Mickiene Gitana, Dalgediene Indre, Dapkunas Zilvinas, Zvirblis Gintautas, Pesliakas Henrikas, Kaupinis Algirdas, Valius Mindaugas, Mistiniene Edita, Pleckaityte Milda
Institute of Biotechnology, Vilnius University, Sauletekio al. 7, 10257, Vilnius, Lithuania.
Profarma UAB, V.A. Graiciuno 6, 02241, Vilnius, Lithuania.
Mol Biotechnol. 2017 Oct;59(9-10):374-384. doi: 10.1007/s12033-017-0026-7.
Granulocyte colony-stimulating factor (G-CSF) has found widespread clinical application, and modified forms with improved biopharmaceutical properties have been marketed as well. PEGylation, the covalent modification of G-CSF with polyethylene glycol (PEG), has a beneficial effect on drug properties, but there are concerns connected to the immunogenicity of PEGylated compounds and bioaccumulation of the synthetic polymer. To overcome challenges connected with chemical modifications, we developed fusion proteins composed of two G-CSF molecules connected via different peptide linkers. Three different homodimeric G-CSF proteins were purified, and their in vitro and in vivo activities were determined. A G-CSF dimer, GCSF-Lα, was constructed using an alpha-helix-forming peptide linker, and it demonstrated an extended half-life in serum with a stronger neutrophil response as compared to the monomeric G-CSF protein. The GCSF-Lα protein, therefore, might be selected for further studies as a potential drug candidate.
粒细胞集落刺激因子（G-CSF）已在临床上广泛应用，具有改善生物制药特性的修饰形式也已上市。聚乙二醇化，即G-CSF与聚乙二醇（PEG）的共价修饰，对药物特性有有益影响，但人们担心聚乙二醇化化合物的免疫原性和合成聚合物的生物蓄积性。为了克服与化学修饰相关的挑战，我们开发了由两个通过不同肽接头连接的G-CSF分子组成的融合蛋白。纯化了三种不同的同型二聚体G-CSF蛋白，并测定了它们的体外和体内活性。使用形成α-螺旋的肽接头构建了一种G-CSF二聚体GCSF-Lα，与单体G-CSF蛋白相比，它在血清中的半衰期延长，中性粒细胞反应更强。因此，GCSF-Lα蛋白可能作为潜在的药物候选物被选用于进一步研究。
