用于糖链抗原CA19-9实时动态活检的免疫双光子荧光标记试剂盒

doi:10.7503/cjcu20150727

高等学校化学学报 ›› 2016, Vol. 37 ›› Issue (4): 638.doi: 10.7503/cjcu20150727

用于糖链抗原CA19-9实时动态活检的免疫双光子荧光标记试剂盒

黄池宝^1,²(), 张道海³, 曾伯平¹, 刘其斌³, 陈华仕¹, 康帅¹, 陈晓远²

1. 遵义师范学院化学化工学院, 遵义 563002
2. 韶关大学农业科学与工程学院, 韶关 512005
3. 贵州大学材料与冶金学院, 贵阳 550025

收稿日期:2015-09-17 出版日期:2016-04-10 发布日期:2016-03-10
基金资助:
国家自然科学基金(批准号: 21562050, 11464052)、贵州省第八批科技创新人才团队建设专项基金[批准号: 黔科合人才团队(2015)4007]、贵州省科学技术基金项目(批准号: 黔科合J字[2015]2146, [2012]2345)、贵州省教育厅自然科学重点研究项目(批准号: 黔教合KY字[2014]296, [2013]171)、贵州省教育厅省级本科教学工程建设项目(批准号: 黔教[2014]JXGChcb)、遵义市“15851人才精英工程”项目[批准号: (2015)4007]和遵义市红花岗区科学技术项目(批准号: 遵红科合社字[2015]18)资助

Two-photon Fluorescent Labeling Kit for the Real-time Dynamic Biopsy of CA19-9^†

HUANG Chibao^1,^2,^*(), ZHANG Daohai³, ZENG Boping¹, LIU Qibin³, CHEN Huashi¹, KANG Shuai¹, CHEN Xiaoyuan²

1. Chemistry and Chemical Engineering College, Zunyi Normal University, Zunyi 563002, China
2. College of Agricultural Science and Engineering, Shaoguan University, Shaoguan 512005, China
3. College of Materials and Metallurgy, Guizhou University, Guiyang 550025, China

Received:2015-09-17 Online:2016-04-10 Published:2016-03-10
Contact: HUANG Chibao E-mail:huangchibao@163.com
Supported by:
† Supported by the National Natural Science Foundation of China(Nos:21562050, 11464052), the Special Fund Project of the Construction of the Eighth Batch of Scientific and Technological Innovation Talent Team in Guizhou Province, China[No:(2015)4007], Guizhou Provincial Science and Technology Fund Project, China(Nos:J[2015]2146, J[2012]2345), the Key Project of Education Department of Guizhou Province(Nos:KY[2014]296), KY(2013)171), the Teaching Contents and Curriculum System Reform Project of Higher Education in Guizhou Province, China(No.KY [2014] JXGChcb), the Project of “15851 Talents Elite Project” in Zunyi City, China[No:(2015)4007] and the Science and Technology Project of Zunyi City Honghuagang District, China(No.[2015]18)

摘要/Abstract

摘要：

采用双光子荧光标记探针LP制备了免疫双光子荧光抗体LP-Ab, 以特异性识别肿瘤标志物CA19-9(Ag)形成抗原-抗体复合物(LP-Ab·Ag), 基于双光子诱导荧光机制用近红外激光(740 nm)激发LP-Ab·Ag, 利用LP-Ab·Ag的双光子荧光实现了CA19-9的高清晰度、高灵敏度及高分辨率的量化检测和实时动态成像, 并制成用于糖链抗原CA19-9检测的简单便捷的双光子荧光标记试剂盒.

关键词: 糖链抗原CA19-9, 抗CA19-9单克隆抗体(Ab19-9), 免疫荧光标记, 双光子, 活检

Abstract:

Carbohydrate antigen CA19-9(Ag) is a tumor marker associated with pancreatic cancer, gallbladder cancer, colon cancer and gastric cancer in early stage of tumor and cancer stage. Antibody Ab19-9(Ab) to carbohydrate antigens were labeled to form probe-labeled antibody LP-Ab using the prepared two-photon fluorescence probe LP. On the basis of the antigen-antibody specificity affinity principle, LP-Ab could combine with carbohydrate antigen Ag into a complex LP-Ab·Ag, which was excited by near infrared laser(740 nm) and emitted strong two-photon fluorescence to accomplish the high sensitive biopsy and real-time dynamic high-resolution imaging of the tumor marker Ag with two-photon fluorescence microscope. Based on this, a simple and convenient two-photon fluorescent labeling kit was prepared to realize the rapid detection of tumor, which is convenient for large-scale clinic testing and screening. The research further broadens the application of LP, and provides a sharp tool for revealing the mechanism of cancer occurrence, development and metastasis. LP-Ab can be used to observe and study the dynamic transfer mechanisms of the tumor markers in cells and the mechanism of carcinogenesis.

Key words: Carbonhydrate antigen CA19-9, Antibody Ab19-9, Immunofluorescence labeling, Two-photon, Biopsy(Ed.: P, H, N, K)

中图分类号:

O657

TrendMD:

黄池宝, 张道海, 曾伯平, 刘其斌, 陈华仕, 康帅, 陈晓远. 用于糖链抗原CA19-9实时动态活检的免疫双光子荧光标记试剂盒. 高等学校化学学报, 2016, 37(4): 638.

HUANG Chibao, ZHANG Daohai, ZENG Boping, LIU Qibin, CHEN Huashi, KANG Shuai, CHEN Xiaoyuan. Two-photon Fluorescent Labeling Kit for the Real-time Dynamic Biopsy of CA19-9^†. Chem. J. Chinese Universities, 2016, 37(4): 638.

图/表 5

Fig.1 Molecular structure of LP

Fig.2 Two-photon action spectra of LP(a, 10 μmol/L) and LP-Ab(b, 10 μmol/L)

Fig.3 TPM images of 1 μmol/L LP-Ab-labeled mouse pancreatic carcinoma cells collected at 450—500 nm^(A) Magnification at 10×; (B) magnification at 40×. The images were collected upon excitation at 740 nm with a femtosecond pulse. Cells shown are representative images from replicate experiments(n=5).

Fig.4 Negative(A) and positive(B) fluorescent images of 1 μmol/L LP-Ab-labeled mouse pancreatic carcinoma and normal cells under the eyepiece

Fig.5 Diagram of two-photon fluorescent labeling kit

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用于糖链抗原CA19-9实时动态活检的免疫双光子荧光标记试剂盒

Two-photon Fluorescent Labeling Kit for the Real-time Dynamic Biopsy of CA19-9^†

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用于糖链抗原CA19-9实时动态活检的免疫双光子荧光标记试剂盒

Two-photon Fluorescent Labeling Kit for the Real-time Dynamic Biopsy of CA19-9†

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Two-photon Fluorescent Labeling Kit for the Real-time Dynamic Biopsy of CA19-9^†