Millex hv pvdf syringe driven filter unit

Example 3

Purified virus particles are resuspended in Hanks Balanced Salt Solution (HBSS) with CaCl₂ and MgCl₂ (Invitrogen) supplemented with 10% sorbitol (Sigma) and stored at −70° C. before being inactivated and injected into a subject.

Purified triple-layered rotavirus particles are diluted to a concentration of 300 micrograms/ml with diluent buffer and sterilized by filtration using a Millex®-HV PVDF Syringe driven filter unit (0.45 micron; Millipore Corporation, Bedford, Mass.). To inactivate rotavirus by heat, virus particles in 3.6 ml cryotubes (NalgeNunc, Rochester, N.Y.) are incubated for 1 hr at 60° C. in a waterbath with re-circulating water (model NesLab Ex10; Thermo Electron Corporation, Newington, N.H.) and then transferred to another fresh tube and incubated for an additional 1 hr at 60° C. A small aliquot is immediately tested for any residual infectivity and the remainder is stored at −70° C. before use in the immunization of a subject.

Example 3

Purified virus particles are resuspended in Hanks Balanced Salt Solution (HBSS) with CaCl₂ and MgCl₂ (Invitrogen) supplemented with 10% sorbitol (Sigma) and stored at −70° C. before being inactivated and injected into a subject.

Purified triple-layered rotavirus particles are diluted to a concentration of 300 micrograms/ml with diluent buffer and sterilized by filtration using a Millex®-HV PVDF Syringe driven filter unit (0.45 micron; Millipore Corporation, Bedford, Mass.). To inactivate rotavirus by heat, virus particles in 3.6 ml cryotubes (NalgeNunc, Rochester, N.Y.) are incubated for 1 hr at 60° C. in a waterbath with re-circulating water (model NesLab Ex10; Thermo Electron Corporation, Newington, N.H.) and then transferred to another fresh tube and incubated for an additional 1 hr at 60° C. A small aliquot is immediately tested for any residual infectivity and the remainder is stored at −70° C. before use in the immunization of a subject.