Freeze-drying of live attenuated Vibrio anguillarum mutant for vaccine preparation.
Vibrio anguillarum MVAV6203 is a mutant strain as a candidate of live attenuated vaccine. In vaccine preparation, the freeze-drying conditions of the strain were investigated to improve the survival after freeze-drying, including the protectant, rehydration medium, freezing temperature, and initial cell concentration. Vibrio anguillarum MVAV6203 is sensitive to freeze-drying and the viability was only 0.03% in the absence of protectant. Of the tested protectants, 5% trehalose with 15% skimmed milk gave the highest viability of 34.2%. Higher cell survival was obtained by quick freezing at -80 degrees C than slow freezing at -20 degrees C. Initial cell concentration was another important factor, preferable for 1-3 x 10(10)CFU/ml. The supplementation of 10% skimmed milk in rehydration medium improved obviously freeze-drying viability. The combination of the optimal conditions achieved 51.4% cell viability after freeze-drying.
Yang
Lu
L
State Key Laboratory of Bioreactor Engineering, East China University of Science and Technology, Shanghai, China.
Ma
Yue
Y
Zhang
Yuanxing
Y
D001428:Bacterial Vaccines / Q000235:genetics* / Q000302:isolation & purification; D015925:Cryopreservation; D003451:Cryoprotective Agents; D005612:Freeze Drying; D005615:Freezing; D005798:Genes, Bacterial; D009154:Mutation; D014613:Vaccines, Attenuated / Q000235:genetics / Q000302:isolation & purification; D014733:Vibrio / Q000235:genetics* / Q000276:immunology* / Q000302:isolation & purification
D001428:Bacterial Vaccines; D003451:Cryoprotective Agents; D014613:Vaccines, Attenuated