Large-scale purification of gp70 from Moloney murine leukemia virus.
The external envelope glycoprotein, gp70, of the Moloney murine leukemia virus was extracted from NIH 3T3 cells utilizing the detergent n-octyl-beta-D-glycopyranoside. The extracted gp70 was sequentially purified utilizing lectin-affinity, anion-exchange, and molecular-exclusion chromatography techniques. Approximately 10 mg of gp70 was purified by this method and shown to be 95% homogeneous, as assessed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. The presence of purified gp70 from Moloney murine leukemia virus was confirmed by amino acid analysis, amino-terminal sequencing, and immunoreactivity with a monoclonal antibody raised against gp70. The procedure is rapid, utilizes commercially available media, and can be used to purify large amounts of retroviral envelope glycoprotein from virus.
Pyle
S W
SW
Program Resources, Inc./Dyn Corp, National Cancer Institute, Frederick Cancer Research and Development Center, MD 21702.
Chabot
D J
DJ
Miller
T L
TL
Serabyn
S A
SA
Bess
J W
JW
Arthur
L O
LO
D000595:Amino Acid Sequence; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D003902:Detergents; D004591:Electrophoresis, Polyacrylamide Gel; D008969:Molecular Sequence Data; D008979:Moloney murine leukemia virus / Q000032:analysis*; D015682:Retroviridae Proteins, Oncogenic / Q000302:isolation & purification*; D014759:Viral Envelope Proteins / Q000302:isolation & purification*
D003902:Detergents; D015682:Retroviridae Proteins, Oncogenic; D014759:Viral Envelope Proteins