Enzyme immunoassay of benzyl penicilloyl (BPO) groups using acetylcholinesterase as label. Application to the study of the BPO-binding sites on albumin.
Benzyl penicilloyl groups (BPO) derive from penicillin G by cleavage of the beta lactam ring; they covalently bind to proteins to give conjugates which have lost all antibiotic properties but are considered as the major allergenic determinants in penicillin allergy. A solid-phase Enzyme Immuno Assay (EIA) of BPO groups in different biological fluids is described. It is a competitive immunoassay using acetylcholinesterase as label. In all biological fluids, very low non-specific binding values are observed. The sensitivity and the precision of the assay are good since ca. 0.5 ng/ml can be measured with a coefficient of variation less than 10%. Cross reactions between BPO and penicillin or penicillin derivatives are nil or very low. This assay is more sensitive, much more rapid and easier to handle than the other methods available and is thus suitable for routine determinations. In association with reversed-phase high performance liquid chromatography this EIA has allowed an initial investigation of the location of BPO-binding sites on micro quantities of serum albumin (ca. 1 mg) from penicillin treated patients.
Wal
J M
JM
Laboratoire des Sciences de la Consommation INRA, CRJ, Jouy-en-Josas, France.
Yvon
M
M
Pradelles
P
P
Grassi
J
J
D000110:Acetylcholinesterase; D001665:Binding Sites; D004342:Drug Hypersensitivity / Q000276:immunology; D006801:Humans; D007124:Immunoenzyme Techniques; D010400:Penicillin G / Q000032:analysis* / Q000276:immunology / Q000378:metabolism; D010446:Peptide Fragments / Q000302:isolation & purification; D012709:Serum Albumin / Q000378:metabolism
D010446:Peptide Fragments; D012709:Serum Albumin; D000110:Acetylcholinesterase; D010400:Penicillin G