深圳大学学报理工版

将自研的基于波长扫描的表面等离子体共振传感系统与动力学分析方法相结合,研究葡萄球菌蛋白A、链球菌蛋白G与免疫球蛋白G(immunoglobulin G, IgG)的相互作用关系.实验测得,蛋白A和蛋白G与IgG的结合速率常数分别为1.3×105 L·mol-1·s-1和5.0×104 L·mol-1·s-1,说明蛋白A与IgG的结合效率更高.同时测得蛋白A和蛋白G与IgG的解离速率常数分别为2.1×10-2 s-1和5.0×10-3 s-1,说明蛋白G与IgG形成的复合物更稳定.此外,由解离速率常数与结合速率常数的比值得出蛋白G对IgG的亲和力较大.实验结果可为优化抗体固定方法提供必要参考和理论支撑.

We study the interactions between staphylococal protein A or streptococcus protein G and immunoglobulin IgG, respectively, based on the self-developed wavelength-based surface plasmon resonance(SPR)sensing system combined with the kinetic analysis. The experimental results show that the association rate constants ka of protein A and protein G with IgG are 1.3×105 L·mol-1·s-1 and 5.0×104 L·mol-1·s-1, indicating that the protein A binds to IgG more efficiently. At the same time, the dissociation rate constants kd of protein A and protein G from IgG are detected to be 2.1×10-2 s-1 and 5.0×10-3 s-1, indicating that the complex formed by protein G and IgG is more stable. In addition, the ratio of kd to ka indicates that protein G has higher affinity for IgG. This study may provide necessary reference and important theoretical support for the optimization of antibody immobilization methods.

引言
1 材料与方法
2 结果与分析
3 结语

图1 实验装置光路图<br/>Fig.1 Light path diagram of experimental device

图1 实验装置光路图
Fig.1 Light path diagram of experimental device

图2 抗体结合蛋白固定过程的共振波长变化监测<br/>Fig.2 Resonance wavelength monitoring of the immobilization of antibody-binding protein

图2 抗体结合蛋白固定过程的共振波长变化监测
Fig.2 Resonance wavelength monitoring of the immobilization of antibody-binding protein

图3 抗体结合蛋白与 IgG动力学反应曲线<br/>Fig.3 Kinetic curves of the interactions between antibody-binding proteins and IgG

图3 抗体结合蛋白与 IgG动力学反应曲线
Fig.3 Kinetic curves of the interactions between antibody-binding proteins and IgG

[1] 杜凯, 张卓玲, 李婷华, 等. 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.
[2] TRILLING A K, BEEKWILDER J, ZUILHOF H. Antibody orientation on biosensor surfaces: a minireview[J]. Analyst, 2013, 138(6): 1619-1627.
[3] QUINTERO-TROCONIS E, BUELVAS N, CARRASCO-LPEZ C, et al. Enolase from Trypanosoma cruzi is inhibited by its interaction with metallocarboxypeptidase-1 and a putative acireductone dioxygenase[J]. Biochimica et Biophysica Acta(BBA)- Proteins & Proteomics, 2018, 1866(5/6): 651- 660.
[4] DAVIDOFF S N, DITTO N T, BROOKS A E, et al. Surface plasmon resonance for therapeutic antibody characterization[J]. Methods in Pharmacology and Toxicology, 2015, 53: 35-76..
[5] YANG Danlin, SINGH A, WU H, et al. Determination of high-affinity antibody-antigen binding kinetics using four biosensor platforms[J]. Journal of Visualized Experiments, 2017, 122: 55659.
[6] HOMOLA J, YEE S S, GAUGLITZ G. Surface plasmon resonance sensors[J]. Sensors and Actuators B: Chemical, 1999, 54(1/2): 3-15.
[7] WANG Dongping, LOO J F C, CHEN Jiajie, et al. Recent advances in surface plasmon resonance imaging sensors[J]. Sensors, 2019, 19(6): 1266.
[8] LIU Le, GUO Jun, HE Yonghong, et al. Study on the despeckle methods in angular surface plasmon resonance imaging sensors[J]. Plasmonics, 2015, 10(3): 729-737.
[9] ZENG Youjun, ZHOU Jie, XIAO Xiaoping, et al. A speckle-free angular interrogation SPR imaging sensor based on galvanometer scan and laser excitation[J]. Plasmonics, 2019, 14: 1497-1504.
[10] GOPINATH S C B. Biosensing applications of surface plasmon resonance-based Biacore technology[J]. Sensors and Actuators B: Chemical, 2010, 150(2): 722-733.
[11] ABDICHE Y N, MALASHOCK D S, PINKERTON A, et al. Exploring blocking assays using Octet, ProteOn, and Biacore biosensors[J]. Analytical Biochemistry, 2009, 386(2): 172-180.
[12] ZENG Youjun, WANG Lei, WU S Y, et al. Wavelength-scanning SPR imaging sensors based on an acousto-optic tunable filter and a white light laser[J]. Sensors, 2017, 17(1): 90.
[13] ZENG Youjun, WANG Lei, WU S Y, et al. High-throughput imaging surface plasmon resonance biosensing based on an adaptive spectral-dip tracking scheme[J]. Optics Express, 2016, 24(25): 28303-28311.
[14] CHONG Xinyuan, LIU Le, LIU Zhiyi, et al. Detect the hybridization of single-stranded DNA by parallel scan spectral surface plasmon resonance imaging[J]. Plasmonics, 2013, 8(2): 1185-1191.
[15] 王霄, 许吉英, 陈义. 生物分子相互作用动力学的表面等离子体共振研究方法[J]. 化学进展, 2015(5): 102-110.
[16] ELIASSON M, OLSSON A, PALMCRANTZ E, et al. Chimeric IgG-binding receptors engineered from staphylococcal protein A and streptococcal protein G[J]. Journal of Biological Chemistry, 1988, 263(9): 4323- 4327.
[17] AYBAY C. Differential binding characteristics of protein G and protein A for Fc fragments of papain-digested mouse IgG[J]. Immunology Letters, 2003, 85(3): 231-235.
[18] ELIASSON M, ANDERSSON R, OLSSON A, et al. Differential IgG-binding characteristics of staphylococcal protein A, streptococcal protein G, and a chimeric protein AG[J]. Journal of Immunology, 1989, 142(2): 575-581.
[19] 张晖, 薛洪宝, 陈飞剑, 等. 表面等离子共振技术快速测定蛋白A与抗人T细胞单克隆抗体结合的反应常数[J]. 分析科学学报, 2019, 35(1): 71-75.

备注

引言

1 材料与方法

2 结果与分析

3 结语

期刊信息

备注

引言

1 材料与方法

2 结果与分析

3 结 语

期刊信息

3 结语