Freeze-dried viral vaccine compositions and method of manufacturing thereof

Abstract

The present disclosure relates to field of lyophilized / freeze-dried viral vaccine composition and methods for manufacturing and obtaining the vaccine composition comprising live attenuated flavivirus as vaccine antigen selected from a group of yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus, Zika virus vaccine antigen; and stabilizers comprising sugar or sugar alcohol; amino acids; lactalbumin hydrolysate and gelatin. Post reconstitution, the composition preserves the desired characteristics of the virus, including immunogenicity and stability. The composition of the present disclosure can be safely administered subcutaneously or intramuscularly such that the vaccine composition induces complete immune response to yellow fever virus infectious agent and meets the criterion for the seroprotection for the said immunogenic components comprising yellow fever virus.

Description

[0001]‘‘FREEZE-DRIED VIRAL VACCINE COMPOSITIONS AND METHOD OF MANUFACTURING THEREOF” TECHNICAL FIELD ϱ^ The present disclosure is related to biotechnology, virology and medicine. It concerns viral vaccines and manufacturing methods. More particularly, the present disclosure relates to a lyophilized / freeze-dried viral vaccine composition / formulation comprising of flavivirus (more specifically yellow fever virus) antigens / immunogens and improved manufacturing methods (providing high virus yield, minimum impurities) for obtaining vaccine exhibiting high ϭϬ^ potency and stability. The present disclosure further relates to an improved methodology in the field of vaccine production and applications of the said vaccine. BACKGROUND OF THE DISCLOSURE All publications herein are incorporated by reference to the same extent as if each individual ϭϱ^ publication or patent application was specifically and individually indicated to be incorporated by reference. The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. ϮϬ^ Yellow fever (YF) is considered one of the deadliest diseases in human history. The yellow fever is an acute viral haemorrhagic disease, characterized in some patients by jaundice, which explains the use of the term "yellow". The featured symptoms of yellow fever may include fever, headache, jaundice, muscle pain, nausea, vomiting and fatigue. Moreover, a small proportion of patients who contract the virus develop severe symptoms and Ϯϱ^ approximately half of those die within 7 to 10 days. The dramatic onset of symptoms and a case fatality rate (CFR) of 30–60% during severe disease is substantially higher than the 1918’s Spanish Influenza (>2.5%), making YF a feared infectious disease since the 18thcentury. Yellow fever decimated the human population in Africa and the Americas until the first half of the 20thcentury. Yellow fever has spread over 50 countries in the tropical areas of ϯϬ^ Africa, Central and South America. While YF vaccines have been a powerful tool to help control YF epidemics, yet the World Health Organization (WHO) still considers 47 countries in Africa and the Americas under high risk of YF epidemics (G. Montalvo Zurbia-Flores et. Al; 2022). YF is endemic in 44 countries in tropical South America and sub-Saharan Africa. Yellow fever is caused by the yellow fever virus (YFV) belonging to the family of ϱ^ flaviviruses, which comprises of Dengue virus (DV), Japanese encephalitis virus (JEV), tick- borne encephalitis virus (TBEV), West Nile virus (WNV) and Zika virus (ZV) as other members. The YFV consists of a lipoprotein envelope surrounding a nucleocapsid composed of the capsid protein and a single-stranded, positive-sense RNA, which has a length of 10862 nucleotides. In between a 5’ untranslated (5’ UTR) and a 3’ untranslated regions (3’ UTR), ϭϬ^ the RNA encodes, from the 5’ end to the 3’ end, three structural proteins, namely a capsid protein (C protein), a premembrane / membrane protein (prM / M protein), an envelope protein (E protein) and eight non-structural (NS) proteins, namely NS1, NS2A, NS2B, NS3, NS4A, P2k peptide, NS4B and NS5 proteins. The virus is maintained in sylvatic (also known as the forest pattern) and urban transmission ϭϱ^ cycles. The sylvatic cycle involves Aedes africanus (Africa), Haemagogus spp., and Sabethes spp. (South America) mosquitoes and NHPs in jungle habitats. Risk of human infection and spillover of the virus into the urban cycle increases as humans get closer in proximity to the forests where the sylvatic cycle is present. Aedes aegypti mosquitoes are responsible for most of the urban spread of the YFV. The R0 of YF can be as high as 5–7 during urban outbreaks. ϮϬ^ As climate change worsens and the Ae. aegypti habitats expand, regions of the world with people who are naïve to YFV infection are in danger of outbreaks. The danger of urban spread can be put into context with the 2015–2016 outbreak in Angola, where there were 4347 suspected cases, 884 confirmed cases, and 377 deaths. During the outbreak in Angola, cases of YF were imported into China, with a total of 11 confirmed cases and one death. While Ϯϱ^ these were the first cases of YF in Asia, it is important to note that all of the cases were imported from Angola. The high R0 and risk of spread to naïve populations highlights the need to control urban outbreaks. Some authors have used the cases in China to suggest the need of vaccination in Asia; however, it is significant to note that none of the cases in China led to secondary cases, and emphasis should be placed on improved vaccination rates in ϯϬ^ current endemic countries to prevent spread to new areas (Hansen, C.A.; Barrett, A.D.T. et. al; 2021). The case fatality rate of YF varies between 5 and 50% depending on the outbreak. In 2018, there were an estimated 109,000 severe infections and 51,000 deaths due to YF. Recent large outbreaks in Brazil (2016–2018), the Democratic Republic of the Congo (DRC) (2016), Angola (2016), Uganda (2016), and Nigeria (2019) demonstrated as to why the YFV is ϱ^ considered a re-emerging pathogen. In Brazil alone, there were more than 2000 confirmed cases, more than 500 deaths, and more than 4000 epizootics (disease in NHPs) between December 2016 and March 2018. This epidemic began in the north of the country and moved to southern coastal areas where the virus had not been detected previously, which was further evidence of its re-emergence. There is a need to have a sufficient vaccine supply to keep YF ϭϬ^ outbreaks under control (Hansen, C.A.; Barrett, A.D.T. et. al; 2021). It is estimated that a total of 900 million people are at risk of YF across the globe, with 610 million people being at risk in Africa alone. The history of YF reveals that it has been a recurrent problem in those endemic regions of Africa and South America. Even in countries that were declared free of YF, re-emergence has been observed (G. Montalvo Zurbia-Flores ϭϱ^ et. Al; 2022). The elimination of YF from endemic areas will surely remain a challenge until the large clusters of the population can access vaccination without further risks linked to the immunization. Recent YF outbreaks have further exhibited the challenging nature of managing infectious diseases in a modern super-connected world, emphasizing the risk of YF importation to non- ϮϬ^ endemic regions via travellers. Moreover, because of the extensive presence of the vector mosquitoes of the genres Aedes, almost half of the world’s population (more than 3 billion people) would be at risk upon YF importation (G. Montalvo Zurbia-Flores et. Al; 2022). Currently, there are no antivirals for yellow fever disease, and vaccination is critical in preventing the disease. In this regard, as early as in the l930’s, two kinds of live-attenuated Ϯϱ^ YFV vaccines (YF-LAVs) were developed. The first one corresponds to the French neurotropic vaccine (FNV), that was prepared from the wild-type French viscerotropic virus (FVV, isolated from Framboise Mayali in Senegal in 1928) and was passaged in mouse brain. However, the FNV proved rapidly to be too neurovirulent, having an exacerbated incidence of post-vaccinal encephalitis in children, and was abandoned in the early l980’s (Barrett, 2017).^ ϯϬ^ The second approach corresponds to the “17D” strain, that was prepared from the wild-type strain Asibi (isolated from a mild human case - “Mr. Asibi” - in Ghana in 1927) and was passaged in mouse and chicken tissues. The vaccine strain 17D has lost both viscerotropism and neurovirulence (Monath, 2005). Currently, six countries are producing live-attenuated YFV vaccine compositions from substrains derived from the 17D strain, namely Brazil (17DD substrain), China (17D-204 ϱ^ substrain), France (17D-204 substrain Stamaril®), Russia (17D-213 substrain), Senegal (17D- 204 substrain) and USA (17D-204 substrain YF- VAX®) (Barrett, 2017).^Nevertheless, only those qualified by the WHO (French, Russian, Senegalese and Brazilian) serve for international purposes and global supply, whilst vaccines manufactured in China are only deployed in domestic markets. The United States was the sixth producer of YF vaccines until ϭϬ^ 2015, when their production was stopped as they transitioned to new manufacturing facilities. To date, YF-LAVs are still produced using traditional manufacturing practices based on the propagation of attenuated YFV in chicken embryos. According to the WHO, the ‘vaccine needs’ to eliminate YF epidemics from 2017 to 2026, including doses for routine immunizations, mass campaigns, and emergencies, is of 1,384 million doses; corresponding ϭϱ^ to 138.4 million doses per year. However, the total vaccine production from all manufacturers running at full capacity only adds up to approximately 80 million doses annually. The unmet requirements of YF-LAV are of almost 60 million doses yearly. Further, the WHO has identified hypersensitivity to egg proteins as a concern for egg based conventional yellow fever vaccines. YF vaccines are also contraindicated in individuals ϮϬ^ suffering from hypersensitivity to egg proteins and their derivatives. Hypersensitivity to chicken eggs is the second most common food allergy in children. Although the prevalence of egg hypersensitivity varies from region to region, it has been suggested to have a prevalence of 0.5–2.5% in young children. As a result of its manufacture using embryonated chicken eggs, individuals suffering of hypersensitivity to egg-derived proteins are not recommended Ϯϱ^ to get full doses of the YF-LAV, otherwise, chances of developing anaphylaxis after its administration are 1.8–3.2 / 100,000.^^ As per pharmacopoeia monograph (IP & Ph. Eur.) and WHO TRS 978 (Annex 5), the concentration of residual protein nitrogen before addition of stabilizers in vaccine shall not be more than 0.25 mg / human dose of the vaccine. Also, concentration of specific egg derived ϯϬ^ protein impurity, ovalbumin, should not be more than 5 μg / human dose. Also, for YF vaccine manufacturing, as per conventional methods given in Pharmacopoeial monographs such as Ph. Eur., I.P and WHO TRS 978, Annex 5 recommends freezing the embryo in PBS post harvesting and utilizing it just before filling procedure. The vaccination strategy can greatly influence the immunogenicity, efficacy, and safety of a vaccine. The many factors impacting the efficacy of a vaccine can be broadly divided into ϱ^ three categories: (1) features of the vaccine itself, including immunogen design, vaccine type, formulation, adjuvant use, and dosing; (2) individual variations among vaccine recipients such as gender, age, developmental stage, nutrition status, and pre-existing immune conditions; and (3) vaccine administration-related parameters including vaccination approach, delivery route, method of administration, number of immunizations, immunization site, ϭϬ^ intervals between administrations and use of prime / boost regimens and vaccine modulators. Conventional route of administration of vaccine approaches include mucosal routes (oral, intranasal, pulmonary, rectal or vaginal) and parenteral routes (subcutaneous (sc), intradermal (id) or intramuscular (im) inoculation), and the choice of one strategy over the other depends on the type of vaccine and protective immunity needed to conquer the disease based on the ϭϱ^ route of infection and transmission. Thermal stability is important for the manufacture, distribution and administration of vaccines, especially in tropical developing countries, where particularly adverse field conditions exist. While many vaccines are efficacious and can be manufactured at low cost, they can be ϮϬ^ unstable which can make them impractical or expensive for widespread use. The efficacy or effectiveness of a vaccine depends in part on its stability during shipping and storage. Vaccines can be prone to degradation, particularly when agitated or exposed to heat. Transportation and storage at ambient temperature can lead to rapid inactivation of viruses. Currently available flavivirus LAV vaccines, including yellow fever virus (YFV) 17D Ϯϱ^ vaccine, are lyophilized in the presence of stabilizers. Nonetheless, these vaccines require storage and shipment at 2 – 8° C, a requirement that is difficult to achieve in the developing world and more remote regions of developed nations. This limited liquid phase physical stability may result in accidental administration of suboptimal doses, prevents use of multi-dose vials for vaccination campaigns, and inhibits ϯϬ^ public health distribution in clinical settings where cold-chains are unavailable. During storage and shipping, vaccines are often inadvertently subjected to potentially detrimental cycles of freezing and thawing. This ultimately leads to expensive wastage, as these vaccines would be discarded according to cGCP guidelines. Developing a formulation which would maintain vaccine viability upon freeze thaw would be advantageous for vaccine ϱ^ manufacturing and commercial distribution. The proteins present within the vaccine can be directly damaged by freeze-thaw cycles by several mechanisms. Larger ice crystals cause more drastic damage, engulfing the proteins and potentially damaging the vaccine container. When thawing, the recrystallization process exerts tension and shear stress on the proteins. Various stabilizers are used to stabilize the vaccine preparation to achieve the desired shelf ϭϬ^ life. Stabilizers such as polyvinylpyrrolidone (PVP), trehalose are used in virus formulations. However, PVP has been reported to destabilize live attenuated virus formulations. (Refer: JA White et al; Development of a stable liquid formulation of live attenuated influenza vaccine; Vaccine Volume 34, Issue 32, 12 July 2016, Pages 3676-3683; 2016). Trehalose is not only costly; but has to also be combined with other sugars and protein ϭϱ^ additives (Gelatin) to achieve stability. Also, other stabilizers are better than trehalose for enhancing shelf-life stability of a lyophilized vaccine. The typical non-ionic surfactants used in pharmaceutical formulations include Triton™ X- 100, Pluronic® F-68, F-88, and F-127 (poloxamers), Brij 35 (polyoxy-ethylene alkyl ether), polyoxyl stearate 40, Cremophor® EL, and alpha-tocopherol TPGS. Each of these surfactants ϮϬ^ have a common fact, in that they all contain polyoxyethylene moieties and thus to a greater or lesser extent, exhibit a similar problem, in that the polyoxyethylene moiety auto oxidizes to produce reactive peroxides, which causes an increase in unwanted protein immunogenicity. (Refer Edward T. Maggio et al; Polysorbates, peroxides, protein aggregation, immunogenicity - a growing concern; Journal of Excipients and Food Chemicals 3(2):46-53; 2012). Ϯϱ^ Polysorbates are prone to degradation by oxidation and hydrolysis, with hydrolysis being induced either chemically or enzymatically. Polysorbate may also be auto-oxidized by temperature, light or transition trace metals, and the resulting peroxide formation may induce protein oxidation, whereas the acid produced may lead to a decrease in solution pH. PS80 degraded via hydrolysis lead to slower surface adsorption rate, and the free fatty acid release ϯϬ^ from hydrolysis also forms insoluble particles, negatively impacting protein quality and stability. Polysorbate 80 has also been causally linked with an increased risk of blood clots, stroke, heart attack, heart failure, and tumor growth recurrence in patients with certain types of cancer. Lyophilization is a common mode of stabilization of vaccines. However, lyophilization causes loss in virus potency. Vaccines lose potency over time and the rate of potency loss is ϱ^ temperature-dependent. Live viruses are susceptible to osmotic, thermal and vacuum shocks. Enveloped viruses possess a lipid bilayer, which is considered as the less stable virus component because of its high fragility. Live viruses are susceptible to various stresses during lyophilization steps like freezing, primary drying, secondary drying that could affect the physico-chemical stability of viruses. Owing to their structure, loss of potency during freeze- ϭϬ^ drying can be due to protein destabilization (e.g., unfolding, degradation and aggregation), nucleic acid degradation, lipid layer alteration (e.g., phase transition, mechanical damage) and stresses related to changes in the internal (ice formation) and external (pH and osmolarity change) virus environment. The dehydration step of lyophilization results in collapse of the hydrogen bond structure of proteins which is accompanied with increased mobility of amino ϭϱ^ acid components of virus epitopes. It has been reported that, in some cases lyophilization causes upto 40% loss in virus potency. Though a lot of information is available on stress mechanisms and stabilization strategies of pharmaceutical peptides, proteins and DNA during lyophilization, due to the molecular complexity of viruses, different destabilization pathways and lack of analytical techniques permitting measurement of physico-chemical changes in the ϮϬ^ antigen’s structure during and after lyophilization mean that viruses constitute a particular lyophilization challenge. The destabilization mechanisms as well as protection mechanisms for live, attenuated viral vaccines during lyophilization are not well known.^There remains a need in the art for formulations suitable for preparing stabilized yellow fever virus preparations for use in immunological formulations and vaccines. It is desirable to provide Ϯϱ^ stabilizing formulations that preserve desired characteristics of said virus, immunological formulation or vaccine, including bioactivity, potency, virus viability and infectivity. Alternatives to lyophilization generally involve adding preservatives to the vaccine solution. This can cause safety concerns. For example, while thiomersal can improve the shelf-life of biologics, it is generally avoided because of its mercury content. ϯϬ^ The need to ensure a long shelf life (mandated by the WHO to be at least three years) in hot climates, vaccine candidates will require a long shelf life & higher stability, which would greatly aid vaccination efforts by allowing for lower doses and increased accessibility. Hence, it is important to develop a stable, potent & safe lyophilized / freeze dried live attenuated viral vaccines for pediatric & adults against yellow fever virus which may prove useful in giving high level, long term immunity in children & adults.^^ůƐŽ^^ the development of effective and efficient processing methods and formulations is of great importance to the development of ϱ^ clinically useful and commercially successful vaccines, including flavivirus vaccines. Applicant accordingly provides improved high virus yielding egg based yellow fever vaccine manufacturing methods (optimized culture conditions, filtration, centrifugation / homogenization) & formulations that - 1) preserve the potency of virus (3 log10IU / human dose of 0.5 ml), 2) reduce the residual egg derived impurity / ovalbumin content ϭϬ^ NMT than 5μg / human dose of 0.5 ml, protein nitrogen content NMT 0.25 mg / human dose, endotoxin content & thereby minimizing the probability of hypersensitivity / allergic reactions, 3) carry out post-harvest homogenization, centrifugation, stabilization and filtration in a continuous operation, devoid of embryo freezing requirement, 4) is devoid of any genetically modified eggs / enzyme / benzonase / trypsin, surfactant / urea / glycerol & additional ϭϱ^ steps like sucrose gradient, protamine precipitation, DFF, TFF, or chromatography 5) use stepwise / cascade 3 step Direct Flow Filtration (DFF) through 5 μm, 1.2 μm and 0.22 μm filters in series for reducing aggregate load & ensuring high virus recovery instead of Two step filtration (5 μm + 0.22 μm) or Two step filtration (1.2 μm + 0.22 μm), and 6) has minimum residual moisture content (not more than 3%), which is devoid of ϮϬ^ complex / costly / animal origin components like human serum albumin, trehalose, mannitol, lactose, sucrose, microcrystalline cellulose, hydroxyethyl starch, dextran, hetastarch, polymer, detergent, Pluronic, PVP, adjuvant, preservative, antibiotic;^wherein said formulations show significant reduction in potency loss with respect to cryoprotection of virus during freeze- thaw cycle. Ϯϱ^ OBJECTS OF THE DISCLOSURE An object of the present disclosure is to ameliorate one or more problems of the prior art or to at least provide a new, useful and non-obvious alternative to existing viral vaccines. An object of the present disclosure is to provide a lyophilized / freeze-dried viral vaccine ϯϬ^ composition / formulation wherein, post-reconstitution the composition preserves the desired characteristics of the virus, including bioactivity, potency, stability and immunogenicity. An object of the present disclosure is to provide a lyophilized viral vaccine composition comprising at least one virus, which meets the criterion for the seroprotection for the said immunogenic components. Another object of the present disclosure is to provide a lyophilized / freeze-dried live ϱ^ attenuated viral vaccine composition / formulation comprising live attenuated flavivirus. Another object of the present disclosure is to provide a lyophilized viral vaccine composition comprising live attenuated yellow fever virus from the genus flavivirus which meets the criterion for the seroprotection for the said immunogenic components. Another object of the present disclosure is to provide methods for manufacturing such ϭϬ^ lyophilized viral vaccine composition. Still another object of the present disclosure is to provide a method of vaccinating a host comprising parenteral immunization. Still another object of the present disclosure is to provide a kit comprising a lyophilized (freeze-dried) viral vaccine composition and an aqueous solution selected from saline or WFI ϭϱ^ (water for injection) for the reconstitution of the lyophilized (freeze-dried) vaccine composition / formulation comprising a live attenuated yellow fever virus from the genus flavivirus. Another object of the present disclosure is to provide improved high virus yielding egg based yellow fever vaccine manufacturing methods (optimized culture conditions, filtration, ϮϬ^ centrifugation / homogenization) & formulations that: 1) preserve the potency of virus (3 log10IU / human dose of 0.5 ml), 2) reduce the residual egg derived impurity / ovalbumin content NMT than 5μg / human dose of 0.5 ml, protein nitrogen content NMT 0.25 mg / human dose, endotoxin content & thereby minimizing the probability of hypersensitivity / allergic reactions, Ϯϱ^ 3) carry out post-harvest homogenization, centrifugation, stabilization and filtration in a continuous operation, devoid of embryo freezing requirement, 4) is devoid of any genetically modified eggs / enzyme / benzonase / trypsin, surfactant / urea / glycerol & additional steps like sucrose gradient, protamine precipitation, DFF, TFF, or chromatography, 5) use stepwise / cascade 3 step Direct Flow Filtration (DFF) through 5 μm, 1.2 μm and 0.22 μm filters in series for reducing aggregate load & ensuring high virus recovery instead of Two step filtration (5 μm + 0.22 μm) or Two step filtration (1.2 μm + 0.22 μm), and 6) has minimum residual moisture content (not more than 3%), which is devoid of ϱ^ complex / costly / animal origin components like human serum albumin, trehalose, mannitol, lactose, sucrose, microcrystalline cellulose, hydroxyethyl starch, dextran, hetastarch, polymer, detergent, Pluronic, PVP, adjuvant, preservative, antibiotic; wherein^said formulation shows significant reduction in potency loss with respect to cryoprotection of virus during freeze- thaw cycle. ϭϬ^ Another object of the present disclosure is to provide improved high virus yielding egg based yellow fever vaccine manufacturing methods which require carrying out post-harvest homogenization, centrifugation, stabilization and filtration in a continuous operation, devoid of embryo freezing requirement. Other objects and advantages of the present disclosure will be more apparent from the following description, which is not intended to limit the scope of ϭϱ^ the present disclosure. SUMMARY OF THE DISCLOSURE The present dislcosure relates to a lyophilized or freeze-dried viral vaccine composition, comprising: ϮϬ^ a) one or more virus particle or antigen thereof; and b) stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; lactalbumin hydrolysate and gelatin. In some embodiments, the vaccine composition of the present disclosure, comprises: a) live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10Ϯϱ^ IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; b) sorbitol at a concentration of about 1 to 20% (w / v); c) tricine at a concentration of about 0.1% to 2% (w / v), L-histidine at a concentration of about 0.1% to 2% (w / v), L-alanine at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride at a concentration of about 0.1% to 5% (w / v); ϯϬ^ d) gelatin at a concentration of about 0.1% to 10% (w / v); and e) lactalbumin hydrolysate at a concentration of about 0.05% to 2% (w / v). In some embodiments, the vaccine composition of the present disclosure, comprises: a) live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10 IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; ϱ^ b) sorbitol at a concentration of about 5% (w / v); c) tricine present at a concentration of about 0.3% (w / v), L-histidine at a concentration of about 0.21% (w / v), L-alanine at a concentration of about 0.1% (w / v) and L-arginine hydrochloride at a concentration of about 1.6% (w / v); d) gelatin at a concentration of about 2.5% (w / v); and ϭϬ^ e) lactalbumin hydrolysate at a concentration of 0.35% (w / v). The present disclosure also provides a method of manufacturing the said lyophilized or freeze-dried viral vaccine composition, the method comprising: a) subjecting at least one live attenuated flavivirus to at least one passaging in an avian leukosis virus (ALV) free embryonated specific pathogen-free (SPF) hen eggs to ϭϱ^ produce flavivirus drug substance; b) inoculating the drug substance into live ALV free embryonated SPF hen eggs and incubating for a period of abou 24 hours to 120 hours at 37 ± 1°C, preferably about 60-80 hours or about 68-76 hours; c) harvesting of the virus infected embryos; ϮϬ^ d) washing and homogenizing of the harvested embryos with phosphate buffer saline to produce embryonic pulp; e) centrifuging of the embryonic pulp and collecting supernatant comprising the virus; f) stabilizing the collected extract with a stabilizer comprising sugar or sugar alcohol; amino acids; gelatin; and lactalbumin hydrolysate; Ϯϱ^ g) clarifying the stabilized extract by direct flow filtration (DFF) through at least one clarification filter to obtain a clarified extract; h) sterilizing the clarified extract by DFF through at least one sterilization grade filter and storing the sterile extract or the drug substance under sub-zero temperature; i) thawing of the drug substance and making up the volume with diluent to achieve a ϯϬ^ predetermined dose, followed by re-sterilizing the blend by DFF through at least one sterilization grade filter; j) filling the sterile drug substance into a container; and k) freeze drying the drug substance containing container obtained in step (j), comprising the steps of freezing, sublimation and secondary drying to obtain the said lyophilized or freeze-dried viral vaccine composition. The present disclosure also relates to a kit comprising: ϱ^ a) a lyophilized or freeze-dried viral vaccine composition comprising: at least one live attenuated virus present at a dose of not less than 3 log10IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; one or more sugar or sugar alcohol at a concentration ranging from about 1 to 20% (w / v); one or more amino acid selected from a group comprising of tricine, leucine, iso-leucine, L-histidine, glycine, ϭϬ^ glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine and Threonine, or any combination thereof, present at a concentration ranging from about 0.01-10% (w / v); lactalbumin hydrolysate at a concentration ranging from about 0.05% to 2% (w / v); and gelatin at a concentration ranging in between 0.1% and ϭϱ^ 10% (w / v); and b) an aqueous solution selected from a group comprising saline, buffer and water for injection (WFI), or any combination thereof, for reconstituting the lyophilized or freeze-dried viral vaccine composition prior to its administration or use as vaccination. The present disclosure also relates to a method of preventing a disease condition caused by a ϮϬ^ virus in a subject, or a method of vaccinating the subject against the virus, said method comprising administering to the subject a reconstituted viral vaccine composition, said composition comprising: a) one or more virus particle or antigen thereof; and b) stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; Ϯϱ^ lactalbumin hydrolysate and gelatin; and wherein the composition is reconstituted in an aqueous solution selected from a group comprising of saline, buffer and water for injection (WFI), or any combination thereof, prior to its administration or use in vaccination. The present disclosure thus provides a live attenuated yellow fever virus vaccine comprisingϯϬ^ live attenuated yellow fever virus strain 17D vaccine virus primary seed (NIBSC Code: 213- 77), which can be safely administered subcutaneously or intramuscularly as a vaccine composition such that it meets the criterion for the seroprotection for the said immunogenic component. The inventors have found that the formulation preferably is a lyophilized formulation comprising of live attenuated yellow fever virus antigens along with sorbitol, gelatin, histidine, alanine, tricine, arginine & lactalbumin hydrolysate, wherein the ϱ^ formulation is stable, immunogenic & safe (with minimum hypersensitivity / allergic reactions). While sorbitol preserves the structural integrity (native like structure) of virus and prevents aggregation of viruses, the gelatin prevents adsorption of the viruses onto the walls of the vessel and promotes desorption done previously. Lactalbumin hydrolysate stabilizes proteins through a variety of mechanisms such as preferential hydration, direct binding, ϭϬ^ buffering, and antioxidation. Further, the said composition is devoid of surfactants like polysorbate. The said formulation shows significant reduction in potency loss with respect to cryoprotection of virus during freeze-thaw cycle. DETAILED DESCRIPTION ϭϱ^ Although the present disclosure may be susceptible to different embodiments, and following detailed discussion, with the understanding that the present disclosure can be considered an exemplification of the principles of the disclosure and is not intended to limit the scope of disclosure to that which is illustrated and disclosed in this description. Embodiments are provided so as to thoroughly and fully convey the scope of the present ϮϬ^ disclosure to the person skilled in the art. Numerous details are set forth, relating to specific components, and methods, to provide a complete understanding of embodiments of the present disclosure. It will be apparent to the person skilled in the art that the details provided in the embodiments should not be construed to limit the scope of the present disclosure. In some embodiments, well-known composition, well-known processes, and well-known Ϯϱ^ techniques are not described in detail. The terminology used, in the present disclosure, is only for the purpose of explaining a particular embodiment and such terminology shall not be considered to limit the scope of the present disclosure. As used in the present disclosure, the forms "a,” "an," and "the" may be intended to include the plural forms as well, unless the context clearly suggests otherwise. ϯϬ^ With respect to the use of substantially any plural and / or singular terms herein, those having skill in the art can translate from the plural to the singular and / or from the singular to the plural as is appropriate to the context and / or application. The various singular / plural permutations may be expressly set forth herein for sake of clarity.^The suffix ‘(s)’ at the end of any term in the present disclosure envisages in scope both the singular and plural forms of said term. ϱ^ The terms "comprises" "comprising" “including” and “having” are open ended transitional phrases and therefore specify the presence of stated features, integers, steps, operations, elements, modules, units and / or components, but do not forbid the presence or addition of one or more other features, integers, steps, operations, elements, components, and / or groups thereof. The particular order of steps disclosed in the process of the present disclosure is not ϭϬ^ to be construed as necessarily requiring their performance as described or illustrated, unless stated otherwise. It is also to be understood that additional or alternative steps may be employed. The terms first, second, third, etc., should not be construed to limit the scope of the present disclosure as the aforementioned terms may be only used to distinguish one element, ϭϱ^ component, region, layer or section from another component, region, layer or section. Terms such as first, second, third etc., when used herein do not imply a specific sequence or order unless clearly suggested by the present disclosure. The present disclosure provides a vaccine composition and a method for manufacturing the same. It is understood that each feature or embodiment, or combination, described herein is a non- ϮϬ^ limiting, illustrative example of any of the aspects of the invention and, as such, is meant to be combinable with any other feature or embodiment, or combination, described herein. For example, where features are described with language such as “one embodiment”, “some embodiments”, “certain embodiments”, “further embodiment”, “specific exemplary embodiments”, and / or “another embodiment”, each of these types of embodiments is a non- Ϯϱ^ limiting example of a feature that is intended to be combined with any other feature, or combination of features, described herein without having to list every possible combination. Such features or combinations of features apply to any of the aspects of the invention. More particularly, as regards the embodiments characterized in this specification, it is intended that each embodiment be read independently as well as in combination with another embodiment. ϯϬ^ For example, in case of an embodiment 1 reciting 3 alternatives A, B and C, an embodiment 2 reciting 3 alternatives D, E and F and an embodiment 3 reciting 3 alternatives G, H and I, it is to be understood that the specification clearly and unambiguously discloses embodiments corresponding to combinations A, D, G; A, D, H; A, D, I; A, E, G; A, E, H; A, E, I; A, F, G; A, F, H; A, F, I; B, D, G; B, D, H; B, D, I; B, E, G; B, E, H; B, E, I; B, F, G; B, F, H; B, F, I; C, D, G; C, D, H; C, D, I; C, E, G; C, E, H; C, E, I; C, F, G; C, F, H; C, F, I, unless specifically mentioned otherwise. ϱ^ The terms "about" or “approximately” are used herein to mean approximately, in the region of, roughly, or around. When the term "about" is used in conjunction with a numerical value / range, it modifies that value / range by extending the boundaries above and below the numerical value(s) set forth. In general, the term "about" is used herein to modify a numerical value(s) or a measurable value(s) such as a parameter, an amount, a temporal duration, and ϭϬ^ the like, above and below the stated value(s) by a variance of + / -20% or less, + / -10% or less, + / -5% or less, + / -1% or less, and + / -0.1% or less of and from the specified value, insofar such variations are appropriate to perform in the disclosed invention, and achieves the desired results and / or advantages as disclosed in the present disclosure. It is to be understood that the value to which the modifier “about” or “approximately” refers is itself also specifically, and ϭϱ^ preferably, disclosed. As used herein the terms "freeze-drying" or “lyophilize” or "lyophilization” or the likes, refers to lyophilization as is commonly known to a person skilled in the art and refers to the method by which a suspension is frozen, after which the water is removed by sublimation at low pressure. ϮϬ^ Within the scope of the present disclosure, all references made to presence or absence of one or more viruses in a vaccine composition / formulation refers to the presence or absence of virus particle as a whole or components thereof or antigens thereof. Based on the virus included in the vaccine composition / formulation within the present disclosure, a person skilled in the art will readily understand as to whether the vaccine comprises the virus as a Ϯϱ^ whole, or by way of any specific component or antigen thereof. The present disclosure provides a lyophilized / freeze-dried viral vaccine composition / formulation wherein, post-reconstitution, the composition preserves the desired characteristics of the virus, including bioactivity, potency, stability and immunogenicity. Accordingly, the present disclosure provides a lyophilized / freeze-dried viral vaccine ϯϬ^ composition / formulation comprising of virus; sugar or sugar alcohol; amino acids; lactalbumin hydrolysate and gelatin. According to some embodiments, the present disclosure provides a lyophilized or freeze-dried viral vaccine composition / formulation, comprising: a) one or more virus particle or antigen thereof; and b) stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; ϱ^ lactalbumin hydrolysate and gelatin. According to some embodiments of the present disclosure, the lyophilized / freeze-dried vaccine composition / formulation comprises of one or more viruses selected from a group comprising poxvirus (e.g. orthopoxviruses; avipoxviruses), morbillivirus (e.g. measles), mumps virus, rubella virus, alphavirus [e.g. sendai virus, sindbis virus and semliki forest ϭϬ^ virus (SFV)], ross river virus, encephalitis virus, flavivirus [e.g. yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus, Zika virus], rhabdovirus [e.g. vesicular stomatitis virus (VSV)], retrovirus (e.g. RNA tumor viruses), adenovirus (e.g. human adenovirus, bovine adenovirus, a canine adenovirus, a non-ϭϱ^ human primate adenovirus, a chicken adenovirus, or a porcine or swine adenovirus), adeno- associated viruses, lentiviral [e.g., human immunodeficiency viruses (HIV), simian immunodeficiency virus (SIV), and feline immunodeficiency virus (FIV)], herpes simplex virus, cytomegalovirus, picornavirus (e.g. Rhinovirus, Poliovirus etc), baculovirus vectors [autographacalifornica multiple nucleopolyhedrovirus (AcMNPV)], hepatitis B virus (HBV), ϮϬ^ rubulavirus (new castle disease virus), parainfluenza virus, influenza virus, respiratory syncytial virus (RSV), human metapneumovirus (hMPV), respiratory Coronavirus (CoV), Ebola, Marburg, Nipah, Chikungunya, Rotavirus, Human papilloma virus, Herpes simplex, Hepatitis A, Hepatitis C, Hepatitis B, Hepatitis E, Poliovirus, Variola Virus (e.g. smallpox, Monkeypox) and Varicella virus antigens. Ϯϱ^ According to some embodiments of the present disclosure, the virus is poxvirus, morbillivirus, mumps virus, rubella virus, alphavirus, ross river virus, encephalitis virus, flavivirus, rhabdovirus, retrovirus, adenovirus, adeno-associated viruses, lentivirus, herpes simplex virus, cytomegalovirus, picornavirus, baculovirus vector, hepatitis B virus (HBV), rubulavirus, parainfluenza virus, influenza virus, respiratory syncytial virus (RSV), human ϯϬ^ metapneumovirus (hMPV), respiratory Coronavirus (CoV), Ebola virus, Marburg virus, Nipah virus, Chikungunya virus, Rotavirus, Human papilloma virus, Herpes simplex virus, Hepatitis A virus, Hepatitis C virus, Hepatitis B virus, Hepatitis E virus, Poliovirus, Variola Virus and Varicella virus antigens. According to some embodiments of the present disclosure, the poxvirus is selected from a group comprising orthopoxviruses and avipoxviruses; the morbillivirus is measles virus, the alphavirus is selected from a group comprising sendai virus, sindbis virus and semliki forest virus (SFV); the flavivirus is selected from a group comprising yellow fever virus, dengue ϱ^ virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus and Zika virus; the rhabdovirus is vesicular stomatitis virus (VSV); the retrovirus is RNA tumor viruse; the adenovirus is selected from a group comprising human adenovirus, bovine adenovirus, canine adenovirus, non-human primate adenovirus, chicken adenovirus, porcine ϭϬ^ adenovirus and swine adenovirus; the lentivirus is selected from a group comprising human immunodeficiency viruses (HIV), simian immunodeficiency virus (SIV), and feline immunodeficiency virus (FIV); the picornavirus is selected from a group comprising Rhinovirus and Poliovirus; the baculovirus vector is autographacalifornica multiple nucleopolyhedrovirus (AcMNPV); the rubulavirus is new castle disease virus; and the Variola ϭϱ^ virus is selected from a group comprising smallpox and monkeypox virus. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of one or more viruses selected from the genus flavivirus. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϮϬ^ vaccine composition / formulation comprises of one or more viruses selected from the genus flavivirus comprising yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus, Zika virus vaccine antigen. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral Ϯϱ^ vaccine composition / formulation comprises of one or more viruses selected from the genus flavivirus consisting of yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus, Zika virus vaccine antigen. In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation ϯϬ^ comprises of virus selected from the genus flavivirus consisting of yellow fever virus. Thus, according to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of yellow fever virus. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition comprises of two or more viruses selected from the genus flavivirus ϱ^ combined together to be given in a single shot, suitable for the prevention and treatment of more than one disease state and which meets the criterion for the seroprotection for each of the said immunogenic components. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises one or more viruses selected from the genus ϭϬ^ flavivirus, along with one or more viruses selected from a group comprising dengue virus, tick-borne encephalitis virus, Rift Valley Fever, Chikungunya virus, Measles virus, Hepatitis A virus and Typhoid virus. In such a case, and as would be apparent to a person skilled in the art, the present disclosure also envisages and anticipates a multivalent viral vaccine, wherein the vaccine comprises at least two viruses, i.e., at least one belonging to the genus of ϭϱ^ flavivirus and the other belonging to a genus which is other than flavivirus. According to some embodiments of the present disclosure, the virus vaccine is a 1) viral vector vaccine; 2) a nucleic acid vaccine (DNA or mRNA based); 3) subunit vaccine; 4) vaccine based on nanoparticles; 5) vaccine based on inactivated whole virus; or 6) live attenuated virus vaccine. ϮϬ^ Thus, according to some embodiments of the present disclosure, the virus is a live attenuated virus (LAV), an inactivated virus, a chimeric virus, or a recombinant virus. In some embodiments, the virus vaccine is a live attenuated virus vaccine generated through various methods of viurus attenuation, including serial passage in egg tissue or cell lines, site directed mutagenesis, deletion of critical regions or through deoptimization of codons etc. Ϯϱ^ Alternatively, the live attenuated virus candidate is inactivated to produce an inactivated virus vaccine, and the methods used for virus inactivation are heat inactivation, UV inactivation or chemical inactivation not limited to formaldehyde, beta-propiolactone etc. The term "live" within the present disclosure is used in its conventional meaning, whereby a live virus is a virus which has not been inactivated, i.e. a virus capable of replicating on ϯϬ^ permissive cells. A “live attenuated virus” is a virus which does not induce the disease caused by the corresponding wild-type virus in animals or humans and which is capable of inducing a specific immune response. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of live attenuated yellow fever virus present at a ϱ^ dose of not less than 1000 IU or 1000 virus particles or 3 log10IU per 0.5 ml. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of live attenuated yellow fever virus strain developed through WHO yellow fever 17D vaccine virus primary seed (NIBSC Code: 213- 77) which was sourced from National Institute for Biological Standards and Control (NIBSC),ϭϬ^ Potters Bar, England and was passaged in avian leukosis virus (ALV) free specific pathogen- free (SPF) Hen eggs to prepare Master Seed Virus (MSV) of passage level 238. The MSV was further passaged in ALV free SPF Hen eggs to prepare Working Seed Virus (WSV) of passage level 239 which was used to generate drug substance of passage level 240 and stored at -60℃ or below temperature. ϭϱ^ According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of sugar or sugar alcohol, amino acid, ϮϬ^ lactalbumin hydrolysate and gelatin, all of which act as stabilizer in the vaccine composition / formulation. In accordance with some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of one or more sugar or sugar alcohol selected from a group comprising, but not limited to, glucose, sucrose, maltose, lactose, Ϯϱ^ fructose, galactose, mannose, maltulose, iso- maltulose, lactulose, mannitol, trehalose, raffinose, lactitol, lactobionic acid, sorbitol, dextrose, fructose, glycerol, and fucose, or any combination thereof. In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of one or more sugar or sugar alcohol selected from a group consisting of glucose, ϯϬ^ sucrose, maltose, lactose, fructose, galactose, mannose, maltulose, iso- maltulose, lactulose, mannitol, trehalose, raffinose, lactitol, lactobionic acid, sorbitol, dextrose, fructose, glycerol, and fucose, or any combination thereof. According to some embodiments of the present disclosure, the sugar or sugar alcohol is sorbitol. ϱ^ In some embodiments, the sugar or sugar alcohol is present at a concentration range of about 1-20% (w / v), preferably in the range of about 1-10% (w / v), and more preferably in the range of about 3-6% (w / v). In some embodiments, the sorbitol is present at a concentration range of about 1-20% (w / v), preferably in the range of about 1-10% (w / v), and more preferably in the range of about 3-6% ϭϬ^ (w / v). In some embodiments, the sorbitol is present at a concentration of about 5% (w / v). Thus, in some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; one or more sugar or ϭϱ^ sugar alcohol selected from a group consisting of glucose, sucrose, maltose, lactose, fructose, galactose, mannose, maltulose, iso- maltulose, lactulose, mannitol, trehalose, raffinose, lactitol, lactobionic acid, sorbitol, dextrose, fructose, glycerol, and fucose, or any combination thereof at a concentration range of about 1-20% (w / v); amino acid; lactalbumin hydrolysate; and gelatin. ϮϬ^ In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; sorbitol at a concentration range of about 1-20% (w / v); amino acid; lactalbumin hydrolysate; and gelatin. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral Ϯϱ^ vaccine composition / formulation comprises of one or more amino acids selected from the group comprising, but not limited to, tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine, Threonine, or any combination thereof. In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of one or more amino acids selected from the group consisting of tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L- alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, ϱ^ Methionine, Proline, Serine and Threonine, or any combination thereof. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of one or more amino acids selected from the group consisting of tricine, L-arginine hydrochloride, L-histidine and L-alanine as suitable amino acids individually or in combination. ϭϬ^ In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises tricine, L-arginine hydrochloride, L-histidine and L-alanine. In some embodiments, the amino acid is present at a concentration range of about 0.01-10% (w / v). In some embodiments, the amino acid tricine is present at a concentration ranging in between ϭϱ^ 0.1% and 2% weight / volume (w / v), preferably in between 0.1-1%, more preferably in between 0.1-0.5%, most preferably equal to 0.3% (w / v). In some embodiments, the amino acid L-histidine is present at a concentration ranging in between 0.1% to 2% (w / v), preferably in between 0.1-1%, more preferably in between 0.1- 0.5%, most preferably equal to 0.21% (w / v). ϮϬ^ In some embodiments, the amino acid L-alanine is present at a concentration ranging in between 0.01% and 1% weight / volume, preferably in between 0.05-0.5%, more preferably in between 0.08-0.2%, most preferably equal to 0.1% (w / v). In some embodiments, the amino acid L-arginine hydrochloride is present at a concentration ranging in between 0.1% and 10% weight / volume, preferably in between 0.1-5%, more Ϯϱ^ preferably in between 0.1-3%, most preferably equal to 1.6% (w / v). Thus, in some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; sorbitol at a concentration range of about 1-20% (w / v); one or more amino acids selected from the group consisting of ϯϬ^ tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine, Threonine, or any combination thereof at a concentration range of about 0.01%-10% (w / v); lactalbumin hydrolysate; and gelatin. In some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of ϱ^ the present disclosure comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; sorbitol at a concentration range of about 1-20% (w / v); amino acids tricine, L-arginine hydrochloride, L-alanine and L-histidine, each at a concentration range of about 0.01%-10% (w / v); lactalbumin hydrolysate; and gelatin. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϭϬ^ vaccine composition / formulation comprises of gelatin at a concentration ranging in between 0.1% and 10% weight / volume (w / v), preferably in between 0.1-5%, more preferably in between 0.1-3%, most preferably equal to 2.5% (w / v). According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprises of lactalbumin hydrolysate at a concentration ϭϱ^ ranging in between 0.05% and 2% weight / volume (w / v), preferably in between 0.1-1%, more preferably in between 0.1-0.5%, most preferably equal to 0.35% (w / v). Thus, in some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises of live attenuated yellow fever virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; sorbitol at a concentrationϮϬ^ range of about 1-20% (w / v); amino acids tricine, L-arginine hydrochloride, L-alanine and L- histidine, each at a concentration range of about 0.01%-10% (w / v); lactalbumin hydrolysate at a concentration ranging in between 0.05% and 2% (w / v); and gelatin at a concentration ranging in between 0.1% and 10% (w / v). As used herein, the term "gelatin" means a sterile non-pyrogenic protein preparation (e.g., Ϯϱ^ fractions) produced by partial acid hydrolysis (type A gelatin) or by partial alkaline hydrolysis (type B gelatin) of animal collagen, most commonly derived from cattle, pig, and fish sources. Gelatin can be obtained in varying molecular weight ranges. Recombinant sources of gelatin may also be used. In some embodiments, the lactalbumin hydrolysate and gelatin are obtained by chemical, ϯϬ^ enzymatic or thermal hydrolysis of protein from either plant or animal sources. Accordingly, in some embodiments, the stabilizers within the composition / formulation of the present disclosure comprise of: a) sorbitol at a concentration of about 1 to 20% (w / v); b) tricine, L-histidine, L-alanine and L-arginine hydrochloride present at a concentration ϱ^ of about 0.01% to 10% (w / v); c) gelatin present at a concentration of about 0.1% to 10% (w / v); and d) lactalbumin hydrolysate present at a concentration of about 0.05% to 2% (w / v). Accordingly, in some embodiments, the stabilizers within the composition / formulation of the present disclosure comprise of: ϭϬ^ a) sorbitol present at a concentration of about 5% (w / v); b) tricine present at a concentration of about 0.3% (w / v), L-histidine present at a concentration of about 0.21% (w / v), L-alanine present at a concentration of about 0.1% (w / v) and L-arginine hydrochloride present at a concentration of about 1.6% (w / v); ϭϱ^ c) gelatin present at a concentration of about 2.5% (w / v); and d) lactalbumin hydrolysate present at a concentration of about 0.35% (w / v). In the lyophilized / freeze-dried vaccine composition / formulation of the present disclosure, the sorbitol preserves the structural integrity (native like structure) of virus and prevents aggregation of viruses; gelatin prevents adsorption of the viruses onto the walls of the vessel ϮϬ^ and promotes desorption done previously & lactalbumin hydrolysate stabilizes proteins through a variety of mechanisms such as preferential hydration, direct binding, buffering, and antioxidation. It is to be noted that while the lyophilized / freeze-dried vaccine composition / formulation of the present disclosure comprises of sorbitol, tricine, L-arginine hydrochloride, L-alanine, L- Ϯϱ^ histidine, lactalbumin hydrolysate and gelatin as stabilizers, combination of all suitable alternatives of these stabilizers with one or more viruses of the present disclosure also fall within the scope of the present disclosure; as long as the composition / formulation provides desired characteristics of said virus, immunological formulation or vaccine, including bioactivity, potency, virus viability and infectivity, as provided herein. ϯϬ^ According to some embodiments of the present disclosure, the lyophilized / freeze-dried vaccine composition / formulation further comprises of an adjuvant selected from the group comprising aluminum hydroxide, aluminum phosphate, aluminum hydroxyphosphate, and potassium aluminum sulfate or a mixture thereof. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation further comprises of an immunostimulatory component ϱ^ selected from the group consisting of Alum, an oil and water emulsion MF-59,a liposome, a lipopolysaccharide, a saponin, lipid A, lipid A derivatives, Monophosphoryl lipid A, 3– deacylated monophosphoryl lipid A, AS01, AS03, an oligonucleotide, an oligonucleotide comprising at least one unmethylated CpG and / or a liposome, Freund’s adjuvant, Freund’s complete adjuvant, Freund’s incomplete adjuvant, polymers, co-polymers such as ϭϬ^ polyoxyethylene-polyoxypropylene copolymers, including block co-polymers, polymer p 1005, CRL-8300 adjuvant, muramyl dipeptide, TLR-4 agonists, imidazoquinolinone, Alhydroxiquim-II flagellin, flagellins derived from gram negative bacteria, TLR-5 agonists, fragments of flagellins capable of binding to TLR-5 receptors, Alpha-C-galactosylceramide, Matrix M, dmLT, Transferrin-Binding Protein, fHbp, Chitosan, Interleukin-2, QS-21, ϭϱ^ ISCOMS, and saponin in combination with sterols and lipids, or any combination thereof. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation is in the form of a single dose composition and is free of preservative. In some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation is in form of a the multi-dose composition and theϮϬ^ multi-dose composition comprises of preservative selected from the group comprising 2- phenoxyethanol, Benzethonium chloride (Phemerol), Phenol, m-cresol, Thiomersal, Formaldehyde, paraben esters (e.g. methyl-, ethyl-, propyl- or butyl- paraben), benzalkonium chloride, benzyl alcohol, chlorobutanol, p-chlor-m-cresol, or benzyl alcohol or a combination thereof. The lyophilized / freeze-dried viral vaccine composition may include material for a Ϯϱ^ single immunization, or may include material for multiple immunizations (i.e., a ‘multidose’ kit). As an alternative (or in addition) to including a preservative in multidose compositions, the compositions are contained in a container having an aseptic adaptor for removal of material. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϯϬ^ vaccine composition / formulation further comprises a pharmaceutically acceptable transporter, excipient, binder, carrier, isotonic agent, emulsifier or humectant, or any combination thereof, wherein the pharmaceutically acceptable excipient is selected from the group comprising of surfactants, polymers and salts. Examples of surfactants may include non-ionic surfactants such as polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 85, nonylphenoxypolyethoxethanol, octylphenoxypolyethoxethanol, oxtoxynol 40, nonoxynol- 9, triethanolamine, triethanolamine polypeptide oleate, polyoxyethylene- 660 ϱ^ hydroxystearate, polyoxyethylene- 35 ricinoleate, soy lecithin and a poloxamer - 0.001%- 0.05%; polymers including dextran, carboxymethylcellulose, hyaluronic acid ad cyclodextrin. Examples of the polymers may include dextran, carboxymethylcellulose, hyaluronic acid, cyclodextrin, etc. Examples of the salts may include NaCl, KCl, KH2PO4, Na2HPO4.2H2O, CaC12, MgC12, etc. ϭϬ^ Optionally in certain embodiments, surfactant agents can include, but are not limited to, a nonionic surfactant such as alkyl poly(ethylene oxide), copolymers of poly(ethylene oxide) and poly(propylene oxide) (EO-PO block copolymers ), poly(vinyl pyrroloidone), alkyl polyglucosides (such as sucrose monostearate, lauryl diglucoside, or sorbitan monolaureate, octyl glucoside and decyl maltoside), fatty alcohols (cetyl alcohol or olelyl alcohol), or ϭϱ^ cocamides (cocamide MEA, cocamide DEA and cocamide TEA). In other embodiments, the surfactants can include, but are not limited to, the Pluronic F127, Pluronic F68, Pluronic P123, or other EO-PO block copolymers of greater than 3,000-4,000 MW, or any combination thereof. In some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine ϮϬ^ composition is reconstituted with an aqueous solution selected from a group consisting of saline, buffer and WFI (water for injection), or any combination thereof, prior to its administration or use of the composition for vaccination. Accordingly, in some embodiments, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprises of live attenuated yellow fever Ϯϱ^ virus strain selected from strain 17D, 17D-204, 17D-213, or 17DD; sorbitol at a concentration range of about 1-20% (w / v); amino acids tricine, L-arginine hydrochloride, L-alanine and L- histidine, each at a concentration range of about 0.01%-10% (w / v); lactalbumin hydrolysate at a concentration ranging in between 0.05% and 2% (w / v); and gelatin at a concentration ranging in between 0.1% and 10% (w / v); which is reconstituted with an aqueous solution ϯϬ^ selected from a group consisting of saline, buffer and WFI (water for injection). According to some embodiments of the present disclosure, the final pH of the reconstituted composition is in the range of pH 6.0 to 7.5. According to some embodiments of the present disclosure, pH of the lyophilized / freeze-dried composition in the container, prior to reconstitution, is also in the range of pH 6.0 to 7.5. According to some embodiments of the present disclosure, the buffer or a buffering agent is selected from a group comprising HEPES, Citrate-phosphate, carbonate, phosphate, citrate, ϱ^ lactate, gluconate, borate, histidine buffer, succinate buffer and tartrate buffering agents, as well as more complex organic buffering agents including a phosphate buffering agent that contains sodium phosphate and / or potassium phosphate in a ratio selected to achieve the desired pH, or any combination of buffering agent thereof. In another example, the buffering agent contains Tris (hydroxymethyl) aminomethane, or "Tris", Tris-Histidine, BIS-Tris, and ϭϬ^ the Good buffers: BIS-Tris-Propane (BTP), PIPES, ACES, MOPS, MOPSO, BES, TES or glycylglycine formulated to achieve the desired pH. Yet in another example, the buffering agent could be the minimum essential medium with Hanks salts. According to some embodiments of the present disclosure, the reconstituted vaccine composition retains the desired characteristics of bioactivity, potency, stability and ϭϱ^ immunogenicity. According to some embodiments of the present disclosure, potency or Log10IU bioactivity or virus concentration of the reconstituted vaccine composition is reduced only by about 0.5 log after storage at 2-8°C for 36 months or after storage at 25°C for 6 months or 37°C for 14 days, in comparison with the bioactivity of a freshly reconstituted vaccine composition. ϮϬ^ The present disclosure also provides a method of manufacturing the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprising of following steps: a) subjecting at least one live attenuated virus to at least one passaging in an avian leukosis virus (ALV) free embryonated specific pathogen-free (SPF) hen eggs to Ϯϱ^ produce viral drug substance; b) inoculating the drug substance into live ALV free embryonated SPF hen eggs and incubating for a period of abou 24 hours to 120 hours at 37 ± 1°C, preferably about 60-80 hours or about 68-76 hours; c) harvesting of the virus infected embryos; ;^ ϯϬ^ d) washing and homogenizing of the harvested embryos with phosphate buffer saline to produce embryonic pulp; e) centrifuging of the embryonic pulp and collecting supernatant comprising the virus; f) stabilizing the collected extract with a stabilizer comprising sugar or sugar alcohol; amino acids; gelatin; and lactalbumin hydrolysate; g) clarifying the stabilized extract by direct flow filtration (DFF) through at least one clarification filter to obtain a clarified extract; ϱ^ h) sterilizing the clarified extract by DFF through at least one sterilization grade filter and storing the sterile extract or the drug substance under sub-zero temperature; i) thawing of the drug substance and making up the volume with diluent to achieve a predetermined dose, followed by re-sterilizing the blend by DFF through at least one sterilization grade filter; ϭϬ^ j) filling the sterile drug substance into a container; and k) freeze drying the drug substance containing container obtained in step (j), comprising the steps of freezing, sublimation and secondary drying to obtain the said lyophilized or freeze-dried viral vaccine composition. ϭϱ^ According to some embodiments of the present disclosure, the general method described above is applicable for manufacturing vaccine composition / formulation with any virus or its antigen, which grows into a chicken (hen) embryo. Thus, the said method is applicable for manufacturing vaccine composition / formulation based on any of such viruses, including flavivirus. ϮϬ^ Accordingly, the present disclosure also provides a method of manufacturing the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprising of following steps: a) subjecting at least one live attenuated flavivirus to at least one passaging in an avian Ϯϱ^ leukosis virus (ALV) free embryonated specific pathogen-free (SPF) hen eggs to produce flavivirus drug substance; b) inoculating the drug substance into live ALV free embryonated SPF hen eggs and incubating for a period of abou 24 hours to 120 hours at 37 ± 1°C, preferably about 60-80 hours or about 68-76 hours; ϯϬ^ c) harvesting of the virus infected embryos; ;^ d) washing and homogenizing of the harvested embryos with phosphate buffer saline to produce embryonic pulp; e) centrifuging of the embryonic pulp and collecting supernatant comprising the virus; f) stabilizing the collected extract with a stabilizer comprising sugar or sugar alcohol; amino acids; gelatin; and lactalbumin hydrolysate; g) clarifying the stabilized extract by direct flow filtration (DFF) through at least one clarification filter to obtain a clarified extract; ϱ^ h) sterilizing the clarified extract by DFF through at least one sterilization grade filter and storing the sterile extract or the drug substance under sub-zero temperature; i) thawing of the drug substance and making up the volume with diluent to achieve a predetermined dose, followed by re-sterilizing the blend by DFF through at least one sterilization grade filter; ϭϬ^ j) filling the sterile drug substance into a container; and k) freeze drying the drug substance containing container obtained in step (j), comprising the steps of freezing, sublimation and secondary drying to obtain the said lyophilized or freeze-dried viral vaccine composition. According to some embodiments of the present disclosure, the live attenuated flavivirus is ϭϱ^ subjected to at least 238 or 239 or 240 passages prior to the subsequent stem of inoculation. According to some embodiments of the present disclosure, the drug substance is inoculated into live ALV free embryonated SPF hen eggs for a period of about 7 to 8 days at strength of about 8000 to 16000 IU per egg; and wherein the post inoculation incubation is carried out for a period of about 72 hours at 37 ± 1°C. ϮϬ^ According to some embodiments of the present disclosure, the washing is carried out with phosphate buffer saline; the homogenization is carried out at about 2000 to 12000 rpm for a period of about 2-3 minutes to produce the embryonic pulp; and wherein the pulp is subjected to centrifugation at about 8000 to 16000 rpm for a period of about 20 minutes. According to some embodiments of the present disclosure, the stabilized embryo extract is Ϯϱ^ clarified and sterilized through filters of decreasing pore sizes in order of 5 μ, 1.2 μ and 0.22 μ to obtain the sterilized drug substance. According to some embodiments of the present disclosure, the embryo extract is stabilized with stabilizer comprises: a) sugar or sugar alcohol or sorbitol at a concentration of about 1 to 20% (w / v); ϯϬ^ b) amino acid or tricine at a concentration of about 0.1% to 2% (w / v), L-histidine present at a concentration of about 0.1% to 2% (w / v), L-alanine present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride present at a concentration of about 0.1% to 5% (w / v); c) gelatin at a concentration of about 0.1% to 5% (w / v); and d) and lactalbumin hydrolysate at a concentration of about 0.05% to 2% (w / v). ϱ^ According to some embodiments of the present disclosure, the freeze-drying comprises: a) a freezing step comprising of: freezing the drug substance containing container at a temperature in a range of about - 40°C to -55°C for about 300 minutes to 500 minutes; and b) a sublimation step comprising of: ϭϬ^ Ramp 1: ramping the temperature of about -55°C to a temperature in a range of about - 18°C to -28°C within about 300 minutes to 500 minutes at about 100 μbar pressure and holding at the temperature of about -18°C to -28°C for 300 minutes to about 1600 minutes at about 100 μbar pressure; and Ramp 2: ramping the temperature of about -18°C to -28°C to a temperature in a range ϭϱ^ of about -4°C to 0°C within about 90 minutes to 300 minutes at about 100 μbar pressure and holding at the temperature of about -4°C to 0°C for about 200 minutes to 500 minutes at about 100 μbar pressure; Ramp 3: ramping the temperature of about -4°C to 0°C to a temperature in a range of about 26°C within about 100 minutes to 300 minutes at about 100 μbar pressure and ϮϬ^ holding at the temperature of about 26°C for about 1 to 5 minutes at about 100 μbar pressure, and c) a secondary drying step comprising of: ramping the temperature to about 26°C within about 1 to 5 minutes and holding at the temperature of about 26°C for about 300 minutes to 500 minutes at about 25 μbar pressure. Ϯϱ^ According to some embodiments of the present disclosure, the flavivirus is a live attenuated yellow fever virus; and wherein strain of the yellow fever virus is selected from strain 17D, 17D-204, 17D-213, or 17DD. Thus, according to some embodiments, the present disclosure provides a method of manufacturing the lyophilized / freeze-dried viral vaccine composition / formulation of the ϯϬ^ present disclosure comprising of following steps: a) The candidate vaccine virus (ex: WHO yellow Fever 17D Primary Seed Virus, Passage No. 237, NIBSC Code: 213-77 or any other virus candidate that is employed as part of the composition / formulation of the present disclosure) is initially passaged in avian leukosis virus (ALV) free SPF embryonated hen eggs producing egg-based ϱ^ Master Seed Virus (MSV) of passage level 238; b) The egg-based Master Seed Virus (MSV) is further passaged in ALV free SPF embryonated hen eggs producing Working Seed Virus (WSV) of passage level 239; c) The Working Seed Virus (WSV) is sub cultured and propagated in ALV free SPF embryonated hen eggs to produce drug substance of passage level 240; ϭϬ^ d) The WSV is inoculated into live ALV free 6 to 9 days old embryonated SPF hen eggs at strength of 1000 to 32000 IU per egg; e) The inoculated eggs are incubated at 37 ± 1°C for 24-120 hours; f) The resulting virus infected embryos are harvested from live eggs and its decapitation; g) The embryos are washed with phosphate buffer saline (PBS; 2 ml per embryo);ϭϱ^ h) The washed embryos are homogenized in PBS (4 to 15 ml PBS per embryo) at 2000- 12000 rpm for 1-10 minutes to produce embryonic pulp; i) The embryonic pulp is centrifuged at RCF value of 6000 to 25000 x g for 10-30 minutes to separate tissue mass and collection of virus rich supernatant (embryo extract); ϮϬ^ j) The virus rich supernatant (embryo extract) is stabilized with stabilizers comprising one or more sugar or sugar alcohol; one or more amino acids; gelatin and lactalbumin hydrolysate; k) The stabilized embryo extract is filtered by direct flow filtration (DFF) through at least one clarification filter to obtain a clarified extract; Ϯϱ^ l) The clarified extract is sterilized by DFF through at least one sterilization grade filter to obtain a sterile drug substance (yellow fever single harvest) and storing of the drug substance at -60°C and below temperature; m) The stored drug substance is thawed and the volume is made up with diluent (Blind Vaccine) to achieve required dose of 0.5ml and again sterilized the blend by DFF ϯϬ^ through at least one sterilization grade filter; n) The sterile formulated drug substance (final bulk) is filled into individual sterile container (such as glass vials) and the containers are partially stoppered under aseptic conditions; and o) The mixture contained in the containers such as glass vials obtained in step (n) is subjected to freeze drying comprising the steps of freezing, sublimation and secondary drying, to obtain the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure. ϱ^ According to some embodiments of the present disclosure, post the infection the virus infected embryos are harvested post incubation period of 40 to 96 hours at 37 ± 1°C; or 68 to 76 hours at 37 ± 1°C, and preferably at 72 hours. According to some embodiments of the present disclosure, the homogenization is carried out at 2000 – 12000 RPM, preferably at 4000 - 10000 RPM, for a period of 2-3 minutes. ϭϬ^ According to some embodiments of the present disclosure, the centrifugation is carried out at RCF value of 6000 to 25000 x g for a period of 10 to 30 minutes. According to some embodiments of the present disclosure, the stabilizers comprise of: a) one or more sugar or sugar alcohol selected from a group consisting of glucose, sucrose, maltose, lactose, fructose, galactose, mannose, maltulose, iso- maltulose, ϭϱ^ lactulose, mannitol, trehalose, raffinose, lactitol, lactobionic acid, sorbitol, dextrose, fructose, glycerol, and fucose, or any combination thereof at a concentration range of about 1-20% (w / v); b) one or more amino acids selected from the group consisting of tricine, leucine, iso- leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, ϮϬ^ L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine, Threonine, or any combination thereof at a concentration range of about 0.01%-10% (w / v); c) gelatin present at a concentration of 0.1% to 10% (w / v); and d) lactalbumin hydrolysate present at a concentration of 0.05% to 2% (w / v). Ϯϱ^ According to some embodiments of the present disclosure, the stabilizers comprise of: a) sorbitol at a concentration of about 1 to 20% (w / v); b) tricine, L-histidine, L-alanine and L-arginine hydrochloride present at a concentration of about 0.01% to 10% (w / v); c) gelatin present at a concentration of about 0.1% to 10% (w / v); and ϯϬ^ d) lactalbumin hydrolysate present at a concentration of about 0.05% to 2% (w / v). According to some embodiments of the present disclosure, the stabilizers comprise of: a) sorbitol present at a concentration of about 5% (w / v); b) tricine present at a concentration of about 0.3% (w / v), L-histidine present at a concentration of about 0.21% (w / v), L-alanine present at a concentration of about ϱ^ 0.1% (w / v) and L-arginine hydrochloride present at a concentration of about 1.6% (w / v); c) gelatin present at a concentration of about 2.5% (w / v); and d) lactalbumin hydrolysate present at a concentration of about 0.35% (w / v). According to some embodiments of the present disclosure, the stabilized monovalent virus ϭϬ^ pool is clarified and sterilized, by direct flow filtration (DFF) typically through filters of decreasing pore sizes (e.g., 6 μ, 5 μ, 1.2 μ, 0.8 μ, 0.65μ, 0.45 μ, 0.22 μ). Suitable commercially available filters and filtration devices are well known in the art and can be selected by those of skill. Exemplary filtration devices could be made of Polypropylene or Cellulose acetate or Polyethersulfone and the commercially available filters could be Millipak ϭϱ^ (Millipore), Kleenpak (Pall), LifeAssure PDA (3M), Betafine PPG / PTG (3M), Sartopure (Sartorius), Sartopore (Sartorius) and Sartobran™ (Sartorius) filtration devices. In some embodiments, the stabilized embryo extract is clarified and sterilized, typically through filters of decreasing pore sizes in order of 5 μ > 1.2 μ > 0.22 μ to obtain a sterilized drug substance. ϮϬ^ According to some embodiments of the present disclosure, additional downstream processing steps may be employed including^ultrafiltration, diafiltration, and chromatography. According to some embodiments of the present disclosure, wherein residual egg derived impurity or ovalbumin content of the composition is in the range from 0.08 to 0.163 μg / 0.5 ml of the vaccine, and protein nitrogen content is in the range from 0.09 to 0.22 mg / 0.5 ml of the Ϯϱ^ vaccine to minimize hypersensitivity / allergic reaction. According to some embodiments of the present disclosure, post sterilization, the adjusted final pH of the sterilized drug substance / vaccine formulation bulk is between pH 6.0 – 7.5. According to some embodiments of the present disclosure, pH of the lyophilized / freeze-dried ϯϬ^ composition in the container, prior to reconstitution, is also in the range of pH 6.0 to 7.5. According to some embodiments of the present disclosure, the sterilized drug substance / vaccine formulation bulk is filled in a container selected from group comprising of a bottle, a vial, an ampule, an IV bag, a wearable injector, a bolus injector, a pre-filled syringe, a pen, a pump, a multidose needle syringe, a multidose vial, a multidose pen, a ϱ^ syrette, an auto-injector or a Vaccine Microarray Patches. In some embodiments, the sterilized drug substance / vaccine formulation bulk is filled in a type-1 glass vials with bromo-butyl rubber stoppers and aluminium seal with coloured polypropylene flip cap. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϭϬ^ vaccine composition / formulation may comprise of virus at a dose of not less than 1000 IU or 1000 virus particles or 3 log10IU per 0.5 ml or not less than of 1000 virus particles per 0.5 ml. According to some embodiments of the present disclosure, the freeze-drying step comprises of: a) the freezing step - freezing the sterile formulated drug substance in the containers such ϭϱ^ as glass vials at a temperature in a range of -40°C to -55°C for 300 minutes to 500 minutes; b) the sublimation step - Ramp 1: ramping the temperature of -55°C to a temperature in a range of -18°C to - 28°C within 300 minutes to 500 minutes at 100 μbar pressure and holding at the ϮϬ^ temperature of -18°C to -28°C for 300 minutes to 1600 minutes at 100 μbar pressure; and Ramp 2: ramping the temperature of -18°C to -28°C to a temperature in a range of - 4°C to 0°C within 90 minutes to 300 minutes at 100 μbar pressure and holding at the temperature of -4°C to 0°C for 200 minutes to 500 minutes at 100 μbar pressure; Ϯϱ^ Ramp 3: ramping the temperature of -4°C to 0°C to a temperature in a range of 26°C within 100 minutes to 300 minutes at 100 μbar pressure and holding at the temperature of 26°C for 1 to 5 minutes at 100 μbar pressure, and c) the secondary drying step - ramping the temperature to 26°C within 1 to 5 minutes and holding at the temperature of 26°C for 300 minutes to 500 minutes at 25 μbar ϯϬ^ pressure. As used herein, the terms "freeze-drying” or “lyophilize” or “lyophilization” involves lyophilization and refers to the method by which a suspension is frozen, and while in the frozen state, the major portion of the water and solvent system is reduced by sublimation and secondary drying (desorption) at low pressure so as to limit biological and chemical reactions ϱ^ at the designated storage temperature. As used herein, the term "sublimation" refers to a change in the physical properties of a composition, wherein the composition changes directly from a solid state to a gaseous state without becoming a liquid. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprising flavivirus, more preferably yellow fever virus, ϭϬ^ obtained by the manufacturing method of the present disclosure is stable at 2-8 deg C for 12 to 36 months; at 25 deg C for 2 to 6 months; at 37 deg C for 1 week to 2 weeks. More preferably, the lyophilized / freeze-dried viral combination vaccine composition - retains 0.5 Log10IU less bioactivity (virus concentration) than the original bioactivity after storage at 2-8°C for 36 months; ϭϱ^ retains 0.5 Log10IU less bioactivity (virus concentration) than the original bioactivity after storage at 25°C for 6 months; and retains 0.5 Log10IU less bioactivity (virus concentration) than the original bioactivity after storage at 37°C for 14 days. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϮϬ^ vaccine composition / formulation comprising flavivirus, more preferably yellow fever virus obtained from the manufacturing method of the present disclosure is stable post freezing at - 60℃ and thawing at room temperature, and wherein the reconstituted composition preserves the desired characteristics of virus, including bioactivity, potency, stability, immunogenicity and no potency loss was observed during repeated cycles of freezing and thawing step. Ϯϱ^ According to some embodiments of the present disclosure, the method allows the vaccine composition to retain Log10IU bioactivity or virus concentration of the vaccine composition after storage of the composition at 2-8°C for 36 months or after storage at 25°C for 6 months or 37°C for 14 days. In an alternative embodiment of the present disclosure, the composition is fully liquid. In an alternative embodiment of the present disclosure, wherein the live attenuated virus candidate is inactivated to produce an inactivated virus vaccine, and the methods used for virus inactivation could be heat inactivation, UV inactivation or chemical inactivation not limited to formaldehyde, beta-propiolactone etc. ϱ^ According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation is formulated for use in a method for reducing the onset of or preventing a health condition caused by flavivirus, such as yellow fever virus infection. The said use involves administration of an immunologically effective amount of the vaccine composition to a human subject via parenteral (subcutaneous or intradermal or intramuscular ϭϬ^ or intraperitoneal or intravenous administration or injectable administration or pulmonary administration, suppositories, needle-less injection, transcutaneous) or sustained release from implants or administration by eye drops or Mucosal (oral, intranasal, pulmonary, rectal or vaginal) or buccal or peroral or intragastric or perlinqual, alveolar or gingival or olfactory or respiratory mucosa administration or interthecally, intralymphatically, via bladder instillation, ϭϱ^ or via scarification or any other routes of immunization. Thus, the present disclosure also provides a method of preventing a disease condition caused by a virus in a subject, or a method of vaccinating the subject against the virus, said method comprising administering to the subject a reconstituted viral vaccine composition, said composition comprising: ϮϬ^ a) one or more virus particle or antigen thereof; and b) stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; lactalbumin hydrolysate and gelatin; and wherein the composition is reconstituted in an aqueous solution selected from a group comprising of saline, buffer and water for injection (WFI), or any combination thereof, prior Ϯϱ^ to its administration or use in vaccination. According to some embodiments of the present disclosure, the virus is a live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml or 1000 IU or 1000 virus particles; the sugar or sugar alcohol is sorbitol present at a concentration of about 5% (w / v); and the amino acid is present as a combination of amino acids such that ϯϬ^ tricine is present at a concentration of about 0.1% to 2% (w / v), L-histidine is present at a concentration of about 0.1% to 2% (w / v), L-alanine is present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride is present at a concentration of about 0.1% to 5% (w / v). According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprising live attenuated yellow ϱ^ fever virus is administered to a human subject via parenteral route. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure comprising live attenuated yellow fever virus is administered to a human subject via parenteral route including subcutaneous or intramuscular administration. ϭϬ^ According to some embodiments of the present disclosure, the administration of the vaccine of the present disclosure is through an intranasal dispensing device, such as a device in the form of an aerosol (intranasal spray) or a drop delivery system. Liquid nasal formulations can be delivered via Instillation and rhinyle catheter, Compressed air nebulizers, Squeezed bottle, Metered-dose pump sprays like multi dose metered dose spray pumps or single / duo dose ϭϱ^ spray pump). Other dosage forms can be selected from Nasal powders (Insufflators, Dry powder inhaler), Nasal Gels, Nasal drops, Solutions, Suspensions, Cosolvent system, Microspheres, Nanoparticles, Microemulsions and Nasal insert, or any combination thereof. The intranasal delivery devices can be selected from but not limited to Becton Dickinson (BD) Accuspray™ delivery device, Bi-Directional™ Optinose nasal device, MAD Intranasal ϮϬ^ Mucosal Atomization device by Teleflex, AeroLife™ and AeroVax™ (AerovectRx Inc., Atlanta, GA), Jet injector - PharmaJet® Stratis®Needle-Free Injector, MUNJIs Multi-use- nozzle jet injectors: Aquapuncture device, Hypospray®, MadaJet®, GentleJet®, Disposable- syringe Jet Injectors: Medi-Jector®, J-Tip®, Injex®, Vitajet™, LectraJet HS, LectraJet® M3, ZetaJet™, PharmaJet®, Aktiv-Dry PuffHaler™ and Nasal spray flu shot device. Ϯϱ^ According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation is formulated for use in a method for reducing the onset of or preventing a health condition such as yellow fever virus infection or its subtypes as disclosed in earlier embodiment of the disclosure. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral ϯϬ^ vaccine composition / formulation is administered via parenteral route in a dose effective for the production of neutralizing antibody and meets the criterion for the seroprotection for each of the said immunogenic components comprising yellow fever virus. The vaccine of the present disclosure is administered in a manner compatible with the dosage formulation, and in such amount as will be prophylactically and / or therapeutically effective. The vaccine composition of the present disclosure can be administered as primary prophylactic agents in ϱ^ adults or children at the risk of infection, or can be used as secondary agents for treating infected patients. For example, the lyophilized live attenuated vaccine composition as disclosed herein can be used in adults or children at risk of flavivirus and more preferably yellow fever virus infection, or can be used as secondary agents for treating yellow fever virus infected patients. ϭϬ^ More preferably the composition is administered parenterally in a dosage volume of about 0.5ml, comprising of not less than 1000 IU or 1000 virus particles or 3 log10IU per 0.5 ml. According to some embodiments of the present disclosure, the lyophilized / freeze-dried viral vaccine composition / formulation comprising flavivirus more preferably yellow fever virus is formulated as single dose vials or multidose vials (2 Dose or 5 Dose or 10 Dose vials) orϭϱ^ multidose kit or as pre-filled syringes or nasal sprays wherein the said lyophilized / freeze- dried vaccine composition / formulation is given in a single dose schedule, or a multiple dose schedule in which a primary course of vaccination is followed by 1-2 separate doses given at subsequent time intervals required to maintain and or reinforce the immune response, for example, at 1-4 months for a second dose, and if needed, a subsequent dose(s) after several ϮϬ^ months or years. The dosage regimen will also, at least in part, be determined on the need of a booster dose required to confer protective immunity. Yet alternatively the lyophilized / freeze-dried yellow fever viral vaccine composition is formulated for administration to a human subject or children 2 years of age or below according to a single dose regimen or two dose regimens consisting of a first dose, and second Ϯϱ^ dose at subsequent time intervals after 1-3 years. Yet alternatively the lyophilized / freeze-dried yellow fever viral vaccine composition is administered concomitantly with other drugs or any other vaccine. In some embodiments, the composition / formulation of the present disclosure is presented in vials, or in pre-filled syringes. The syringes may be supplied with or without needles. A ϯϬ^ syringe includes a single dose of the composition, whereas a vial includes a single dose or multiple doses (e.g., 2 doses). In one embodiment the dose is for human. In a further embodiment the dose is for an adult, adolescent, toddler, infant or less than one-year old human and is administered by injection. Vaccines of the invention are packaged in unit dose form or in multiple dose form (e.g., 2 doses). The said multidose composition can be selected from a group consisting of 2 dose, 5 ϱ^ dose and 10 doses. For multiple dose forms, vials are preferred to pre-filled syringes. Effective dosage volumes can be routinely established, but a typical human dose of the composition for injection has a volume of 0.5mL. In order to facilitate the commercialization of the vaccine composition / formulation of the present disclosure, and allow the reconstitution of the lyophilized composition, the present ϭϬ^ disclosure also provides a kit comprising: a) a lyophilized or freeze-dried viral vaccine composition comprising: at least one live attenuated virus present at a dose of not less than 3 log10IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; one or more sugar or sugar alcohol at a concentration ranging from about 1 to 20% (w / v); one or more amino acid selected ϭϱ^ from a group comprising tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine and Threonine, or any combination thereof, present at a concentration ranging from about 0.01-10% (w / v); lactalbumin hydrolysate at a concentration ranging from about 0.05% ϮϬ^ to 2% (w / v); and gelatin at a concentration ranging in between 0.1% and 10% (w / v); and b) an aqueous solution selected from a group comprising saline, buffer and water for injection (WFI), or any combination thereof, for reconstituting the lyophilized or freeze-dried viral vaccine composition prior to its administration or use as vaccination. Ϯϱ^ According to some embodiments of the present disclosure, the virus is a live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml or 1000 IU or 1000 virus particles; the sugar or sugar alcohol is sorbitol present at a concentration of about 5% (w / v); and the amino acid is present as a combination of amino acids such that tricine is present at a concentration of about 0.1% to 2% (w / v), L-histidine is present at a ϯϬ^ concentration of about 0.1% to 2% (w / v), L-alanine is present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride is present at a concentration of about 0.1% to 5% (w / v). According to some embodiments of the present disclosure, the composition and the aqueous solution are contained in a single or different containers selected from a group comprising a bottle, a vial, an ampule, an IV bag, a wearable injector, a bolus injector, a pre-filled syringe, a pen, a pump, a multidose needle syringe, a multidose vial, a multidose pen, a syrette, an ϱ^ auto-injector or a Vaccine Microarray Patches; wherein when the composition and the aqueous solution are contained in a single container, the container comprises a partition or mechanism to prevent the composition and the aqueous solution from being mixed, as part of the kit. Thus, a person skilled in the art will readily appreciate that the composition / formulation of the ϭϬ^ present disclosure can be commercialized in two ways: x first, where the reconstituted final composition / formulation ready for administration to a human subject is directly commercialized; or x second, where the lyophilized or free-dried composition / formulation of the present disclosure and the solution required for its reconstitution before administration to a ϭϱ^ human subject, are both part of a commercial kit, but not mixed together at the time of commercialization. Thus, in some embodiments, the present disclosure provides a vaccine kit, wherein the kit comprises of: a) a first container containing a lyophilized / freeze-dried viral vaccine ϮϬ^ composition / formulation comprising: flavivirus selected from a group consisting of live attenuated yellow fever virus present at a dose of not less than 3 log10 IU per 0.5 ml; sorbitol at a concentration range of about 1-20% (w / v); amino acids tricine, L- arginine hydrochloride, L-alanine and L-histidine, each at a concentration range of about 0.01%-10% (w / v); lactalbumin hydrolysate at a concentration ranging in Ϯϱ^ between 0.05% and 2% (w / v); and gelatin at a concentration ranging in between 0.1% and 10% (w / v); and b) a second container containing an aqueous solution selected from saline or water for injection (WFI) for the reconstitution of the lyophilized (freeze-dried) viral vaccine composition. ϯϬ^ In some embodiments, the vaccine kit comprises of: a) a first container containing a lyophilized / freeze-dried viral vaccine composition / formulation comprising: flavivirus selected from a group consisting of live attenuated yellow fever virus present at a dose of not less than 3 log10 IU per 0.5 ml; sorbitol present at a concentration of about 5% (w / v); tricine present at a ϱ^ concentration of about 0.3% (w / v), L-histidine present at a concentration of about 0.21% (w / v), L-alanine present at a concentration of about 0.1% (w / v) and L-arginine hydrochloride present at a concentration of about 1.6% (w / v); gelatin present at a concentration of about 2.5% (w / v); and lactalbumin hydrolysate present at a concentration of about 0.35% (w / v); and ϭϬ^ b) a second container containing an aqueous solution selected from saline or water for injection (WFI) for the reconstitution of the lyophilized (freeze-dried) viral vaccine composition. Throughout this specification the word “comprise”, or variations such as “comprises” or “comprising”, will be understood to imply the inclusion of a stated element, integer or step, or ϭϱ^ group of elements, integers or steps, but not the exclusion of any other element, integer or step, or group of elements, integers or steps and can mean "includes," "including," and the like; "consisting essentially of' or "consists essentially" likewise has the meaning ascribed in U.S. Patent law and the term is open-ended, allowing for the presence of more than that which is recited so long as basic or novel characteristics of that which is recited is not changed by ϮϬ^ the presence of more than that which is recited, but excludes prior art embodiments. As used herein the term “comprising” or “comprises” is used in reference to compositions, methods, and respective component(s) thereof, that are useful to an embodiment, yet open to the inclusion of unspecified elements, whether useful or not. It will be understood by those within the art that, in general, terms used herein are generally intended as “open” terms (e.g., Ϯϱ^ the term “including” should be interpreted as “including but not limited to,” the term “having” should be interpreted as “having at least,” the term “includes” should be interpreted as “includes but is not limited to,” etc.). Although the open-ended term “comprising,” as a synonym of terms such as including, containing, or having, is used herein to describe and claim the invention, the present invention, or embodiments thereof, may alternatively be ϯϬ^ described using alternative terms such as “consisting of’ or “consisting essentially of”. Throughout this specification the word, “lyophilized / freeze-dried vaccine composition / formulation" or "lyophilized live attenuated vaccine composition / formulation" covers any composition that elicits an immune response against the antigen or immunogen of interest; for instance, after administration into a subject, elicits an immune response against the targeted immunogen or antigen of interest. The word “lyophilized / freeze-dried vaccine composition / formulation" covers: use of single virus or virus antigen or combination of more ϱ^ than one virus or virus antigens mixed together to form a combination vaccine. The terms "vaccine composition" and "vaccine" covers any composition that induces a protective immune response against the antigen of interest, or which efficaciously protects against the antigen; for instance, after administration or injection into the subject, elicits a protective immune response against the targeted antigen or immunogen or provides efficacious ϭϬ^ protection against the antigen or immunogen. The use of the expression “one or more” or “two or more” or “at least one” suggests the use of one or more elements or ingredients or quantities, as the use may be in the embodiment of the invention to achieve one or more of the desired objects or results. It may suggest comprising: one element or combination of more than one element mixed together. While ϭϱ^ certain embodiments of the inventions have been described, these embodiments have been presented by way of example only, and are not intended to limit the scope of the inventions. Variations or modifications to the composition of this invention, within the scope of the invention, may occur to those skilled in the art upon reviewing the disclosure herein. Such variations or modifications are well within the spirit of this disclosure. ϮϬ^ The numerical values given for various physical parameters, dimensions and quantities are only approximate values and it is envisaged that the values higher than the numerical value assigned to the physical parameters, dimensions and quantities fall within the scope of the invention unless there is a statement in the specification to the contrary. Numerical ranges stated in the form ‘from x to y’ or 'between x and y' include the values mentioned and those Ϯϱ^ values that lie within the range of the respective measurement accuracy as known to the skilled person. If several preferred numerical ranges are stated in this form, of course, all the ranges formed by a combination of the different end points are also included, including decimal or fractional values. Similarly, the components used in purification, e.g., filters, columns, are not intended to be in ϯϬ^ any way limiting or exclusionary, and can be substituted for other components to achieve the same purpose at the discretion of the practitioner. While considerable emphasis has been placed herein on the specific features of the preferred embodiment, it will be appreciated that many additional features can be added and that many changes can be made in the preferred embodiment without departing from the principles of the disclosure. These and other changes in the preferred embodiment of the disclosure will be ϱ^ apparent to those skilled in the art from the disclosure herein, whereby it is to be distinctly understood that the foregoing descriptive matter is to be interpreted merely as illustration of the disclosure and not as a limitation. All patent applications / publications referred herein are incorporated by reference to the same extent as if each individual publication or patent application was specifically and individually ϭϬ^ indicated to be incorporated by reference. The following description includes information that may be useful in understanding the present invention. It is not an admission that any of the information provided herein is prior art or relevant to the presently claimed invention, or that any publication specifically or implicitly referenced is prior art. ϭϱ^ Technical Advantages: The lyophilized / freeze-dried viral vaccine composition / formulation and method of the present disclosure described herein above has several technical advantages including, but not limited to the following: x Applicant provides an improved high virus yielding egg based yellow fever vaccine ϮϬ^ manufacturing methods (optimized culture conditions, filtration, centrifugation / homogenization) & formulations. x Applicant’s method / formulation preserves the potency of virus (3 log IU / human dose of 0.5 ml). x Applicant’s manufacturing method reduces the residual egg derived Ϯϱ^ impurity / ovalbumin content in the range from about 0.08 to 0.163 μg / human dose of 0.5 ml, more preferably 0.087 μg / dose of 0.5 ml (recommended NMT than 5μg / human dose of 0.5 ml), protein nitrogen content in the range from about 0.09 to 0.22 mg / human dose of 0.5 ml, more preferably about 0.057 mg / human dose of 0.5 ml (recommended NMT than 0.25 mg / human dose), and the endotoxin content, ϯϬ^ thereby minimizing the probability of hypersensitivity / allergic reactions. x Applicant’s manufacturing method requires carrying out post-harvest homogenization, centrifugation, stabilization and filtration in a continuous operation. x Applicant’s manufacturing method provides high virus recovery & is cost effective since it is devoid of any genetically modified eggs / enzyme / benzonase / trypsin, ϱ^ surfactant / urea / glycerol & additional steps like sucrose gradient, protamine precipitation, DFF, TFF, or chromatography. x Applicant’s manufacturing method utilizes stepwise / cascade 3 step Direct Flow Filtration (DFF) through 5 μm, 1.2 μm and 0.22 μm filters in series for reducing aggregate load, assuring sterility & ensuring high virus recovery instead of Two step ϭϬ^ filtration (5 μm + 0.22 μm) or Two step filtration (1.2 μm + 0.22 μm). x Applicant’s composition / formulation provides minimum residual moisture content (not more than 3%) devoid of complex / costly / animal origin components like human serum albumin, trehalose, mannitol, lactose, sucrose, microcrystalline cellulose, hydroxyethyl starch, dextran, hetastarch, polymer, detergent, Pluronic, PVP, adjuvant, ϭϱ^ preservative, antibiotic. x Simple and effective method for inducing complete immune response to yellow fever virus infectious agent and meets the criterion for the seroprotection for the said immunogenic components comprising yellow fever virus in a single dose. x Improved immunological memory, and long-term memory cellular immune response. ϮϬ^ x The lyophilized presentation of the vaccine improves stability of the vaccine composition for longer periods at 2-8 deg C for 12 to 36 months; at 25 deg C for 2 to 6 months; at 37 deg C for 1 week to 4 weeks and the reconstituted vaccine preserves desired characteristics of a virus including bioactivity, potency, virus viability, immunogenicity and stability. Ϯϱ^ x The formulation is preferably a lyophilized formulation comprising of live attenuated yellow fever virus along with sorbitol, gelatin, histidine, alanine, tricine, arginine & lactalbumin hydrolysate and is stable and immunogenic, wherein the sorbitol preserves the structural integrity (native like structure) of virus and prevents aggregation of viruses. Gelatin prevents adsorption of the viruses onto the walls of the ϯϬ^ vessel and promotes desorption done previously & lactalbumin hydrolysate stabilizes proteins through a variety of mechanisms such as preferential hydration, direct binding, buffering, and antioxidation. x Said vaccine formulation is devoid of stabilizer like sucrose, surfactants like polysorbates. ϱ^ x The said formulation shows significant reduction in potency loss with respect to cryoprotection of virus during freeze-thaw cycle. x The method of preparing said formulation does not require use of enzymes, chromatography columns and antibiotics. ϭϬ^ EXAMPLES The following examples are included to demonstrate preferred embodiments of the invention. It should be appreciated by those of skill in the art that the compositions and techniques disclosed in the examples which follow represent techniques discovered by the inventor to function well in the practice of the invention, and thus can be considered to constitute ϭϱ^ preferred modes for its practice. However, those of skill in the art should, in light of the present disclosure, appreciate that many changes can be made in the specific embodiments which are disclosed and still obtain a like or similar result without departing from the spirit and scope of the invention. Yellow fever virus: The strain used for the development of live attenuated yellow fever virus ϮϬ^ vaccine is WHO yellow fever 17D vaccine virus primary seed (NIBSC code: 213-77) of passage level 237 sourced from National Institute for Biological Standards and Control (NIBSC), Potters Bar, England. The WHO seed was used to prepare master seed (P-238), working seed (P-239) and final product (YF17D vaccine, P-240). Ϯϱ^ Example 1: Details of final live attenuated yellow fever virus vaccine composition The following 4 vaccine compositions / formulations were prepared in accordance with the present disclosure. The method employed for manufacturing these compositions / formulations is provided in Example 2 below: Table 1: Yellow fever virus vaccine composition: 1 dose vial No. Components Yellow fever virus 1. Active component YF17D, P-240 Not less than 3.0 log10 IU / dose of 0.5 ml2. StabilizerGelatin - 0.1-5% (w / v)Sorbitol - 1-10% (w / v) Amino acid comprising tricine present at a concentration of 0.1% to 2% (w / v), L-histidine present at a concentration of 0.1% to 2% (w / v), L-alanine present at a concentration of 0.01% to 1% (w / v) and L-arginine hydrochloride present at a concentration of 0.1% to 5% (w / v) Lactalbumin hydrolysate - 0.1-2% Phosphate buffered saline (PBS) q.s. Table 2: Yellow fever virus vaccine composition: 1 dose vial No. Components Yellow fever virus 1. Active component YF17D, P-240 Not less than 3.0 log10IU / dose of 0.5 ml 2. Stabilizer Gelatin - 2.5% (w / v) Sorbitol - 5% (w / v) L-histidine - 0.21% (w / v) L-alanine - 0.1% (w / v) Tricine - 0.3% (w / v) L-arginine hydrochloride - 1.6% (w / v) Lactalbumin hydrolysate - 0.35% (w / v) Phosphate buffered saline (PBS) q.s. Table 3: Yellow fever virus vaccine composition: 5 dose vial No. Components Yellow fever virus 1. Active component YF17D, P-240 Not less than 3.0 log10IU / dose of 0.5 ml 2. Stabilizer Gelatin - 0.5% (w / v) Sorbitol – 1% (w / v) L-histidine - 0.04% (w / v) L-alanine - 0.02% (w / v) Tricine - 0.06% (w / v) L-arginine hydrochloride – 0.32% (w / v) Lactalbumin hydrolysate - 0.07% (w / v) Phosphate buffered saline (PBS) q.s. Table 4: Yellow fever virus vaccine composition: 10 dose vial Components Yellow fever virus Active component YF17D, P-240 Not less than 3.0 log 10IU / dose of 2. Stabilizer Gelatin - 0.5% (w / v) Sorbitol – 1% (w / v) L-histidine - 0.04% (w / v) L-alanine - 0.02% (w / v) Tricine - 0.06% (w / v) L-arginine hydrochloride – 0.32% (w / v) Lactalbumin hydrolysate - 0.07% (w / v) Phosphate buffered saline (PBS) q.s. Example 2: Method of manufacturing the lyophilized / freeze-dried vaccine composition / formulation Method of manufacturing a lyophilized / freeze-dried vaccine composition / formulation comprises of: ϱ^ a) The candidate vaccine virus (WHO yellow Fever 17D Primary Seed Virus, Passage No. 237, NIBSC Code: 213-77) is initially passaged in avian leukosis virus (ALV) free SPF embryonated hen eggs producing egg-based Master Seed Virus (MSV) of passage level 238; b) The egg-based Master Seed Virus (MSV) is further passaged in ALV free SPF ϭϬ^ embryonated hen eggs producing Working Seed Virus (WSV) of passage level 239; c) The Working Seed Virus (WSV) is sub cultured and propagated in ALV free SPF embryonated hen eggs to produce drug substance of passage level 240; d) The WSV is inoculated into live ALV free 6 to 9 days old embryonated SPF hen eggs at strength of 8000 to 16000 IU per egg; ϭϱ^ e) The inoculated eggs are incubated at 37 ± 1°C for 72 hours; f) The resulting virus infected embryos are harvested from live eggs and is decapitated; g) The embryos are washed with phosphate buffer saline (PBS; 2 ml per embryo); h) The washed embryos are homogenized in PBS (6 to 8 ml PBS per embryo) at 4000- 10000 rpm for 2-3 minutes to produce embryonic pulp; ϮϬ^ i) The embryonic pulp is centrifuged at RCF value of 8000 to 16000 x g for 20 minutes to separate tissue mass and collection of virus rich supernatant (embryo extract); j) The virus rich supernatant (embryo extract) is stabilized with stabilizers comprising sugar or sugar alcohol; amino acids; gelatin; and lactalbumin hydrolysate; k) The stabilized embryo extract is filtered by direct flow filtration (DFF) through Ϯϱ^ clarification filters of decreasing pore sizes in order of 5 μ > 1.2 μ to obtain clarified extract; l) The clarified extract is sterilized by DFF through at least one sterilization grade filter of pore size 0.22 μ to obtain a sterile drug substance (yellow fever single harvest) and storing of the drug substance at -60°C and below temperature; ϯϬ^ m) The stored drug substance is thawed and the volume is made up with diluent (Blind Vaccine) to achieve required dose of 0.5ml and again sterilized the blend by DFF through at least one sterilization grade filter; n) The sterile formulated drug substance (final bulk) is filled into individual sterile type-1 glass vials and the vials are partially stoppered under aseptic conditions with bromo- butyl rubber stoppers; and o) The mixture contained in the glass vials obtained in step (n) is subjected to freeze ϱ^ drying comprising the steps of freezing, sublimation and secondary drying, to obtain the lyophilized / freeze-dried viral vaccine composition / formulation of the present disclosure. The freeze-drying step for 1 dose and 5 dose vaccine comprises of: a) the freezing step - freezing the sterile formulated drug substance in the glass vial at a ϭϬ^ temperature of -55°C for 420 minutes; b) the sublimation step - Ramp 1: ramping the temperature of -55°C to a temperature of -18°C within 420 minutes at 100 μbar pressure and holding at the temperature of -18°C for 360 minutes at 100 μbar pressure; and ϭϱ^ Ramp 2: ramping the temperature of -18°C to a temperature of 0°C within 190 minutes at 100 μbar pressure and holding at the temperature of 0°C for 200 minutes at 100 μbar pressure; Ramp 3: ramping the temperature of 0°C to a temperature in a range of 26°C within 140 minutes at 100 μbar pressure and holding at the temperature of 26°C for 1 minutes ϮϬ^ at 100 μbar pressure, and c) the secondary drying step - ramping the temperature to 26°C within 1 minute and holding at the temperature of 26°C for 300 minutes at 25 μbar pressure. The freeze-drying step for 10 dose vaccine comprises of: a) the freezing step - freezing the sterile formulated drug substance in the glass vial at a Ϯϱ^ temperature of -55°C for 420 minutes; b) the sublimation step - Ramp 1: ramping the temperature of -55°C to a temperature of -18°C within 420 minutes at 100 μbar pressure and holding at the temperature of -18°C for 1600 minutes at 100 μbar pressure; and ϯϬ^ Ramp 2: ramping the temperature of -18°C to a temperature of -4°C within 90 minutes at 100 μbar pressure and holding at the temperature of -4°C for 500 minutes at 100 μbar pressure; Ramp 3: ramping the temperature of -4°C to a temperature in a range of 26°C within 300 minutes at 100 μbar pressure and holding at the temperature of 26°C for 1 minutes at 100 μbar pressure. c) the secondary drying step - ramping the temperature to 26°C within 1 minute and ϱ^ holding at the temperature of 26°C for 420 minutes at 25 μbar pressure. Example 3: Selection of Stabilizer for inclusion in the vaccine composition / formulation of the present disclosure Stability studies were conducted on a live attenuated yellow fever vaccine with a combination ϭϬ^ of stabilizer chosen based on the process knowledge in order to select the best combination empirically. Stabilizers were added to live attenuated viruses for enhancing the virus stability. Gelatin Sorbitol (GS) stabilizer system and Sucrose Glycine (SG) stabilizer system have been used in Serum Institute of India Pvt. Ltd. for viral vaccine production and hence, were considered for stability evaluation for yellow fever vaccine virus. As the efficacy depends on ϭϱ^ their ability for thermal stability, yellow fever vaccine (drug product) with two formulations - one each with Gelatin Sorbitol (GS) stabilizer system and Sucrose Glycine (SG) stabilizer system - were exposed at 37 °C for 2 weeks and each combination was tested for vaccine potency. And at the end of the exposure period, the virus potency of drug product should not have decreased by more than 1.0 log10. ϮϬ^ Gelatin Sorbitol (GS) stabilizer Formulation 1 contains 2.5% hydrolysed gelatin, 5.0% sorbitol, 0.21% L-Histidine, 0.1% L-Alanine, 0.3% Tricine, 1.6% L-Arginine and 0.35% Lactalbumin hydrolysate. Sucrose Glycine (SG) stabilizer Formulation 2 contains 5 % sucrose and 5 % Glycine. Table 5: Thermostability results Average potency loss at Details Stabilizer System 37°C for 14 days (log10IU) Yellow Fever Vaccine Sucrose Glycine stabilizer 1.07 YF17D, P-240 (Drug Product) Gelatin Sorbitol stabilizer 0.468 Ϯϱ^ Conclusion: Formulation with SG stabilizer system has unacceptable potency loss during thermal stability as the virus potency decreased by more than 1.0 log10while, formulation with GS stabiliser system showed potency loss significantly less than 1.0 log10.Hence, Gelatin Sorbitol (GS) stabilizer system was selected as a potential stabilizer system candidate ϱ^ over Sucrose Glycine (SG) stabilizer system. Example 4: Cryoprotection study for Yellow Fever Single Harvest (Drug Substance) Yellow Fever Single Harvest (YF-SH) formulations were prepared using different stabilizer components and were studied for potency loss during freezing and thawing steps. Three types ϭϬ^ of stabilizers [2-component (2C) stabilizer, 3-component (3C) stabilizer and 7-component (7C) stabilizer] formulations were frozen at -60°C temperature in deep freezer. Post completion of 7 days storage in deep freezer, all the YF-SH formulations (2C, 3C and 7C) were thawed and were checked for potency loss during freeze-thaw cycle. ϭϱ^ Table 6: Results Sr. YF-SH Pre-freezing Post-thawing Freeze-Thaw No. Formulation Details potency potency potency loss (log10IU / ml) (log10IU / ml) (log10IU / ml) Gelatin, Sorbitol, Histidine, 7C b Alanine, Tricine, 1 ased formulation Arginine and 6.019 5.950 0.069 LAH (as provided in table 2 above) Gelatin, Sorbitol, Alanine 2 3C based (respective formulation concentrations are 5.890 5.647 0.243 as provided in table 2 above) Gelatin and Sorbitol 3 2C based (respective formulation concentrations are 5.905 5.616 0.289 as provided in table 2 above) Conclusion: During freezing and thawing process, no virus loss was observed for 7C based YF-SH formulation while, 3C and 2C based YF-SH formulations showed virus loss of 0.243 and 0.289 log10IU / ml respectively. Hence, primarily it can be inferred that 7-component (7C) ϱ^ (Table 2 formulation) stabilizer is better with respect to cryoprotection of virus during freeze- thaw cycle. Example 5: Stability & Immunogenicity results for Yellow Fever Vaccine Formulation - Stability data of yellow fever vaccine formulation (Drug Product) at different ϭϬ^ temperatures (2-8°C, 25°C and 37°C): Following are stability results at different temperatures and time intervals. Table 7: Stability study data At 2-8°C for 36 Months: 0 Month 12 Months 24 Months 36 Months Yellow Fever (log10IU / (log10IU / (log1IU / (log IU / Potency loss Vaccine0 100.5 ml) 0.5 ml) 0.5 ml) 0.5 ml) (log10IU) YF17D, P-240 (Drug Product) 4.214 3.830 3.770 3.688 0.562 At 25°C for 6 Months: Yellow Fever 0 Month (log IU / 0.5 ml) 6 Months (log10IU / 0.5 Potency loss Vaccine10ml) (log10IU) YF17D, P-240 (Drug Product) 4.008 3.431 0.577 At 37°C for 14 days: Yellow Fever 0 Day (log accine10IU / 0.5 ml) 14 days (log1IU / 0.5 ml) Potency loss V0(log10IU) YF17D, P-240 (Drug Product) 4.008 3.585 0.423 ^ Accordingly, the Log10IU bioactivity or virus concentration of the reconstituted vaccine composition is reduced only by about 0.5 log than the original bioactivity after storage at 2- ϭϱ^ 8°C for 36 months or after storage at 25°C for 6 months or 37°C for 14 days, in comparison with the bioactivity of a freshly reconstituted vaccine composition. Immunogenicity Results: A Phase I study was conducted to evaluate the safety and immunogenicity in healthy adult volunteers. And YF-neutralizing antibodies were measured at baseline and at intervals. There was a significant increase in geometric mean titer (GMTs) post vaccination as ϱ^ compared to the baseline on Day 28. Seroconversion was reported in all participants on Day 28, except 1 participant in the SII YFV (SC) group. All participants reported seroprotection on Day 28. Table 8: Immunogenicity Result details Groups SII YFV (SC) SII YFV (IM) N 18 18 GMT (95% CI) Day 1 5.49 (4.51, 6.69) 6.62 (4.64, 9.46)  Day 28 7022.68 (2925.21, 16859.64) 14468.62 (10259.31, 20404.98) Seroconversion N (%) (95% CI)  Day 28 17 (94.44) (72.71, 99.86) 18 (100) (81.47, 100) Seroprotection N (%) (95% CI) Day 1 1 (5.56) (0.14, 27.29) 3 (16.67) (3.58, 41.42)  Day 28 18 (100) (81.47, 100) 18 (100) (81.47, 100) Note: N is number of participants, CI is confidence intervals IM-Intra-muscular injection; SC-Subcutaneous injection ϭϬ^ Interpretation: The new YFV was found safe and immunogenic by IM as well as SC routes. The foregoing description of the specific embodiments fully reveals the general nature of the embodiments herein that others can, by applying current knowledge, readily modify and / or ϭϱ^ adapt for various applications such specific embodiments without departing from the generic concept, and, therefore, such adaptations and modifications should and are intended to be comprehended within the meaning and range of equivalents of the disclosed embodiments. It is to be understood that the phraseology or terminology employed herein is for the purpose of description and not of limitation. Therefore, while the embodiments in this disclosure have ϮϬ^ been described in terms of preferred embodiments, those skilled in the art will recognize that the embodiments herein can be practiced with modification within the spirit and scope of the embodiments as described herein.

Claims

We claim:

1. A lyophilized or freeze-dried viral vaccine composition, comprising: a) one or more virus particle or antigen thereof; and b) stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; ϱ^ lactalbumin hydrolysate and gelatin.

2. The vaccine composition according to claim 1, wherein the virus is poxvirus selected from a group comprising orthopoxviruses and avipoxviruses, morbillivirus or measles virus, mumps virus, rubella virus, alphavirus selected from a group comprising sendai virus, ϭϬ^ sindbis virus and semliki forest virus (SFV), ross river virus, encephalitis virus, flavivirus selected from a group comprising yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus and Zika virus, rhabdovirus or vesicular stomatitis virus (VSV), retrovirus or RNA tumor virus, adenovirus selected from ϭϱ^ a group comprising human adenovirus, bovine adenovirus, canine adenovirus, non-human primate adenovirus, chicken adenovirus, porcine adenovirus and swine adenovirus, adeno- associated viruses, lentivirus selected from a group comprising human immunodeficiency viruses (HIV), simian immunodeficiency virus (SIV), and feline immunodeficiency virus (FIV), herpes simplex virus, cytomegalovirus, picornavirus selected from a group ϮϬ^ comprising Rhinovirus and Poliovirus, baculovirus vector or autographacalifornica multiple nucleopolyhedrovirus (AcMNPV), hepatitis B virus (HBV), rubulavirus or new castle disease virus, parainfluenza virus, influenza virus, respiratory syncytial virus (RSV), human metapneumovirus (hMPV), respiratory Coronavirus (CoV), Ebola virus, Marburg virus, Nipah virus, Chikungunya virus, Rotavirus, Human papilloma virus, Ϯϱ^ Herpes simplex virus, Hepatitis A virus, Hepatitis C virus, Hepatitis B virus, Hepatitis E virus, Poliovirus, Variola virus selected from a group comprising smallpox and monkeypox virus, and Varicella virus antigens.

3. The vaccine composition according to any one of claims 1 to 2, wherein the virus is ϯϬ^ flavivirus selected from a group comprising yellow fever virus, dengue virus, Japanese encephalitis (JE) virus, Kunjin virus, West Nile (WN) virus, tick-borne encephalitis (TBE) virus, St. Louis encephalitis virus, Murray Valley encephalitis virus, and Zika virus.

4. The vaccine composition according to any one of claims 1 to 3, wherein the virus is a live attenuated virus (LAV), an inactivated virus, a chimeric virus, or a recombinant virus.

5. The vaccine composition according to any one of claims 1 to 4, wherein the virus is a live ϱ^ attenuated yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml or 1000 IU or 1000 virus particles.

6. The vaccine composition according to claim 5, wherein strain of the yellow fever virus is selected from 17D, 17D-204, 17D-213, or 17DD. ϭϬ^ 7. The vaccine composition according to any one of claims 1 to 6, wherein residual egg derived impurity or ovalbumin content is in the range from 0.08 to 0.163 μg / 0.5 ml of the vaccine, and protein nitrogen content is in the range from 0.09 to 0.22 mg / 0.5 ml of the vaccine to minimize hypersensitivity / allergic reaction. ϭϱ^ 8. The vaccine composition according to any one of claims 1 to 7, wherein the stabilizer comprises of one or more sugar or sugar alcohol selected from a group comprising glucose, sucrose, maltose, lactose, fructose, galactose, mannose, maltulose, iso- maltulose, lactulose, mannitol, trehalose, raffinose, lactitol, lactobionic acid, sorbitol, dextrose, ϮϬ^ fructose, glycerol, and fucose, or any combination thereof, present at a concentration ranging from about 1 to 20% (w / v).

9. The vaccine composition according to claim 8, wherein the sugar or sugar alcohol is sorbitol present at a concentration of about 5% (w / v). Ϯϱ^ 10. The vaccine composition according to any one of claims 1 to 9, wherein the stabilizer comprises of one or more amino acid selected from a group comprising tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, ϯϬ^ Methionine, Proline, Serine and Threonine, or any combination thereof, present at a concentration ranging from about 0.01-10% (w / v).

11. The vaccine composition according to claim 10, wherein the composition comprises a combination of amino acids tricine present at a concentration of about 0.1% to 2% (w / v),L-histidine present at a concentration of about 0.1% to 2% (w / v), L-alanine present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride present at a concentration of about 0.1% to 5% (w / v). ϱ^ 12. The vaccine composition according to any one of claims 1 to 11, wherein the stabilizer comprises of gelatin at a concentration ranging from about 0.1% to 10% (w / v) and lactalbumin hydrolysate at a concentration ranging from about 0.05% to 2% (w / v).

13. The vaccine composition according to claim 12, wherein the lactalbumin hydrolysate and ϭϬ^ gelatin are obtained by chemical, enzymatic or thermal hydrolysis of protein from either plant or animal sources.

14. The vaccine composition according to any one of claims 1 to 13, wherein the composition further comprises of an adjuvant selected from a group comprising aluminum hydroxide, ϭϱ^ aluminum phosphate, aluminum hydroxyphosphate, and potassium aluminum sulfate or a mixture thereof.

15. The vaccine composition according to any one of claims 1 to 14, wherein the composition further comprises of an immunostimulatory component selected from a group comprising ϮϬ^ an oil and water emulsion, MF-59, a liposome, a lipopolysaccharide, a saponin, lipid A, lipid A derivatives, Monophosphoryl lipid A, 3–deacylated monophosphoryl lipid A, AS01, AS03, an oligonucleotide, an oligonucleotide comprising at least one unmethylated CpG and / or a liposome, Freund’s adjuvant, Freund’s complete adjuvant, Freund’s incomplete adjuvant, CRL-8300 adjuvant, muramyl dipeptide, TLR-4 agonists, Ϯϱ^ flagellin, flagellins derived from gram negative bacteria, TLR-5 agonists, fragments of flagellins capable of binding to TLR-5 receptors, QS-21, ISCOMS, Matrix M, dmLT, Transferrin-Binding Protein, fHbp, Chitosan and saponin in combination with sterols and lipids, or any combination thereof. ϯϬ^ 16. The vaccine composition according to any one of claims 1 to 15, wherein the composition further comprises of a pharmaceutically acceptable additive selected from a group comprising transporter, excipient, binder, carrier, isotonic agent, emulsifier and humectant, or any combination thereof.

17. The vaccine composition according to claim 16, wherein the excipient is selected from a group comprising salt including NaCl, KCl, KH2PO4, Na2HPO4.2H2O, CaC12, and MgCl2; the non-ionic surfactant is selected from a group comprising polysorbate 20, polysorbate 40, polysorbate 60, polysorbate 65, polysorbate 80, polysorbate 85, ϱ^ nonylphenoxypolyethoxethanol, octylphenoxypolyethoxethanol, oxtoxynol 40, nonoxynol- 9, triethanolamine, triethanolamine polypeptide oleate, polyoxyethylene- 660 hydroxystearate, polyoxyethylene- 35 ricinoleate, soy lecithin, alkyl poly(ethylene oxide), copolymers of poly(ethylene oxide) and poly(propylene oxide) (EO-PO block copolymers), poly(vinyl pyrroloidone), alkyl polyglucosides (such as sucrose ϭϬ^ monostearate, lauryl diglucoside, or sorbitan monolaureate, octyl glucoside and decyl maltoside), fatty alcohols (cetyl alcohol or olelyl alcohol), or cocamides (cocamide MEA, cocamide DEA and cocamide TEA and a poloxamer - 0.001%-0.05%; polymers including dextran, carboxymethylcellulose, hyaluronic acid, cyclodextrin, Pluronic F127, Pluronic F68, Pluronic P123, and EO-PO block copolymers of greater than 3,000-4,000 MW, or ϭϱ^ any combination thereof.

18. The vaccine composition according to any one of claims 1 to 17, wherein the lyophilized or freeze-dried viral vaccine composition is reconstituted with an aqueous solution selected from a group comprising saline, buffer and water for injection (WFI), or any ϮϬ^ combination thereof, prior to its administration or use in vaccination.

19. The vaccine composition according to claim 18, wherein the buffer is selected from a group comprising sodium chloride, acetate, carbonate, citrate, lactate, gluconate, tartrate, phosphate buffer saline, borate, histidine buffer, succinate buffer, HEPES, TRIS and Ϯϱ^ Citrate-phosphate, or any combination thereof.

20. The vaccine composition according to claim 18, wherein pH of the reconstituted composition is in the range of pH 6.0 to 7.

5. ϯϬ^ 21. The vaccine composition as claimed in claim 1, comprising: a) live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; b) sorbitol at a concentration of about 1 to 20% (w / v);c) tricine at a concentration of about 0.1% to 2% (w / v), L-histidine at a concentration of about 0.1% to 2% (w / v), L-alanine at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride at a concentration of about 0.1% to 5% (w / v); d) gelatin at a concentration of about 0.1% to 10% (w / v); and ϱ^ e) lactalbumin hydrolysate at a concentration of about 0.05% to 2% (w / v).

22. The vaccine composition as claimed in claim 21, comprising: a) live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; ϭϬ^ b) sorbitol at a concentration of about 5% (w / v); c) tricine present at a concentration of about 0.3% (w / v), L-histidine at a concentration of about 0.21% (w / v), L-alanine at a concentration of about 0.1% (w / v) and L-arginine hydrochloride at a concentration of about 1.6% (w / v); d) gelatin at a concentration of about 2.5% (w / v); and ϭϱ^ e) lactalbumin hydrolysate at a concentration of 0.35% (w / v).

23. The vaccine composition according to claim 18, wherein the reconstituted composition retains the desired characteristics of bioactivity, potency, stability and immunogenicity for at least a period of about 14 days to about 36 months; and wherein Log10IU bioactivity or ϮϬ^ virus concentration of the reconstituted vaccine composition: is reduced only by about 0.5 log after storage at 2-8°C for 36 months or after storage at 25°C for 6 months or 37°C for 14 days, in comparison with the bioactivity of a freshly reconstituted vaccine composition. Ϯϱ^ 24. A method of manufacturing the lyophilized or freeze-dried viral vaccine composition according to claim 1, the method comprising: a) subjecting at least one live attenuated flavivirus to at least one passaging in an avian leukosis virus (ALV) free embryonated specific pathogen-free (SPF) hen eggs to produce flavivirus drug substance; ϯϬ^ b) inoculating the drug substance into live ALV free embryonated SPF hen eggs and incubating for a period of abou 24 hours to 120 hours at 37 ± 1°C, preferably about 60-80 hours or about 68-76 hours; c) harvesting of the virus infected embryos; ;^d) washing and homogenizing of the harvested embryos with phosphate buffer saline to produce embryonic pulp; e) centrifuging of the embryonic pulp and collecting supernatant comprising the virus; f) stabilizing the collected extract with a stabilizer comprising sugar or sugar alcohol; ϱ^ amino acids; gelatin; and lactalbumin hydrolysate; g) clarifying the stabilized extract by direct flow filtration (DFF) through at least one clarification filter to obtain a clarified extract; h) sterilizing the clarified extract by DFF through at least one sterilization grade filter and storing the sterile extract or the drug substance under sub-zero temperature; ϭϬ^ i) thawing of the drug substance and making up the volume with diluent to achieve a predetermined dose, followed by re-sterilizing the blend by DFF through at least one sterilization grade filter; j) filling the sterile drug substance into a container; and k) freeze drying the drug substance containing container obtained in step (j), comprising ϭϱ^ the steps of freezing, sublimation and secondary drying to obtain the said lyophilized or freeze-dried viral vaccine composition.

25. The method according to claim 24, wherein the live attenuated flavivirus is subjected to at least 238 or 239 or 240 passages prior to the subsequent stem of inoculation. ϮϬ^ 26. The method according to any one of claims 24 to 25, wherein the drug substance is inoculated into live ALV free embryonated SPF hen eggs for a period of about 7 to 8 days at strength of about 8000 to 16000 IU per egg; and wherein the post inoculation incubation is carried out for a period of about 72 hours at 37 ± 1°C. Ϯϱ^ 27. The method according to any one of claims 24 to 26, wherein the washing is carried out with phosphate buffer saline; the homogenization is carried out at about 2000 to 12000 rpm for a period of about 2-3 minutes to produce the embryonic pulp; and wherein the pulp is subjected to centrifugation at about 8000 to 16000 rpm for a period of about 20 ϯϬ^ minutes.

28. The method according to any one of claims 24 to 26, wherein the stabilized embryo extract is clarified and sterilized through filters of decreasing pore sizes in order of 5 μ, 1.2 μ and 0.22 μ to obtain the sterilized drug substance.

29. The method according to any one of claims 24 to 28, wherein the embryo extract is stabilized with stabilizer comprises: a) sugar or sugar alcohol or sorbitol at a concentration of about 1 to 20% (w / v); ϱ^ b) amino acid or tricine at a concentration of about 0.1% to 2% (w / v), L-histidine present at a concentration of about 0.1% to 2% (w / v), L-alanine present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride present at a concentration of about 0.1% to 5% (w / v); c) gelatin at a concentration of about 0.1% to 5% (w / v); ϭϬ^ d) and lactalbumin hydrolysate at a concentration of about 0.05% to 2% (w / v).

30. The method according to claim 24, wherein the freeze-drying comprises: a) a freezing step comprising of: freezing the drug substance containing container at a temperature in a range of about - ϭϱ^ 40°C to -55°C for about 300 minutes to 500 minutes; and b) a sublimation step comprising of: Ramp 1: ramping the temperature of about -55°C to a temperature in a range of about - 18°C to -28°C within about 300 minutes to 500 minutes at about 100 μbar pressure and holding at the temperature of about -18°C to -28°C for 300 minutes to about 1600 ϮϬ^ minutes at about 100 μbar pressure; and Ramp 2: ramping the temperature of about -18°C to -28°C to a temperature in a range of about -4°C to 0°C within about 90 minutes to 300 minutes at about 100 μbar pressure and holding at the temperature of about -4°C to 0°C for about 200 minutes to 500 minutes at about 100 μbar pressure; Ϯϱ^ Ramp 3: ramping the temperature of about -4°C to 0°C to a temperature in a range of about 26°C within about 100 minutes to 300 minutes at about 100 μbar pressure and holding at the temperature of about 26°C for about 1 to 5 minutes at about 100 μbar pressure, and c) a secondary drying step comprising of: ramping the temperature to about 26°C within ϯϬ^ about 1 to 5 minutes and holding at the temperature of about 26°C for about 300 minutes to 500 minutes at about 25 μbar pressure.

31. The method according to any one of claims 24 to 30, wherein the flavivirus is a live attenuated yellow fever virus; and wherein strain of the yellow fever virus is selected from strain 17D, 17D-204, 17D-213, or 17DD. ϱ^ 32. The method according to claim 24, wherein the method allows the vaccine composition to retain Log10IU bioactivity or virus concentration of the vaccine composition after storage of the composition at 2-8°C for 36 months or after storage at 25°C for 6 months or 37°C for 14 days. ϭϬ^ 33. A kit comprising: a) a lyophilized or freeze-dried viral vaccine composition comprising: at least one live attenuated virus present at a dose of not less than 3 log10IU per 0.5 ml of the composition or 1000 IU or 1000 virus particles; one or more sugar or sugar alcohol at a concentration ranging from about 1 to 20% (w / v); one or more amino acid selected ϭϱ^ from a group comprising tricine, leucine, iso-leucine, L-histidine, glycine, glutamine, L-arginine, L-arginine hydrochloride, lysine, L-alanine, Tryptophan, Phenylalanine, Tyrosine, Valine, Cysteine, Glycine, Histidine, Methionine, Proline, Serine and Threonine, or any combination thereof, present at a concentration ranging from about 0.01-10% (w / v); lactalbumin hydrolysate at a concentration ranging from about 0.05% ϮϬ^ to 2% (w / v); and gelatin at a concentration ranging in between 0.1% and 10% (w / v); and b) an aqueous solution selected from a group comprising saline, buffer and water for injection (WFI), or any combination thereof, for reconstituting the lyophilized or freeze-dried viral vaccine composition prior to its administration or use as vaccination. Ϯϱ^ 34. The kit as claimed in claim 33, wherein the virus is a live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml or 1000 IU or 1000 virus particles; the sugar or sugar alcohol is sorbitol present at a concentration of about 5% (w / v); and the amino acid is present as a combination of amino acids such that tricine ϯϬ^ is present at a concentration of about 0.1% to 2% (w / v), L-histidine is present at a concentration of about 0.1% to 2% (w / v), L-alanine is present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride is present at a concentration of about 0.1% to 5% (w / v).

35. The kit as claimed in any one of claims 33 to 34, wherein the composition and the aqueous solution are contained in a single or different containers selected from a group comprising a bottle, a vial, an ampule, an IV bag, a wearable injector, a bolus injector, a pre-filled syringe, a pen, a pump, a multidose needle syringe, a multidose vial, a multidose ϱ^ pen, a syrette, an auto-injector or a Vaccine Microarray Patches; wherein when the composition and the aqueous solution are contained in a single container, the container comprises a partition or mechanism to prevent the composition and the aqueous solution from being mixed, as part of the kit. ϭϬ^ 36. A method of preventing a disease condition caused by a virus in a subject, or a method of vaccinating the subject against the virus, said method comprising administering to the subject a reconstituted viral vaccine composition, said composition comprising: a. one or more virus particle or antigen thereof; and b. stabilizer comprising one or more sugar or sugar alcohol; one or more amino acid; ϭϱ^ lactalbumin hydrolysate and gelatin; and wherein the composition is reconstituted in an aqueous solution selected from a group comprising saline, buffer and water for injection (WFI), or any combination thereof, prior to its administration or use in vaccination. ϮϬ^ 37. The method as claimed in claim 36, wherein the virus is a live attenuated flavivirus or yellow fever virus present at a dose of not less than 3 log10IU per 0.5 ml or 1000 IU or 1000 virus particles; the sugar or sugar alcohol is sorbitol present at a concentration of about 5% (w / v); and the amino acid is present as a combination of amino acids such that tricine is present at a concentration of about 0.1% to 2% (w / v), L-histidine is present at a Ϯϱ^ concentration of about 0.1% to 2% (w / v), L-alanine is present at a concentration of about 0.01% to 1% (w / v) and L-arginine hydrochloride is present at a concentration of about 0.1% to 5% (w / v).

38. The method as claimed in claim 36, wherein the reconstituted viral vaccine composition is ϯϬ^ administered to a human subject via parenteral route selected from subcutaneous or intramuscular administration.

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