Ezan E, Tarrade T, Cazenave C, Ardouin T, Genet R, Grassi J, Grognet J M, Pradelles P
CEA, Service de Pharmacologie et d'Immunologie, Gif-sur-Yvette, France.
Peptides. 1995;16(3):449-55. doi: 10.1016/0196-9781(94)00218-u.
A novel type of enzyme immunometric assay has been developed for a heptapeptide, BN 52080. This compound is a short C-terminal analogue of sorbin and is under clinical evaluation for treatment of chronic diarrhea. In this solid-phase immobilized epitope immunoassay (SPIE-IA), the peptide is first immunologically bound to polyclonal antibodies adsorbed to a solid phase and then, after covalent immobilization with glutaraldehyde, is released from the antibody paratope by NaOH. The peptide linked to the solid phase is further quantified with a tracer consisting of the same antibodies purified by affinity chromatography and coupled to acetylcholinesterase. This assay has a detection limit of 10 pg/ml and is therefore five times more sensitive than competitive enzyme immunoassay using the same antibodies and BN 52080 coupled to acetylcholinesterase as tracer. The assay is specific and allows direct measurement of peptide in human plasma after subcutaneous or intravenous administration of 200 micrograms of BN 52080 to volunteers.
已开发出一种针对七肽BN 52080的新型酶免疫测定法。该化合物是山梨醇的一种短C端类似物,正在进行治疗慢性腹泻的临床评估。在这种固相固定表位免疫测定法(SPIE-IA)中,首先将肽免疫结合到吸附在固相上的多克隆抗体上,然后在用戊二醛进行共价固定后,通过氢氧化钠从抗体互补位释放出来。与固相相连的肽再用一种示踪剂进行定量,该示踪剂由通过亲和层析纯化并与乙酰胆碱酯酶偶联的相同抗体组成。该测定法的检测限为10 pg/ml,因此比使用相同抗体且以与乙酰胆碱酯酶偶联的BN 52080作为示踪剂的竞争性酶免疫测定法灵敏五倍。该测定法具有特异性,在向志愿者皮下或静脉注射200微克BN 52080后,可直接测量人血浆中的肽。