Improved expression of recombinant sweet-tasting brazzein using codon optimization and host change as new strategies

Jafarian, Vahab; Bagheri, Khadijeh; Zarei, Jabraeil; Karami, Shima; Ghanavatian, Parisa

doi:10.1080/08905436.2019.1711113

Cited by 4 publications

(2 citation statements)

References 22 publications

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“…Host change is a promising strategy to improve recombinant proteins’ production and making cost objects more efficient. 26 Different expression systems have been introduced to produce Fab fragments, but none is regarded as optimal. 27 The study scope herein described was to produce the ranibizumab Fab fragments in the new host, P. pastoris .…”

Section: Discussionmentioning

confidence: 99%

Expression of functional eGFP-fused antigen-binding fragment of ranibizumab in Pichia pastoris

et al. 2021

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Introduction: Ranibizumab is a mouse monoclonal antibody fragment antigen-binding (Fab) against human vascular endothelial growth factor-A (VEGF-A), inhibiting angiogenesis. This antibody is commercially produced in Escherichia coli host and used to treat wet age-related macular degeneration (AMD).Methods: In this study, the heavy and light chains of ranibizumab were expressed in Pichia pastoris. The expressed chains were incubated overnight at 4°C for interaction. The formation of an active structure was evaluated based on the interaction with substrate VEGF-A using an indirect ELISA, and an electrochemical setup. Furthermore, reconstruction of split enhanced green fluorescent protein (eGFP) reporter, chimerized at the C-terminus of the heavy and light chains, was used to characterize chains’ interaction. Results: P. pastoris efficiently expressed designed constructs and secreted them into the culture medium. The anti-Fab antibody detected the constructed Fab structure in western blot analysis. Reconstruction of the split reporter confirmed the interaction between heavy and light chains. The designed ELISA and electrochemical setup results verified the binding activity of the recombinant Fab structure against VEGF-A. Conclusion: In this work, we indicated that the heavy and light chains of ranibizumab Fab fragments (with or without linkage to split parts of eGFP protein) were produced in P. pastoris. The fluorescence of reconstructed eGFP was detected after incubating the equal ratio of chimeric-heavy and light chains. Immunoassay and electrochemical tests verified the bioactivity of constructed Fab. The data suggested that P. pastoris could be considered a potential efficient eukaryotic host for ranibizumab production.

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Section: Discussionmentioning

confidence: 99%

Expression of functional eGFP-fused antigen-binding fragment of ranibizumab in Pichia pastoris

et al. 2021

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“…A purification procedure was established for recombinant brazzein produced by B. licheniformis, and approximately 5 mg/L brazzein of high purity was obtained [39]. In another, in order to optimize the expression of brazzein protein from E. coli, codonoptimized gene was cloned into two different strains and results showed that the strain used had an effect on protein yield [41]. Use of different lactic acid bacteria were also investigated for expression of recombinant brazzein, but low amounts of recombinant protein were produced [43], [46], [66].…”

Section: Recombinant Sweet Plant Proteins Production Studiesmentioning

confidence: 99%

Tatlı Bitkisel Proteinler ve Rekombinant Üretimleri

ASR>

Yüksel

İÇİER>

et al. 2022

Türk Doğa Ve Fen Dergisi

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Tüketicilerin doğal gıda ürünlerine karşı olan ilgisi ve artan bilinçleri nedeniyle, gıda endüstrisi doğal içeriklere sahip gıdalar üretmeye yönelmiştir. Öte yandan gıda endüstrisinde yüksek miktarda tatlandırıcı kullanımı da bir diğer sağlık sorunudur. Tatlı proteinler, sakkarozdan yüzlerce/binlerce kat daha fazla tatlılığa sahip ilgi çekici doğal tatlandırıcılardır.Tatlı proteinler, yüksek tatlılığa ancak düşük kalori değerlerine sahiptir ve doğal veya yapay tatlandırıcılara sağlıklı alternatifler olarak kullanım potansiyelleri yüksektir. Bilinen bitki tatlı proteinleri tropik bitkiler tarafından üretilir ve bu, elde edilebilecek protein miktarını sınırlar. Protein miktarını arttırmak için farklı ekspresyon sistemleri kullanılarak bitkisel tatlı proteinlerin rekombinant üretimi üzerine birçok çalışma yapılmıştır. Bu makalede, tatlı bitki proteinlerinin kaynakları, türleri, fizikokimyasal ve yapısal özellikleri ve rekombinant üretimi ile ilgili çalışmalar derlenmiş ve yeni yapılabilecek çalışmalar üzerinde durulmuştur.

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Detection of insecticides by Tetronarce californica acetylcholinesterase via expression and in silico analysis

Jiang,

Gu,

2023

Appl Microbiol Biotechnol

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Improved expression of recombinant sweet-tasting brazzein using codon optimization and host change as new strategies

Cited by 4 publications

References 22 publications

Expression of functional eGFP-fused antigen-binding fragment of ranibizumab in Pichia pastoris

Expression of functional eGFP-fused antigen-binding fragment of ranibizumab in Pichia pastoris

Tatlı Bitkisel Proteinler ve Rekombinant Üretimleri

Detection of insecticides by Tetronarce californica acetylcholinesterase via expression and in silico analysis

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