Expression of human recombinant CD23 in insect cells.
Human CD23 (low affinity receptor for IgE) has been expressed in insect cells (Sf9) using the baculovirus expression system and the baculovirus transfer vector pAc373. Insect cells infected with a recombinant baculovirus coding for CD23 synthesized a polypeptide not found in wild-type infected insect cells that had antigenic properties similar to natural CD23 produced in RPMI 8866 cells. Surface expression of recombinant CD23 was demonstrated by its ability to bind IgE. Recombinant CD23 expressed in insect cells had a slightly lower molecular weight (43 kDa) than that of natural CD23 (45 kDa) from RPMI 8866 cells as detected by SDS-PAGE followed by Western-blotting. Affinity-purified recombinant CD23 from infected insect cells showed B-cell growth promoting activitiy. These observations demonstrate for the first time that biologically active recombinant CD23 can be produced by the baculovirus expression system, thus providing a useful source of recombinant material to elucidate the biological functions of CD23.
Jansen
K U
KU
Glaxo Institute for Molecular Biology S.A., Geneva, Switzerland.
Shields
J
J
Gordon
J
J
Cairns
J
J
Graber
P
P
Bonnefoy
J Y
JY
D000818:Animals; D015703:Antigens, CD / Q000096:biosynthesis*; D000944:Antigens, Differentiation, B-Lymphocyte / Q000096:biosynthesis*; D001402:B-Lymphocytes / Q000166:cytology; D016367:Baculoviridae / Q000235:genetics; D001483:Base Sequence; D015153:Blotting, Western; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D002846:Chromatography, Affinity; D005434:Flow Cytometry; D005822:Genetic Vectors; D006801:Humans; D007915:Lepidoptera / Q000378:metabolism*; D008969:Molecular Sequence Data; D011961:Receptors, Fc / Q000096:biosynthesis*; D017455:Receptors, IgE; D011994:Recombinant Proteins / Q000096:biosynthesis; D014407:Tumor Cells, Cultured
D015703:Antigens, CD; D000944:Antigens, Differentiation, B-Lymphocyte; D011961:Receptors, Fc; D017455:Receptors, IgE; D011994:Recombinant Proteins