Escherichia coli expression, purification, and biological activity of a truncated soluble CD4.
A truncated molecule containing the N-terminal 183 amino acid residues of CD4 (sCD4-183) has been produced in Escherichia coli at high levels, using the trp promoter and an AT-rich ribosome binding site to direct expression in a pBR322-derived vector. A culture has been selected which allows large-scale fermentation and production of this material as an insoluble inclusion body protein. Procedures which solubilize, refold, and purify sCD4-183 have been developed. The purified sCD4-183 binds gp120 in solution and blocks human immunodeficiency virus (HIV) infection of human peripheral blood lymphocytes in vitro.
Bioprocess Research & Development, Upjohn Company, Kalamazoo, MI 49001.
R L
RL
Garlick
R J
RJ
Kirschner
F M
FM
Eckenrode
W G
WG
Tarpley
C S
CS
Tomich
D000596:Amino Acids / Q000032:analysis; D015704:CD4 Antigens / Q000096:biosynthesis* / Q000276:immunology / Q000302:isolation & purification; D004926:Escherichia coli / Q000235:genetics*; D006678:HIV / Q000276:immunology; D006801:Humans; D010957:Plasmids; D011994:Recombinant Proteins / Q000096:biosynthesis*
D000596:Amino Acids; D015704:CD4 Antigens; D011994:Recombinant Proteins