Immunofluorescent Labeling of Human HepG2 Cells with CdTe Quantum Dot Probe Conjugated with Anti-pan CK MAb

Chemical Research in Chinese Universities ›› 2011, Vol. 27 ›› Issue (2): 277-281.

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Immunofluorescent Labeling of Human HepG2 Cells with CdTe Quantum Dot Probe Conjugated with Anti-pan CK MAb

SUI Yu-jie^1,2, ZHANG Gui-zhen¹, WANG Qian³, WANG Ya-li¹, WU Mei¹, DU Zhen-wu¹, ZHANG Jie⁴ and JIANG Ri-hua^1*

1. Central Laboratory, China-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China;
2. Central Laboratory, the Second Hospital of Jilin University, Changchun 130041, P. R. China;
3. Department of Urinary Surgery, China-Japan Union Hospital of Jilin University, Changchun 130033, P. R. China;
4. College of Chemistry, Jilin University, Changchun 130012, P. R. China

Received:2010-03-26 Revised:2010-05-16 Online:2011-03-25 Published:2011-03-09
Contact: JIANG Ri-hua E-mail:jrh1963@163.com
Supported by:
Supported by the Major Project Fund of Jilin Provincial Science and Technology Department, China(No.20082123) and the Province University Union Fund of Jilin Province, China(No.20082011).

Abstract

Abstract: A relatively sensitive, specific, and photostable method for the detection of cytokeratin of cancer cells via conjugation with cadmium telluride quantum dots(CdTe QDs) was described. Water soluble CdTe QDs were conjugated to anti-pan-cytokeratin(CK) monoclonal antibody(MAb) through coupling reagent [1-ethyl-3-(3-dimethyla- mino propyl)carbodiimide, EDC] and the conjugates were purified by dialysis. The expression of pan CK protein in HepG2 cells was observed by immunocytochemistry and direct immunofluorescence via QDs-Ab conjugates respectively. Fluorescence intensity and photostability of QDs were compared with those of FITC(fluorescein isothiocyanate). The results show that the QDs-Ab conjugates recognized specifically pan CK protein in HepG2 cells. Compared with FITC, CdTe QDs had higher brightness and photostability without obvious photobleaching under continuous exciting light illumination for 30 min and after the placement at room temperature for 3 d. The results indicate that conjugates of CdTe quantum dot with anti-pan CK MAb can be used for labeling cancer cells derived from epithelial tissues, which provides the basis for the detection of circulating tumor cells(CTCs).

Key words: Quantum dot, Pan-cytokeratin, Immunofluorescence labeling, Fluorescence photostability

SUI Yu-jie, ZHANG Gui-zhen, WANG Qian, WANG Ya-li, WU Mei, DU Zhen-wu, .... Immunofluorescent Labeling of Human HepG2 Cells with CdTe Quantum Dot Probe Conjugated with Anti-pan CK MAb[J]. Chemical Research in Chinese Universities, 2011, 27(2): 277-281.

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Immunofluorescent Labeling of Human HepG2 Cells with CdTe Quantum Dot Probe Conjugated with Anti-pan CK MAb

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