Artificial immunoglobulin G-binding protein mimetic to staphylococcal protein A. Its production and application to affinity purification of immunoglobulin G.
Staphylococcal protein A consists of a single polypeptide with five immunoglobulin G (IgG)-binding domains, which are linked as E-D-A-B-C in this order from the amino terminal. The DNA coding domains A-B were polymerized one to six times linearly, taking advantage of the non-palindromic nucleotide sequence of the AccI recognition site and the resultant DNAs were inserted in pTRP vector carrying trp promoter. The artificial IgG-binding proteins [pA(AB)1-6], which had been expressed in Escherichia coli JM109, were purified by methods involving IgG-Sepharose affinity chromatography. Among pA(AB)1-6 immobilized on cyanogen bromide-Sepharose, pA(AB)4-Sepharose was the highest in IgG-binding capacity at the same level of mg protein per ml gel, about 30% higher than protein A-Sepharose. At 8 mg protein per ml gel, it bound and eluted about 24 mg of IgG from rabbit serum. Its IgG-binding capacities were the highest with porcine, rabbit, human and guinea pig sera, intermediate with bovine, horse and sheep sera and the lowest with mouse, goat, rat and chicken sera.
Kihira
Y
Y
Nagahama Institute for Biochemical Science, Oriental Yeast Co., Ltd., Shiga-ken, Japan.
Aiba
S
S
D000595:Amino Acid Sequence; D001483:Base Sequence; D002846:Chromatography, Affinity; D004247:DNA / Q000138:chemical synthesis; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D007074:Immunoglobulin G / Q000302:isolation & purification*; D008222:Lymphokines / Q000032:analysis / Q000737:chemistry* / Q000302:isolation & purification; D008969:Molecular Sequence Data; D029584:Prostatic Secretory Proteins; D011994:Recombinant Proteins / Q000032:analysis / Q000302:isolation & purification; D013205:Staphylococcal Protein A / Q000032:analysis*; D014170:Transformation, Genetic
D007074:Immunoglobulin G; D008222:Lymphokines; D029584:Prostatic Secretory Proteins; D011994:Recombinant Proteins; D013205:Staphylococcal Protein A; C043076:beta-microseminoprotein; C013059:immunoglobulin-binding factors; D004247:DNA