一种用于测量血浆中神经肽Y的免疫放射分析方法。

An immunoradiometric assay for the measurement of neuropeptide Y in plasma.

作者信息

Corder R, Lowry P J

出版信息

Peptides. 1985 Nov-Dec;6(6):1195-200. doi: 10.1016/0196-9781(85)90450-4.

Abstract

The development of an immunoradiometric assay (IRMA) for the direct measurement of neuropeptide Y (NPY) concentrations in plasma is reported. The assay employs simultaneous addition of 125I-labelled affinity purified sheep anti-(NPY 31-36) immunoglobulin (IgG) and a rabbit anti-NPY serum to 0.25 ml volumes of standard or unknown. After 16 hr incubation at 4 degrees C NPY-bound labelled IgG is precipitated using sheep anti-(rabbit IgG Fc region) IgG coupled to Dynospheres solid phase. Precipitated counts are proportional to the NPY concentration in samples. Using this methodology it is possible to measure basal levels in normal human subjects (range 1-5 fmol/ml). Technical difficulties encountered in raising "site-specific" antisera to NPY during the establishment of this assay are outlined.

摘要

本文报道了一种用于直接测定血浆中神经肽Y（NPY）浓度的免疫放射分析（IRMA）方法的建立。该分析方法是将125I标记的亲和纯化羊抗（NPY 31 - 36）免疫球蛋白（IgG）和兔抗NPY血清同时加入0.25 ml的标准品或未知样品中。在4℃孵育16小时后，使用与Dynospheres固相偶联的羊抗（兔IgG Fc区）IgG沉淀与NPY结合的标记IgG。沉淀计数与样品中NPY浓度成正比。采用这种方法能够测定正常人类受试者的基础水平（范围为1 - 5 fmol/ml）。文中概述了在建立该分析方法过程中，制备针对NPY的“位点特异性”抗血清时遇到的技术难题。