Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM

Lv, Miaomiao; Qiu, Feng; Li, Tingting; Sun, Yuanjie; Zhang, Chunmei; Zhu, Ping; Qi, Xiaokun; Wan, Jun; Yang, Kun; Zhang, Kui

doi:10.1155/2015/460264

Cited by 11 publications

(10 citation statements)

References 33 publications

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“…The severe side effects of the conventional cancer therapies encouraged the scientists to look for new the innovate strategies. Immunotoxins are the novel class of the therapeutics in which the therapeutic agents directly target the cancer cells with no or minimal effect on the normal cells (22). The success of this method is highly dependent on the specificity of the biomarker on the target cancer cells.…”

Section: Discussionmentioning

confidence: 99%

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Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers

et al. 2018

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Background Epidermal growth factor receptor (EGFR) plays an important role in the progression and tumorigenesis of the various cancers. In this regards, anti-EGFR antibodies are valuable approved therapeutics for the EGFR over-expressing cancers. However, the occurrence of mutations in the EGFR and/or KRAS genes; a common phenomenon which is seen in many cancers, lead to the resistance to the EGFR-directed antibodies. EGFR based immunotoxins are capable of overcoming this limitation by directing the toxin moieties to the cancer cells resulting in cell death. Objectives In the present study, a novel immunotoxin consisting of the truncated Pseudomonas exotoxin A (PE-40) and anti-EGFR huscFv was developed and evaluated for the induction of cell death in EGFR positive A431tumoral cells. Materials and Methods PE-40 fragment of the exotoxin A was amplified by using PCR and ligated to pET22b-huscFv. The reaction was confirmed by PCR and restriction digestion. The immunotoxin was expressed in E. coli BL21 (plysS) and then was purified by Ni-NTA affinity column. Subsequently, the toxicity of the purified immunotoxin was evaluated on EGFR over-expressing epidermoid carcinoma of skin, A431 cell line. Results PCR and restriction digestion experiments have verified the integrity of the immunotoxin construct. Purification by affinity column resulted in a highly purified recombinant immunotoxin. MTT assay revealed the growth inhibitory effect of the huscFv-PE40 immunotoxin on EGFR-over-expressing A431 cells with an IC50 value of 250 ng.mL -1 . Conclusion In conclusion, the results indicated that the immunotoxin developed in this study has a high toxicity on the EGFR-over-expressing tumor cells and could be considered as a promising candidate for the treatment of the EGFR positive cancers.

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Section: Discussionmentioning

confidence: 99%

“…The immunogenicity and penetration potency are two important limitations in the immunotoxins therapy. Humanized or human versions of small antibody fragments such as scFv have been suggested as a solution for these limitations (22). Zhou et al (2012) was used an anti-EGFR EGF scFv for treating non-small cell lung cancer (NSCLC) (23).…”

Section: Discussionmentioning

confidence: 99%

Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers

et al. 2018

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“…A number of studies have been carried out to isolate scFvs capable of specifically targeting signature proteins of a particular cancer cell, mainly exploiting the phage display technology [124]. Many scFvs have been generated against known proteins overexpressed by a number of different kinds of cancer cells, such as HER [125] and EpCAM [126], or against target proteins that are essential for cancer growth and spreading, such as VEGF [127] (involved in neo-angiogenesis). Recently, a novel scFv able to bind CD24, a target overexpressed by hepatocellular carcinoma (HCC), has been developed, showing a higher accumulation in the hepatocellular carcinoma xenograft mouse model and proving its potential as a therapeutic and diagnostic agent [128].…”

Section: Scfv To Targetmentioning

confidence: 99%

The Dual Role of the Liver in Nanomedicine as an Actor in the Elimination of Nanostructures or a Therapeutic Target

Baboci

Capolla

Cintio

et al. 2020

Journal of Oncology

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e development of nanostructures for therapeutic purpose is rapidly growing, following the results obtained in vivo in animal models and in the clinical trials. Unfortunately, the potential therapeutic efficacy is not completely exploited, yet. is is mainly due to the fast clearance of the nanostructures in the body. Nanoparticles and the liver have a unique interaction because the liver represents one of the major barriers for drug delivery. is interaction becomes even more relevant and complex when the drug delivery strategies employing nanostructures are proposed for the therapy of liver diseases, such as hepatocellular carcinoma (HCC). In this case, the selective delivery of therapeutic nanoparticles to the tumor microenvironment collides with the tendency of nanostructures to be quickly eliminated by the organ. e design of a new therapeutic approach based on nanoparticles to treat HCC has to particularly take into consideration passive and active mechanisms to avoid or delay liver elimination and to specifically address cancer cells or the cancer microenvironment. is review will analyze the different aspects concerning the dual role of the liver, both as an organ carrying out a clearance activity for the nanostructures and as target for therapeutic strategies for HCC treatment.

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“…VB4-845 (or oportuzumab monatox, Proxinium) is a recombinant fusion protein that combines the specificity of an anti-EpCAM ScFv with the toxicity of PE 40. [61][62][63] Mesothelin, expressed on mesotheliomas, ovarian, pancreatic, and lung cancers, has been targeted with PE toxin and was found to have a synergistic effect with chemotherapeutic agents in a mice model. 44,49,64 One of the more interesting ITs is SS1P or antimesothelin (Fv)-PE38.…”

Section: Growth Factor Receptors As Targetsmentioning

confidence: 99%

Immunotoxins: Targeted Toxin Delivery for Cancer Therapy

Xin

Chen

Wang

et al. 2019

Pharmaceutical Fronts

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Immunotoxins are proteins that consist of a protein toxin conjugated to a specific targeting moiety. The targeting moiety is usually an antibody or ligand, such as monoclonal antibody, antibody fragment, a cytokine, or a growth factor. The toxins usually come from plant toxins, bacterial toxins, or human-origin cytotoxic elements. Nearly all toxins work by enzymatically inhibiting protein synthesis. After binding to antigens or receptors on target cell surfaces, immunotoxins are internalized and translocated to the cytosol where they can kill the cells. Immunotoxins have demonstrated high cytotoxicity to cancer cells and to date two immunotoxins have been approved by U.S. Food and Drug Administration on the markets for the treatment of hematological tumors: Lumoxiti and Ontak. Many other molecules are under development or clinical trials for different forms of cancer. Although immunotoxins exhibit great potency in xenograft model systems and early clinical trials, there are obstacles that limit successful treatments, including immunogenicity, nonspecific toxicity, and poor penetration. However, efforts are underway to address these problems. In this review, we summarize immunotoxins currently in clinical trials for either hematological tumors or solid tumors, outline the design of immunotoxins utilizing variety of components, and discuss the prominent examples of redesigned immunotoxins with reduced immunogenicity and nonspecific toxicity, as well as the strategies in manufacturing immunotoxins. With further improvements, it is anticipated that immunotoxins will play an increasing role in cancer therapy.

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Construction, Expression, and Characterization of a Recombinant Immunotoxin Targeting EpCAM

Cited by 11 publications

References 33 publications

Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers

Expression and Evaluation of HuscFv Antibody -PE40 Immunotoxin for Target Therapy of EGFR-Overexpressing Cancers

The Dual Role of the Liver in Nanomedicine as an Actor in the Elimination of Nanostructures or a Therapeutic Target

Immunotoxins: Targeted Toxin Delivery for Cancer Therapy

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