Mirbaha Samira, Rezaei Marzieh, Emamzadeh Rahman, Zarkesh Esfahani Sayyed Hamid
Department of Cell and Molecular Biology and Microbiology, Faculty of Biological Science and Technology, University of Isfahan, Isfahan, I.R. Iran.
Department of Immunology, School of Medicine, Isfahan University of Medical Sciences, Isfahan, I.R. Iran.
Res Pharm Sci. 2022 Apr 18;17(3):284-293. doi: 10.4103/1735-5362.343082. eCollection 2022 Jun.
Growth hormone (GH) has been known as a crucial metabolic hormone expressed at the pituitary and the other number of cells and tissues and responsible for body growth. Because of the short half-life of GH, daily subcutaneous injections were shown to be more effective for GH therapy. This represents a burden for patients. So, there is a strong effort from the industry to create a long-acting form of GH and lots of technologies like GH fusion proteins are used to increase GH half-life.
In this study, the Fc domain of human IgG1 with serine-glycine linkers was attached to the C-terminal of a GH superagonist molecular cloning. The presence of recombinant vector in host was confirmed by PCR. SDS-PAGE and western blot analysis showed the expression of recombinant proteins in the bacterial lysate. The binding ability to growth hormone receptors is determined by ELISA.
FINDINGS / RESULTS: Our results showed that the novel SupGH-Fc has a good binding affinity to its receptor in ELISA in comparison to standard GH, although it has a big size.
Our data in this study clearly demonstrated the expression of the SupGH-Fc in a recombinant protein expression system. It is an introduction to the production of the new recombinant GH, which can bind to its receptor more effectively than commercial growth hormones and also might have a longer half-life.
生长激素（GH）是一种重要的代谢激素，由垂体以及其他多种细胞和组织分泌，负责身体生长。由于生长激素半衰期短，每日皮下注射对生长激素治疗更为有效。这给患者带来了负担。因此，业界大力致力于研发长效生长激素，许多技术如生长激素融合蛋白被用于延长生长激素的半衰期。
在本研究中，将带有丝氨酸 - 甘氨酸连接子的人IgG1的Fc结构域连接到生长激素超级激动剂的C末端进行分子克隆。通过PCR确认宿主中重组载体的存在。SDS - PAGE和蛋白质印迹分析显示重组蛋白在细菌裂解物中的表达。通过ELISA测定与生长激素受体的结合能力。
我们的结果表明，尽管新型SupGH - Fc体积较大，但在ELISA中与标准生长激素相比，它与其受体具有良好的结合亲和力。
我们在本研究中的数据清楚地证明了SupGH - Fc在重组蛋白表达系统中的表达。这为新型重组生长激素的生产奠定了基础，该重组生长激素与商业生长激素相比能更有效地与其受体结合，并且可能具有更长的半衰期。
