To improve the efficacy of HER2 ADCs with enhancing the target specificity and internalization, we constructed a PRLR/HER2-targeting bispecific ADC (BsADC). We evaluated the characterization of PRLR × HER2 BsADC from the affinity and internalization, and further assessed its in vitro cytotoxicity in human breast-cancer cell lines (BT474, T47D, and MDA-MB-231) using Cell Count Kit-8 analysis. Our data demonstrated that PRLR × HER2 BsADC kept the affinity to two targeting antigens after conjugating drugs and exhibited higher internalization efficiency in comparison to HER2 ADC. Furthermore, PRLR × HER2 BsADC demonstrated to have superior antitumor activity in human breast cancer in vitro. In conclusion, our findings indicate that it is feasible through increasing the internalization of target antibody to enhance the antitumor activity and therapeutic potential that could be further evaluated in in vivo animal model.

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Generating a Novel Bispecific Nanobody to Enhance Antitumor Activity

Cited by 2 publications

References 38 publications

Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4

Tumour inhibitory activity on pancreatic cancer by bispecific nanobody targeting PD-L1 and CXCR4

Generating a Bispecific Antibody Drug Conjugate Targeting PRLR and HER2 with Improving the Internalization

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