Purification of 120,000 dalton envelope glycoprotein from culture fluids of human immunodeficiency virus (HIV)-infected H9 cells.
The outer envelope glycoprotein, gp120, has been purified from large volumes (greater than 100 liters) of HTLV-IIIB-infected H9 cell culture fluids using immunoaffinity chromatography resins prepared from immunoglobulins of AIDS patients plasma. By using a single-step immunoaffinity purification, between 7 and 28 micrograms of gp120 was recovered from each liter of culture fluid which represented between 60 and 95% of the total envelope glycoprotein present in the fluid. Envelope glycoprotein in culture media was concentrated more than 10,000 times over the starting material. The water-soluble gp120, containing trace contaminating proteins, was purified to apparent homogeneity by preparative polyacrylamide gel electrophoresis (PAGE). Envelope glycoprotein purified from culture fluids was immunogenic in laboratory animals in both native and PAGE-purified forms and was reactive with AIDS patient sera in immunoassays.
Pyle
S W
SW
Program Resources, Inc., Frederick Cancer Research Facility (NCI-FCRF), MD 21701.
Bess
J W
JW
Robey
W G
WG
Fischinger
P J
PJ
Gilden
R V
RV
Arthur
L O
LO
D000163:Acquired Immunodeficiency Syndrome / Q000276:immunology; D002460:Cell Line; D004797:Enzyme-Linked Immunosorbent Assay; D006678:HIV / Q000235:genetics* / Q000302:isolation & purification; D015699:HIV Envelope Protein gp120; D006801:Humans; D008970:Molecular Weight; D011863:Radioimmunoassay; D012191:Retroviridae Proteins / Q000302:isolation & purification*; D014759:Viral Envelope Proteins / Q000302:isolation & purification*
D015699:HIV Envelope Protein gp120; D012191:Retroviridae Proteins; D014759:Viral Envelope Proteins