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Multivalent conjugation of antibody to dendrimers for the enhanced capture and regulation on colon cancer cells.

Xie J, Wang J, Chen H, Shen W, Sinko PJ, Dong H, Zhao R, Lu Y, Zhu Y, Jia L - Sci Rep (2015)

Bottom Line: G6-5aSlex-FITC conjugate showed capture efficiency better than FITC-G6-COOH-5aSlex conjugate.The capture resulted in a concentration-dependent restraint of the cell activity.In conclusion, the aSlex-coated dendrimer conjugate displayed the great potential in capturing and restraining colorectal CTCs in blood.

View Article: PubMed Central - PubMed

Affiliation: Cancer Metastasis Alert and Prevention Center, and Biopharmaceutical Photocatalysis of State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.

ABSTRACT
Circulation tumor cells (CTCs) in the bloodstream of early-stage cancer patients carry the important information about valuable biomarkers and biological properties of primary tumor. However, detection and capture of CTCs are challenging owing to their low concentrations. Traditional technologies have the limited detection sensitivity and the low capture efficiency. We, herein, report an effective approach to specifically bind and capture colon cancer HT29 cells by using multiple Sialyl Lewis X antibodies (aSlex)-conjugated PAMAM dendrimers. The conjugation was characterized by using atom force microscope, UV and fluorescence measurements. The capturing and regulating HT29 cells by the aSlex-coated dendrimer conjugate were analyzed by microscopy and flow cytometry. The results indicated that the conjugate showed the enhanced capture of HT29 cells in a concentration-dependent manner and the maximum capture efficiency of 77.88% was obtained within 1 h-exposure. G6-5aSlex-FITC conjugate showed capture efficiency better than FITC-G6-COOH-5aSlex conjugate. G6-5aSlex-FITC conjugate could specifically capture HT29 cells even when the target HT29 cells were diluted with the interfering cells (e.g., RBCs) to a low concentration. The capture resulted in a concentration-dependent restraint of the cell activity. In conclusion, the aSlex-coated dendrimer conjugate displayed the great potential in capturing and restraining colorectal CTCs in blood.

No MeSH data available.


Related in: MedlinePlus

Flow cytometric analysis of the capture behaviors of G6-5aSlex-FITC and FITC-G6-COOH-5aSlex conjugates to HT29 cells in the absence or presence of aSlex.(A), Comparison of the capture efficacy between two conjugates. (B), The difference in capture efficacy between two conjugates in the presence of aSlex.
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f4: Flow cytometric analysis of the capture behaviors of G6-5aSlex-FITC and FITC-G6-COOH-5aSlex conjugates to HT29 cells in the absence or presence of aSlex.(A), Comparison of the capture efficacy between two conjugates. (B), The difference in capture efficacy between two conjugates in the presence of aSlex.

Mentions: Cell-capture efficiency of conjugates to HT29 cells was evaluated by flow cytometry. The effect exerted by aSlex-FITC was investigated. After exclusion of the non-specific binding and autofluorescence both using 1% BSA and IgG-FITC isotype control, the capture efficiencies of two conjugates (G6-5aSlex-FITC and FITC-G6-COOH-5aSlex) were shown. Compared to control, the captured cells by aSlex-FITC alone demonstrated the specific antigen-antibody interaction as well as the high expression level of Slex on HT29 cell surface. G6-5aSlex-FITC conjugate had the capture efficiency up to 3-fold more than FITC-G6-COOH-5aSlex conjugate at the same exposure time of 1 h and concentration of 20 μg/ml (Fig. 4A). 30 min of pre-incubation with aSlex-FITC led to an increase in capture efficiency of 19.5% for G6-5aSlex-FITC conjugate and 4.5% for FITC-G6-COOH-5aSlex conjugate, respectively. Moreover, G6-5aSlex-FITC captured HT29 cells 24.9% more than FITC-G6-COOH-5aSlex did (Fig. 4B). The presence of aSlex-FITC significantly improved the capture efficiency of conjugates in a cooperative manner. G6-5aSlex-FITC conjugate was selected for the following experiments owing to its better capture capability than FITC-G6-COOH-5aSlex conjugate.


Multivalent conjugation of antibody to dendrimers for the enhanced capture and regulation on colon cancer cells.

Xie J, Wang J, Chen H, Shen W, Sinko PJ, Dong H, Zhao R, Lu Y, Zhu Y, Jia L - Sci Rep (2015)

Flow cytometric analysis of the capture behaviors of G6-5aSlex-FITC and FITC-G6-COOH-5aSlex conjugates to HT29 cells in the absence or presence of aSlex.(A), Comparison of the capture efficacy between two conjugates. (B), The difference in capture efficacy between two conjugates in the presence of aSlex.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4377633&req=5

f4: Flow cytometric analysis of the capture behaviors of G6-5aSlex-FITC and FITC-G6-COOH-5aSlex conjugates to HT29 cells in the absence or presence of aSlex.(A), Comparison of the capture efficacy between two conjugates. (B), The difference in capture efficacy between two conjugates in the presence of aSlex.
Mentions: Cell-capture efficiency of conjugates to HT29 cells was evaluated by flow cytometry. The effect exerted by aSlex-FITC was investigated. After exclusion of the non-specific binding and autofluorescence both using 1% BSA and IgG-FITC isotype control, the capture efficiencies of two conjugates (G6-5aSlex-FITC and FITC-G6-COOH-5aSlex) were shown. Compared to control, the captured cells by aSlex-FITC alone demonstrated the specific antigen-antibody interaction as well as the high expression level of Slex on HT29 cell surface. G6-5aSlex-FITC conjugate had the capture efficiency up to 3-fold more than FITC-G6-COOH-5aSlex conjugate at the same exposure time of 1 h and concentration of 20 μg/ml (Fig. 4A). 30 min of pre-incubation with aSlex-FITC led to an increase in capture efficiency of 19.5% for G6-5aSlex-FITC conjugate and 4.5% for FITC-G6-COOH-5aSlex conjugate, respectively. Moreover, G6-5aSlex-FITC captured HT29 cells 24.9% more than FITC-G6-COOH-5aSlex did (Fig. 4B). The presence of aSlex-FITC significantly improved the capture efficiency of conjugates in a cooperative manner. G6-5aSlex-FITC conjugate was selected for the following experiments owing to its better capture capability than FITC-G6-COOH-5aSlex conjugate.

Bottom Line: G6-5aSlex-FITC conjugate showed capture efficiency better than FITC-G6-COOH-5aSlex conjugate.The capture resulted in a concentration-dependent restraint of the cell activity.In conclusion, the aSlex-coated dendrimer conjugate displayed the great potential in capturing and restraining colorectal CTCs in blood.

View Article: PubMed Central - PubMed

Affiliation: Cancer Metastasis Alert and Prevention Center, and Biopharmaceutical Photocatalysis of State Key Laboratory of Photocatalysis on Energy and Environment, College of Chemistry, Fuzhou University, Fuzhou 350002, China.

ABSTRACT
Circulation tumor cells (CTCs) in the bloodstream of early-stage cancer patients carry the important information about valuable biomarkers and biological properties of primary tumor. However, detection and capture of CTCs are challenging owing to their low concentrations. Traditional technologies have the limited detection sensitivity and the low capture efficiency. We, herein, report an effective approach to specifically bind and capture colon cancer HT29 cells by using multiple Sialyl Lewis X antibodies (aSlex)-conjugated PAMAM dendrimers. The conjugation was characterized by using atom force microscope, UV and fluorescence measurements. The capturing and regulating HT29 cells by the aSlex-coated dendrimer conjugate were analyzed by microscopy and flow cytometry. The results indicated that the conjugate showed the enhanced capture of HT29 cells in a concentration-dependent manner and the maximum capture efficiency of 77.88% was obtained within 1 h-exposure. G6-5aSlex-FITC conjugate showed capture efficiency better than FITC-G6-COOH-5aSlex conjugate. G6-5aSlex-FITC conjugate could specifically capture HT29 cells even when the target HT29 cells were diluted with the interfering cells (e.g., RBCs) to a low concentration. The capture resulted in a concentration-dependent restraint of the cell activity. In conclusion, the aSlex-coated dendrimer conjugate displayed the great potential in capturing and restraining colorectal CTCs in blood.

No MeSH data available.


Related in: MedlinePlus