Electrodissociation of high molecular weight complexes of interferon alpha during recycling isoelectric focusing.
Natural human interferon alpha has been separated by selective ultrafiltration into low molecular weight components and the molecules exceeding 100K daltons. Interferon associated with a higher molecular weight fraction showed partial pH sensitivity and resisted dissociation after treatment with urea, mercaptoethanol, sodium chloride or significant changes in pH. However, interferon activity was released from high molecular weight components during recycling isoelectric focusing. Electrodissociation was carried out in 1% ampholytes for 574 watt-hours. The interferon activity was concentrated in a pH range of 6-6.5, whereas, the majority of proteins were generally found in a more acidic position. The dissociated interferon was neutralized by polyclonal antibody to human interferon alpha (IFN alpha) and showed no presence of pH labile form. A pH sensitivity of high molecular weight interferon (HMW-IFN) may reflect an aggregation phenomenon rather than intrinsic structural differences.
Kruzel
M L
ML
Novaferon Labs, Inc., Stafford, Texas.
Rodkey
L S
LS
Georgiades
J A
JA
D002460:Cell Line; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007370:Interferon Type I / Q000032:analysis / Q000302:isolation & purification*; D007525:Isoelectric Focusing; D008970:Molecular Weight; D014462:Ultrafiltration; D014762:Viral Interference
D007370:Interferon Type I