Method for detecting biological activity of RANKL (receptor activator for nuclear factor-KB ligands) targeted therapy medicines

Abstract

The invention provides a method for detecting the biological activity of RANKL (receptor activator for nuclear factor-KB ligands) targeted therapy medicines. The method includes steps of firstly, constructing transcription factor NF-kB (nuclear factor-kB) binding sequences, cloning the NF-kB binding sequences into luciferase reporter gene carriers, and guiding luciferase reporter gene transcription by the aid of the NF-kB binding sequences used as components of starters so as to obtain plasmids with the NF-kB binding sequences and luciferase reporter genes; secondly, transfecting cells by theplasmids with the NF-kB binding sequences and the luciferase reporter genes; thirdly, vaccinating the cells, adding RANKL and the RANKL targeted therapy medicines into the cells and then culturing thecells; fourthly, lysing the cells, adding luciferase substrates into the cells and detecting the fluorescence intensity so as to determine the biological activity of the RANKL targeted therapy medicines. The method has the advantage that the biological activity of the denosumab RANKL targeted therapy medicines can be accurately, easily, feasibly and quickly detected by the aid of the method.

Description

Technical field [0001] The invention belongs to the field of biological detection, and specifically relates to a method for detecting the biological activity of RANKL targeted therapy drugs. Background technique [0002] The monoclonal antibody Denosumab (Denosumab hereinafter referred to as Denosumab, trade name Erbitux, a product listed by Amgen) for the treatment of osteoporosis and bone cancer metastasis is achieved by blocking RANKL (Receptor Activator for NuclearFactor-kB Ligand, The biological activity of the nuclear factor kB receptor activator ligand) protein plays a role. When RNAKL binds to its receptor RANK protein, it activates a series of transcription factors and induces gene transcription through signal transduction. Some of these genes induced to be transcribed are involved in inducing the differentiation and maturation of osteoclasts and in inducing the metastasis of bone cancer cells. [0003] The transcription factor NF-kB is a key transcription factor that ha...

AI Technical Summary

Problems solved by technology

The main disadvantages of this detection method are: 1. Time-consuming, it needs to be induced for up to 5 to 10 days, and the cells need to be replaced and various pharmaceutical ingredients are added in the middle; In terms of percentage, the difference between negative control and positive control is between 100%-300%, and the detection window is small

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