Transient expression of human serum albumin (HSA) in tobacco leaves
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ORIGINAL ARTICLE
Transient expression of human serum albumin (HSA) in tobacco leaves Behnam Sedaghati1,2 · Raheem Haddad1 · Mojgan Bandehpour2 Received: 12 May 2020 / Accepted: 1 July 2020 © The Author(s) 2020
Abstract Today, recombinant human proteins make up a considerable part of FDA-approved biotechnological drugs. The selection of proper expression platform for manufacturing recombinant protein is a vital factor in achieving the optimal yield and quality of a biopharmaceutical in a timely fashion. This experiment was aimed to compare the transient expression level of human serum albumin gene in different tobacco genotype. For this, the Agrobacterium tumefaciens strains LB4404 and GV3101 harboring pBI121-HSA binary vector were infiltered in leaves of three tobacco genotypes, including Nicotiana benthamiana and N. tabacum cv Xanthi and Samsun. The qRT-PCR, SDS-PAGE, western blotting and ELISA analysis were performed to evaluate the expression of HSA gene in transgenic plantlets. Our results illustrated that the expression level of rHSA in tobacco leaves was highly dependent on Agrobacterium strains, plant genotypes and harvesting time. The highest production of recombinant HSA protein was obtained in Samsun leaves infected with A. tumefaciens strain GV3101 after 3 days of infiltration. Keywords Biopharmaceutical · Expression system · Transgenic plants · Agroinfiltration · Tobacco
Introduction Human serum albumin (HSA) is one of the most plentiful proteins in human body, which constitute more than half of blood plasma protein. It is a monomeric multi-domain macromolecule with a molecular mass of 66.5 kDa. This non-glycosylated polypeptide has 585 amino acids folding into a single chain. HSA is an important multi-functional protein stabilizing plasma oncotic pressure, carrying a wide range of substances, and provisioning the bulk of plasma antioxidant activity. Besides, HSA is clinically used to treat several diseases such as hypovolemia, shock, burns and hemorrhage [1]. Like other pharmaceutical proteins, HSA is initially attained by fractionating the donated human blood. However, blood is a finite and insecure source for the extraction of human therapeutic drugs. Moreover, extracted proteins * Raheem Haddad [email protected] 1
Department of Biotechnology, Faculty of Agriculture and Natural Resources, Imam Khomeini International University, Qazvin, Iran
Cellular and Molecular Biology Research Center, Shahid Beheshti University of Medical Sciences, Tehran, Iran
2
can be contaminated with different blood-derived pathogens such as human immunodeficiency virus (HIV) and hepatitis. Thereby, it is necessary to achieve a simple and efficient approach to produce safe therapeutic proteins for scale-up to commercial levels. The production of human pharmaceuticals in genetically modified organism (GMO) has alleviated these problems and made plenty of biological drugs available for patients [2]. Manufacturing of pharmaceutical products and industrial enzymes from genetically engineered organisms is a mu
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