Hyclone hypure water

Samples of monovalent, bivalent, and trivalent vaccine formulations were lyophilized using an FTS Systems LyoStar lyophilizer (Warminster, PA) using a protocol as previously described [21 (link)]. Briefly, the lyophilizer shelves were pre-cooled to −10°C. Half-stoppered sample vials were surrounded with unstoppered dummy vials containing 1 mL of water. During freezing, the shelf temperature was decreased to −40°C at a rate of 0.5°C/min then held at temperature for 1 h. The primary drying phase (at 60 mTorr) started with a temperature ramp of 1°C/min to −20°C followed by a hold for 20 h. Then, during secondary drying, the shelf temperature was increased to 0°C at 0.2 °C/min, then to 30°C at 0.5°C/min, and finally was held at 30°C for 5 h. After the lyophilization cycle was complete, the lyophilizer was back-filled with filtered nitrogen and the vials were fully stoppered automatically by moving the lyophilizer shelves. The vials were removed from the lyophilizer shelves and the stoppers immediately secured with aluminum caps. All samples were kept at 4°C until analysis. Prior to use for analytical characterization or administration to mice or primates, lyophilized vaccine formulations were reconstituted with sterile, deionized (MilliQ^®) water or HyClone HyPure water for injection (GE Healthcare Life Sciences, Chicago, IL) to 1.0 mL total volume.