Current Issue : April - June Volume : 2015 Issue Number : 2 Articles : 5 Articles
Current human biologics are most commonly produced by mammalian cell culture-based fermentation technologies. However, its limited scalability and high cost prevent this platform from meeting the ever increasing global demand. Plants offer a novel alternative system for the production of pharmaceutical proteins that is more scalable, cost-effective, and safer than current expression paradigms. The recent development of deconstructed virus-based vectors has allowed rapid and high-level transient expression of recombinant proteins, and in turn, provided a preferred plant based production platform. One of the remaining challenges for the commercial application of this platform was the lack of a scalable technology to deliver the transgene into plant cells. Therefore, this review focuses on the development of an effective and scalable technology for gene delivery in plants. Direct and indirect gene delivery strategies for plant cells are first presented, and the two major gene delivery technologies based on agroinfiltration are subsequently discussed. Furthermore, the advantages of syringe and vacuum infiltration as gene delivery methodologies are extensively discussed, in context of their applications and scalability for commercial production of human pharmaceutical proteins in plants. The important steps and critical parameters for the successful implementation of these strategies are also detailed in the review. Overall, agroinfiltration based on syringe and vacuum infiltration provides an efficient, robust and scalable gene-delivery technology for the transient expression of recombinant proteins in plants. The development of this technology will greatly facilitate the realization of plant transient expression systems as a premier platform for commercial production of pharmaceutical proteins....
Lectins are present in microorganisms, plants and animals and have attracted great interest due to their varied physiological\nroles in cell agglutination, anti-tumour, immunomodulatory, antifungal and antiviral effects. Legume lectins are important to\nthe pharmaceuticals but they are produced in low amounts in the plant seeds. Moreover, the genes controlling these proteins are\nconditionally active, i.e., they work under specific circumstances and not in regular manner. Looking into these limitations, we aimed\nthis work to produce a recombinant lectin for pharmaceutical use especially for the treatment of cancer. Three different legume plant\nspecies were collected from Aseer region of Kingdom of Saudi Arabia viz., Acacia seyal, Pisum sativum (wild type) and Pisum sativum\n(Pea). The plant tissues were subjected to RNA extraction, the extracted RNA was used for lectin gene amplification using specific\nprimers. Cloning, subcloning of the Acacia 400bp gene was carried out and in vitro transcription, combined with protein purification\nwas undertaken. The cytotoxicity of the recombinant lectin was performed on two cell lines such as breast cancer MCF-7 and liver\ncancer HepG-2 cells. Our study resulted in the observation of two amplicones with all the three examined species, the amplicone\nmolecular sizes were 800 and 400bp. The 400bp amplicone was excised from the agarose gel, purified and sequenced. The sequence\nanalysis revealed that the nucleotide sequence belongs to lectin gene. The sequence analysis revealed that the lectin gene isolated\nfrom Pisum sativum (wild plant) is similar to the Pisum sativum lect1 with identity 90%, whereas, lectin isolated from Pisum (pea)\nshowed similarity of 91% with the other lectins. On the other hand, Acacia lectin showed similarity with Lotus japonicus nod factor\nbinding lectin gene with identity of 95%. Thus we conclude that new lectin protein of 17 and 15 kDa was produced that can be used by\npharmaceutical industries....
Chloroplast transformation in the photosynthetic alga Chlamydomonas reinhardtii has been used to explore the\npotential to use it as an inexpensive and easily scalable system for the production of therapeutic recombinant\nproteins. Diverse proteins, such as bacterial and viral antigens, antibodies and, immunotoxins have been successfully\nexpressed in the chloroplast using endogenous and chimeric promoter sequences. In some cases, proteins have\naccumulated to high level, demonstrating that this technology could compete with current production platforms.\nThis review focuses on the works that have engineered the chloroplast of C. reinhardtii with the aim of producing\nrecombinant proteins intended for therapeutical use in humans or animals....
Background: Recombinant proteins are usually required in laboratories interested in the protein but not in the\nproduction process itself. Thus, technical equipment which is easy to handle and straight forward protein\nproduction procedures are of great benefit to those laboratories. Companies selling single use cultivation bags and\nbioreactors are trying to satisfy at least part of these needs. However, single-use systems can contribute to major\ncosts which might be acceptable when ââ?¬Å?good manufacturing practicesââ?¬Â are required but not acceptable for most\nlaboratories facing tight funding.\nResults: The assembly and application of a simple self-made ââ?¬Å?smart sustainable bottleââ?¬Â (SSB) system for E. coli based\nprotein production is presented. The core of the SSB system is a 2-L glass bottle which is operated at constant\ntemperature, air flow, and stirrer speed without measurement and control of pH and dissolved oxygen. Oxygen\ntransfer capacities are in the range as in conventional bioreactors operated at intermediate aeration rates and by far\nexceed those found in conventional shaking flasks and disposable bioreactors. The SSB system was applied for the\nproduction of various recombinant proteins using T7-based expression systems and a defined autoinduction\nmedium. The production performance regarding amount and solubility of proteins with robust and delicate properties\nwas as good as in state-of-the-art stirred tank commercial bioreactors.\nConclusions: The SSB system represents a low cost protein production device applicable for easy, effective, and\nreproducible recombinant protein production....
an application for regulatory approval of biosimilar products. Unlike small molecule drug products, biosimilars are\nnot exact copies of their brand-name counterpart, and they are usually very sensitive to changes in environmental\nfactors and have greater variabilities due to their complexity and sensitivity to variation in manufacturing processes.\nFacing these challenges, a biosimilarity index based on reproducibility probability is proposed to assess biosimilarity.\nIn this article, we have demonstrated how to assess biosimilarity between the test and reference product in relative\nto a reference standard that is established in a study where reference product is compared with itself. Biosimilairty\nindex approach is robust against biosimilarity criteria and has the advantage of allowing the assessment of the\ndegree of similarity....
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