US 6,878,541 B2Granted

Papilloma pseudo-virus and preparation

OwnerLiang Qiao, Wei Shi +2inventors

InventorsLiang Qiao, Wei Shi, Yujun Huang, Jianzhong Liu

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Abstract

The invention involves a papilloma pseudo-virus that can induce immune response after oral intake as well as its preparation. It is characterized in that HPV or BPV pseudo-virus are made by disrupting HPV-VLP or BPV-VLP, mixing them with plasmids (plasmids or DNA vaccine), and reassembling them into the pseudo-viruses (VLPs with plasmids inside). Oral administration of the pseudo-viruses will result in delivery to mucosal and systemic lymphoid tissues and induce immune responses for disease prevention and treatment. The pseudo-virus induces stronger immune response than DNA vaccines. Additionally, the pseudo-virus can be applied in gene therapy by bringing the therapeutic genes into lymphoid tissues in the human body.

Claims18

Dependent claims

A method for treating a patient with a papillomavirus pseudo-virus including plasmids, said method comprising:

making a papillomavirus pseudo-virus by
disrupting papillomavirus-VLPs by mixing said papillomavirus-VLPs with a disruption buffer, said disruption buffer including EGTA 20mM, DTT 40 mM, NaCl 300mM, and Tris-hydrochloric acid (Tris-HCl) (pH 8.0) 100mM,
incubating the mixture for 60 minutes at room temperature,
mixing said disrupted papillomavirus-VLPs with said plasmids by adding said plasmids to the mixture in 1/10 ratio, said plasmids being at a concentration 0.5-1.0 microgram/microliter,
adding a stop buffer gradually to reassemble said papillomavirus-VLPs and said plasmids into pseudo-viruses of VLPs with said plasmids inside, said stop buffer including CaCl, 25mM and DMSO 20% by volume, and
incubating the mixture at 4 centigrade from 4 to 12 hours;
and
administering said pseudo-virus via the mucosa of said patient to induce a mucosal response.

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