Polyclonal Antibody Preparation Of HSV-1Capsid Protein VP23and Pharmacokinetics Of Anti-HSV Drug PGG In Rats

Objective：To construct the recombinant plasmid pET-28a(+)-UL18, and utilizeprokaryotic expressing system E. coli to express large quantity of VP23. The VP23was purified and for preparation of polyclonal antibodies. The pharmacokineticsparameters and bioavailability of anti-HSV-1drug PGG in male and female SD ratswere also explored.Methods: UL18gene was amplified by PCR and was inserted into plasmidpET-28a(+). Recombinant plasmids were transformed into prokaryotic expressingsystem E.coli BL21(DE3) to express recombinant VP23which attaching with a6-Histag. Then the optimized expressing conditions of VP23were carried out. VP23can bepurified with its His-tag in affinity chromatography column. After injected into rabbits,we finally harvested its antibody.The PGG was given to rats by intravenous injection and intragastricadministration (20mg/Kg), respectively. Then the concentrations of PGG in plasmawere analysed by HPLC and the pharmacokinetics parameters and bioavailability ofPGG were calcmLated by PKSolver software.Results: The recombinant vectors containing UL18gene were successfullyconstructed to express its encoding capsid protein VP23that attaching with a6-Histag in prokaryotic expressing system, and then the fermentation conditions wereoptimized to acquire large quantity of VP23. The recombinant VP23finally waspurified and injected into New Zealand rabbits to obtain its polyclonal antibodies.The analytical method in HPLC was confirmed and the pharmacokineticsparameters of i.v. and i.g. administrations in SD rats were acquired. After intravenousinjection, the drug concentration-time curve showed a trend of hyperbolic decline andthe pharmacokinetic profile fitted two-compartment model. The half-life ofdistribution phases for male and female rats respectively were0.15±0.02h and0.21±0.03h, which was far less than the elimination half-life (male:3.29±0.34h,female:3.83±0.59h). After intragastric administration, the Tmaxfor male and female respectively were0.08±0.03h and1.00±0.44h with the peak concentrationrespectively at0.02±0.001μg/mL for male and0.02±0.0002μg/mL for female. Thebioavailability of PGG in male rats was2.88%and in female was5.20%.Conclusion: The polyclonal antibodies of VP23provide a tool for screeninganti-HSV-1drugs which targeting virus assembly. Along with pharmacokineticsparameters of anti-HSV-1drug PGG, this study provides an application methods and abasis for anti-virus therapy of HSV-1in vivo.