Martin K, Talukder R, Hay F C, Axford J S
Department of Biochemistry and Immunology, St. George's Hospital Medical School, London, United Kingdom.
J Rheumatol. 2001 Jul;28(7):1531-6.
To investigate fluorophore linked carbohydrate electrophoresis (FCE) as a method of analyzing serum immunoglobulin G (IgG) oligosaccharides in healthy individuals and those with rheumatic disease and compare with lectin binding assays of carbohydrate composition.
IgG was isolated from patients with rheumatoid arthritis (RA) (n = 21), ankylosing spondylitis (AS) (n = 20), psoriatic arthritis (PsA) (n = 20), and healthy adults (n = 36). IgG oligosaccharides were released enzymatically, fluorescently labelled using 8 aminonaphthalene-136 trisulfonic acid; and identification of the oligosaccharide bands was by stepwise enzymatic degradation. Comparison of FCE was made with lectin binding analysis in which the lectins Ricinus communis (RCA1) and Bandeiraea simplicifolia (BSII) were used to detect galactose (Gal) and N-acetylglucosamine (GlcNAc), respectively.
Each disease could be differentiated from healthy adults on the basis of Band 1 asialodigalacto core fucosylated oligosaccharide (gf2) intensity (p = 0.001), but not from each other. Reduced levels of different sugars were associated with specific diseases: reduced gf2 with RA (p < 0.001), PsA (p < 0.001) and AS (p < 0.02), reduced Band 5 disialo-digalacto core fucosylated (a2f) oligosaccharide with AS (p < 0.001), reduced Band 6 disialo-digalacto (a2) oligosaccharide with AS (p < 0.001) and PsA (p = 0.021). All diseases were associated with a significant increase in Band 4 asialo-agalacto core fucosylated oligosaccharide (g0f) (p < 0.001). In RA, FCE band intensities correlated with sugar quantity when identified using lectin binding analysis (p < 0.003). In contrast, there was no correlation between the same bands in healthy individuals.
FCE is an accurate method of analyzing IgG associated oligosaccharides and reveals unique band patterns or sugar prints associated with healthy adults and patients with RA, PsA, and AS, and comparison with lectin binding analysis suggests undetected RA glycoprotein structural differences. FCE has potential in the early diagnosis and differentiation of rheumatic diseases.
研究荧光团连接碳水化合物电泳（FCE）作为分析健康个体和风湿性疾病患者血清免疫球蛋白G（IgG）寡糖的方法，并与碳水化合物组成的凝集素结合测定法进行比较。
从类风湿关节炎（RA）患者（n = 21）、强直性脊柱炎（AS）患者（n = 20）、银屑病关节炎（PsA）患者（n = 20）和健康成年人（n = 36）中分离IgG。IgG寡糖通过酶法释放，用8-氨基萘-1,3,6-三磺酸进行荧光标记；寡糖条带的鉴定采用逐步酶解。将FCE与凝集素结合分析进行比较，其中分别使用蓖麻凝集素（RCA1）和简单半刀豆凝集素（BSII）检测半乳糖（Gal）和N-乙酰葡糖胺（GlcNAc）。
根据1号带无唾液酸二半乳糖核心岩藻糖基化寡糖（gf2）强度，每种疾病都可与健康成年人区分开来（p = .001），但彼此之间无法区分。不同糖类水平的降低与特定疾病相关：RA（p < .001）、PsA（p < .001）和AS（p < .02）中gf2降低，AS中5号带双唾液酸-二半乳糖核心岩藻糖基化（a2f）寡糖降低（p < .001），AS（p < .001）和PsA（p = .021）中6号带双唾液酸-二半乳糖（a2）寡糖降低。所有疾病均与4号带无唾液酸-无半乳糖核心岩藻糖基化寡糖（g0f）显著增加相关（p < .001）。在RA中，当使用凝集素结合分析鉴定时FCE条带强度与糖量相关（p < .003）。相比之下，健康个体中相同条带之间无相关性。
FCE是分析IgG相关寡糖的准确方法，揭示了与健康成年人以及RA、PsA和AS患者相关的独特条带模式或糖谱，与凝集素结合分析的比较表明存在未检测到的RA糖蛋白结构差异。FCE在风湿性疾病的早期诊断和鉴别方面具有潜力。
