High-level production of murine interleukin-5 (IL-5) utilizing recombinant baculovirus expression. Purification of the rIL-5 and its use in assessing the biologic role of IL-5 glycosylation.
We describe here a recombinant baculovirus expression system useful for high level production of murine recombinant interleukin-5 (rIL-5). In addition, we describe a single-step technique of purification of the rIL-5 from the baculovirus-infected Sf9 cell supernatants, using an anti-IL-5 affinity column. The baculovirus-derived rIL-5 has physical properties and functional activities in various lymphoid cell assays similar to those of natural T cell-derived IL-5 and reacts with anti-IL-5 antibodies. Finally, the rIL-5 is similar to natural T cell-derived IL-5 in manifesting heterogeneous glycosylation; however, glycosylation does not appear to be necessary for biologic function, at least in a lymphoid cell proliferation assay.
Kunimoto
D Y
DY
Department of Medicine, University of Alberta, Edmonton, Canada.
Allison
K C
KC
Watson
C
C
Fuerst
T
T
Armstrong
G D
GD
Paul
W
W
Strober
W
W
D000818:Animals; D000917:Antibody Formation / Q000187:drug effects; D016367:Baculoviridae / Q000235:genetics*; D015153:Blotting, Western; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002846:Chromatography, Affinity; D006031:Glycosylation; D007070:Immunoglobulin A / Q000096:biosynthesis; D007313:Insecta; D015848:Interleukin-5 / Q000235:genetics* / Q000302:isolation & purification / Q000494:pharmacology; D051379:Mice; D008807:Mice, Inbred BALB C; D010957:Plasmids; D011994:Recombinant Proteins / Q000302:isolation & purification* / Q000494:pharmacology; D015183:Restriction Mapping; D013154:Spleen / Q000276:immunology; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology; D014162:Transfection
D007070:Immunoglobulin A; D015848:Interleukin-5; D011994:Recombinant Proteins