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中国生物工程杂志

CHINA BIOTECHNOLOGY
中国生物工程杂志
中国生物工程杂志  2018, Vol. 38 Issue (4): 78-89    DOI: 10.13523/j.cb.20180411
综述     
抗体固定化方法研究进展
杜凯,张卓玲,李婷华,饶微()
深圳市新产业生物医学工程股份有限公司 深圳 518118
The Research Progress of Antibody Immobilization
Kai DU,Zhuo-ling ZHANG,Ting-hua LI,Wei RAO()
Shenzhen New Industries Biomedical Engineering Co., Ltd., Shenzhen 518118, China
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摘要：

在免疫分析和生物芯片中,抗原-抗体特异性结合被广泛应用,其中抗体的固定化是研发高效诊断和分离工具的关键环节。生物分子工程、材料化学与交联剂化学的进步极大地促进了抗体固定化技术的发展。 抗体可以通过物理吸附、共价偶联和亲和相互作用固定到不同类型的固相表面。 抗体固定化的目标是以一种正确的空间取向将抗体固定到固相表面,在完全保留抗体构象和活性的同时最大化抗原的结合能力,这对固相化抗体的分析性能至关重要。 对固定抗体到固相载体表面的各种最新方法进行了阐述,包括物理吸附法,通过羧基、氨基、巯基、糖基和点击化学的共价结合法以及基于生物亲和作用的固定法,并对固定化抗体的表征方法进行了归纳,最后对抗体固定化方法的发展方向进行了展望。
关键词： 抗体;  固定化;  免疫分析    
Abstract:

The antigen-antibody specific binding reaction is widely used in immunoassays and biochips in which antibody immobilization play a key role in the development of efficient diagnostics and separation tools. Progress in the field of biomolecular engineering, material chemistry and crosslinker chemistry have greatly promoted the development of antibody immobilization techniques. Antibody can be immobilized on different types of solid-phase surfaces by physical adsorption, covalent attachment and affinity-based interaction. The aim of antibody immobilization is to full retain antibody comformation and activity while maximize the antigen binding capacities by immobilizing antibody to the surface in the right orientation which is critical to the analytical performance. The most recent methods for immobilization of antibody on solid-phase surface, including physical adsorption, covalent binding through carboxyl, amine, thiol, carbohydrates as well as click chemistries, and through bioaffinity techniques. The characterization methods for the investigation of immobilized antibodies are summarized. In addition, future perspectives for methods of antibody immobilization are also discussed.
Key words: Antibody    Immobilization    Immunoassays
收稿日期: 2017-11-06 出版日期: 2018-05-08
ZTFLH:  Q511O629.7  
基金资助: 深圳市战略新兴产业发展专项资助项目(JSGG20160225150949068)
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杜凯
张卓玲
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引用本文:

杜凯,张卓玲,李婷华,饶微. 抗体固定化方法研究进展[J]. 中国生物工程杂志, 2018, 38(4): 78-89.

Kai DU,Zhuo-ling ZHANG,Ting-hua LI,Wei RAO. The Research Progress of Antibody Immobilization. China Biotechnology, 2018, 38(4): 78-89.

链接本文:

https://manu60.magtech.com.cn/biotech/CN/10.13523/j.cb.20180411    或    https://manu60.magtech.com.cn/biotech/CN/Y2018/V38/I4/78

图1  抗体的结构及固定化抗体的空间取向
图2  EDC/NHS活化羧基表面的反应机理
图3  光反应性探针辅助的亚硼酸连接法用于抗体固定
图4  通过Diels-Alder反应制备Herceptin 单抗偶联纳米粒
图5  末端含有硫酯的EYFP及连接有EYFP的脂质体的制备
图6  位点特异性定向共价偶联生物素化IgG的策略
图7  紫外线辅助下的核苷酸结合位点的抗体固定
图8  紫外线辅助下基于金属鳌合的抗体固定
技术方法 输入 输出 信息 应用
XPS[60,61] 单色X射线 光电子 元素和化学成分 定量载体表面抗体的密度
SE[62,63] 椭圆偏振光 光的相位和强度的变化 厚度、折射率、表面粗糙度 模式化分析、推断抗体形态
DPI[64,65] 激光 衰逝波的变化 质量、膜厚度、折射率、密度 通过质量和膜厚度推断抗体形态
SPR[66,67] 多角度单色激光 反射光和吸收光的变化 折射率、膜厚度 通过抗体和抗原的光吸收特性推断其形态
NR[68,69] 中子束 中子束反射后角度和波长的变化 折射率、膜厚度、表面粗糙度 模式化分析、推断抗体形态
AFM[70,71,72,73] 反馈驱动悬臂式纳米尖端 抗体在载体表面的高度、表面张力 表面粗糙度、相位信息、图像 精确定位高于载体表面14nm的抗体
QCM[74,75] 微量天平的共振频率 频率和振幅的变化 质量吸收系数、生物亲和性 通过压电石英晶体对抗体的吸收性和质量推断抗体形态
ToF-SIMS[76,77] 电离金属簇,“一级离子” 电离片段,“二级离子” 元素半定量、化学成分、分子质量 分辨F(ab')2和Fc片段氨基酸的种类及数量
表1  应用于抗体偶联与固相表面的分析技术
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doi: 10.1021/acs.langmuir.6b02312 pmid: 27494212
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