Weber Julia, Sachse Janine, Bergmann Sarah, Sparwaßer Andrea, Struck Joachim, Bergmann Andreas
sphingotec GmbH, Hennigsdorf, Germany.
Adrenomed AG, Hennigsdorf, Germany.
J Appl Lab Med. 2017 Sep 1;2(2):222-233. doi: 10.1373/jalm.2017.023655.
Adrenomedullin (ADM) is a circulating peptide known to regulate vasodilation and vascular integrity. Increased plasma ADM concentrations have been described for several life-threatening conditions, including cardiovascular diseases and septic shock. Reliable methods for the simple quantification of bioactive ADM (bio-ADM) are lacking.
Monoclonal antibodies against the amidated C-terminus and middle portion of bio-ADM were generated and used for the development of a 1-step immunometric assay for the specific quantification of bio-ADM in plasma. The assay was developed in a microtiter plate/chemiluminescence label format with a significantly reduced incubation time. Precision, linearity, specimen stability, and distribution of results in healthy subjects were evaluated.
The use of monoclonal antibodies against predetermined epitopes of bio-ADM enabled the development of an assay for the determination of bio-ADM directly in EDTA plasma. Plasma samples were stable for up to 24 h at ambient temperature and over multiple freeze-thaw cycles without loss of immunoreactivity. The assay had a limit of detection of 3 pg/mL and a limit of quantification of 11 pg/mL. The assay exhibited acceptable linearity characteristics and was not influenced by complement factor H, a putative ADM-binding protein. In healthy subjects, bio-ADM concentrations were all above the limit of detection, and approximately half of them were above the limit of quantification.
By using monoclonal antibodies with defined epitope specificities, we have developed a simple, rapid, accurate, and sensitive sandwich immunoassay for bio-ADM. The assay is a potentially novel tool to support patient management, particularly in acute care in the field of sepsis and other indications, which are currently being investigated, such as acute heart failure.
肾上腺髓质素（ADM）是一种循环肽，已知其可调节血管舒张和血管完整性。在包括心血管疾病和感染性休克在内的几种危及生命的病症中，血浆ADM浓度会升高。目前缺乏简单定量生物活性ADM（生物ADM）的可靠方法。
制备了针对生物ADM酰胺化C末端和中间部分的单克隆抗体，并用于开发一步免疫测定法，以特异性定量血浆中的生物ADM。该测定法采用微孔板/化学发光标记形式开发，孵育时间显著缩短。评估了该测定法的精密度、线性、样本稳定性以及健康受试者的结果分布。
使用针对生物ADM预定表位的单克隆抗体能够开发一种直接在EDTA血浆中测定生物ADM的测定法。血浆样本在室温下长达24小时以及经过多次冻融循环后仍保持稳定，且免疫反应性无损失。该测定法的检测限为3 pg/mL，定量限为11 pg/mL。该测定法表现出可接受的线性特征，且不受补体因子H（一种假定的ADM结合蛋白）的影响。在健康受试者中，生物ADM浓度均高于检测限，其中约一半高于定量限。
通过使用具有明确表位特异性的单克隆抗体，我们开发了一种用于生物ADM的简单、快速、准确且灵敏的夹心免疫测定法。该测定法是一种潜在的新型工具，可支持患者管理，特别是在脓毒症领域的急性护理以及其他正在研究的适应症（如急性心力衰竭）中。
