Method for eliciting an immune response to an immunogen
A technology for immunogens and uses, applied in the field of eliciting immune responses to immunogens, which can solve problems such as expensive, unrecognizable target proteins, and lengthy
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Embodiment 1
[0058] A method for using alpha-hemolysin to present polymer vesicles to elicit an immune response to alpha-hemolysin in mice is now described in the following paragraphs.
[0059] Materials & methods
[0060] Poly(butadiene-b-ethylene oxide) (PBd 21 -PEO 14 ) BD21 amphiphilic block copolymer was purchased from Polymer Source (Polymer Source, Canada). Alpha-hemolysin comes from Staphylococcus aureus, 3-(N-morpholino)propanesulfonic acid (MOPS), tris(hydroxymethyl)aminomethane hydrochloride (Tris), magnesium chloride and sodium chloride were purchased from Sigma Aldrich Corporation (Singapore). Tetrahydrofuran (THF) was purchased from Tedia (Ohio, USA).
[0061] Preparation of polymer vesicles
[0062] The polymer vesicles are prepared by a membrane rehydration method. The BD21 polymer was dissolved in THF and dried into a thin film on the wall of the conical bottom glass tube under nitrogen flow. The polymer film is further dried under vacuum. Subsequently, ultrapure water was ...
Embodiment 2
[0077] A method of using hemagglutinin to present polymer vesicles to elicit an immune response in mice is now described in the following paragraphs.
[0078] Preparation of hemagglutinin polymer vesicles
[0079] The polymer vesicles were prepared by the membrane rehydration method. BD21 polymer is dissolved in chloroform and dried into a film. Subsequently, ultrapure water was added to rehydrate the membrane and allow the spontaneous formation of polymer vesicles. The resulting vesicle dispersion was then extruded with 0.45 μm and 0.22 μm membranes, and dialyzed against ultrapure water to remove any remaining solvent.
[0080] Recombinant hemagglutinin (HA) protein (influenza A virus H3N2 Wisconsin 67 / 05 , MyBioSource) is provided as a sterile filtered solution in 10 mM sodium phosphate, pH 7.4, 150 mM NaCl and 0.005% Tween-20.
[0081] HA polymer vesicles are formed by adding an aliquot of the HA solution to the polymer vesicle dispersion. The mixture is then incubated to allo...
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