Preparation and Characterization of Quantum Dots-anti-GSH Human scFvs Bioconjugations

• 研究论文 • Previous Articles Next Articles

Preparation and Characterization of Quantum Dots-anti-GSH Human scFvs Bioconjugations

XU Jun-Jie¹, WANG Shi-Wen¹, ZHAO Hong⁴, CHEN Gui-Qiu⁵, HUO Rui¹, TIAN Li¹, DUAN Yu-Jing¹, LI Min-Jie³, YANG Bai³, WEI Jing-Yan^1,2*

1. College of Pharmaceutical Science,
2. Key Laboratory for Molecular Enzymology and Engineering of Ministry of Education, College of Life Science, Jilin University, Changchun 130021, China;
3. State Key Lab for Supramolecular Structure and Materials, College of Chemistry, Jilin University, Changchun 130012, China;
4. Central Laboratory,
5. Pathology Department, China-Japan Union Hospital, Jilin University, Changchun 130031, China

Received:2008-06-20 Revised:1900-01-01 Online:2009-03-10 Published:2009-03-10
Contact: WEI Jing-Yan

Abstract

Abstract: In order to generate catalytic antibodies with glutathione peroxidase(GPX) activity, the clone B3 that bound specifically to glutathione(GSH) was selected from the phage display antibody library[human synthetic VH+VL single-chain Fv fragment(scFv) library] and the expression vector of scFv-B3 was constructed in previous study. The expression vector pPELB-B3 constructed was transformed into the Escherichia coli Rosetta to express the human anti-GSH single chain fragments variable(scFvs) antibodies. The scFv-B3 was purified by Ni2+-immobilized metal affinity chromatography(IMAC). 3-Mercaptopropyl acid-stabilized CdTe quantum dots synthesized in aqueous solution were used to conjugate with scFvs. Spectra analysis show that the fluorescent of QD-scFvs undergo a blue-shift in the emission peak, which suggest that the scFvs have been effectively bound to the scFvs, via covalent conjugation. It is also suggested by the result of the simple new method of dot blot. The results of membrane dot indicate that the QD-scFvs can specifically recognize the GSH. The status of the QD-scFvs acting on the MCF-7 cells was observed under fluorescence microscope. The results show that QD-scFvs can enter the cytoplasm across the membrane.

Key words: Quantum dot, Human single chain fragments variable(scFvs), Glutathione peroxidase(GPX), Covalent conjugation, Dot blot

CLC Number:

O629.72
Q511

TrendMD:

XU Jun-Jie¹, WANG Shi-Wen¹, ZHAO Hong⁴, CHEN Gui-Qiu⁵, HUO Rui¹, TIAN Li¹, DUAN Yu-Jing¹, LI Min-Jie³, YANG Bai³, WEI Jing-Yan^1,2*
. Preparation and Characterization of Quantum Dots-anti-GSH Human scFvs Bioconjugations
[J]. Chem. J. Chinese Universities, doi: .

References

Related Articles 15

[1]	WANG Ruina, SUN Ruifen, ZHONG Tianhua, CHI Yuwu. Fabrication of a Dispersible Large-sized Graphene Quantum Dot Assemblies from Graphene Oxide and Its Electrogenerated Chemiluminescence Behaviors [J]. Chem. J. Chinese Universities, 2022, 43(8): 20220161.
[2]	XIA Wu, REN Yingyi, LIU Jing, WANG Feng. Chitosan Encapsulated CdSe QDs Assemblies for Visible Light-induced CO₂ Reduction in an Aqueous Solution [J]. Chem. J. Chinese Universities, 2022, 43(7): 20220192.
[3]	HUANG Shan, YAO Jiandong, NING Gan, XIAO Qi, LIU Yi. Efficient Determination of Alkaline Phosphatase Activity Based on Graphene Quantum Dots Fluorescent Probes [J]. Chem. J. Chinese Universities, 2021, 42(8): 2412.
[4]	ZHOU Jieqiong, HUANG Yan, ZHANG Zhiling, PANG Daiwen, TIAN Zhiquan. Water-soluble Ag₂Te Quantum Dots with Emission in the Second Near-infrared Window [J]. Chem. J. Chinese Universities, 2021, 42(6): 2072.
[5]	SHU Xin, WANG Di, YUAN Chunling, QIN Xiu, WANG Yilin. Colorimetric Determination of Sarcosine with Carbon Quantum Dots as Mimetic Peroxidase [J]. Chem. J. Chinese Universities, 2021, 42(6): 1761.
[6]	CAO Zhiyuan, SUN Hui, SU Bin. Electrochemiluminescence of Quantum Dots: Research Progress and Future Perspectives [J]. Chem. J. Chinese Universities, 2020, 41(9): 1945.
[7]	WANG Zhiqing, CHEN Binbin, SHEN Jie, CHEN Wen, LIU Yueli, GONG Shaokang, ZHOU Jing. Light Assisted Resistive Switching Characteristics of Cu₁₂Sb₄S₁₃ Quantum Dots [J]. Chem. J. Chinese Universities, 2020, 41(8): 1908.
[8]	HUANG Jialing,LIU Fengjiao,WANG Tingting,LIU Cuie,ZHENG Fengying,WANG Zhenhong,LI Shunxing. Nitrogen and Sulfur co-Doped Carbon Quantum Dots for Accurate Detection of pH in Gastric Juice^† [J]. Chem. J. Chinese Universities, 2020, 41(7): 1513.
[9]	ZHANG Min, LIU Huan. Research Processes in Fabricating Micro-patterned Quantum Dot Films by Solution Processes ^† [J]. Chem. J. Chinese Universities, 2020, 41(3): 401.
[10]	LI Ming,CUI Xiaoqian,WANG Xuan,LI Zaijun. Synthesis of Amphiphilic Graphene Quantum Dots and Their Sustained Release Effect on L-Menthol ^† [J]. Chem. J. Chinese Universities, 2020, 41(2): 324.
[11]	MU Yaxin,A Li,ZHUANG Qianfen,WANG Yong,NI Yongnian. Hydrothermal Synthesis of Ultrasmall Fluorescent Tungsten Disulfide Quantum Dots and Their Application in Cell imaging^† [J]. Chem. J. Chinese Universities, 2019, 40(4): 693.
[12]	YU Zhaochuan, MA Wenhui, WU Tao, WEN Jing, ZHANG Yong, WANG Liyan, CHU Hongtao. Preparation of B, N, S co-Doped Graphene Quantum Dots for Fluorescence Detection of Fe ³⁺ and H₂P $O 4 - †$ [J]. Chem. J. Chinese Universities, 2019, 40(11): 2286.
[13]	LU Simin, YU Rujia, LONG Yitao. Single Nanoparticle Sizing Based on the Confined Glass Nanopore ^† [J]. Chem. J. Chinese Universities, 2019, 40(11): 2281.
[14]	ZHANG Yanan, YANG Lingling, TU Jiawei, CUI Ran, PANG Daiwen. Live-cell Synthesis of ZnSe Quantum Dots in Staphylococcus aureus^† [J]. Chem. J. Chinese Universities, 2018, 39(6): 1158.
[15]	ZHANG Dan, YU Jinhai, LI Dongze, ZHOU Miao, ZHANG Ying. Preparation of High Quality Ag₂S Quantum Dots with Near-Infrared Emission and Their Applications in Bioimaging^† [J]. Chem. J. Chinese Universities, 2018, 39(4): 623.