Design and construction of scFv-PE35KDEL as a novel immunotoxin against human epidermal growth factor receptor 2 for cancer therapy

Vaziri, Zahra Shariaty; Shafiee, Fatemeh; Akbari, Vajihe

doi:10.1007/s10529-023-03360-4

Cited by 3 publications

(1 citation statement)

References 43 publications

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“…This fusion protein (i.e., scFv-PE35KDEL) was expressed using the pET28a vector containing the T7 promoter, resulting in signi cant formation of inclusion bodies with only 10% of the resulting proteins being soluble. Adjusting the culture conditions (e.g., temperature and inducer concentration) barely improved the soluble expression of the recombinant protein (Shariaty Vaziri, Sha ee et al 2023). In the current study, the immunotoxin was expressed under the regulation of the trc promoter.…”

Section: Introductionmentioning

confidence: 99%

Integrated bioprocessing and genetic strategies to enhance soluble expression of anti-HER2 immunotoxin in E. coli

Mani,

Arab,

Akbari

et al. 2024

Preprint

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Immunotoxins are widely applied for cancer therapy. However, bacterial expression of immunotoxins usually leads to the formation of insoluble and non-functional recombinant proteins. In this study, we aimed to improve soluble expression of a novel anti-HER2 immunotoxin under the regulation of the trc promoter in Escherichia coli by optimization of the cultivation conditions using response surface methodology (RSM). To conduct RSM, four cultivation variables (i.e., inducer concentration, post-induction time, post-induction temperature, and medium recipe), were selected for statistical characterization and optimization using the Box-Behnken design and Design Expert software. Based on the developed model using the Box-Behnken design, the optimal cultivation conditions for soluble expression of anti-HER2 immunotoxin were determined to be 0.1 mM IPTG for induction in the LB medium at 33 °C for 18 h. The expressed immunotoxin was successfully purified using affinity chromatography with more than 90% purity and its bioactivity was confirmed using cell-based ELISA. Technical approach developed in this study can be generally applied to enhance the production yield and quality of recombinant proteins using E. coli as the gene expression system.

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Section: Introductionmentioning

confidence: 99%

Integrated bioprocessing and genetic strategies to enhance soluble expression of anti-HER2 immunotoxin in E. coli

Mani,

Arab,

Akbari

et al. 2024

Preprint

Self Cite

View full text Add to dashboard Cite

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Targeting HER2 in solid tumors: Unveiling the structure and novel epitopes

Liu,

Song,

Cheng

et al. 2024

Cancer Treatment Reviews

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The Potential of Single‐Chain Variable Fragment Antibody: Role in Future Therapeutic and Diagnostic Biologics

Gezehagn Kussia,

Tessema

2024

Journal of Immunology Research

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The advancement of genetic engineering has revolutionized the field of immunology by allowing the utilization of intrinsic antibody structures. One of the biologics that are being produced by recombinant antibody technology is single‐chain fragments variable (scFv). Genes of variable regions, the heavy and light chains that are genetically linked into a single transcript by a short flexible linker peptide, are used to generate this fragment from cellular and synthetic libraries. The specificity and affinity of these molecules are comparable to those of parental antibodies. Fusion with marker proteins and other potent molecules improves their stability, circulation half‐life, activity, and efficient purification. Besides, this review comprises construction protocols, therapeutics, and diagnostic applications of scFv, as well as related challenges. Nonetheless, there are still issues with efficacy, stability, safety, intracellular administration, and production costs that need to be addressed.

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Design and construction of scFv-PE35KDEL as a novel immunotoxin against human epidermal growth factor receptor 2 for cancer therapy

Cited by 3 publications

References 43 publications

Integrated bioprocessing and genetic strategies to enhance soluble expression of anti-HER2 immunotoxin in E. coli

Integrated bioprocessing and genetic strategies to enhance soluble expression of anti-HER2 immunotoxin in E. coli

Targeting HER2 in solid tumors: Unveiling the structure and novel epitopes

The Potential of Single‐Chain Variable Fragment Antibody: Role in Future Therapeutic and Diagnostic Biologics

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