Boarder M R, Weber E, Evans C J, Erdelyi E, Barchas J
J Neurochem. 1983 Jun;40(6):1517-22. doi: 10.1111/j.1471-4159.1983.tb08120.x.
A sensitive assay, which cross-reacts with and is specific for diverse opioid peptides, is described. This is based on the prior acetylation of samples and subsequent radioimmunoassay with an antiserum highly specific for the acetylated NH2 terminus of opioid peptides. The result is a procedure that can be used to investigate multiple forms of opioid peptides in extracts of biological material. The sensitivity of the assay is approximately 15 fmol of beta-endorphin per incubation tube, i.e., approximately 100-fold greater sensitivity than the radioreceptor assay used in our laboratory. The peptide concentration required for 50% displacement of trace ranged from 0.65 nM (beta-endorphin) to 1.6 nM (Met-enkephalin). The assay apparently shows an absolute requirement for a free (or acetylated) NH2 terminus corresponding to either a Leu- or Met-enkephalin sequence. Use of the assay with and without prior acetylation of sample provides a method for estimation of the ratio of acetylated:nonacetylated opioid peptides in crude or fractionated extracts. The procedure is used to investigate the forms of opioid peptide found in rat brain and pituitary.
本文描述了一种灵敏的检测方法，该方法能与多种阿片肽发生交叉反应且具有特异性。此方法基于样品的预先乙酰化处理，随后采用针对阿片肽乙酰化氨基末端的高度特异性抗血清进行放射免疫测定。结果得到一种可用于研究生物材料提取物中多种形式阿片肽的方法。该检测方法的灵敏度约为每孵育管15 fmol的β-内啡肽，即比我们实验室使用的放射受体检测法灵敏度高约100倍。使微量物质50%被置换所需的肽浓度范围为0.65 nM（β-内啡肽）至1.6 nM（甲硫氨酸脑啡肽）。该检测方法显然对与亮氨酸脑啡肽或甲硫氨酸脑啡肽序列相对应的游离（或乙酰化）氨基末端有绝对要求。对样品进行预先乙酰化和不进行预先乙酰化处理的情况下使用该检测方法，可提供一种估算粗提物或分级提取物中乙酰化阿片肽与非乙酰化阿片肽比例的方法。该方法用于研究大鼠脑和垂体中发现的阿片肽形式。
