A HER2 bispecific antibody can be efficiently expressed in Escherichiaï¿½coli with potent cytotoxicity

Lin, Lehang; Li, Li; Zhou, Changhua; Li, Jing; Liu, Jiayu; Shu, Rui; Dong, Bin; Li, Qing; Wang, Zhong

doi:10.3892/ol.2018.8698

Cited by 13 publications

(16 citation statements)

References 51 publications

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“…Also for patients whose tumors are or become resistant to trastuzumab(−based) therapy, anti-HER2 nanobody-targeted PDT could be a viable option. In the particular case of trastuzumab-resistant patients with metastasis, one could foresee the usage of nanobodies targeting HER2 as carriers of drugs or toxic compounds (such as nanobody-drug conjugates) [ 77 – 81 ], although this remains to be further investigated.…”

Section: Discussionmentioning

confidence: 99%

Nanobody-targeted photodynamic therapy induces significant tumor regression of trastuzumab-resistant HER2-positive breast cancer, after a single treatment session

Deken

Kijanka

Hernández

et al. 2020

Journal of Controlled Release

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Rationale A substantial number of breast cancer patients with an overexpression of the human epidermal growth factor receptor 2 (HER2) have residual disease after neoadjuvant therapy or become resistant to trastuzumab. Photodynamic therapy (PDT) using nanobodies targeted to HER2 is a promising treatment option for these patients. Here we investigate the in vitro and in vivo antitumor efficacy of HER2-targeted nanobody-photosensitizer (PS) conjugate PDT. Methods Nanobodies targeting HER2 were obtained from phage display selections. Monovalent nanobodies were engineered into a biparatopic construct. The specificity of selected nanobodies was tested in immunofluorescence assays and their affinity was evaluated in binding studies, both performed in a panel of breast cancer cells varying in HER2 expression levels. The selected HER2-targeted nanobodies 1D5 and 1D5-18A12 were conjugated to the photosensitizer IRDye700DX and tested in in vitro PDT assays. Mice bearing orthotopic HCC1954 trastuzumab-resistant tumors with high HER2 expression or MCF-7 tumors with low HER2 expression were intravenously injected with nanobody-PS conjugates. Quantitative fluorescence spectroscopy was performed for the determination of the local pharmacokinetics of the fluorescence conjugates. After nanobody-PS administration, tumors were illuminated to a fluence of 100 J∙cm -2 , with a fluence rate of 50 mW∙cm -2 , and thereafter tumor growth was measured with a follow-up until 30 days. Results The selected nanobodies remained functional after conjugation to the PS, binding specifically and with high affinity to HER2-positive cells. Both nanobody-PS conjugates potently and selectively induced cell death of HER2 overexpressing cells, either sensitive or resistant to trastuzumab, with low nanomolar LD 50 values. In vivo , quantitative fluorescence spectroscopy showed specific accumulation of nanobody-PS conjugates in HCC1954 tumors and indicated 2 h post injection as the most suitable time point to apply light. Nanobody-targeted PDT with 1D5-PS and 1D5-18A12-PS induced significant tumor regression of trastuzumab-resistant high HER2 expressing tumors, whereas in low HER2 expressing tumors only a slight growth delay was observed. Conclusion Nanobody-PS conjugates accumulated selectively in vivo and their fluorescence could be detected through optical imaging. Upon illumination, they selectively induced significant tumor regression of HER2 overexpressing tumors with a single treatment session. Nanobody-targeted PDT is therefore suggested as a new additional treatment for HER2-positive breast cancer, particularly of interest for trastuzumab-resistant HER2-positive breast cancer. Further studies are now needed to assess the value of t...

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Section: Discussionmentioning

confidence: 99%

Nanobody-targeted photodynamic therapy induces significant tumor regression of trastuzumab-resistant HER2-positive breast cancer, after a single treatment session

Deken

Kijanka

Hernández

et al. 2020

Journal of Controlled Release

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“…In this regard, smaller sized BiTEs can be generated by substituting one or both the scFv with nanobodies. An example is the Her2 × CD3 bispecific antibody using anti-CD3 clone UTCH1 scFv and anti-Her2 nanobody, which showed strong anti-tumour effect in vitro and inhibits tumour growth in vivo [101,102]. A similar approach was used by Mølgaard et al with the generation of the bispecific light T-cell engager (LiTE) consisting of anti-CD3 UTCH1 scFv and anti-EGFR nanobody [103].…”

Section: Nanobodies As Therapeuticsmentioning

confidence: 99%

The Therapeutic Potential of Nanobodies

2019

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Today, bio-medical efforts are entering the subcellular level, which is witnessed with the fast-developing fields of nanomedicine, nanodiagnostics and nanotherapy in conjunction with the implementation of nanoparticles for disease prevention, diagnosis, therapy and follow-up. Nanoparticles or nanocontainers offer advantages including high sensitivity, lower toxicity and improved safety-characteristics that are especially valued in the oncology field. Cancer cells develop and proliferate in complex microenvironments leading to heterogeneous diseases, often with a fatal outcome for the patient. Although antibody-based therapy is widely used in the clinical care of patients with solid tumours, its efficiency definitely needs improvement. Limitations of antibodies result mainly from their big size and poor penetration in solid tissues. Nanobodies are a novel and unique class of antigen-binding fragments, derived from naturally occurring heavy-chain-only antibodies present in the serum of camelids. Their superior properties such as small size, high stability, strong antigen-binding affinity, water solubility and natural origin make them suitable for development into next-generation biodrugs. Less than 30 years after the discovery of functional heavy-chain-only antibodies, the nanobody derivatives are already extensively used by the biotechnology research community. Moreover, a number of nanobodies are under clinical investigation for a wide spectrum of human diseases including inflammation, breast cancer, brain tumours, lung diseases and infectious diseases. Recently, caplacizumab, a bivalent nanobody, received approval from the European Medicines Agency (EMA) and the US Food and Drug Administration (FDA) for treatment of patients with thrombotic thrombocytopenic purpura.

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“…Regarding the fact that the anti-PD-1 scFv is purified from E. coli , since the 1980's there have been a number of recombinant biopharmaceuticals produced in E. coli (reviewed [ 95 ]) and biopharmaceuticals produced in E. coli include antibodies (e.g. [ 96 ]). Finally, the high cost of production of therapeutic antibodies in mammalian cell culture can limit their availability to patients (e.g.…”

Section: Discussionmentioning

confidence: 99%

Characterization of a single chain variable fragment of nivolumab that targets PD-1 and blocks PD-L1 binding

Shin

Phelan

Gjoerup³

et al. 2021

Protein Expression and Purification

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Activated T-cells express Programmed cell Death protein 1 (PD-1), a key immune checkpoint receptor. PD-1 functions primarily in peripheral tissues, where T cells may encounter tumor-derived immunosuppressive ligands. Monoclonal antibodies that disrupt the interaction between T-cell derived PD-1 and immunosuppressive ligands, such as PD-L1, have revolutionized approaches to cancer therapy. For instance, Nivolumab is a monoclonal Ab that targets human PD-1 and has played an important role in immune checkpoint therapy. Herein we report the purification and initial characterization of a ∼ 27 kDa single chain variable fragment (scFv) of Nivolumab that targets human PD-1 and blocks binding by PD-L1. The possibility that the anti-PD-1 scFv can serve as both an anti-tumor agent and as an anti-viral agent is discussed. Importance The clinical significance of anti-PD-1 antibodies for treatment of a range of solid tumors is well documented (reviewed in [1-4]). In this report, we describe the results of studies that establish that an anti-PD-1 scFv purified from E. coli binds tightly to human PD-1. Furthermore, we demonstrate that upon binding, the anti-PD-1 scFv disrupts the interaction between PD-1 and PD-L1. Thus, the properties of this scFv, including its small size, stability and affinity for human PD-1, suggest that it has the potential to be a useful reagent in subsequent immunotherapeutic, diagnostic and anti-viral applications.

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A HER2 bispecific antibody can be efficiently expressed in Escherichiaï¿½coli with potent cytotoxicity

Cited by 13 publications

References 51 publications

Nanobody-targeted photodynamic therapy induces significant tumor regression of trastuzumab-resistant HER2-positive breast cancer, after a single treatment session

Nanobody-targeted photodynamic therapy induces significant tumor regression of trastuzumab-resistant HER2-positive breast cancer, after a single treatment session

The Therapeutic Potential of Nanobodies

Characterization of a single chain variable fragment of nivolumab that targets PD-1 and blocks PD-L1 binding

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