Cell-ELA-based determination of binding affinity of DNA aptamer against U87-EGFRvIII cell / 生物工程学报

A15, a DNA aptamer with binding specificity for U87 glioma cells stably overexpressing the epidermal growth factor receptor variant III (U87-EGFRvIII), was generated by cell systematic evolution of ligands by exponential enrichment (cell-SELEX) using a random nucleotide library. Subsequently, we established a cell enzyme-linked assay (cell-ELA) to detect the affinity of A15 compared to an EGFR antibody. We used A15 as a detection probe and cultured U87-EGFRvIII cells as targets. Our data indicate that the equilibrium dissociation constants (K(d)) for A15 were below 100 nmol/L and had similar affinity compared to an EGFR antibody for U87-EGFRvIII. We demonstrated that the cell-ELA was a useful method to determine the equilibrium dissociation constants (K(d)) of aptamers generated by cell-SELEX.

Preliminary studies on the andrographolide transformation in the baking process of the extract of Andrographis paniculata Nees / 中成药

Objective: To study the changes of andrographolides in the drying process of the extract of Andrographis paniculata Nees. Methods: HPLC method was applied to analyze andrographolide and 14 deoxy 11, 12 didehydroandrographolide in the process. Results: The content of andrographolide descended rapidly in the whole drying process, while the content of 14 deoxy 11, 12 didehydroandrographolide ascended at first 12 hours, declined in content was slowly to follow. Conclusion: Baking temperature is not the only main factor to stimulate the transformation of andrographolide and 14 deoxy 11, 12 didehydroandrographolide.