ABSTRACT
Hybrid protein, cancer necrosis factor thymosin-alpha1 (CNF-T), when synthesizing in strain-producer of Escherichia coli SG200-50 with plasmid pThy315, was a part of "inclusion bodies" mostly in the form of a high-molecular complex with other proteins due to the S-S bonds formation. An approach of purification of CNF-T has been proposed, which is based on the destruction of the complex in the presence of sodium dodecylsulfate (DDS-NA) and dithiotreitol (DDT) followed by gel-filtration on Sephadex G-100 and renaturation by ultrafiltration on hollow fibers. The method allows the isolation of electrophoretically homogeneous CNF-T containing no DDS-Na and having high cytotoxic activity against cancer cells of mouse adenocarcinome L-929. The yield of CNF-T achieved 80% relative its content in biomass and 30% relative the total protein.
Subject(s)
Chromatography, Gel/methods , Recombinant Proteins/isolation & purification , Thymosin/analogs & derivatives , Animals , Cell Line, Tumor , Dithiothreitol/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Inclusion Bodies/chemistry , Mice , Recombinant Proteins/pharmacology , Sodium Dodecyl Sulfate/chemistry , Thymalfasin , Thymosin/isolation & purification , Thymosin/pharmacology , UltrafiltrationSubject(s)
Hemostasis/drug effects , Anticoagulants/administration & dosage , Anticoagulants/pharmacology , Antifibrinolytic Agents/administration & dosage , Antifibrinolytic Agents/pharmacology , Blood Transfusion , Fibrinolytic Agents/administration & dosage , Fibrinolytic Agents/pharmacology , Hemostatics/administration & dosage , Hemostatics/pharmacology , Humans , Platelet Aggregation Inhibitors/administration & dosage , Platelet Aggregation Inhibitors/pharmacologyABSTRACT
Overall 76 histories of births, case reports of neonates and protocols of autopsies of the dead because of infection were analyzed. It has been established that the overwhelming majority of the children had contracted infection antenatally or during birth, with the ascending pathway of infection being predominant. Attention is drawn to the clinical and postnatal diagnosis with the aid of the screening tests.