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The co-expression of denileukin diftitox immunotoxin with Artemin: soluble and aggregation analysis in presence of an efficient protein chaperone

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Abstract

Denileukin diftitox is the first Food and Drug Administration (FDA) approved recombinant immunotoxin that has been prescribed in the treatment of some leukemia and lymphoma such as cutaneous T-cell lymphoma, chronic lymphocytic leukemia, and B-cell non-Hodgkin’s lymphoma in which the IL-2 receptor is overexpressed. This smart and directed toxin was named DT389IL-2 because it contains truncated diphtheria toxin (DT387), which is fused to human interleukin 2 (IL2). Today, since this protein is not stable for a long time, its production is dependent on customer demand. Aggregation is an important obstacle in the recombinant production of heterologous proteins, especially those with therapeutic properties. This study aimed to synthesize denileukin diftitox in the presence of Artemin as an efficient chaperone in the E. coli expression system, as well as to assess the soluble/aggregation of the synthesized protein. Two plasmids, one containing the denileukin diftitox protein-coding gene and the other carrying artemin gene were simultaneously transferred into E. coli BL21 (DE3). The transformed bacteria were then induced to overexpress the mentioned proteins. The soluble expression of the proteins was confirmed by the native PAGE technique. The results confirmed that the soluble expression of the denileukin diftitox protein significantly increased in the presence of Artemin as a new chaperon protein. To conclude, this strategy can be utilized for the soluble expression and production of this protein as a smart and directed toxin for the treatment of recurrent cutaneous T-cell lymphoma (CTCL).

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Abbreviations

FDA:

U.S. Food and Drug Administration

IL2:

Interleukin 2

CTCL:

Cutaneous T-Cell Lymphoma

DT:

Diphtheria Toxin

HD:

Hodgkin’s Disease

ATL:

Adult T-Cell Leukemia

HSP60:

Heat Shock Proteins

IPTG:

Isopropyl-ß-thiogalactopyranoside

EDTA:

Ethylenediaminetetraacetic acid

SDS:

Sodium Dodecyl Sulfate

PMSF:

Phenylmethylsulfonyl Fluoride

PAGE:

Polyacrylamide Gel Electrophoresis

APS:

Ammonium Persulfate

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Acknowledgements

We would like to thank the research council of Malek Ashtar Universities of Technology (MUT) for the instrument support of this investigation.

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Authors and Affiliations

  1. Faculty of Chemistry and Chemical Engineering, Malek Ashtar University of Technology, Tehran, Iran

    Mohamad Najarasl, Mehdi Zeinoddini & Ali Reza Saeeidinia

  2. Department of Biochemistry, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, Iran

    Reza Hasan Sajedi

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  1. Mohamad Najarasl
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  2. Mehdi Zeinoddini
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  3. Ali Reza Saeeidinia
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  4. Reza Hasan Sajedi
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Contributions

The authors confirm contribution to the paper as follows: study conception and design: Mehdi Zeinoddini, Ali Reza Saeeidinia, Reza Hasan Sajedi; experiments and data collection: Mohamad Najarasl; analysis and interpretation of results: Mehdi Zeinoddini, Ali Reza Saeeidinia, Mohamad Najarasl, Reza Hasan Sajedi; draft manuscript preparation: Mohamad Najarasl, Mehdi Zeinoddini. All authors reviewed the results and approved the final version of the manuscript.

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Correspondence to Mehdi Zeinoddini.

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Najarasl, M., Zeinoddini, M., Saeeidinia, A.R. et al. The co-expression of denileukin diftitox immunotoxin with Artemin: soluble and aggregation analysis in presence of an efficient protein chaperone. Biologia 76, 3421–3428 (2021). https://doi.org/10.1007/s11756-021-00846-2

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