Department of Clinical Biochemistry, Faculty of Health Sciences, Bispebjerg and Frederiksberg Hospital, University of Copenhagen, Nielsine Nielsens Vej 4, 2400 Copenhagen, NV, Denmark.
Int J Mol Sci. 2023 Oct 12;24(20):15102. doi: 10.3390/ijms242015102.
Pituitary adenylate cyclase-activating polypeptide (PACAP) is a naturally occurring neuropeptide found in both the central and peripheral nervous systems of vertebrates. Recent studies have revealed the presence of PACAP and its corresponding receptors, namely, the pituitary adenylate cyclase-activating polypeptide type I receptor (PAC1R), vasoactive intestinal peptide receptor 1 (VIPR1), and vasoactive intestinal peptide receptor 2 (VIPR2), in various structures implicated in migraine pathophysiology, including sensory trigeminal neurons. Human studies have demonstrated that when infused, PACAP can cause dilation of cranial vessels and result in delayed migraine-like attacks. In light of this, we present a novel ELISA assay that has been validated for quantifying PACAP in tissue extracts and human plasma. Using two well characterized antibodies specifically targeting PACAP, we successfully developed a sandwich ELISA assay, capable of detecting and accurately quantifying PACAP without any cross-reactivity to closely related peptides. The quantification range was between 5.2 pmol/L and 400 pmol/L. The recovery in plasma ranged from 98.2% to 100%. The increasing evidence pointing to the crucial role of PACAP in migraine pathophysiology necessitates the availability of tools capable of detecting changes in the circulatory levels of PACAP and its potential application as a reliable biomarker.
垂体腺苷酸环化酶激活肽（PACAP）是一种在脊椎动物中枢和外周神经系统中均存在的天然存在的神经肽。最近的研究表明，PACAP 及其相应的受体，即垂体腺苷酸环化酶激活肽 I 型受体（PAC1R）、血管活性肠肽受体 1（VIPR1）和血管活性肠肽受体 2（VIPR2），存在于偏头痛发病机制中涉及的各种结构中，包括感觉三叉神经神经元。人体研究表明，当输注时，PACAP 可引起颅血管扩张，并导致延迟性偏头痛样发作。鉴于此，我们提出了一种新的 ELISA 检测法，该方法已被验证可用于定量组织提取物和人血浆中的 PACAP。使用两种专门针对 PACAP 的特征明确的抗体，我们成功开发了一种夹心 ELISA 检测法，能够检测和准确定量 PACAP，而与密切相关的肽没有任何交叉反应性。定量范围为 5.2pmol/L 至 400pmol/L。在血浆中的回收率在 98.2%至 100%之间。越来越多的证据表明 PACAP 在偏头痛发病机制中起着至关重要的作用，这就需要有能够检测到 PACAP 循环水平变化的工具，以及将其作为可靠的生物标志物的潜在应用。
