Comparison of in vitro and in vivo labeling of virus-induced L-cell interferon.
Preparations of NDV(uv)-induced L-cell interferon were labeled in vitro with (125)I and (3)H gas, or in vivo through incorporation of amino acids-(3)H during synthesis. Prior to purification, more than 90% of the interferon titer was lost during in vitro labeling by either procedure, whereas 34% of the initial activity of in vivo-labeled material was preserved during preparatory handling. Purification by carboxymethyl-Sephadex chromatography and electrophoresis in polyacrylamide gels was about 100-fold, and electrophoretic profiles revealed close concordance between isotopes and interferon titers in all instances. Noninterferon proteins from control cells, although less extensively labeled with tritium during synthesis than proteins from interferon-producing cells and released in lesser amounts, also contained components of identical electrophoretic mobility and distribution in acrylamide gels as interferon. The highest specific activity (6 x 10(6) U/mg protein) but lowest cpm per interferon unit ratio (0.3) were exhibited by in vivo-labeled interferon. The advantage of better isotope incorporation through in vitro labeling techniques was largely offset by extensive losses in interferon activity.
Stancek
D
D
Virus Laboratories, Children's Hospital of Philadelphia and School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19146.
Golgher
R R
RR
Paucker
K
K