Technical Field
[0001] CROSS-REFERENCE
This application claims the benefit of U.S. provisional application Ser. No. 61/324,542,
filed April 15, 2010, which is incorporated herein by reference in its entirety.
Background Art
[0002]
Influenza vaccines formulated as liquids can be subject to chemical degradation, e.g.,
aggregation, denaturation, hydrolysis, and oxidation that can result in their inactivation.
Liquid vaccine formulations can also be sensitive to temperature: high temperatures can
increase inactivation, and freezing temperatures can result in ice that can damage
antigen in the vaccine. Thus, to prevent inactivation, liquid vaccines are often stored and
distributed in a temperature range between 2 and 8 degrees C. Such storage can be
costly, both for long-term storage and transportation of vaccines, and from loss of
vaccine due to expiration. Generation of vaccines that are stable at room temperature
would result in savings with respect to storage and would facilitate stockpiling. There is
a need for means of generating vaccine formulations that are stable at room temperature,
such as dry powder vaccines.
[0003]
Several methods of freeze-drying vaccines have been described. For example,
lyophilization (freeze-drying) of influenza vaccine solution can be used to generate a
vaccine powder. However, the influenza vaccine powder produced by this method can
be a hard cake, which does not facilitate consistent and reliable administration. Spray -
freeze-drying (SFD) of an influenza vaccine solution can provide fine particles of
influenza vaccine powder; however, SFD is a high-cost method. Thus, there is a need
for low-cost methods of generating fine powder vaccines with relatively high
flowability and relatively low hygroscopicity.
[0004]
The mode of administration of a vaccine can play a role in its efficacy. One mode of

administration, nonparental administration (e.g., nasal), can induce and promote mucosal and systemic humoral and cell mediated immune responses. Mucosal vaccination can result in induction of secretory IgA (slgA) responses in the respiratory tract and oropharyngeal region. One feature of mucosal slgA antibodies is that they can provide cross-protection against antigenically distinct viruses; thus, mucosal slgA responses have the potential to provide protection against a viral strain that has drifted from the strain used to generate the vaccine (for example, influenza virus H1N1 can drift to H2N1 or H1N2). Furthermore, slgA can help bind a virus or other pathogen at the mucosal surface, preventing access of the pathogen to deeper tissues and/or decreasing the likelihood of full-blown infection. Described herein are novel methods for generating an slgA inducing vaccine, for example, a powder vaccine formulation for nonparental administration. Summary of Invention [0005]
Disclosed herein is a dry vaccine powder formulation comprising: one or more antigens, one or more saccharides, one or more buffers; and microcrystalline cellulose. An antigen in a vaccine powder formulation described herein can be a viral antigen. A viral antigen can be live attenuated virus, whole inactivated virus, split-inactivated virus, subunit antigens, virosome, or cold-adapted live influenza virus. A viral antigen can be influenza virus; for example, an antigen could H1N1; or H5N1; or a mixture of H1N1, H3N2 and Influenza type B. An antigen in a vaccine powder formulation described herein can be a bacterial antigen. A bacterial antigen can be killed whole bacteria, attenuated bacteria, toxoids, purified surface protein, or purified recombinant surface protein. A bacterial antigen can be tetanus toxoid or diphtheria toxoid. An antigen in the dry vaccine powder formulation can also be a protist. An antigen could also be protein. The saccharide used can be trehalose, mannitol, or lactose. The saccharide used can be trehalose. The buffer used can be a phosphate buffer. A vaccine powder formulation described herein can be stable at room temperature and 60% relative humidity for at least 12 months. [0006]
Also provided herein is a method for generating a dry vaccine powder formulation comprising: preparing a liquid formulation comprising an antigen; quick freezing said

liquid formulation, wherein the quick freezing does not comprise spray freezing; blending the freeze-dried sample with one or more excipients to generate the dry vaccine powder formulation. A viral antigen can be live attenuated virus, whole inactivated virus, split-inactivated virus, subunit antigens, virosome, or cold-adapted live influenza virus. A viral antigen can be influenza virus; for example, an antigen could H1N1; or H5N1; or a mixture of H1N1, H3N2 and Influenza type B. An antigen in a vaccine powder formulation described herein can be a bacterial antigen. A bacterial antigen can be killed whole bacteria, attenuated bacteria, toxoids, purified surface protein, or purified recombinant surface protein. A bacterial antigen can be tetanus toxoid or diphtheria toxoid. An antigen in the dry vaccine powder formulation can also be a protist. An antigen could also be protein. The preparation of a liquid formulation can comprise addition of a saccharide, for example trehalose, mannitol, or lactose. Preparation of a liquid formulation can also comprise addition of a buffer, such as a phosphate buffer. The powder can comprise fine particles. The powder can be stable at room temperature and 60% relative humidity for at least 12 months. Excipients useful in methods described herein can comprise one or more nasal carriers, such as microcrystalline cellulose and tribasic calcium phosphate. An excipient can improve flowability of the powder and/or reduce hygroscopicity of the powder. Some vaccine powders produced by a method herein do not comprise an adjuvant. Quick freezing can comprise the use of liquid nitrogen. [0007]
Another method provided herein is a method of stimulating an slgA response in a subject to an antigen comprising administering a dry vaccine powder formulation to a subject, wherein the dry powder formulation comprises the antigen and wherein the dry powder formulation is generated by quick freezing a liquid vaccine formulation, wherein the quick freezing does not comprise spray-freezing. In some instances, an IgG response is also stimulated. slgA production can be stimulated at the site of administration and/or at a mucosal site other than the site of administration. Administration can be intranasal. An antigen in a vaccine powder formulation described herein can be a viral antigen. A viral antigen can be live attenuated virus, whole inactivated virus, split-inactivated virus, subunit antigens, virosome, or cold-adapted live influenza virus. A viral antigen can be influenza virus; for example, an antigen

could H1N1; or H5N1; or a mixture of H1N1, H3N2 and Influenza type B. An antigen
in a vaccine powder formulation described herein can be a bacterial antigen. A bacterial
antigen can be killed whole bacteria, attenuated bacteria, toxoids, purified surface
protein, or purified recombinant surface protein. A bacterial antigen can be tetanus
toxoid or diphtheria toxoid. An antigen in the dry vaccine powder formulation can also
be a protist. An antigen could also be protein. The preparation of a liquid formulation
can comprise addition of a saccharide, for example trehalose, mannitol or lactose.
Preparation of a liquid formulation can also comprise addition of a buffer, such as a
phosphate buffer. The powder can comprise fine particles. The powder can be stable at
room temperature and 60% relative humidity for at least 12 months. Excipients useful in
methods described herein can comprise one or more nasal carriers, such as
microcrystalline cellulose and tribasic calcium phosphate. An excipient can improve
flowability of the powder and/or reduce hygroscopicity of the powder.
[0008]
Also provided herein is a device for administration of a vaccine powder formulation
disclosed herein. Such a device can be configured for a single use.
[0009]
INCORPORATION BY REFERENCE
All publications, patents, and patent applications mentioned in this specification are
herein incorporated by reference to the same extent as if each individual publication,
patent, or patent application was specifically and individually indicated to be
incorporated by reference.
Brief Description of Drawings
[0010]
The novel features of the invention are set forth with particularity in the appended
claims. A better understanding of the features and advantages of the present invention
will be obtained by reference to the following detailed description that sets forth
illustrative embodiments, in which the principles of the invention are utilized, and the
accompanying drawings of which:
[Fig. 1]
Figure 1 illustrates properties of influenza vaccine powders generated using
conventional slow freezing and freeze-drying processing with trehalose, mannitol and

lactose.
[Fig. 2]
Figure 2 illustrates a process for preparing a dry nasal vaccine powder formulation by
quick freezing with liquid nitrogen. Exemplary properties of powders before and after
addition of nasal carriers are also described.
[Fig. 3]
Figure 3 illustrates an embodiment of a manufacturing process of the provided
invention.
[Fig. 4]
Figure 4 illustrates a study design for testing a H1N1 nasal influenza vaccine powder
formulation.
[Fig. 5A]
Figure 5 A tabulates HI titers measured in serum samples collected during a test of a
H1N1 nasal influenza vaccine powder formulation.
[Fig. 5B]
Figure 5B tabulates HI titers measured in nasal wash samples collected during a test of a
H1N1 nasal influenza vaccine powder formulation.
[Fig. 6A]
Figure 6A tabulates serum IgG antibody titers measured in samples collected during a
test of a H1N1 nasal influenza vaccine powder formulation.
[Fig. 6B]
Figure 6B tabulates nasal wash sIGA antibody titers measured in samples collected
during a test of a H1N1 nasal influenza vaccine powder formulation.
[Fig. 7]
Figure 7 graphically illustrates IgG and sIgA antibody titers measured during a test of a
H1N1 nasal influenza vaccine powder formulation.
[Fig. 8]
Figure 8 tabulates HI titers measured in serum and nasal wash samples collected during
a test of a H1N1 nasal influenza vaccine powder formulation.
[Fig. 9]
Figure 9 tabulates serum IgG and nasal wash sIgA antibody titers measured in samples
collected during a test of a H1N1 nasal influenza vaccine powder formulation.

[Fig. 10]
Figure 10 illustrates a study design for testing a H5N1 nasal influenza vaccine powder
formulation.
[Fig. 11 A]
Figure 11A tabulates serum IgG antibody titers measured in samples collected during a
test of a H5N1 nasal influenza vaccine powder formulation.
[Fig. 1 IB]
Figure 1 IB tabulates nasal wash slgA antibody titers measured in samples collected
during a test of a H5N1 nasal influenza vaccine powder formulation.
[Fig. 12]
Figure 12 graphically illustrates IgG and slgA antibody titers measured during a test of
a H5N1 nasal influenza vaccine powder formulation.
[Fig. 13]
Figure 13 illustrates a study design for testing a Tetanus toxoid nasal vaccine powder
formulation.
[Fig. 14A]
Figure 14A tabulates the absorbance ratio of serum IgG measured in samples collected
during a test of a Tetanus toxoid nasal vaccine powder formulation
[Fig. 14B]
Figure 14B graphically illustrates the absorbance ratio of serum IgG measured in
samples collected during a test of a Tetanus toxoid nasal vaccine powder formulation
[Fig. 15]
Figure 15 tabulates IFN gamma levels measured in samples collected during a test of a
Tetanus toxoid nasal vaccine powder formulation.
[Fig. 16]
Figure 16 illustrates a study design for testing a Diphtheria toxoid nasal vaccine powder
formulation.
[Fig. 17A]
Figure 17A tabulates serum IgG antibody titers measured in samples collected during a
test of a Diphtheria toxoid nasal vaccine powder formulation.
[Fig. 17B]
Figure 17B graphically illustrates serum IgG antibody titers measured in samples

collected during a test of a Diphtheria toxoid nasal vaccine powder formulation. [Fig. 18]
Figure 18 illustrates a study design for testing a homogenized ovalbumin nasal vaccine powder formulation. [Fig. 19A]
Figure 19A tabulates serum IgG antibody titers measured in samples collected during a test of a homogenized ovalbumin nasal vaccine powder formulation. [Fig. 19B]
Figure 19B graphically illustrates serum IgG antibody titers measured in samples collected during a test of a homogenized ovalbumin nasal vaccine powder formulation. [Fig. 20A]
Figure 20A tabulates nasal wash slgA antibody titers measured in samples collected during a test of a homogenized ovalbumin nasal vaccine powder formulation. [Fig. 20B]
Figure 20B graphically illustrates nasal wash slgA antibody titers measured in samples collected during a test of a homogenized ovalbumin nasal vaccine powder formulation. Description of Embodiments [0011]
DETAILED DESCRIPTION OF THE INVENTION I. OVERVIEW
Conventional freeze-drying processes for liquid influenza vaccine formulations, such as cooling from room temperature to -40 degrees C over 24 hr, can lead to suboptimal particle properties or loss of antigenic (e.g. influenza hemagglutinin (HA)) potency (Figure 1). For example, liquid influenza vaccine formulations with trehalose that are subjected to a conventional freeze-drying process can form a partially caked powder (Figure 1). Liquid influenza vaccine formulations with mannitol that are subjected to a conventional freeze-drying process can have reduced HA potency (Figure 1). Liquid influenza vaccine formulations with lactose that are subjected to a conventional freeze-drying process can form a partially caked powder and can have reduced HA potency (Figure 1). [0012] The present disclosure provides methods comprising a quick freezing step for

generating a dry vaccine powder formulation (see e.g., Figures 2 and 3) which overcomes the limitations of previous freeze drying methods, resulting in high potency powdered vaccines with high flowability. The methods can comprise a step of generating a liquid formulation containing one or more antigens, such as a pathogen or a component thereof (e.g., a whole inactivated influenza virus) with one or more agents (e.g., a saccharide and/or buffer, e.g., phosphate buffer). A liquid vaccine formulation can be freeze-dried (e.g., comprising quick freezing in liquid nitrogen) to generate a powder (e.g., a vaccine powder). The powder can comprise fine particles and can be stable at room temperature. If the antigen is an influenza virus, the powder can have high HA potency (e.g., at least about 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%), 99%), or 100%>). After freeze drying, the powder can be blended (e.g., by vortexing) with one or more excipients (e.g., nasal carriers and/or flowability agents) to form a dry vaccine powder formulation. [0013]
A dry vaccine powder formulation described herein can be stable at room temperature. This is an advance over liquid influenza vaccines, which are unstable at room temperature and can require expensive storage and distribution under refrigerated conditions (e.g., cold-chain distribution). In some vaccine preparations, a liquid formulation containing disaccharides, for example, trehalose or lactose, is prepared. Such additives generally allow for maintenance of HA potency of a dry influenza vaccine powder formulation. Although the use of such component saccharides is known, the methods described herein can provide a dry vaccine form which does not form hard cakes using these saccharide components. Hard caking can be avoided using the buffers and quick freezing techniques described herein. The powders produced from quick frozen and dried antigen preparations can then be combined with one or more excipients, such as a nasal carrier (e.g., microcrystalline cellulose) and/or a flowability agent (e.g., tribasic calcium phosphate). The present formulations can result in dry powder vaccines suitable for intranasal delivery which can be stable at room temperature and under accelerated conditions. A dry vaccine powder formulation provided herein can afford complete and consistent delivery from a nasal delivery device and result in stimulation of the recipient's immune response to the antigen/pathogen to which the vaccine is directed

[0014]
The methods provided herein can allow for reducing hygroscopicity and improving the
flowability of a dry vaccine powder formulation provided herein. The methods can
include addition of a physiologically acceptable agent (e.g., microcrystalline cellulose)
to a powder formulation to reduce hygroscopicity and improve flowability of a dry
vaccine powder formulation.
[0015]
Methods provided herein can allow for improving the efficacy of a vaccine. The
methods can comprise steps for generating a dry vaccine powder compositions that can
stimulate a local immune response, for example, a mucosal immune response (e.g.,
involving mucosal slgA). slgA can provide cross-protection against mutated influenza
viruses (e.g., a dry vaccine powder formulation can be used as a pandemic influenza
vaccine) and/or viruses which have undergone genetic drift. A dry vaccine powder
formulation, e.g., a dry nasal influenza powder formulation, can induce protection in
distal mucosal sites. For example, introduction of a vaccine of the present disclosure at
the nasal mucosa can lead to protection (e.g., slgA production in the upper respiratory
tract, the lower respiratory tract, the gastrointestinal tract, and vagina). A dry vaccine
powder formulation can stimulate a systemic immune response (e.g., producing serum
IgG). Dry vaccine powder compositions can comprise microcrystalline cellulose. In
some embodiments, a dry vaccine powder formulation does not comprise adjuvant.
[0016]
II. LIQUID FORMULATIONS FOR USE IN GENERATING A POWDER
FORMULATION
To generate a dry vaccine powder formulation, a liquid formulation can be first
generated. The liquid formulation can comprise one or more antigens (e.g., one or more
pathogens or components of pathogens), one or more saccharides, one or more buffers,
and one or more other components. Typically, the liquid formulation is subjected to
quick freezing (e.g., by immersion in liquid nitrogen) and freeze-drying prior to
producing the dry vaccine powder formulation.
[0017]
The volume of the liquid formulation can be about 0.1 mL, 1.0 mL, 10 mL, 25 mL, 50
mL, 100 mL, 250 mL, 500 mL, 1 L, 10 L, 50 L, 100 L, 250 L, 500 L, or 1000 L. The

volume of the liquid formulation can be more than about 0.1 mL, 1.0 mL, 10 mL, 25
mL, 50 mL, 100 mL, 250 mL, 500 mL, 1 L, 10 L, 50 L, 100 L, 250 L, 500 L, or 1000 L.
The volume of the liquid formulation can be about 0.01 -1 mL, about 1-10 mL, about
10-50 mL, about 50-100 mL, about 1-1000 mL, about 100-1000 mL, about 1-10 L,
about 10-50 L, about 50-100 L, about 100-500 L, about 100-1000 L, or about 1-1000 L.
Following freeze drying, the amount of dry vaccine produced can be between about
0.05 mg to 500 mg, about 0.0.05 mg to 1 mg, about 1 mg to about 100 mg, or about 100
mg to about 500 mg.
[0018]
A. VIRAL VACCINE COMPONENTS
The methods of generating a dry vaccine powder formulation described herein can be
used to produce a vaccine with a live attenuated virus, whole inactivated virus, split
virus, subunit antigen, virosome, or cold-adapted live influenza virus.
[0019]
The methods of generating a dry vaccine powder formulation described herein can be
used to produce a vaccine with a live attenuated virus. Live attenuated vaccines can be
derived from serial passage in cultured cells, including, for example, human diploid
cells (e.g. fetal lung tissue, other fibroblasts), monkey kidney cells, and chick embryos.
Adaptation of a virus to growth in the cultured cells can be accompanied by a gradual
loss of virulence for the natural host. Avirulence can be conferred, e.g., by accumulation
of point mutations. Genetic engineering can be used to achieve viral attenuation by, e.g.,
generating temperature sensitive mutants, generating deletion mutants, site-directed
mutagenesis, or generating live recombinant viruses.
[0020]
The methods of generating a dry vaccine powder formulation described herein can be
used to produce a vaccine with a whole inactivated virus. Inactivated viruses can be
generated, for example, by using ultraviolet light, low pH (e.g., acid, e.g., caprylic acid),
pasteurization, solvents/detergents, sodium thiocyanate, formalin, beta-propiolactone, or
ethylenimines. UV rays can damage DNA through by creating nucleic acid dimers,
which can inactivate viruses by preventing the replication of genetic material. Some
viruses denature upon exposure to low pH solutions. This method can be particularly
effective when employed verses enveloped viruses. Pasteurization can inactivate viruses

by means of temperature induced denaturation. Solvent/detergent inactivation is only effective against viruses enveloped in a lipid coat. The detergent used is typically Triton-X 100. Sodium thiocyanate can denature the protein coat of viruses, rendering the virus inactive. Formalin can chemically modify the surface proteins of the viral coat, which can prevent infection. Ethylenimines and beta-propiolactone can act on the nucleic acids of the virus while leaving the protein coat mostly unmodified. Inactivation can destroy infectivity of the virus while maintaining its immunogenicity. Multiple applications of inactivated virus can be administered to a subject. [0021]
The methods of generating a dry vaccine powder formulation described herein can be used to produce a vaccine with one or more antigenic proteins (vaccine proteins) from one or more pathogens. An antigenic protein can be from any pathogen to which a vaccine is to be produced. For example, where the vaccine is to target influenza virus, an antigenic protein can be hemagglutinin (HA) and/or neuraminidase (NA). Hemagglutinin is an antigenic glycoprotein and a major surface protein of the influenza A virus. It mediates the biding between an influenza virus and the cell to be infected by binding to sialic acid-containing receptors on the surface of the cell. Viral particles bound to the surface of the cell are engulfed into endosomes. Inside the endosome, HA mediates a fusion of the viral membrane and the endosomal membrane, releasing the viral genome into the cell. Structurally, HA consists of three identical monomers organized into a helical coil. A function blocking antibody could inhibit either the cell binding or membrane fusing functions of HA. Neuraminidase is another glycoprotein found on the surface of an influenza virus. NAs are enzymes that function by cleaving sialic acid groups from glycoproteins. This cleavage seems to serve two functions: to prevent viral clumping and to release progeny viruses from the surface of a cell. [0022]
There are at least 16 known HA subtypes. A vaccine antigen can be HA1, HA2, HA3, HA4, HA5, HA6, HA7, HA8, HA9, HA10, HA11, HA12, HA13, HA14, HA15, or HA16. There are 9 known NA subtypes. A vaccine antigen can be NA1, NA2, NA3, NA4, NA5, NA6, NA7, NA8, or NA9. A vaccine prepared from a HA and/or NA subtype can be used individually or in any combination. For example, two or more of the various HA and NA antigens can be mixed during manufacture of a dry vaccine

powder formulation, or dry powder formulations of individual HA and NA antigens can be combined. An antigenic protein can be surface proteins from the pathogen. An antigenic protein can be produced recombinantly. For example, nucleic acid encoding an antigen of interest can be introduced in a prokaryotic cell (e.g. bacteria), eukaryotic cells (e.g., yeast cells and insect cells), and the protein can be expressed and purified from the cells. Where the pathogen is a virus, nonessential components of a virion can be removed (e.g., using ether and detergents). [0023]
The methods of generating a dry vaccine powder formulation described herein can be used to produce a virosomal vaccine. A virosomal vaccine comprises virus-like particles of reconstituted virus envelopes with no genetic material of the native virus. Influenza virosomes are vesicles consisting of a unilamellar phospholipid bilayer with intercalated HA and NA proteins. Because they have no genetic material, virosomes are not infectious. [0024]
The concentration of a vaccine protein (e.g., antigen or antigen containing component) in a liquid vaccine formulation can be from about 0.05 mg/mL to 10 mg/mL, about 0.1 mg/mL to 10 mg/mL, about 0.1 mg/mL to 5 mg/mL, about 0.1 mg/mL to 2.5 mg/mL, about 0.1 mg/mL to 1 mg/mL, about 0.1 mg/mL to 0.5 mg/ML, about 0.5 mg/mL to 1 mg/mL, about 0.05 mg/mL to 1 mg/mL, or about 0.05 mg/mL to 2.5 mg/mL. The concentration of a vaccine protein (e.g., antigen or antigen containing component) in a liquid vaccine formulation can be about 0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL, 0.4 mg/mL, 0.5 mg/mL, 0.6 mg/mL, 0.7 mg/mL, 0.8 mg/mL, 0.9 mg/mL, 1.0 mg/mL, 1.1 mg/mL, 1.2 mg/mL, 1.3 mg/mL, 1.4 mg/mL, 1.5 mg/mL, 1.6 mg/mL, 1.7 mg/mL, 1.8 mg/mL, 1.9 mg/mL, 2.0 mg/mL, 2.5 mg/mL, 3 mg/mL, 3.5 mg/mL, 4 mg/mL, 4.5 mg/mL, 5 mg/mL, 5.5 mg/mL, 6 mg/mL, 6.5 mg/mL, 7.0 mg/mL, 8.0 mg/mL, 8.5 mg/mL, 9 mg/mL, or 10 mg/mL. The concentration of a vaccine protein (e.g. antigen or antigen-containing component) in a liquid vaccine formulation can be more than about 0.05 mg/mL, 0.1 mg/mL, 0.2 mg/mL, 0.3 mg/mL, 0.4 mg/mL, 0.5 mg/mL, 0.6 mg/mL, 0.7 mg/mL, 0.8 mg/mL, 0.9 mg/mL, 1.0 mg/mL, 1.1 mg/mL, 1.2 mg/mL, 1.3 mg/mL, 1.4 mg/mL, 1.5 mg/mL, 1.6 mg/mL, 1.7 mg/mL, 1.8 mg/mL, 1.9 mg/mL, 2.0 mg/mL, 2.5 mg/mL, 3 mg/mL, 3.5 mg/mL, 4 mg/mL, 4.5 mg/mL, 5 mg/mL,

5.5 mg/mL, 6 mg/mL, 6.5 mg/mL, 7.0 mg/mL, 8.0 mg/mL, 8.5 mg/mL, 9 mg/mL, or 10 mg/mL. [0025]
A dry vaccine powder formulation can be used to prevent and/or treat infection by one or more influenza viruses. Influenza viruses belong to the Orthomyxoviridae family of viruses, which includes five genera: Influenzavirus A, Influenzavirus B, Influenzavirus C, Isavirus, and Thogotovirus. Dhori virus is a species of the genus Thogotovirus. An influenza virus can infect humans and other species. Influenza type A viruses can infect humans, birds, pigs, horses, seals and other animals. Wild birds can be natural hosts for these viruses. Influenza type A viruses can be divided into subtypes and named on the basis of two proteins on the surface of the virus: hemagglutinin (HA) and neuraminidase (NA). For example, an "H7N2 virus" designates an influenza A subtype that has an HA7 protein and an NA2 protein. Similarly an "H5N1" virus has an HA 5 protein and an NA 1 protein. There are 16 known HA subtypes and 9 known NA subtypes. Many different combinations of HA and NA proteins are possible. Any number of the known HA subtypes (HA1, HA2, HA3, HA4, HA5, HA6, HA7, HA8, HA9, HA10, HA11, HA12, HA13, HA14, HA15, and HA16) can be combined with any number of the known NA subtypes (NA1, NA2, NA3, NA4, NA5, NA6, NA7, NA8, and NA9) to produce a vaccine to prevent or treat an infection. The HA and NA subtypes can also be used individually in a vaccine to prevent or treat infection. Different subtype vaccines can be combined at the point of use, either sequentially or simultaneously, to prevent or treat an infection. Some influenza A subtypes (e.g., H1N1, H1N2, and H3N2) are currently in general circulation among people. Other subtypes can be found in other animal species. For example, H7N7 and H3N8 viruses can cause illness in horses, and H3N8 also has recently been shown to cause illness in dogs (http://www.cdc.gov/flu/avian/gen-info/flu-viruses). [0026]
Antiviral agents can be used to protect high-risk groups (e.g., individuals in a hospital unit, individuals at an institute caring for the elderly, or immuno-suppressed individuals). A potential use for an antiviral agent is to limit the spread and severity of the future pandemics whether caused by, e.g. avian H5N1 or another strains of influenza virus (e.g., H1N1). Avian influenza A viruses of the subtypes H5 and H7, including

H5N1, H7N7, and H7N3 viruses, have been associated with high pathogenicity, and human infection with these viruses have ranged from mild (e.g., H7N3, H7N7) to severe and fatal disease (e.g., H7N7, H5N1). Human illness due to infection with low pathogenicity viruses has been documented, including very mild symptoms (e.g., conjunctivitis) to influenza-like illness. Examples of low pathogenicity viruses that have infected humans include H7N7, H9N2, and H7N2. (http://www.cdc.gov/flu/avian/gen-info/flu-viruses). [0027]
Influenza B viruses can be found in humans and can also infect seals. Unlike influenza A viruses, these viruses are not classified according to subtype. Influenza B viruses can cause morbidity and mortality among humans, but in general are associated with less severe epidemics than influenza A viruses. Although influenza type B viruses can cause human epidemics, they have not caused pandemics, (http://www.cdc.gov/flu/avian/gen-info/flu-viruses). [0028]
Influenza type C viruses can cause mild illness in humans and do not cause epidemics or pandemics. These viruses can also infect dogs and pigs. These viruses are not classified according to subtype, (http://www.cdc.gov/flu/avian/gen-info/flu-viruses). [0029]
The methods and compositions described herein can be useful for the prevention and/or treatment of infection by any virus, including, for example, Abelson leukemia virus, Abelson murine leukemia virus, Abelson's virus, Acute laryngotracheobronchitis virus, Adelaide River virus, Adeno associated virus group, Adenovirus, African horse sickness virus, African swine fever virus, ADDS virus, Aleutian mink disease parvovirus, Alpharetrovirus, Alphavirus, ALV related virus, Amapari virus, Aphthovirus, Aquareovirus, Arbovirus, Arbovirus C, arbovirus group A, arbovirus group B, Arenavirus group, Argentine hemorrhagic fever virus, Argentine hemorrhagic fever virus, Arterivirus, Astrovirus, Ateline herpesvirus group, Aujezky's disease virus, Aura virus, Ausduk disease virus, Australian bat lyssavirus, Aviadenovirus, avian erythroblastosis virus, avian infectious bronchitis virus , avian leukemia virus, avian leukosis virus, avian lymphomatosis virus, avian myeloblastosis virus, avian paramyxovirus, avian pneumoencephalitis virus, avian reticuloendotheliosis virus, avian

sarcoma virus, avian type C retrovirus group, Avihepadnavirus, Avipoxvirus, B virus, B19 virus, Babanki virus, baboon herpesvirus, baculovirus, Barmah Forest virus, Bebaru virus, Berrimah virus, Betaretrovirus, Birnavirus, Bittner virus, BK virus, Black Creek Canal virus, bluetongue virus, Bolivian hemorrhagic fever virus, Boma disease virus, border disease of sheep virus, borna virus, bovine alphaherpesvirus 1, bovine alphaherpesvirus 2, bovine coronavirus, bovine ephemeral fever virus, bovine immunodeficiency virus, bovine leukemia virus, bovine leukosis virus, bovine mammillitis virus, bovine papillomavirus, bovine papular stomatitis virus, bovine parvovirus, bovine syncytial virus, bovine type C oncovirus, bovine viral diarrhea virus, Buggy Creek virus, bullet shaped virus group, Bunyamwera virus supergroup, Bunyavirus, Burkitt's lymphoma virus, Bwamba Fever, CA virus, Calicivirus, California encephalitis virus, camelpox virus, canarypox virus, canid herpesvirus, canine coronavirus, canine distemper virus, canine herpesvirus , canine minute virus, canine parvovirus, Cano Delgadito virus, caprine arthritis virus, caprine encephalitis virus, Caprine Herpes Virus, Capripox virus, Cardiovirus, caviid herpesvirus 1, Cercopithecid herpesvirus 1, cercopithecine herpesvirus 1, Cercopithecine herpesvirus 2, Chandipura virus, Changuinola virus, channel catfish virus, Charleville virus, chickenpox virus, Chikungunya virus, chimpanzee herpesvirus, chub reovirus, chum salmon virus, Cocal virus, Coho salmon reovirus, coital exanthema virus, Colorado tick fever virus, Coltivirus, Columbia SK virus, common cold virus, contagious ecthyma virus, contagious pustular dermatitis virus, Coronavirus, Corriparta virus, coryza virus, cowpox virus, coxsackie virus, CPV (cytoplasmic polyhedrosis virus), cricket paralysis virus, Crimean-Congo hemorrhagic fever virus, croup associated virus, Cryptovirus, Cypovirus, Cytomegalovirus, cytomegalovirus group, cytoplasmic polyhedrosis virus, deer papillomavirus, deltaretrovirus, dengue virus, Densovirus, Dependovirus, Dhori virus, diploma virus, Drosophila C virus, duck hepatitis B virus, duck hepatitis virus 1, duck hepatitis virus 2, duovirus, Duvenhage virus, Deformed wing virus DWV, eastern equine encephalitis virus, eastern equine encephalomyelitis virus, EB virus, Ebola virus, Ebola-like virus, echo virus, echovirus, echovirus 10, echovirus 28, echovirus 9, ectromelia virus, EEE virus, EIA virus, EIA virus, encephalitis virus, encephalomyocarditis group virus, encephalomyocarditis virus, Enterovirus, enzyme elevating virus, enzyme elevating virus (LDH), epidemic hemorrhagic fever virus,

epizootic hemorrhagic disease virus, Epstein-Barr virus, equid alphaherpesvirus 1, equid alphaherpesvirus 4, equid herpesvirus 2, equine abortion virus, equine arteritis virus, equine encephalosis virus, equine infectious anemia virus, equine morbillivirus, equine rhinopneumonitis virus, equine rhinovirus, Eubenangu virus, European elk papillomavirus, European swine fever virus, Everglades virus, Eyach virus, felid herpesvirus 1, feline calicivirus, feline fibrosarcoma virus, feline herpesvirus, feline immunodeficiency virus, feline infectious peritonitis virus, feline leukemia /sarcoma virus, feline leukemia virus, feline panleukopenia virus, feline parvovirus, feline sarcoma virus, feline syncytial virus, Filovirus, Flanders virus, Flavivirus, foot and mouth disease virus, Fort Morgan virus, Four Corners hantavirus, fowl adenovirus 1, fowlpox virus, Friend virus, Gammaretrovirus, GB hepatitis virus, GB virus, German measles virus, Getah virus, gibbon ape leukemia virus, glandular fever virus, goatpox virus, golden shinner virus, Gonometa virus, goose parvovirus, granulosis virus, Gross' virus, ground squirrel hepatitis B virus, group A arbovirus, Guanarito virus, guinea pig cytomegalovirus, guinea pig type C virus, Hantaan virus, Hantavirus, hard clam reovirus, hare fibroma virus, HCMV (human cytomegalovirus), hemadsorption virus 2, hemagglutinating virus of Japan, hemorrhagic fever virus, hendra virus, Henipaviruses, Hepadnavirus, hepatitis A virus, hepatitis B virus group, hepatitis C virus, hepatitis D virus, hepatitis delta virus, hepatitis E virus, hepatitis F virus, hepatitis G virus, hepatitis nonA nonB virus, hepatitis virus, hepatitis virus (nonhuman), hepatoencephalomyelitis reovirus 3, Hepatovirus, heron hepatitis B virus, herpes B virus, herpes simplex virus, herpes simplex virus 1, herpes simplex virus 2, herpesvirus, herpesvirus 7, Herpesvirus ateles, Herpesvirus hominis, Herpesvirus infection, Herpesvirus saimiri, Herpesvirus suis, Herpesvirus varicellae, Highlands J virus, Hirame rhabdovirus, hog cholera virus, human adenovirus 2, human alphaherpesvirus 1, human alphaherpesvirus 2, human alphaherpesvirus 3, human B lymphotropic virus, human betaherpesvirus 5, human coronavirus, human cytomegalovirus group, human foamy virus, human gammaherpesvirus 4, human gammaherpesvirus 6, human hepatitis A virus, human herpesvirus 1 group, human herpesvirus 2 group, human herpesvirus 3 group, human herpesvirus 4 group, human herpesvirus 6, human herpesvirus 8, human immunodeficiency virus, human immunodeficiency virus 1, human immunodeficiency virus 2, human papillomavirus, human T cell leukemia virus, human T cell leukemia

virus I, human T cell leukemia virus II, human T cell leukemia virus III, human T cell lymphoma virus I, human T cell lymphoma virus II, human T cell lymphotropic virus type 1, human T cell lymphotropic virus type 2, human T lymphotropic virus I, human T lymphotropic virus II, human T lymphotropic virus III, Ichnovirus, infantile gastroenteritis virus, infectious bovine rhinotracheitis virus, infectious haematopoietic necrosis virus, infectious pancreatic necrosis virus, influenza virus A, influenza virus B, influenza virus C, influenza virus D, influenza virus pr8, insect iridescent virus, insect virus, iridovirus, Japanese B virus , Japanese encephalitis virus, JC virus, Junin virus, Kaposi's sarcoma-associated herpesvirus, Kemerovo virus, Kilham's rat virus, Klamath virus, Kolongo virus, Korean hemorrhagic fever virus, kumba virus, Kysanur forest disease virus, Kyzylagach virus, La Crosse virus, lactic dehydrogenase elevating virus, lactic dehydrogenase virus, Lagos bat virus, Langur virus, lapine parvovirus, Lassa fever virus, Lassa virus, latent rat virus, LCM virus, Leaky virus, Lentivirus, Leporipoxvirus, leukemia virus, leukovirus, lumpy skin disease virus, lymphadenopathy associated virus, Lymphocryptovirus, lymphocytic choriomeningitis virus, lymphoproliferative virus group, Machupo virus, mad itch virus, mammalian type B oncovirus group, mammalian type B retroviruses, mammalian type C retrovirus group, mammalian type D retroviruses, mammary tumor virus, Mapuera virus, Marburg virus, Marburg-like virus, Mason Pfizer monkey virus, Mastadenovirus, Mayaro virus, ME virus, measles virus, Menangle virus, Mengo virus, Mengovirus, Middelburg virus, milkers nodule virus, mink enteritis virus, minute virus of mice, MLV related virus, MM virus, Mokola virus, Molluscipoxvirus, Molluscum contagiosum virus, monkey B virus, monkeypox virus, Mononegavirales, Morbillivirus, Mount Elgon bat virus, mouse cytomegalovirus, mouse encephalomyelitis virus, mouse hepatitis virus, mouse K virus, mouse leukemia virus, mouse mammary tumor virus, mouse minute virus, mouse pneumonia virus, mouse poliomyelitis virus, mouse polyomavirus, mouse sarcoma virus, mousepox virus, Mozambique virus, Mucambo virus, mucosal disease virus, mumps virus, murid betaherpesvirus 1, murid cytomegalovirus 2, murine cytomegalovirus group, murine encephalomyelitis virus, murine hepatitis virus, murine leukemia virus, murine nodule inducing virus, murine polyomavirus, murine sarcoma virus, Muromegalovirus, Murray Valley encephalitis virus, myxoma virus, Myxovirus, Myxovirus multiforme, Myxovirus parotitidis, Nairobi sheep disease virus, Nairovirus,

Nanirnavirus, Nariva virus, Ndumo virus, Neethling virus, Nelson Bay virus, neurotropic virus, New World Arenavirus, newborn pneumonitis virus, Newcastle disease virus, Nipah virus, noncytopathogenic virus, Norwalk virus, nuclear polyhedrosis virus (NPV), nipple neck virus, O'nyong'nyong virus, Ockelbo virus, oncogenic virus, oncogenic viruslike particle, oncornavirus, Orbivirus, Orf virus, Oropouche virus, Orthohepadnavirus, Orthomyxovirus, Orthopoxvirus, Orthoreovirus, Orungo, ovine papillomavirus, ovine catarrhal fever virus, owl monkey herpesvirus, Palyam virus, Papillomavirus, Papillomavirus sylvilagi, Papovavirus, parainfluenza virus, parainfluenza virus type 1, parainfluenza virus type 2, parainfluenza virus type 3, parainfluenza virus type 4, Paramyxovirus, Parapoxvirus, paravaccinia virus, Parvovirus, Parvovirus B19, parvovirus group, Pestivirus, Phlebovirus, phocine distemper virus, Picodnavirus, Picornavirus, pig cytomegalovirus - pigeonpox virus, Piry virus, Pixuna virus, pneumonia virus of mice, Pneumovirus, poliomyelitis virus, poliovirus, Polydnavirus, polyhedral virus, polyoma virus, Polyomavirus, Polyomavirus bovis, Polyomavirus cercopitheci, Polyomavirus hominis 2, Polyomavirus maccacae 1, Polyomavirus muris 1, Polyomavirus muris 2, Polyomavirus papionis 1, Polyomavirus papionis 2, Polyomavirus sylvilagi, Pongine herpesvirus 1, porcine epidemic diarrhea virus, porcine hemagglutinating encephalomyelitis virus, porcine parvovirus, porcine transmissible gastroenteritis virus, porcine type C virus, pox virus, poxvirus, poxvirus variolae, Prospect Hill virus, Provirus, pseudocowpox virus, pseudorabies virus, psittacinepox virus, quailpox virus, rabbit fibroma virus, rabbit kidney vacuolating virus, rabbit papillomavirus, rabies virus, raccoon parvovirus, raccoonpox virus, Ranikhet virus, rat cytomegalovirus, rat parvovirus, rat virus, Rauscher's virus, recombinant vaccinia virus, recombinant virus, reovirus, reovirus 1, reovirus 2, reovirus 3, reptilian type C virus, respiratory infection virus, respiratory syncytial virus, respiratory virus, reticuloendotheliosis virus, Rhabdovirus, Rhabdovirus carpia, Rhadinovirus, Rhinovirus, Rhizidiovirus, Rift Valley fever virus, Riley's virus, rinderpest virus, RNA tumor virus, Ross River virus, Rotavirus, rougeole virus, Rous sarcoma virus, rubella virus, rubeola virus, Rubivirus, Russian autumn encephalitis virus, SA 11 simian virus, SA2 virus, Sabia virus, Sagiyama virus, Saimirine herpesvirus 1, salivary gland virus, sandfly fever virus group, Sandjimba virus, SARS virus, SDAV (sialodacryoadenitis virus), sealpox virus, Semliki Forest Virus, Seoul

virus, sheeppox virus, Shope fibroma virus, Shope papilloma virus, simian foamy virus, simian hepatitis A virus, simian human immunodeficiency virus, simian immunodeficiency virus, simian parainfluenza virus, simian T cell lymphotrophic virus, simian virus, simian virus 40, Simplexvirus, Sin Nombre virus, Sindbis virus, smallpox virus, South American hemorrhagic fever viruses, sparrowpox virus, Spumavirus, squirrel fibroma virus, squirrel monkey retrovirus, SSV 1 virus group, STLV (simian T lymphotropic virus) type I, STLV (simian T lymphotropic virus) type II, STLV (simian T lymphotropic virus) type III, stomatitis papulosa virus, submaxillary virus, suid alphaherpesvirus 1, suid herpesvirus 2, Suipoxvirus, swamp fever virus, swinepox virus, Swiss mouse leukemia virus, TAC virus, Tacaribe complex virus, Tacaribe virus, Tanapox virus, Taterapox virus, Tench reovirus, Theiler's encephalomyelitis virus, Theiler's virus, Thogoto virus, Thottapalayam virus, Tick borne encephalitis virus, Tioman virus, Togavirus, Torovirus, tumor virus, Tupaia virus, turkey rhinotracheitis virus, turkeypox virus, type C retroviruses, type D oncovirus, type D retrovirus group, ulcerative disease rhabdovirus, Una virus, Uukuniemi virus group, vaccinia virus, vacuolating virus, varicella zoster virus, Varicellovirus, Varicola virus, variola major virus, variola virus, Vasin Gishu disease virus, VEE virus, Venezuelan equine encephalitis virus, Venezuelan equine encephalomyelitis virus, Venezuelan hemorrhagic fever virus, vesicular stomatitis virus, Vesiculovirus, Vilyuisk virus, viper retrovirus, viral haemorrhagic septicemia virus, Visna Maedi virus, Visna virus, volepox virus, VSV (vesicular stomatitis virus), Wallal virus, Warrego virus, wart virus, WEE virus, West Nile virus, western equine encephalitis virus, western equine encephalomyelitis virus, Whataroa virus, Winter Vomiting Virus, woodchuck hepatitis B virus, woolly monkey sarcoma virus, wound tumor virus, WRSV virus, Yaba monkey tumor virus, Yaba virus, Yatapoxvirus, yellow fever virus, and the Yug Bogdanovac virus. [0030]
B. NON-VIRAL PATHOGEN VACCINE COMPONENTS A vaccine described herein can comprise bacterial, fungal, or protist cells or components thereof. For example, a vaccine to a bacterial pathogen can comprise a killed bacterium or a purified antigenic determinant thereof. Attenuated bacteria can also be used as an antigen. In some instances, a vaccine to a toxin produced by a cellular

pathogen (e.g., cholera toxin) can be produced by combining the inactivated toxin (toxoid) with one or more of the vaccine components described herein. An antigenic peptide from a target pathogen can be purified from a source pathogen and/or produced recombinantly prior to combining with the one or more of the components of the vaccine. Conjugate antigens can also be used. In a conjugate antigen, the poorly antigenic polysaccharide outer coat of a bacterial pathogen is attached to toxic protein that can stimulate an immune response. Typically, vaccines to non-viral pathogens will be designed to produce immune responses (e.g., slgA production) to pathogens which affect mucosal surfaces, or gain access to the body via mucosal surfaces. Non-limiting examples of such pathogens include Cryptococcus neoformans, Shigella spp., Salmonella typhi, Sa. paratyphi, enterotoxigenic Escherischia coli, Yersinia pestis, Mycobacterium tubercolosis, Ureaplasma urealyticum, Cryptosporidium spp., Clostridium tetani, Corynebacterium diphtheriae, Neisseria meningitidis, Bordetella pertussis, Streptococcus pneumoniae, Bacillus anthracis, Leptospira interrogans, Leptospira kirschneri, Leptospira noguchii, Leptospira alexanderi, Leptospira weilii, Leptospira borgpetersenii, Leptospira santarosai, Leptospira kmetyi, Borrelia burgdorferi, Brucella abortus, Brucella canis, Brucella melitensis, Brucella suis, Campylobacter jejuni, Chlamydia pneumoniae, Chlamydia trachomatis, Chlamydophila psittaci, Clostridium botulinum, Clostridium difficile, Clostridium perfringens, Enterococcus faecalis, Enterococcus faecium, Francisella tularensis, Haemophilus influenzae, Helicobacter pylori, Legionella pneumophila, Leptospira interrogans, Listeria monocytogenes, Mycobacterium leprae, Mycobacterium ulcerans, Mycoplasma pneumoniae, Neisseria gonorrhoeae, Neisseria meningitidis, Pseudomonas aeruginosa, Rickettsia rickettsii, Salmonella typhi, Salmonella typhimurium, Shigella sonnei, Staphylococcus aureus, Staphylococcus epidermidis, Staphylococcus saprophyticus, Streptococcus agalactiae, Streptococcus pneumoniae, Streptococcus pyogenes, Treponema pallidum, Vibrio cholerae, Candida albicans, Aspergillus fumigatus, Aspergillus flavus, Cryptococcus gattii, Histoplasma capsulatum, Pneumocystis jirovecii, Stachybotrys chartarum, Plasmodium falciparum, etc.

I/We Claim:
1 . An intranasal dry vaccine powder formulation comprising:
a. one or more antigens;
b. one or more saccharides; and
c. microcrystalline cellulose.
2. An intranasal dry vaccine powder formulation for use in stimulating an slgA response in a subject to an antigen, wherein the intranasal dry vaccine powder formulation comprises one or more antigens, and wherein the intranasal dry vaccine powder formulation is generated by quick freezing a liquid vaccine formulation, wherein the quick freezing does not comprise spray-freezing.
3. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein at least one of the one or more antigens is a viral antigen, optionally wherein i) at least one of the one or more antigens is live attenuated virus, whole inactivated virus, split virus, subunit antigen, virosome, cold-adapted live influenza virus, or any combination thereof; or ii) at least one of the one or more antigens is influenza viral antigen.
4. The intranasal dry vaccine powder formulation of claim I or formulation for use of claim 2, wherein the one or more antigens comprises H1N1 influenza virus, H3N2 influenza virus, H5N1 influenza virus, Influenza B virus, or any combination thereof.
5. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein at least one of the one or more antigens is a bacterial antigen, optionally wherein at least one of the one or more antigens is killed whole bacteria, attenuated bacteria, toxoids, purified surface protein, purified recombinant surface protein, or any combination thereof.
6. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein at least one of the one or more antigens is tetanus toxoid, diphtheria toxoid, a protist antigen, a protein, or any combination thereof.

7. The intranasal dry vaccine powder formulation for use of claim 2. wherein the liquid vaccine formulation comprises one or more saccharides.
8. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 7, wherein at least one of the one or more saccharides is trehalose, mannitol, or lactose.
9. The intranasal dry vaccine powder formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation comprises microcrystalline cellulose.
10. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 9, wherein the microcrystalline cellulose has a mean particle diameter of from 10 micro m to 100 micro m and/or a specific surface area from 1.3 m2/g to 20 m2/g, and/or a bulk density from 0.1 g/cm3 to
1.0 g/cm3.
11. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation further comprises one or more buffers, optionally wherein at least one of the one or more buffers is a phosphate buffer.
12. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation is stable at room temperature and 60% relative humidity for at least 12 months.
13. The intranasal dry vaccine powder formulation for use of claim 2, wherein an IgG response is also stimulated.
14. The intranasal dry vaccine powder formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation is capable of inducing sigA production at a mucosal site other than the site of administration.

15. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation does not comprise an adjuvant.
16. The intranasal dry vaccine powder formulation for use of claim 2, wherein the quick freezing comprises using liquid nitrogen.
17. The intranasal dry vaccine powder formulation of claim 1 or formulation for use of claim 2, wherein the intranasal dry vaccine powder formulation further comprises one or more flowability agents that improve flowability and/or reduces hygroscopicity, optionally wherein the one or more flowability agents comprise tribasic calcium phosphate (TCP).
18. A device for administration of an intranasal dry vaccine powder formulation, comprising the intranasal dry vaccine powder formulation of any one of claims 1-17, optionally wherein the device is configured for a single use.
19. A method for generating an intranasal dry vaccine powder formulation comprising:
a. preparing a liquid formulation comprising one or more antigens;
b. quick freezing the liquid formulation, wherein the quick freezing does not comprise
spray-freezing; and,
c. blending the freeze-dried sample with one or more excipients to generate the intranasal
dry vaccine powder formulation.
20. The method of claim 19. wherein at least one of the one or more antigens is a viral antigen or bacterial antigen.
21. The method of claim 19, wherein at least one of the one or more antigens is influenza virus, optionally wherein the influenza virus is HIN1 influenza virus, H5N1 influenza virus, H3N2 influenza virus. Influenza B virus, or any combination thereof.

22. The method of claim 19, wherein at least one of the one or more antigens is live attenuated virus, whole inactivated virus, split virus, subunit antigen, virosome, cold adapted live influenza virus, or any combination thereof.
23. The method of claim 19, wherein at least one of the one or more antigens is killed whole bacteria, attenuated bacteria, toxoids, purified surface protein, purified recombinant surface protein, or any combination thereof.
24. The method of claim 19, wherein at least one of the one or more antigens is tetanus toxoid, diphtheria toxoid, a protist antigen, a protein, or any combination thereof.
25. The method of claim 19, wherein preparing the liquid formulation further comprises adding one or more saccharides, optionally wherein at least one of the one or more saccharides is lactose, trehalose, or mannitol.
26. The method of claim 19, wherein preparing the liquid formulation further comprises adding
one or more buffers, optionally wherein at least one of the one or more buffers is a phosphate buffer.
27. The method of claim 19, wherein the one or more excipients comprise microcrystalline
cellulose or tribasic calcium phosphate (TCP), optionally wherein the microcrystalline cellulose or
TCP has a mean particle diameter of from 10 micro m to 100 micro m and/ora specific surface area
from 1.3 m /g to 20 m /g, and/or a bulk density from 0.1 g/cm to 1.0 g/cm .