Experimental study on affinity identification of aptamers in acute promyelocytic leukemia HL 60 cells
Objective To identify the affinity of aptamer single-stranded DNA(ssDNA)of HL60 cells by different detection methods,and compared the identification results of different methods.Methods Fluorescence microscope,chemical method(microplate reader)and flow cytometer were used to identify the affinity of 5 ssDNA by microscopic observation,OD value,fluorescence intensity and equilibrium dissociation constant.Results Fluorescence microscopy were observed the green light emitted by the labeled aptamer(5'-FAM-ssDNA)bound to HL60 cells under the blue background,The fluorescence results were analyzed using Image J,the affinity of L36P was the highest;chemical meth-od is used to determine the OD value,the histogram is made by GraphPad Prism 7 and statistically analyzed,and 5 ssD-NA and the affinity of HL60 cells was significantly higher than that of the control group(lymphocytes),and the affinity of L36P was the highest;the fluorescence intensity was measured by flow cytometry,the equilibrium dissociation con-stant was calculated,and the affinity of 5 ssDNA to HL60 cells was determined from high to low,L36P,L24P,L139P,L2P,L135P.Conclusion Fluorescence microscopy can directly observe the binding of ssDNA and cells,the chemical method(microplate reader)and flow cytometer can be quantitatively analyzed,the detection results of the three meth-ods are basical consistent.
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