Subject(s)
Antineoplastic Agents/toxicity , Cyclophosphamide/toxicity , Granulocyte Colony-Stimulating Factor/pharmacology , Hematopoiesis/drug effects , Animals , Cells, Cultured , Colony-Forming Units Assay , Granulocytes/cytology , Granulocytes/drug effects , Humans , Male , Mice , Mice, Inbred CBA , Recombinant ProteinsABSTRACT
The process of isolation of highly purified human lymphotoxin was studied and optimized. A set of methods providing an increase in the content of the target product in the biomass (plasmid DNA amplification and selection of clones of transformed cells) was applied at the stage of cultivation. A two-step purification scheme (ion-exchange chromatography on DEAE-Biotone A1 and hydroxylapatite) was developed. A number of physical characteristics were studied.
Subject(s)
Lymphotoxin-alpha/genetics , Chromatography, Ion Exchange , Cloning, Molecular , Electrophoresis, Polyacrylamide Gel , Escherichia coli/genetics , Humans , Lymphotoxin-alpha/isolation & purification , Lymphotoxin-alpha/metabolism , Recombinant Proteins/genetics , Recombinant Proteins/isolation & purification , Recombinant Proteins/metabolismABSTRACT
Study of the biosynthesis of human alpha 2-interferon and tumor necrosis factor in derivatives of E. coli SG200-50 recipient strain containing pIF16, pTNF311, and pLT21 plasmids demonstrated elimination of the plasmids from the cells without degradation thereof in the course of culturing. Methods increasing the content of the end products in the biomass are proposed.
