title: Efficient extraction of vaccines formulated in aluminum hydroxide gel by including surfactants in the extraction buffer.
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 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.
authors: Laboratory of Malaria Immunology and Vaccinology, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Rockville, MD 20852, USA. dzhu@niaid.nih.gov
authors: Daming
authors: D
authors: Zhu
authors: Shuhui
authors: S
authors: Huang
authors: Holly
authors: H
authors: McClellan
authors: Weili
authors: W
authors: Dai
authors: Najam R
authors: NR
authors: Syed
authors: Elizabeth
authors: E
authors: Gebregeorgis
authors: Kelly M
authors: KM
authors: Rausch
authors: Gregory E D
authors: GE
authors: Mullen
authors: Carole
authors: C
authors: Long
authors: Laura B
authors: LB
authors: Martin
authors: David
authors: D
authors: Narum
authors: Patrick
authors: P
authors: Duffy
authors: Louis H
authors: LH
authors: Miller
authors: Allan
authors: A
authors: Saul
mesh_terms: D000276:Adjuvants, Immunologic / Q000737:chemistry*; D000536:Aluminum Hydroxide / Q000737:chemistry*; D000818:Animals; D000953:Antigens, Protozoan / Q000032:analysis* / Q000302:isolation & purification*; D002021:Buffers; D004356:Drug Storage / Q000379:methods; D006801:Humans; D017780:Malaria Vaccines / Q000737:chemistry* / Q000276:immunology / Q000592:standards; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification*; D015800:Protozoan Proteins / Q000032:analysis* / Q000302:isolation & purification*; D011786:Quality Control; D013501:Surface-Active Agents / Q000737:chemistry*; D013678:Technology, Pharmaceutical / Q000592:standards*
chemical_list: D000276:Adjuvants, Immunologic; D000953:Antigens, Protozoan; D002021:Buffers; D017780:Malaria Vaccines; D008565:Membrane Proteins; D015800:Protozoan Proteins; D013501:Surface-Active Agents; C090077:apical membrane antigen I, Plasmodium; D000536:Aluminum Hydroxide