Efficient extraction of vaccines formulated in aluminum hydroxide gel by including surfactants in the extraction buffer

TY - JOUR

T1 - Efficient extraction of vaccines formulated in aluminum hydroxide gel by including surfactants in the extraction buffer

AU - Zhu, Daming

AU - Huang, Shuhui

AU - McClellan, Holly

AU - Dai, Weili

AU - Syed, Najam R

AU - Gebregeorgis, Elizabeth

AU - Rausch, Kelly M

AU - Mullen, Gregory E D

AU - Long, Carole

AU - Martin, Laura B

AU - Narum, David

AU - Duffy, Patrick

AU - Miller, Louis H

AU - Saul, Allan

PY - 2012/1/5

Y1 - 2012/1/5

N2 - 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 20mM 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.

AB - 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 20mM 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.

KW - Adjuvants, Immunologic

KW - Aluminum Hydroxide

KW - Animals

KW - Antigens, Protozoan

KW - Buffers

KW - Drug Storage

KW - Humans

KW - Malaria Vaccines

KW - Membrane Proteins

KW - Protozoan Proteins

KW - Quality Control

KW - Surface-Active Agents

KW - Technology, Pharmaceutical

U2 - 10.1016/j.vaccine.2011.11.025

DO - 10.1016/j.vaccine.2011.11.025

M3 - Article

C2 - 22107848

SN - 0264-410X

VL - 30

SP - 189

EP - 194

JO - Vaccine

JF - Vaccine

IS - 2

ER -