Nuti Francesca, Fernandez Feliciana Real, Sabatino Giuseppina, Peroni Elisa, Mulinacci Barbara, Paolini Ilaria, Pisa Margherita Di, Tiberi Caterina, Lolli Francesco, Petruzzo Martina, Lanzillo Roberta, Morra Vincenzo Brescia, Rovero Paolo, Papini Anna Maria
Interdepartmental Research Unit of Peptide and Protein Chemistry and Biology, Department of Chemistry "Ugo Schiff", University of Florence, Via della Lastruccia 13, 50019 Sesto Fiorentino, Italy.
CNR-IC Istituto di Cristallografia, Via Paolo Gaifami 18, 95126 Catania, Italy.
Brain Sci. 2020 Jul 15;10(7):453. doi: 10.3390/brainsci10070453.
Diagnostics of Multiple Sclerosis (MS) are essentially based on the gold standard magnetic resonance imaging. Few alternative simple assays are available to follow up disease activity. Considering that the disease can remain elusive for years, identification of antibodies fluctuating in biological fluids as relevant biomarkers of immune response is a challenge. In previous studies, we reported that anti--glucosylated (-Glc) peptide antibodies that can be easily detected in Solid-Phase Enzyme-Linked ImmunoSorbent Assays (SP-ELISA) on MS patients' sera preferentially recognize hyperglucosylated adhesin of non-typeable . Since multivalency can be useful for diagnostic purposes to allow an efficient coating in ELISA, we report herein the development of a collection of Multiple -glucosylated Peptide Epitopes (-Glc MEPs) to detect anti--Glc antibodies in MS. To this aim, a series of -Glc peptide antigens to be represented in the -GlcMEPs were tested in competitive ELISA. We confirmed that the epitope recognized by antibodies shall contain at least 5-mer sequences including the fundamental -Glc moiety. Using a 4-branched dendrimeric lysine scaffold, we selected the -Glc MEP , carrying the minimal epitope Asn(Glc) anchored to a polyethylene glycol-based spacer (PEG) containing a 19-atoms chain, as an efficient multivalent probe to reveal specific and high affinity anti--Glc antibodies in MS.
多发性硬化症（MS）的诊断主要基于金标准磁共振成像。几乎没有其他简单的检测方法可用于跟踪疾病活动。鉴于该疾病可能多年难以捉摸，识别生物体液中波动的抗体作为免疫反应的相关生物标志物是一项挑战。在先前的研究中，我们报告称，在MS患者血清的固相酶联免疫吸附测定（SP-ELISA）中可以轻松检测到的抗糖基化（-Glc）肽抗体优先识别不可分型的高糖基化粘附素。由于多价性对于诊断目的可能有用，以允许在ELISA中进行有效包被，我们在此报告了一系列多糖基化肽表位（-Glc MEPs）的开发，用于检测MS中的抗-Glc抗体。为此，在竞争性ELISA中测试了一系列将在-GlcMEPs中呈现的-Glc肽抗原。我们证实，抗体识别的表位应包含至少5聚体序列，包括基本的-Glc部分。使用4分支树枝状赖氨酸支架，我们选择了-Glc MEP，它携带锚定在含有19个原子链的基于聚乙二醇的间隔物（PEG）上的最小表位Asn(Glc)，作为一种有效的多价探针，以揭示MS中特异性和高亲和力的抗-Glc抗体。
