Expression of single-chain antibody-barstar fusion in plants

Semenyuk, Ekaterina G.; Stremovskiy, Oleg A.; Edelweiss, Evelina; Shirshikova, Olga V.; Balandin, Taras; Buryanov, Yа. I.; Deyev, Sergey M.

doi:10.1016/j.biochi.2006.07.012

Cited by 35 publications

(22 citation statements)

References 33 publications

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“…To meet these requirements, small antibody fragments, such as the single-chain variable fragment (scFv), have been developed to deliver toxins to cancer cells. The scFv 4D5 was derived from the humanized monoclonal anti-HER2 antibody (Herceptin), and has been successfully used previously [11][12][13][14][15][16][17][18][19][20][21][22]. The scFv 4D5 antibody shows above-average thermodynamic and serum stability, and binds selectively with high affinity to the HER2 antigen [15].…”

Section: Introductionmentioning

confidence: 99%

Antitumor activity and toxicity of anti-HER2 immunoRNase scFv 4D5-dibarnase in mice bearing human breast cancer xenografts

et al. 2009

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Ribonucleases (RNases) are a non-mutagenic alternative to harmful DNA-damaging anticancer drugs. Targeting of RNases with antibodies to surface antigens that are selectively expressed on tumor cells endows specificity to the cytotoxic actions of RNases. Barnase, a ribonuclease from Bacillus amyloliquefaciens, is a promising candidate for targeted delivery to cancer cells because of its insusceptibility to the ubiquitous cytoplasmic ribonuclease inhibitor, and its high stability and catalytic activity. Here, we characterized in vitro and in vivo an immunoRNase, scFv 4D5-dibarnase, which consists of two barnase molecules that are fused serially to the single-chain variable fragment (scFv) of humanized 4D5 antibody. The latter is directed against the extracellular domain of human epidermal growth factor receptor 2 (HER2), a cancer marker that is overexpressed in many human carcinomas. The scFv 4D5-dibarnase exerted a specific cytotoxic effect on HER2-overexpressing SKBR-3 and BT-474 human breast carcinoma cells (IC(50) = 4.1 and 2.4 nM, respectively) via induction of apoptosis. Ten doses of 0.7 mg/kg scFv 4D5-dibarnase to BALB/c nude mice that bore SKBR-3 human breast cancer xenografts resulted in a 76% reduction in tumor growth. A single injection of scFv 4D5-dibarnase at a total course dose of 7 mg/kg did not cause severe side effects in BALB/c nude or BDF1 mice. The cytotoxicity and selectivity of scFv 4D5-dibarnase merit consideration of this immunoRNase as a potent anticancer agent.

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Section: Introductionmentioning

confidence: 99%

Antitumor activity and toxicity of anti-HER2 immunoRNase scFv 4D5-dibarnase in mice bearing human breast cancer xenografts

et al. 2009

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“…Plant-based production of monoclonal antibody and scFv has already been reported by several groups (Larrick et al 2001;Peeters et al 2001;Semenyuk et al 2007). There was notable variability in the expression levels of 3D8 scFv in individual transgenic lines from the same mother K. pinnata plant (Fig.…”

Section: Discussionmentioning

confidence: 93%

Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

et al. 2009

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We developed an asexual reproductive plant, Kalanchoe pinnata, as a new bioreactor for plant-based molecular farming using a newly developed transformation method. Leaf crenate margins were pin-pricked to infect the plant with the Agrobacterium strain LBA4404 and vacuum infiltration was also applied to introduce the target gene into the plants. Subsequently, the young mother leaf produced new clones at the leaf crenate margins without the need for time-and labor-consuming tissue culture procedures. The average transformation rates were approximately 77 and 84% for pin-prickling and vacuum-infiltration methods, respectively. To functionally characterize an introduced target protein, a nucleic acid hydrolyzing recombinant 3D8 scFv was selected and the plant based 3D8 scFv proteins were purified and analyzed. Based on abzyme analysis, the purified protein expressed with this system had catalytic activity and exhibited all of properties of the protein produced in an E. coli system. This result suggested that vegetatively reproductive K. pinnata can be a novel and potent bioreactor for bio-pharmaceutical proteins.

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“…Способность барназы формировать высокоаффинный комплекс с барстаром является классическим примером белок-белкового взаимодействия с очень высоким срод-ством субстратов [51,52]. Это позволяет использовать барназу или барстар в качестве метки (или вектора) для конъюгации с другими белками и пептидами по типу «молекулярного конструктора» для достижения разных целей, а также облегчения очистки фермента [53].…”

Section: барназаunclassified

Ribonucleases with Antiproliferative Properties: Molecular Biological and Biochemical Characteristics

Покровский¹,

Трещалина²,

Nv³

et al. 2016

Клиническая онкогематология

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The article dwells on ribonucleases (RNAses) whose cytotoxic activity depends on the enzymatic activity, i.e. the ability to catalyze the cleavage of phosphodiester bonds of RNA. It presents both well-known information and our own data on RNAses of different origins with antitumor properties; it investigates the relation between the mechanism of cytotoxicity and biochemical and molecular biological characteristics. The analysis of published data demonstrates that all above characteristics contribute to the antiproliferative activity of RNAses. The major challenge for this group of enzymes is the achieving of selective bioavailability. This problem can be solved by creating conjugates as in case with ranpirnase and barnase. Based on their major pharmacological properties, active antitumor RNAses have great perspectives for treatment of not only oncohematological, but also solid malignancies.

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Expression of single-chain antibody-barstar fusion in plants

Cited by 35 publications

References 33 publications

Antitumor activity and toxicity of anti-HER2 immunoRNase scFv 4D5-dibarnase in mice bearing human breast cancer xenografts

Antitumor activity and toxicity of anti-HER2 immunoRNase scFv 4D5-dibarnase in mice bearing human breast cancer xenografts

Production of recombinant single chain antibodies (scFv) in vegetatively reproductive Kalanchoe pinnata by in planta transformation

Ribonucleases with Antiproliferative Properties: Molecular Biological and Biochemical Characteristics

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