Efficient extraction of vaccines formulated in aluminum hydroxide gel by including surfactants in the extraction buffer |
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Authors: | Zhu Daming Huang Shuhui McClellan Holly Dai Weili Syed Najam R Gebregeorgis Elizabeth Mullen Gregory E D Long Carole Martin Laura B Narum David Duffy Patrick Miller Louis H Saul Allan |
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Affiliation: | Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852, USA |
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Abstract: | Efficient antigen extraction from vaccines formulated on aluminum hydroxide gels is a critical step for the evaluation of the quality of vaccines following formulation. It has been shown in our laboratory that the efficiency of antigen extraction from vaccines formulated on Alhydrogel decreased significantly with increased storage time. To increase antigen extraction efficiency, the present study determined the effect of surfactants on antigen recovery from vaccine formulations. The Plasmodium falciparum apical membrane antigen 1 (AMA1) formulated on Alhydrogel and stored at 2-8 °C for 3 years was used as a model in this study. The AMA1 on Alhydrogel was extracted in the presence or absence of 30 mM sodium dodecyl sulfate (SDS) or 20 mM cetylpyridinium chloride in the extraction buffer (0.60 M citrate, 0.55 M phosphate, pH 8.5) using our standard antigen extraction protocols. Extracted AMA1 antigen was analyzed by 4-20% Tris-glycine SDS-PAGE followed by silver staining or western blotting. The results showed that inclusion of SDS or cetylpyridinium chloride in extraction buffer increased the antigen recovery dramatically and can be used for efficient characterization of Alhydrogel vaccines. |
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Keywords: | AMA1, apical membrane antigen 1 SDS, sodium dodecyl sulfate CPC, cetylpyridinium chloride pI, isoelectric point |
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