Expression and secretion of human recombinant LIF by genetically modified mammalian cells

• Main Authors: Rymar, S.Yu., Ruban, T.A., Irodov, D.M., Kordium, V.A.
• Format: Article
• Language: English
• Published: Інститут молекулярної біології і генетики НАН України 2011
• Series: Вiopolymers and Cell
• Subjects: Molecular and Cell Biotechnologies
• Online Access: http://dspace.nbuv.gov.ua/handle/123456789/156336

Aim. The aim of this work was to express the human LIF gene in mammalian cells and to study the secretion of recombinant LIF into culture medium. Methods. Recombinant LIF was detected by Western blot analysis and immunoprecipitation in culture medium of CHO-K1, L-M (TK– ) (ins+ ), 293Ò cells, transfected with recombinant plasmids containing human LIF gene. Results. The recombi- nant plasmids. containing human gene LIF, were constructed. The cells of three (CHO-K1, L-M (TK– ) (ins+ ), 293Ò) mammalian lines were transfected with these plasmids. It was shown that the transfected mammalian cells secreted recombinant human LIF which was characterized by variable degree of glycosylation including completely glycosylated form (approximately 68 kD). Ñonclusions. The conditioned medium of developed cell lines can be used as a sourñe of human recombinant LIF for different purposes, including purification of human recombinant LIF and as an additional supplement for cell culturing. Keywords: recombinant LIF, expression, secretion, transfection, cell lines.