A Fast-detection System Of HBV Capsid Like Particle

Objective:Screening for drugs targeting the capsid of hepatitis B virus(HBV)is currently lacking a simple and effective screening cell model.Relatively simple and reliable in the method of directly detecting the capsid is Native agarose gel electrophoresis(NAGE).For drug screening,NAGE has problems with low throughput and cumbersome operation.In response to these problems,a new system that reflect HBV capsid-like particle was established.Methods: Two near-infra fluorescent proteins were fused to the core protein of HBV with a long glycine-serine linker respectively.The expression of the infra fluorescent proteins were detected by an infra fluorescence scanner.Stable cell line 293miRFP709-HBc which expresses miRFP709-HBc in a tetracycline-controlled manner was established by lentivirus transduction.Bay 41-4109 and Arbidol were used to treat 293miRFP709-HBc cells,and intracellular capsid like particles were tested after treatment.Results:(1)The infra fluorescence signal of miRFP709-HBc was higher than that of IMRFP-HBc and miRFP709-HBc supported the formation of capsid like particles.(2)Stable cell line 293 tTA and 293miRFP709-HBc were successfully established.(3)The capsid like particle in 293miRFP709-HBc cells was inhibited by Bay 41-4109 and Arbidol in a dose-dependent manner,respectively.Conclusion: A new system that reflect HBV capsid-like particle was successfully constructed.This system saves 20 hours compared with the traditional method,completely eliminates the transfer and immune hybridization steps,and requires no antibodies?transfer and hybridization related reagents.Therefore,the system can be used in a 2nd-round screening of HBV capsid-targeting compounds.The system can also be used as an independent screening model for direct screening of compounds that interfere with the formation of capsids.In addition to screening for compounds that cause changes in the amount of capsid formation,compounds that only result in a change in the capsid configuration(change in migration rate)can be screened.