Immunotoxin Screening System: A Rapid and Direct Approach to Obtain Functional Antibodies with Internalization Capacities

Hamamichi, Shusei; Fukuhara, Takeshi

doi:10.3390/toxins12100658

Cited by 15 publications

(9 citation statements)

References 111 publications

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“…The affinity of immunotoxins likely needs to be optimized in an optimal range to achieve selective tumor targeting for better anti-tumor efficacy and to avoid on- and off-target toxicities. Cell-based immunotoxin screening system was established to provide a rapid and direct approach for screening functional antibodies with internalization capacities [ 59 ]. Such a screening system, when applied to cell lines with different target protein densities, helps to identify functional antibody hits with selective tumor targeting and cytotoxicity.…”

Section: Modulating the Affinity Of Binding Moiety For Selective Tumo...mentioning

confidence: 99%

Strategies to mitigate the on- and off-target toxicities of recombinant immunotoxins: an antibody engineering perspective

Mei

Yang

et al. 2022

Antibody Therapeutics

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Targeted cancer therapies using immunotoxins has achieved remarkable efficacies in hematological malignancies. However, the clinical development of immunotoxins is also faced with many challenges like anti-drug antibodies and dose-limiting toxicity issues. Such a poor efficacy/safety ratio is also the major hurdle in the research and development of antibody-drug conjugates. From an antibody engineering perspective, various strategies were summarized/proposed to tackle the notorious on target off tumor toxicity issues, including passive strategy (XTENylation of immunotoxins) and active strategies (modulating the affinity and valency of the targeting moiety of immunotoxins, conditionally activating immunotoxins in the tumor microenvironments and reconstituting split toxin to reduce systemic toxicity etc.). By modulating the functional characteristics of the targeting moiety and the toxic moiety of immunotoxins, selective tumor targeting can be augmented while sparing the healthy cells in normal tissues expressing the same target of interest. If successful, the improved therapeutic index will likely help to address the dose-limiting toxicities commonly observed in the clinical trials of various immunotoxins.

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Section: Modulating the Affinity Of Binding Moiety For Selective Tumo...mentioning

confidence: 99%

Strategies to mitigate the on- and off-target toxicities of recombinant immunotoxins: an antibody engineering perspective

Mei

Yang

et al. 2022

Antibody Therapeutics

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“…Immunotoxins can bind to the cancerous cells, internalized, and kill them by the toxic active segment ( 7 8 ). Till now, the denileukin diftitox, tagraxofusp, and moxetumomab pasudotox have been approved by the US Food and Drug Administration ( 9 ).…”

Section: Introductionmentioning

confidence: 99%

Design of two immunotoxins based rovalpituzumab antibody against DLL3 receptor; a promising potential opportunity

et al. 2022

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Background and purpose: The lack of a new effective treatment for small cell lung cancer (SCLC) is an unresolved problem. Due to the new identification of delta-like ligand 3 (DLL3) and its high expression in SCLC patients, the use of DLL3 in target therapy can be effective. The use of bacterial toxins belonging to the ADP-ribosyl transferase toxins family and human enzymes to remove cancerous cells has been effective in the structure of immunotoxins. In this study, single-chain fragment variable of rovalpituzumab antibody fused to granzyme B (Rova-GrB) and PltA of typhoid toxin (Rova-Typh) as immunotoxins were designed, and bioinformatics analysis was done. Experimental approach: In silico analysis including the physicochemical properties, evaluation of the secondary and tertiary structure, refinement and validation of 3D models, and docking were performed. Immunotoxin genes were cloned and expressed in the Escherichia coli BL21 (DE3) host, purified, subsequently confirmed by western blotting and their secondary structure was evaluated by the circular dichroism method. Findings/Results: The bioinformatics analysis showed that Rova-GrB and Rova-Typh had hydrophilic properties, their codon optimization parameters were standard, validation parameters were improved after immunotoxin refinement, and docking analysis showed that the binding domain of immunotoxins could bind the N-terminal region of DLL3. immunotoxins had high expression and after purification under denaturing condition by Ni-NTA column, the immunotoxins were dialyzed against PBS buffer. Conclusion and implications: The immunotoxins had the right structure and can be produced in a prokaryotic host. The recombinant immunotoxins against DLL3 can be promising therapeutic agents for SCLC cancer.

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“…10 Although a few immunotoxins have been successful in the clinic, improved designs are needed that exhibit enhanced efficacy and target-recognition affinity, tunable in vivo lifetimes, and reduced immunogenic and other adverse effects. 11 Unfortunately, developing new variants is particularly challenging. Insufficient selectivity of receptor-binding proteins has been associated with off-target toxicity.…”

Section: ■ Introductionmentioning

confidence: 99%

IgG-Engineered Protective Antigen for Cytosolic Delivery of Proteins into Cancer Cells

Truex

Melo

et al. 2021

ACS Cent. Sci.

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Therapeutic immunotoxins composed of antibodies and bacterial toxins provide potent activity against malignant cells, but joining them with a defined covalent bond while maintaining the desired function is challenging. Here, we develop novel immunotoxins by dovetailing full-length immunoglobulin G (IgG) antibodies and nontoxic anthrax proteins, in which the C terminus of the IgG heavy chain is connected to the side chain of anthrax toxin protective antigen. This strategy enabled efficient conjugation of protective antigen variants to trastuzumab (Tmab) and cetuximab (Cmab) antibodies. The conjugates effectively perform intracellular delivery of edema factor and N terminus of lethal factor (LF N ) fused with diphtheria toxin and Ras/Rap1-specific endopeptidase. Each conjugate shows high specificity for cells expressing human epidermal growth factor receptor 2 (HER2) and epidermal growth factor receptor (EGFR), respectively, and potent activity across six Tmab- and Cmab-resistant cell lines. The conjugates also exhibit increased pharmacokinetics and pronounced in vivo safety, which shows promise for further therapeutic development.

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Immunotoxin Screening System: A Rapid and Direct Approach to Obtain Functional Antibodies with Internalization Capacities

Cited by 15 publications

References 111 publications

Strategies to mitigate the on- and off-target toxicities of recombinant immunotoxins: an antibody engineering perspective

Strategies to mitigate the on- and off-target toxicities of recombinant immunotoxins: an antibody engineering perspective

Design of two immunotoxins based rovalpituzumab antibody against DLL3 receptor; a promising potential opportunity

IgG-Engineered Protective Antigen for Cytosolic Delivery of Proteins into Cancer Cells

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