Tandem immunoaffinity and reversed-phase high-performance liquid chromatography for the identification of the specific binding sites of a hapten on a proteic carrier.
Immunoaffinity (IA) and reversed-phase (RP) high-performance liquid chromatography were combined for the identification of the specific binding sites of benzylpenicilloyl (BPO) groups on human serum albumin (HSA). Tryptic hydrolysates of BPO-HSA were loaded on the IA column. BPO-peptides were desorbed and concentrated directly on the RP column, coupled via a switching valve, then separated by using gradient elution and identified by the amino acid sequences. This single-step procedure permitted more than 95% recovery of the BPO-peptides present in minute amounts, with good specificity.
Yvon
M
M
Station de Recherches Laitières, CRJ-INRA, Jouy-en-Josas, France.
Wal
J M
JM
D000595:Amino Acid Sequence; D044482:Benzeneacetamides; D002352:Carrier Proteins / Q000378:metabolism*; D002846:Chromatography, Affinity / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D006241:Haptens / Q000032:analysis* / Q000737:chemistry / Q000378:metabolism; D006801:Humans; D008969:Molecular Sequence Data; D010400:Penicillin G / Q000031:analogs & derivatives / Q000737:chemistry / Q000302:isolation & purification; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification
D044482:Benzeneacetamides; D002352:Carrier Proteins; D006241:Haptens; D010455:Peptides; C015026:benzylpenicilloyl G; D010400:Penicillin G