喹啉腈荧光微球构建及在荧光免疫层析中对SAA的检测应用

李强; 徐建新; 郭志前

doi:10.14135/j.cnki.1006-2080.20220414001

喹啉腈荧光微球构建及在荧光免疫层析中对SAA的检测应用

doi: 10.14135/j.cnki.1006-2080.20220414001

李强^{1, 2,},
徐建新¹,
郭志前^2, ,

1.
上海奥普生物医药股份有限公司, 上海即时检测医学工程技术研究中心, 上海, 200201
2.
华东理工大学化学与分子工程学院, 精细化工研究所, 上海, 200237

基金项目: 国家自然科学基金面上项目(21878087 )

详细信息

作者简介:
李强（1990-），男，上海人，博士，主要从事功能性荧光染料微球的研究。E-mail: liq0311@163.com

通讯作者:
郭志前，E-mail：guozq@ecust.edu.cn

中图分类号: O69
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- 文章访问数: 20
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- PDF下载量: 4
- 被引次数: 0
出版历程
- 收稿日期: 2022-04-14
- 网络出版日期: 2022-05-09

Construction of Quinoline-malononitrile Fluorescent Microspheres for the Detection of SAA via Fluorescent Immunochromatography

LI Qiang^{1, 2
,},
XU Jianxin¹,
GUO Zhiqian^{2
, ,}

1.
Shanghai UPPER Bio-Tech Pharma Co., LTD., Shanghai Engineering Research Center of iPOCT Medicine, Shanghai 200201, China
2.
Institute of Fine Chemicals, School of Chemistry and Molecular Engineering, East China University of Science and Technology, Shanghai 200237, China

摘要

摘要: 基于聚集诱导发光（AIE）喹啉腈（QM）染料的衍生物QM-OH制备了乳胶荧光微球QM-OH@PS-COOH（羧基聚苯乙烯），通过扫描电子显微镜等表征仪器研究了微球的形貌结构，并进一步将该荧光微球应用于荧光免疫层析法检测中，实现了对临床样本中血清淀粉样蛋白A（SAA）含量变化的灵敏检测。研究结果表明：QM-OH@PS-COOH荧光微球大小均一、形状规整，与市售荧光微球对比，其密度、固含量、羧基含量基本一致；该荧光微球体系不仅可以定性区分炎症感染，而且能对SAA含量进行定量分析，实现了对炎症感染的有效评估和监控。因此，基于QM-OH@PS-COOH的检测平台可用于早期疾病检测的筛查，特别是用于即时检测领域中的自我检测。
- 聚集诱导发光 /
- 喹啉腈 /
- 荧光微球 /
- 荧光免疫层析 /
- 血清淀粉样蛋白A
Abstract: Serum amyloid A (SAA) is an acute-phase protein mainly produced by the liver in response to proinflammatory cytokines. SAA genes and proteins are significantly activated during the acute phase response, which comprises a number of phenomena that occur in the presence of inflammation and infection, increased temperature and hormonal and metabolic alterations. Therefore SAA is a sensitive indicator of inflammation in the early stage of infectious diseases, which is important for diagnosis, evaluation, monitoring and treatment of inflammation. Fluorescent immunochromatography is one of the most popular strategies for point-of-care testing (POCT), which is capable of rapid screening for disease detection. Fluorescent microspheres QM-OH@PS-COOH were obtained from aggregation-induced emission (AIE) quinoline-malononitrile (QM) derivatives QM-OH. The morphology and structure of these QM fluorescent microspheres were characterized by scanning electron microscopy et al. Finally, these fluorescent microspheres were utilized to detect for SAA concertation in clinical samples via fluorescent immunochromatography. The results showed that QM-OH@PS-COOH has uniform sizes with regular shapes. Compared with commercial fluorescent microspheres, these AIE microspheres had similar density, solid content and carboxyl content. The QM-OH@PS-COOH system exhibited the detection of SAA with high sensitivity in clinical samples via fluorescent immunochromatography. Thus, this new type of QM fluorescent microspheres could be employed as an important tool for clinical diagnosis, enabling quantitatively analyze and monitor the concentration of SAA during the inflammation process. Therefore, we believe that these detection platforms based on QM-OH@PS-COOH can serve as a screening platform for early disease detection, especially self-testing in POCT.
- aggregation-induced emission /
- quinoline-malononitril /
- fluorescent microspheres /
- fluorescent immunochromatography /
- serum amyloid A

HTML全文

图 1 激发波长440 nm下化合物QM-OH在水和四氢呋喃体系中（不同水质量分数）的紫外吸收图（a）和荧光光谱图（b）

Figure 1. Absorption (a) and emission (b) spectra of QM-OH in H₂O/THF mixtures with different volume fraction of water (f_w) under λ _ex = 440 nm

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图 2 AIE荧光微球QM-OH@PS-COOH的TEM图，插图是QM-OH@PS-COOH的乳液图片

Figure 2. TEM image of QM-OH@PS-COOH, illustrated by emulsion image of QM-OH@PS-COOH

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图 3 AIE荧光微球QM-OH@PS-COOH的表面羧基电导滴定变化图

Figure 3. Surface carboxyl conductance titration of QM-OH@PS-COOH

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图 4 喹啉腈AIE荧光微球QM-OH@PS-COOH荧光免疫层析检测SAA原理示意图

Figure 4. Schematic diagram of quinoline-malononitrile AIE fluorescent microsphere QM-OH@PS-COOH for SAA detection in fluorescent immunochromato graphy

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图 5 SAA质控品定标的曲线

Figure 5. Calibration curve of SAA quality control product

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图 6 （a）MAYA荧光免疫定量分析仪；（b）日光灯下，阴性、阳性低值、阳性高值样本的检测条；（c）荧光灯下，阴性、阳性低值、阳性高值样本的检测条

Figure 6. (a) MAYA fluorescence immunoquantitative analyzer; (b) Test strips for negative, low positive and high positive samples under fluorescent lamps; (c) Test strips for negative, low positive and high positive samples under fluorescent lamps

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表 1 SAA质控品定标的数据

Table 1. SAA quality control product calibration data

ρ（SAA） /(mg·L⁻¹)	Fluorescene/（a.u.）
	Data 1		Data 2		Data average
	A Line	B Line	A Line	B Line	A Line	B Line
0	1145	9	1150	9	1147.5	9
6.75	1260	109	1262	109	1261	109
13.5	1355	193	1350	187	1352.5	190
27	1540	354	1536	345	1538	349.5
54	1881	638	1899	662	1890	650
108	2638	1298	2619	1275	2628.5	1286.5
216	3627	2022	3665	2038	3646	2030

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表 2 本文和龙华医院测试临床样本SAA测值比较

Table 2. Comparation of SAA value between this study and Longhua Hospital

Sample	ρ（SAA）/（mg·L⁻¹）
Sample	This study	Longhua Hospital	Deviation/%
1	1.61	1.72	−6.40
2	2.30	2.13	7.98
3	3.47	3.75	−7.47
4	3.73	3.98	−6.28
5	11.88	10.38	14.45
6	22.58	20.89	8.09
7	23.73	26.63	−10.89
8	41.49	36.25	14.46
9	59.72	53.76	11.09
10	115.14	100.52	14.54

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