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Abstract
Efficient antigen entrapment is a key factor in preparation of poly (lactide-co-glycolide) acid (PLGA) vaccine formulations when the antigen is of short supply. This study presents a systematic approach in the testing of formulation variables with the objective to increase antigen entrapment in particles when the antigen stock concentration was low. Some of the experimental variables tested were poly (vinyl) alcohol (PVA) concentration in the inner (W1) and outer (W2) aqueous phase, W1/oil (O) phase ratio and choice of organic solvent. The double emulsion solvent evaporation technique was applied to prepare PLGA particles with sonication as the emulsifying force. To measure antigen entrapment efficiency, the antigen (bovine serum albumin, BSA) was isotope labeled with 125iodine (125I). Our results demonstrated that a low PVA concentration in the inner aqueous (W1) phase was beneficial to achieve a high encapsulation efficiency of antigen. On the contrary, in the outer aqueous (W2) phase, a high PVA concentration favored antigen entrapment. We also demonstrated that decreasing the W1 to O/polymer ratio contributed to increased entrapment efficiency. Testing different organic solvents (ethyl acetate, dichloromethane and chloroform), either alone or in combination, revealed that using chloroform as solvent resulted in the highest encapsulation of antigen and the highest production yield. Some of the results presented in this work are in disagreement with well-established formulation variables from previous studies.
ACKNOWLEDGMENTS
This project was funded by the Research Council of Norway (contract nos. 172508, 182035, and 183204) and by the University of Tromsø (“Tromsø forskningsstiftelse”). The authors would like to acknowledge Dr. Natasha Hynes for improving the quality of the manuscript.