A novel approach has been developed that enables sterile pharmaceutical products to be freeze-dried in the open laboratory without specialist facilities. The product is filled into vials, semi-stoppered and sealed inside one, followed by a second, sterilization pouch under class 100 conditions. The product is then freeze-dried in the laboratory where the vials are shelf-stoppered before being returned to class 100, unwrapped and crimped. The sterilization pouches increased the resistance to water vapor movement during sublimation, thereby increasing the sublimation time and product temperature. Ovine immunoglobulins were double wrapped and lyophilized (as above) adjusting the primary drying time and shelf temperature for increased product temperature and, therefore, prevention of collapse. Ovine immunoglobulin G formulations freeze-dried to ≤ 1.1% residual moisture with no effect on protein aggregation or biological activity. The process was simulated with tryptone soya broth and no growth of contaminating microbial cells was observed after incubation at 35 °C for 2 weeks. Although increasing lyophilization time, this approach offers significant plant and validation cost savings when sterile freeze-drying small numbers of vials thereby making the manufacture of treatments for neglected and orphan diseases more viable economically.

This study describes the use of containment systems to prevent escape of microorganisms during lyophilization, thereby avoiding contamination of freeze-drying equipment. Cultures of Escherichia coli (JM 109) of an approximate cell concentration of 10 9 cfu/mL were suspended in 0.9% saline, aseptically dispensed into vials, double-wrapped in either medical-grade paper or Tyvek sterilization pouches and freeze-dried. An intentional collapse phenomenon was observed during the freeze-drying process, ejecting debris and aerosols from the vials, thus representing a worst-case challenge for containment. Following freeze drying, the layers of the pouches were tested for microbial contamination using 3 M Clean-Trace surface ATP analyzer swabs, surface swabs, and tryptone soya agar contact plates. The paper and Tyvek pouches were able to contain a maximum cell concentration of 1 x 10 6 cfu/mL of E. coli recovered from ejected debris. Some penetration through the first paper pouch layer was observed (although not Tyvek); however, this was successfully retained by the second, outer layer preventing contamination of the lyophilization apparatus and laboratory environment.