Arenavirus formulations, methods and uses thereof

Abstract

The present disclosure relates to improved liquid and lyophilized arenavirus formulations, and pharmaceutical compositions derived thereof. The present disclosure further relates to methods of making such arenavirus formulations, and using such arenavirus formulations and pharmaceutical compositions derived thereof for treating or preventing diseases or disorders (e.g., cancers and infectious diseases).

Description

ARENAVIRUS FORMULATIONS, METHODS AND USES THEREOFCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to United States Provisional Application No. 63 / 582,913 filed September 15, 2023, the content of which is incorporated by reference in its entirety herein.SEQUENCE LISTING

[0002] This application contains a computer readable Sequence Listing which has been submitted in XML file format with this application, the entire content of which is incorporated by reference herein in its entirety. The Sequence Listing XML file submitted with this application is entitled “13194-086-228_SEQLISTING.xml”, was created on September 11, 2024, and is 54,146 bytes in size.1. FIELD

[0003] The present disclosure relates to improved liquid and lyophilized arenavirus formulations, and pharmaceutical compositions comprising thereof. The present disclosure further relates to methods of using said arenavirus formulations and pharmaceutical compositions comprising thereof for treating or preventing diseases.2. BACKGROUND

[0004] One main challenge in developing engineered arenavirus vector-based therapeutics is the design of virus formulations to ensure optimal vector safety, efficacy, and stability. To maintain arenavirus vector activity during purification and storage, conditions and excipients of the formulations need to be thoroughly selected to stabilize viral particles against mechanical or thermal stress (e.g., filtration steps or freeze / thaw cycles), and to avoid loss of vector infectivity through inactivation or aggregation of viral particles and / or non-specific binding to contact materials used during the purification process. The buffer conditions have to be carefully adapted throughout the manufacturing process to provide a stabilizing environment with minimal interactions among viral particles.

[0005] Thus, there exist needs for improved virus formulations that maintain stability, infectivity and viral titer during manufacturing, purification, filtration, and storage.3. SUMMARY

[0006] The present disclosure provides improved arenavirus formulations that significantly increase arenavirus based viral vector stability in frozen and liquid state and greatly reduce the formation of vector-derived aggregates and subvisible particles, thereby facilitating filterability and increasing viral recovery during final filtration steps. In particular, as compared to the previously known formulations, the presently disclosed arenavirus formulations advantageously keep the viral vectors dispersed and thereby significantly reduce substantial filtration losses (in particular during sterile filtration) seen with many enveloped vectors. In addition, as particle formation is commonly seen in the viral vector field, minimizing particle (aggregate) formation by the presently disclosed formulations is considered an advantage with regard to product safety. Finally, increasing the stability of the formulation when stored refrigerated (2°C - 8°C) and at ambient conditions (e.g., 20°C - 25°C) by the presently disclosed formulations allows a broader use and intermediate storage of the arenavirus in hospitals and regions lacking ultralow freezer equipment. The stability at -20°C storage could also be increased by the use of appropriate carbohydrates disclosed herein. Furthermore, the presently disclosed liquid formulations can use excipients that are compatible with freeze-drying, which further increases storage stability at elevated temperatures.

[0007] In one aspect, the present disclosure provides a liquid arenavirus formulation comprising: (a) an arenavirus; and (b) a polyvalent anion at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

[0008] In certain embodiments, the polyvalent anion is at a concentration of about 50 mM or about 25 mM. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

[0009] In one aspect, the present disclosure provides a liquid arenavirus formulation comprising: (a) an arenavirus; and (b) a polyvalent anion, wherein the polyvalent anion is at a concentration that (i) reduces the formation of subvisible particles during manufacturing, and / or (ii) reduces the loss of viral titer during filtration, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process.

[0010] In certain embodiments, the polyvalent anion is at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

[0011] In certain embodiments, the polyvalent anion is at a concentration of about 50 mM or about 25 mM. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

[0012] In certain embodiments, the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.

[0013] In certain embodiments, the polyvalent anion is citrate. In certain embodiments, the citrate is at a concentration of about 50 mM or about 25 mM. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and the citrate is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

[0014] In certain embodiments, the liquid arenavirus formulation further comprises at least one of: (c) a carbohydrate; (d) a salt; (e) a buffer; (f) a surfactant; and (g) a stabilizer.

[0015] In certain embodiments, (c) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v); (d) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM; (e) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM; (f) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or (g) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v), optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).

[0016] In certain embodiments, (c) the carbohydrate is at a concentration of about 5% (w / v); (d) the salt is at a concentration of about 125 mM or about 75 mM; (e) the buffer is at a concentration of about 10 mM; (f) the surfactant is at a concentration of about 0.03% (w / v); and / or (g) the stabilizer is at a concentration of about 0.1% (w / v).

[0017] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and (c) the carbohydrate is at a concentration of about 5% (w / v); (d) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (e)the buffer is at a concentration of about 10 mM; (f) the surfactant is at a concentration of about 0.03% (w / v); and / or (g) the stabilizer is at a concentration of about 0.1% (w / v).

[0018] In certain embodiments, (c) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or a combination thereof; (d) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride; (e) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS; (f) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is pol oxamer 188; and / or (g) the stabilizer is selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the stabilizer is rHSA.

[0019] In certain embodiments, (c) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v); (d) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM; (e) the buffer is TRIS at a concentration of about 10 mM; (f) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or (g) the stabilizer is rHSA at a concentration of about 0.1%.

[0020] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and: (c) the carbohydrate is trehalose at a concentration of about 5% (w / v); (d) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (e) the buffer is TRIS at a concentration of about 10 mM; (f) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or (g) the stabilizer is rHSA at a concentration of about 0.1%.

[0021] In certain embodiments, the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0. In certain embodiments, the pH is about 8.0.

[0022] In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality ofabout 530 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0023] In certain embodiments, (c)-(g) are present at concentrations to reduce the loss of viral titer during filtration, freezing and thawing, and / or storage; and / or reduce the formation of subvisible particles during manufacturing, freezing and thawing, and / or storage.

[0024] In certain embodiments, the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C and about 8°C, above about 15°C, about - 20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.

[0025] In one aspect, the present disclosure provides a liquid arenavirus formulation comprising: (a) an arenavirus; (b) citrate at a concentration of about 50 mM; (c) sucrose at a concentration of about 5% (w / v); (d) NaCl at a concentration of about 125 mM; (e) TRIS at a concentration of about 10 mM; (f) poloxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

[0026] In one aspect, the present disclosure provides a liquid arenavirus formulation comprising: (a) an arenavirus; (b) citrate at a concentration of about 25 mM; (c) trehalose at a concentration of about 5% (w / v); (d) NaCl at a concentration of about 75 mM; (e) TRIS at a concentration of about 10 mM; (f) poloxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

[0027] In one aspect, the present disclosure provides a liquid arenavirus formulation suitable for lyophilization comprising: (a) an arenavirus; (b) citrate at a concentration of between about 20 mM and about 50 mM;(c) trehalose at a concentration of about 5% (w / v); (d) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (e) TRIS at a concentration of about 10 mM; (f) poloxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0028] In certain embodiments, the concentration of the arenavirus is between about 102RCV-FFU / ml and about IO10RCV-FFU / ml, or between about 2* 105RCV-FFU / ml and about 5* 108RCV-FFU / ml.

[0029] In certain embodiments, the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.

[0030] In certain embodiments, the arenavirus is a genetically engineered arenavirus.

[0031] In certain embodiments, the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of a tumor antigen, a tumor associated antigen, an antigen of a pathogen that causes an infectious disease, or an antigenic fragment of any of the foregoing. In certain embodiments, the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.

[0032] In certain embodiments, the antigen comprises a cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of a CMV glycoprotein gB with a truncation of the carboxy-terminus (i.e., gB(dCt)), a CMV tegument protein pp65, and combinations thereof. In certain embodiments, the antigen comprises a human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7 / E6 fusion protein. In certain embodiments, the antigen comprises a prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof. In certain embodiments, the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide and combinations thereof. In certain embodiments, the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef. In certain embodiments, the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the at least one antigenic fragment from N- to C-terminus comprise the mutations G13D, G12V, G12C, G12D, and G12R respectively. In certain embodiments, the antigen comprises two or more cancer testis antigen (CTA) fragments (e.g., antigenic fragments or epitopes). In certain embodiments, the two or more CTA fragmentsare selected from the group consisting of epitopes or antigenic fragments of preferentially expressed antigen in melanoma (PRAME), melanoma-associated antigen 3 (MAGE-A3), melanoma-associated antigen 4 (MAGE-A4), and melanoma-associated antigen 6 (MAGE- A6).

[0033] In certain embodiments, the arenavirus is a non-replicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles. In certain embodiments, the arenavirus is a non-replicating arenavirus, and the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen. In certain embodiments, the arenavirus is a replicating tri-segmented arenavirus. In certain embodiments, the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF. In certain embodiments, the tri- segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3’ UTR. In certain embodiments, the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.

[0034] In one aspect, the present disclosure provides a lyophilized arenavirus formulation prepared from the presently disclosed liquid arenavirus formulation.

[0035] In one aspect, the present disclosure provides a method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

[0036] In certain embodiments, the polyvalent anion is at a concentration of about 50 mM or about 25 mM.

[0037] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.

[0038] In one aspect, the present disclosure provides method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at aconcentration that reduces the formation of subvisible particles during manufacturing, and / or reduces the loss of viral titer during filtration.

[0039] In certain embodiments, the polyvalent anion is at a concentration of between about 5 mM and about 150 mM. In certain embodiments, the polyvalent anion is at a concentration of about 50 mM or about 25 mM.

[0040] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.

[0041] In certain embodiments, the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.

[0042] In certain embodiments, the polyvalent anion is citrate. In certain embodiments, the citrate is at a concentration of about 50 mM or about 25 mM. In certain embodiments, the citrate is at a concentration of between about 10 mM and about 60 mM.

[0043] In certain embodiments, the liquid arenavirus formulation further comprises at least one of: (b) a carbohydrate; (c) a salt; (d) a buffer; (e) a surfactant; and (f) a stabilizer.

[0044] In certain embodiments, (b) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v); (c) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM; (d) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM; (e) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or (f) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v) , optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).

[0045] In certain embodiments, (b) the carbohydrate is at a concentration of about 5% (w / v); (c) the salt is at a concentration of about 125 mM or about 75 mM; (d) the buffer is at a concentration of about 10 mM; (e) the surfactant is at a concentration of about 0.03% (w / v); and / or (f) the stabilizer is at a concentration of about 0.1% (w / v).

[0046] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and (b) the carbohydrate is at a concentration of about 5% (w / v); (c) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (d)the buffer is at a concentration of about 10 mM; (e) the surfactant is at a concentration of about 0.03% (w / v); and / or (f) the stabilizer is at a concentration of about 0.1% (w / v).

[0047] In certain embodiments, (b) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or a combination thereof; (c) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride; (d) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS; (e) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is poloxamer 188; and / or (f) the stabilizer is a protein selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the protein is rHSA.

[0048] In certain embodiments, (b) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v); (c) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM; (d) the buffer is TRIS at a concentration of about 10 mM; (e) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or (f) the stabilizer is rHSA at a concentration of about 0.1%.

[0049] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization, and: (b) the carbohydrate is trehalose at a concentration of about 5% (w / v); (c) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (d) the buffer is TRIS at a concentration of about 10 mM; (e) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or (f) the stabilizer is rHSA at a concentration of about 0.1%.

[0050] In certain embodiments, the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0, optionally wherein the pH is about 8.0.

[0051] In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality ofabout 530 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg. In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0052] In certain embodiments, (b)-(f) are present at concentrations to reduce the loss of viral titer during filtration, reduce the formation of subvisible particles, maintain biophysical stability of the arenavirus during freeze and thaw process, and / or maintain biophysical stability of the arenavirus during storage.

[0053] In certain embodiments, the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C and about 8°C, above about 15°C, about - 20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.

[0054] In one aspect, the present disclosure provides a method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises: (a) citrate at a concentration of about 50 mM; (b) sucrose at a concentration of about 5% (w / v); (c) sodium chloride at a concentration of about 125 mM; (d) TRIS at a concentration of about 10 mM; (e) pol oxamer 188 at a concentration of about 0.03% (w / v); and (f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

[0055] In one aspect, the present disclosure provides a method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises: (a) citrate at a concentration of about 25 mM; (b) trehalose at a concentration of about 5% (w / v); (c) sodium chloride at a concentration of about 75 mM; (d) TRIS at a concentration of about 10 mM; (e) pol oxamer 188 at a concentration of about 0.03% (w / v); and (f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

[0056] In one aspect, the present disclosure provides a method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation is suitable for lyophilization andcomprises: (a) citrate at a concentration of between 10 mM and about 60 mM; (b) trehalose at a concentration of about 5% (w / v); (c) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (d) TRIS at a concentration of about 10 mM; (e) poloxamer 188 at a concentration of about 0.03% (w / v); and (f) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0057] In certain embodiments, the concentration of the arenavirus is between about 102RCV-FFU / ml and about 1010RCV-FFU / ml, or between about 2* 105RCV-FFU / ml and about 5x l08RCV-FFU / ml.

[0058] In certain embodiments, the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.

[0059] In certain embodiments, the arenavirus is a genetically engineered arenavirus.

[0060] In certain embodiments, the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of a tumor antigen, a tumor associated antigen, an antigen of a pathogen that causes an infectious disease, or an antigenic fragment of any of the foregoing. In certain embodiments, the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.

[0061] In certain embodiments, the antigen comprises at least one cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of gB(dCt), pp65, and combinations thereof. In certain embodiments, the antigen comprises at least one human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7ZE6 fusion protein. In certain embodiments, the antigen comprises at least one prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof. In certain embodiments, the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide, and combinations thereof. In certain embodiments, the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected fromthe group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef. In certain embodiments, the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the antigenic fragment of the mutant KRAS from N- to C- terminus comprises the mutations G13D, G12V, G12C, G12D, and G12R, respectively. In certain embodiments, the antigen comprises two or more CTA fragments (e.g., antigenic fragments or epitopes). In certain embodiments, the two or more CTA fragments are selected from the group consisting of epitopes or antigenic fragments of PRAME, MAGE-A3, MAGE-A4, and MAGE-A6.

[0062] In certain embodiments, the arenavirus is a non-replicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles. In certain embodiments, the arenavirus is a non-replicating arenavirus, and the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen.

[0063] In certain embodiments, the arenavirus is a replicating tri-segmented arenavirus.

[0064] In certain embodiments, the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF.

[0065] In certain embodiments, the tri-segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3’ UTR.

[0066] In certain embodiments, the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.

[0067] In certain embodiments, the method further comprises lyophilizing the liquid arenavirus formulation.

[0068] In one aspect, the present disclosure provides a method of producing a lyophilized arenavirus formulation, comprising lyophilizing the liquid arenavirus formulation disclosed herein.

[0069] In one aspect, the present disclosure provides a method of treating or preventing a disease, comprising administering to a subject in need thereof the liquid arenavirus formation disclosed herein, optionally wherein the liquid arenavirus formulation is further diluted prior to the administration.

[0070] In one aspect, the present disclosure provides a method of treating or preventing a disease, comprising administering to a subject in need thereof a liquid arenavirus formulationreconstituted from a lyophilized formulation disclosed herein, wherein the lyophilized formulation is reconstituted prior to the administration.

[0071] In certain embodiments, the liquid arenavirus formulation is administered parenterally, optionally wherein the liquid arenavirus formulation is administered intravenously, intramuscularly, intratumorally, or subcutaneously.

[0072] In certain embodiments, the disease is a neoplastic disease or an infectious disease.3.1 Illustrative Embodiments

[0073] Illustrative embodiments of the present disclosure are provided in the paragraphs below.1. A liquid arenavirus formulation comprising:(a) an arenavirus; and(b) a polyvalent anion at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.2. The liquid arenavirus formulation of embodiment 1, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.3. The liquid arenavirus formulation of embodiment 1, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.4. A liquid arenavirus formulation comprising:(a) an arenavirus; and(b) a polyvalent anion, wherein the polyvalent anion is at a concentration that (i) reduces the formation of subvisible particles during manufacturing, and / or (ii) reduces the loss of viral titer during filtration, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process.5. The liquid arenavirus formulation of embodiment 4, wherein the polyvalent anion is at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.6. The liquid arenavirus formulation of embodiment 5, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.7. The liquid arenavirus formulation of embodiment 5, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.8. The liquid arenavirus formulation of any one of embodiments 1-7, wherein the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.9. The liquid arenavirus formulation of embodiment 8, wherein the polyvalent anion is citrate.10. The liquid arenavirus formulation of embodiment 9, wherein the citrate is at a concentration of about 50 mM or about 25 mM.11. The liquid arenavirus formulation of embodiment 9, wherein the liquid arenavirus formulation is suitable for lyophilization, and the citrate is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.12. The liquid arenavirus formulation of any one of embodiments 1-11, further comprising at least one of:(c) a carbohydrate;(d) a salt;(e) a buffer;(f) a surfactant; and(g) a stabilizer.13. The liquid arenavirus formulation of embodiment 12, wherein:(c) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v);(d) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM;(e) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM;(f) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or(g) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v), optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).14. The liquid arenavirus formulation of embodiment 12 or 13, wherein(c) the carbohydrate is at a concentration of about 5% (w / v);(d) the salt is at a concentration of about 125 mM or about 75 mM;(e) the buffer is at a concentration of about 10 mM;(f) the surfactant is at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is at a concentration of about 0.1% (w / v).15. The liquid arenavirus formulation of embodiment 12 or 13, wherein the liquid arenavirus formulation is suitable for lyophilization, and(c) the carbohydrate is at a concentration of about 5% (w / v);(d) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) the buffer is at a concentration of about 10 mM;(f) the surfactant is at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is at a concentration of about 0.1% (w / v).16. The liquid arenavirus formulation of any one of embodiments 12-15, wherein:(c) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or a combination thereof;(d) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride;(e) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS;(f) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is pol oxamer 188; and / or(g) the stabilizer is selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the stabilizer is rHSA.17. The liquid arenavirus formulation of embodiment 16, wherein:(c) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v);(d) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM;(e) the buffer is TRIS at a concentration of about 10 mM;(f) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is rHSA at a concentration of about 0.1%.18. The liquid arenavirus formulation of embodiment 16, wherein the liquid arenavirus formulation is suitable for lyophilization, and:(c) the carbohydrate is trehalose at a concentration of about 5% (w / v);(d) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) the buffer is TRIS at a concentration of about 10 mM;(f) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is rHSA at a concentration of about 0.1%.19. The liquid arenavirus formulation of any one of embodiments 1-18, wherein the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0.20. The liquid arenavirus formulation of embodiment 19, wherein the pH is about 8.0.21. The liquid arenavirus formulation of any one of embodiments 1-20, wherein the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg.22. The liquid arenavirus formulation of embodiment 21, wherein the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.23. The liquid arenavirus formulation of embodiment 21, wherein the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.24. The liquid arenavirus formulation of any one of embodiments 21, wherein the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.25. The liquid arenavirus formulation of any one of embodiments 12-24, wherein (c)-(g) are present at concentrations to reduce the loss of viral titer during filtration, freezing and thawing, and / or storage; and / or reduce the formation of subvisible particles during manufacturing, freezing and thawing, and / or storage.26. The liquid arenavirus formulation of embodiment 25, wherein the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C andabout 8°C, above aboutl5°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.27. A liquid arenavirus formulation comprising:(a) an arenavirus;(b) citrate at a concentration of about 50 mM;(c) sucrose at a concentration of about 5% (w / v);(d) NaCl at a concentration of about 125 mM;(e) TRIS at a concentration of about 10 mM; and(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.28. A liquid arenavirus formulation comprising:(a) an arenavirus;(b) citrate at a concentration of about 25 mM;(c) trehalose at a concentration of about 5% (w / v);(d) NaCl at a concentration of about 75 mM;(e) TRIS at a concentration of about 10 mM; and(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.29. A liquid arenavirus formulation suitable for lyophilization comprising:(a) an arenavirus;(b) citrate at a concentration of between about 20 mM and about 50 mM;(c) trehalose at a concentration of about 5% (w / v);(d) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) TRIS at a concentration of about 10 mM;(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.30. The liquid arenavirus formulation of any one of embodiments 1-29, wherein the concentration of the arenavirus is between about 102RCV-FFU / ml and about IO10RCV- FFU / ml, or between about 2* 105RCV-FFU / ml and about 5* 108RCV-FFU / ml.31. The liquid arenavirus formulation of any one of embodiments 1-30, wherein the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.32. The liquid arenavirus formulation of any one of embodiments 1-31, wherein the arenavirus is a genetically engineered arenavirus.33. The liquid arenavirus formulation of any one of embodiments 1-32, wherein the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.34. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises a cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of gB(dCt), pp65, and combinations thereof.35. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises a human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7ZE6 fusion protein.36. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises a prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof.37. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide and combinations thereof.38. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef.39. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the at least one antigenic fragment from N- to C-terminus comprises the mutations G13D, G12V, G12C, G12D, and G12R respectively.40. The liquid arenavirus formulation of embodiment 33, wherein the antigen comprises two or more cancer testis antigen (CTA) fragments, optionally wherein the two or more CTA fragments are selected from the group consisting of epitopes or antigenic fragments of preferentially expressed antigen in melanoma (PRAME), melanoma-associated antigen 3 (MAGE-A3), melanoma-associated antigen 4 (MAGE-A4), and melanoma-associated antigen 6 (MAGE-A6).41. The liquid arenavirus formulation of any one of embodiments 1-40, wherein the arenavirus is a non-replicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles, further optionally wherein the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen.42. The liquid arenavirus formulation of any one of embodiments 1-40, wherein the arenavirus is a replicating tri-segmented arenavirus.43. The liquid arenavirus formulation of embodiment 42, wherein the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF.44. The liquid arenavirus formulation of embodiment 43, wherein the tri-segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the arenaviral glycoprotein (GP) is under control of an arenavirus 3 ’ UTR.45. The liquid arenavirus formulation of embodiment 42, wherein the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.46. A lyophilized arenavirus formulation prepared from the liquid arenavirus formulation of any one of embodiments 1-45.47. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at a concentration of between about 5mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.48. The method of embodiment 47, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.49. The method of embodiment 47, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.50. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at a concentration that reduces the formation of subvisible particles during manufacturing, and / or reduces the loss of viral titer during filtration.51. The method of embodiment 50, wherein the polyvalent anion is at a concentration of between about 5 mM and about 150 mM.52. The method of embodiment 51, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.53. The method of embodiment 51, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.54. The method of any one of embodiments 47-53, wherein the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.55. The method of embodiment 54, wherein the polyvalent anion is citrate.56. The method of embodiment 55, wherein the citrate is at a concentration of about 50 mM or about 25 mM.57. The method of embodiment 55, wherein the citrate is at a concentration of between about 10 mM and about 60 mM.58. The method of any one of embodiments 47-57, wherein the liquid arenavirus formulation further comprises at least one of:(b) a carbohydrate;(c) a salt;(d) a buffer;(e) a surfactant; and(f) a stabilizer.59. The method of embodiment 58, wherein:(b) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v);(c) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM;(d) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM;(e) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or(f) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v) , optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).60. The method of embodiment 58 or 59, wherein(b) the carbohydrate is at a concentration of about 5% (w / v);(c) the salt is at a concentration of about 125 mM or about 75 mM;(d) the buffer is at a concentration of about 10 mM;(e) the surfactant is at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is at a concentration of about 0.1% (w / v).61. The method of embodiment 58 or 59, wherein the liquid arenavirus formulation is suitable for lyophilization, and(b) the carbohydrate is at a concentration of about 5% (w / v);(c) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) the buffer is at a concentration of about 10 mM;(e) the surfactant is at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is at a concentration of about 0.1% (w / v).62. The method of any one of embodiments 58-61, wherein:(b) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or acombination thereof; (c) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride;(d) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS;(e) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is pol oxamer 188; and / or(f) the stabilizer is a protein selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the protein is rHSA.63. The method of embodiment 62, wherein:(b) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v);(c) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM;(d) the buffer is TRIS at a concentration of about 10 mM;(e) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is rHSA at a concentration of about 0.1%.64. The method of embodiment 62, wherein the liquid arenavirus formulation is suitable for lyophilization, and:(b) the carbohydrate is trehalose at a concentration of about 5% (w / v);(c) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) the buffer is TRIS at a concentration of about 10 mM;(e) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is rHSA at a concentration of about 0.1%.65. The method of any one of embodiments 47-64, wherein the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0, optionally wherein the pH is about 8.0.66. The method of any one of embodiments 47-65, wherein the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg.67. The method of embodiment 66, wherein the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.68. The method of embodiment 66, wherein the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.69. The method of embodiments 66, wherein the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.70. The method of any one of embodiments 58-69, wherein (b)-(f) are present at concentrations to reduce the loss of viral titer during filtration, reduce the formation of subvisible particles, maintain biophysical stability of the arenavirus during freeze and thaw process, and / or maintain biophysical stability of the arenavirus during storage.71. The method of embodiment 70, wherein the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about - 80°C.72. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises:(a) citrate at a concentration of about 50 mM;(b) sucrose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of about 125 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0 optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.73. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises:(a) citrate at a concentration of about 25 mM;(b) trehalose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of about 75 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg74. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation is suitable for lyophilization and comprises:(a) citrate at a concentration of between 10 mM and about 60 mM;(b) trehalose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process, optionally wherein the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.75. The method of any one of embodiments 47-74, wherein the concentration of the arenavirus is between about 102RCV-FFU / ml and about 1010RCV-FFU / ml, or between about 2* 105RCV-FFU / ml and about 5 108RCV-FFU / ml.76. The method of embodiments 47-75, wherein the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.77. The method of any one of embodiments 47-76, wherein the arenavirus is a genetically engineered arenavirus.78. The method of any one of embodiments 47-77, wherein the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.79. The method of embodiment 78, wherein the antigen comprises at least one cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of gB(dCt), pp65, and combinations thereof.80. The method of embodiment 78, wherein the antigen comprises at least one human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7 / E6 fusion protein.81. The method of embodiment 78, wherein the antigen comprises at least one prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof.82. The method of embodiment 78, wherein the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide, and combinations thereof.83. The method of embodiment 78, wherein the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef.84. The method of embodiment 78, wherein the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the antigenic fragment of the mutant KRAS from N- to C-terminus comprises the mutations G13D, G12V, G12C, G12D, and G12R, respectively.85. The method of embodiment 78, wherein the antigen comprises two or more cancer testis antigen (CTA) fragments, optionally wherein the two or more CTA fragments are selected from the group consisting of epitopes or antigenic fragments of preferentially expressed antigen in melanoma (PRAME), melanoma-associated antigen 3 (MAGE-A3), melanoma- associated antigen 4 (MAGE-A4), and melanoma-associated antigen 6 (MAGE-A6)86. The method of any one of embodiments 47-85, wherein the arenavirus is a nonreplicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles, further optionally wherein the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen.87. The method of any one of embodiments 47-85, wherein the arenavirus is a replicating trisegmented arenavirus.88. The method of embodiment 87, wherein the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF.89. The method of embodiment 88, wherein the tri-segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3’ UTR.90. The method of embodiment 87, wherein the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.91. The method of any one of embodiments 47-90, further comprising lyophilizing the liquid arenavirus formulation.92. A method of producing a lyophilized arenavirus formulation, comprising lyophilizing the liquid arenavirus formulation of any one of embodiments 1-45.93. A method of treating or preventing a disease, comprising administering to a subject in need thereof the liquid arenavirus formation of any one of embodiments 1-45, optionally wherein the liquid arenavirus formulation is further diluted prior to the administration.94. A method of treating or preventing a disease, comprising administering to a subject in need thereof a liquid arenavirus formulation reconstituted from the lyophilized formulation of embodiment 46, wherein the lyophilized formulation is reconstituted prior to the administration.95. The method of embodiment 93 or 94, wherein the liquid arenavirus formulation is administered parenterally, optionally wherein the liquid arenavirus formulation is administered intravenously, intramuscularly, intratumorally, or subcutaneously.96. The method of embodiments 93-95, wherein the disease is a neoplastic disease or an infectious disease.4. BRIEF DESCRIPTION OF THE FIGURES

[0074] FIG. 1 depicts the effects plot (computed as twice the multiple linear regression (MLR) coefficients) of polyvalent anions on 0.22 pm sterile filtration recovery of virus particles from arenavirus formulations having a TRIS-based formulation. Error bars represent the 95% confidence level. TSP=TSPP; Cit=Citrate; Mal= Malate; Suc=Succinate; Tar=Tartrate. Evaluation was performed by MODDE Pro software and data were fitted to a multiple linear regression (MLR) model.

[0075] FIG. 2 depicts the recovery of virus particles [%] determined by FFU and RCV-FFU prior to and after sterile filtration as a function of sodium chloride (NaCl) and citrate concentrations in the arenavirus formulation. Evaluation was performed by MODDE Pro software and data were fitted to a multiple linear regression (MLR) model.

[0076] FIGS. 3A-3B depict virus titers determined during the manufacturing process and filtration recovery after bioburden reduction filtration for batches of arenavirus formulations processed with a HEPES-based formulation (10 mM HEPES, 150 mM NaCl, 20 mM glycine, 1% rHSA; pH 7.4). Virus titers were analyzed by quantification of RCV-FFU (FIG. 3A) and FFU (FIG. 3B). HEPES I-III correspond to the different 20 L laboratory scale batches of arenavirus productions. Processing Step (PS)1 : harvest; PS2: nuclease treatment; PS3: chromatography 1; PS4: chromatography 2; PS5: final formulation; PS6: bioburden reduction filtration.

[0077] FIGS. 4A-4B depict virus titers determined during the manufacturing process and filtration recovery after bioburden reduction filtration (PS6) for batches of arenavirus formulations processed with a TRIS-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 5% sucrose, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). Virus titers were analyzed by quantification of RCV-FFU (FIG. 4A) and FFU (FIG. 4B). TRIS / Citrate I-V correspond to the different 20 L laboratory scale batches of arenavirus productions. PSI : harvest; PS2: nuclease treatment; PS3: chromatography 1; PS4: chromatography 2; PS5: final formulation; PS6: bioburden reduction filtration.

[0078] FIG. 5 depicts subvisible particle concentrations, as measured by flow imaging microscopy (FIM), for arenavirus formulations prepared in a TRIS-buffered formulation containing different concentrations (25 mM (first bar) and 50 mM (second bar)) of polyanions (TSPP, succinate, tartrate, and malate).

[0079] FIG. 6 depicts subvisible particle concentrations, as measured by FIM, in arenavirus formulations containing different concentrations of NaCl, citrate, and poloxamer 188. Evaluation was performed by MODDE Pro software and data were fitted to a MLR model.

[0080] FIGS. 7A-7C depict subvisible particle concentrations of total subvisible particles (i.e., particles >1 pm) (FIG. 7A), particles >10 pm (FIG. 7B), and particles >25 pm (FIG. 78C) determined during the manufacturing process and bioburden reduction filtration (PS6) of arenavirus formulations prepared in either a TRIS-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 5% (w / v) sucrose, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0) or a HEPES-based formulation (10 mM HEPES, 150 mM NaCl, 20 mM glycine, 1% rHSA; pH 7.4). HEPES I-III correspond to the different batches of HEPES-based arenavirus purifications; Citrate I-V correspond to the different batches of TRIS-based arenavirus purifications. For each of the tested process intermediates derived after PS3-PS6 in the graphs, the bars from left to right are HEPES I, HEPES II, HEPES III, Citrate I, Citrate II, Citrate III, Citrate IV, and Citrate V. All samples underwent one freeze / thaw cycle beforesubvisible particle measurement. PS3: chromatography 1; PS4: chromatography 2; PS5: final formulation; PS6: bioburden reduction filtration.

[0081] FIG. 8 depicts viral titers of arenavirus formulations comprising different polyanions (20 mM TSPP; 50 mM succinate; or 50 mM citrate) in combination with different carbohydrates (sucrose, trehalose, or sorbitol at a concentration of 5% (w / v) each) prepared in a TRIS-based formulation (10 mM Tris, 125 mM NaCl, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). Viral titers were measured by RCV-FFU assays performed prior to freezing (w / o F / T), after three (3) freeze / thaw cycles (3x F / T), and after subsequent sterile filtration (sterile filtered) of the formulations. As a reference, viral titers of an arenavirus formulation lacking polyanions and carbohydrates prepared in a HEPES-based formulation (10 mM HEPES, 20 mM glycine, 150 mM NaCl, 1% rHSA; pH 7.4) were also measured prior to freezing (w / o F / T), after three (3) freeze / thaw cycles (3x F / T), and after subsequent sterile filtration (sterile filtered) of the reference formulation. For each tested combination, the bars from left to right are w / o F / T, 3* F / T, and sterile filtered.

[0082] FIG. 9 depicts viral titers of arenavirus formulations having different concentrations of sucrose (0 - 10% (w / v)) prepared in a TRIS-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). Viral titers were measured by RCV-FFU assays performed prior to and after five (5) freeze / thaw cycles, z.e., prior to sterile filtration (RCV-FFU before 0.22 pm), after sterile filtration (RCV-FFU after 0.22 pm), and after five freeze / thaw cycles (5x F / T). For each tested sucrose concentration, the bars from left to right are RCV-FFU before 0.22 pm, RCV-FFU after 0.22 pm, and 5 F / T.

[0083] FIG. 10 depicts virus titer recovery from arenavirus formulations having different concentrations of sucrose (0 - 10% (w / v)) prepared in a TRIS-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0) after five (5) freeze / thaw cycles and filtration (0.22 pm).

[0084] FIG. 11 depicts subvisible particle concentrations of particles >1 pm, particles >10 pm, and particles >25 pm in arenavirus formulations prepared in 10 mM Tris, 50 mM citrate, 125 mM NaCl, 5% sucrose, 0.03% poloxamer 188, and 0.1% rHSA (pH 8.0). Particle concentrations were measured in the formulations by FIM prior to (0*FT) and after each of three (3) consecutive freeze / thaw cycles (1XFT, 2><FT, and 3><FT). For each of the tested F / T condition, the bars from left to right are particles >1 pm, particles >10 pm, particles >25 pm.

[0085] FIG. 12 depicts subvisible particle concentrations (>1 pm) in arenavirus formulations prepared in 10 mM Tris, 20 mM TSPP, 250 mM NaCl, and 0.03% poloxamer 188 (pH 8.0)with and without 5% sucrose. Particle concentrations were measured in the formulations by FIM after each of five (5) consecutive freeze / thaw cycles.

[0086] FIG. 13 depicts viral titers of arenavirus formulations having different rHSA concentrations (0 - 1% (w / v)) prepared in a TRIS-based formulation (10 mM Tris, 20 mM TSPP, 125 mM NaCl, 5% sucrose, 0.015% poloxamer 188; pH 8.0) after storing the formulations in 15 mL polypropylene (PP) tubes at 2°C - 8°C for five weeks. Viral titer was analyzed by quantification of RCV-FFU.

[0087] FIGS. 14A-14B depict viral titers in arenavirus formulations having either no carbohydrate additive (w / o sugar), 5% sucrose, 5% trehalose, or 5% sorbitol prepared in a TRIS-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 0.03% (w / v) poloxamer 188, 0.1% (w / v) rHSA; pH 8.0) before and after one (1) to eighteen (18) months of storage at -20°C. Viral titers were analyzed by quantification of RCV-FFU (FIG. 14A) and FFU (FIG.14B)

[0088] FIG. 15 depicts subvisible particle concentrations (>1 pm) in arenavirus formulations having either no carbohydrate additive (w / o sugar), 5% sucrose, 5% trehalose, or 5% sorbitol before and after one (1) to eighteen (18) months of storage at -20°C. The concentrations of subvisible particles were analyzed in the formulations by FIM prior to freezing as well as after 1, 2, 3, and 6 months of storage. For each of the storage length, the bars from left to right are w / o sugar, trehalose, sorbitol, and sucrose.

[0089] FIG. 16 depicts virus particle diameters in arenavirus formulations having either no carbohydrate additive (w / o sugar), 5% sucrose, 5% trehalose, or 5% sorbitol before and after one (1) to three (3) months of storage at -20°C. The diameter of the virus particles was analyzed by Nanoparticle Tracking Analysis (NTA) prior to freezing as well as after one, two, and three months of storage. For each of the storage length, the bars from left to right are w / o sugar, trehalose, sorbitol, and sucrose.

[0090] FIGS. 17A-17B depict particle diameters of arenavirus in arenavirus formulations having either no carbohydrate additive (FIG. 17A) or being supplemented with 5% trehalose (FIG. 17B) after three (3) months of storage at -20°C.5. DETAILED DESCRIPTION

[0091] The present disclosure provides improved arenavirus formulations (e.g., liquid formulations, and lyophilized formulations). In certain embodiments, the arenavirus formulations can maintain the biophysical stability and viral titer during manufacturing,purification, and storage of formulations comprising the arenaviruses. In particular, the arenavirus formulations disclosed herein can minimize the loss of viral titer during the purification process (e.g., sterile filtration), minimize the formation of aggregates and subvisible particles, maintain the biophysical stability of viral particles during freeze / thaw process(es), and / or maintain the biophysical stability of viral particles during storage.

[0092] The present disclosure is based, in part, on the discovery that certain arenavirus formulations significantly increased arenavirus based viral vector stability in frozen and liquid state and greatly reduced the formation of vector-derived aggregates and subvisible particles, thereby facilitating filterability and increasing viral recovery during final filtration steps. As compared to the previously known formulations, the presently disclosed arenavirus formulations advantageously keep the viral vectors dispersed and thereby significantly reduce substantial filtration losses (in particular during sterile filtration) seen with many enveloped vectors. In addition, as particle (aggregate) formation is commonly seen in the viral vector field, minimizing particle formation by the presently disclosed formulations is considered an advantage with regard to product safety. Finally, increasing the stability of the formulation when stored refrigerated (2°C - 8°C) and at ambient conditions (e.g., 20°C - 25°C) by the presently disclosed formulations allows a broader use and intermediate storage of the arenavirus in hospitals and regions lacking ultralow freezer equipment. The stability at -20°C storage could also be increased by the use of appropriate carbohydrates disclosed herein. Furthermore, the presently disclosed liquid formulations can use excipients that are compatible with freeze-drying, which further increases storage stability at elevated temperatures.

[0093] In certain embodiments, the arenavirus formulation disclosed herein is a liquid arenavirus formulation (see Section 5.1). In certain embodiments, the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (e.g., citrate) (see Section 5.1.1). In certain embodiments, the liquid arenavirus formulation further comprises at least one of a carbohydrate (e.g., sucrose) (see Section 5.1.2), a salt (e.g., sodium chloride) (see Section 5.1.3), a buffer (e.g., TRIS) (see Section 5.1.4), a surfactant (e.g., poloxamer 188) (see Section 5.1.5), and a stabilizer (e.g., rHSA) (see Section 5.1.6). In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization.

[0094] In certain embodiments, the arenavirus formulation disclosed herein is a lyophilized arenavirus formulation (see Section 5.2). In certain embodiments, the lyophilized arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (e.g., citrate) (see Section 5.1.1). In certain embodiments, the lyophilized arenavirus formulation furthercomprises at least one of a carbohydrate (e.g., trehalose) (see Section 5.1.2), a salt (e.g., sodium chloride) (see Section 5.1.3), a buffer (e.g., TRIS) (see Section 5.1.4), a surfactant (e.g., poloxamer 188) (see Section 5.1.5), and a stabilizer (e.g., rHSA) (see Section 5.1.6). In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation disclosed herein (see Section 5.1).

[0095] The present disclosure also provides pharmaceutical compositions (see Section 5.4) comprising a presently disclosed liquid arenavirus formulation (see Section 5.1) or a lyophilized arenavirus formulation (see Section 5.2).

[0096] The present disclosure also provides methods for reducing the formation of subvisible particles and / or reducing the loss of viral titer by combining an arenavirus with a solution to form a presently disclosed liquid arenavirus formulation (see Section 5.5).

[0097] The present disclosure further provides methods for treating or preventing a disease (see Section 5.6), comprising administering to a subject in need thereof a presently disclosed liquid arenavirus formulation (see Section 5.1), or a presently disclosed lyophilized arenavirus formulation (see Section 5.2).

[0098] The term “about” means within 20%, within 15%, within 10%, within 9%, within 8%, within 7%, within 6%, within 5%, within 4%, within 3%, within 2%, within 1%, or less variation of a given value or range.

[0099] For purposes of clarity of disclosure and not by way of limitation, the detailed description is divided into the following subsections:5.1 Liquid Arenavirus Formulations;5.2 Lyophilized Arenavirus Formulations;5.3 Arenaviruses;5.4 Pharmaceutical Compositions;5.5 Methods of Reducing Formation of Subvisible Particles or Loss of Viral Titer; and5.6 Methods of Treatment.5.1 Liquid Arenavirus Formulations

[0100] The present disclosure provides liquid arenavirus formulations having various improved properties. In certain embodiments, the presently disclosed liquid arenavirus formulations can facilitate the manufacturing, purification, and storage of formulations comprising arenaviruses, wherein the formulations can be used for treating or preventing a disease.

[0101] In certain embodiments, the liquid arenavirus formulation comprises an arenavirus and a polyvalent anion. In certain embodiments, the liquid arenavirus formulation further comprises at least one of a stabilizer, a carbohydrate, a salt, a buffer, and a surfactant.

[0102] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation is suitable for preparation of a lyophilized arenavirus formulation disclosed in Section 5.2.

[0103] In certain embodiments, prior to the administration to the subject, a liquid arenavirus formulation is reconstituted from a lyophilized arenavirus formulation disclosed herein (see Section 5.2), and thus can comprise the same components as the liquid arenavirus formulation from which the lyophilized arenavirus formulation was prepared but different concentrations thereof.

[0104] In certain embodiments, prior to the administration to the subject, a liquid arenavirus formulation is diluted from a liquid arenavirus formulation disclosed herein (see Section 5.1), and thus can comprise the same components as the liquid arenavirus formulation disclosed herein but different concentrations thereof.5.1.1 Polyvalent Anions

[0105] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises a polyvalent anion. Without being bound by theory, the polyvalent anion can reduce the aggregation of virus particles, and thus minimize the formation of aggregates and subvisible particles. Such properties enable filtration (e.g., bioburden reduction filtration or sterile filtration) without substantial loss of viral titer.

[0106] In certain embodiments, the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof. In certain embodiments, the polyvalent anion is selected from the group consisting of citrate, TSPP, malate, succinate, tartrate, and combinations thereof. In certain embodiments, the polyvalent anion is citrate.

[0107] Citrate (CeHsO?3), malate (C4H4O52), succinate (C4H4O42'), tartrate (C4H4O62), fumarate (C4H2O42), and maleate (C4H2O42) refer to any salt or ester of the corresponding di- and tricarboxylic acids. Pyrophosphate (P2O74) refers to any salt of pyrophosphoric acid such as di- or tetrasodium pyrophosphate (TSPP).

[0108] In certain embodiments, the liquid arenavirus formulation comprises a polyvalent anion as disclosed herein at a concentration of up to about 150 mM, up to about 125 mM, up to about 100 mM, up to about 80 mM, up to about 70 mM, up to about 60 mM, up to about 50 mM, up to about 40 mM, up to about 30 mM, up to about 20 mM, or up to about 10 mM.In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of at least about 5 mM, at least about 10 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, at least about 50 mM, at least about 60 mM, at least about 70 mM, at least about 80 mM, at least about 100 mM, or at least about 125 mM. In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of between about 5 mM and about 150 mM, between about 10 mM and about 150 mM, between about 20 mM and about 150 mM, between about 30 mM and about 150 mM, between about 40 mM and about 150 mM, between about 50 mM and about 150 mM, between about 60 mM and about 150 mM, between about 70 mM and about 150 mM, between about 80 mM and about 150 mM, between about 100 mM and about 150 mM, between about 125 mM and about 150 mM, between about 5 mM and about 125 mM, between about 10 mM and about 125 mM, between about 20 mM and about 125 mM, between about 30 mM and about 125 mM, between about 40 mM and about 125 mM, between about 50 mM and about 125 mM, between about 60 mM and about 125 mM, between about 70 mM and about 125 mM, between about 80 mM and about 125 mM, between about 100 mM and about 125 mM, between about 5 mM and about 100 mM, between about 10 mM and about 100 mM, between about 20 mM and about 100 mM, between about 30 mM and about 100 mM, between about 40 mM and about 100 mM, between about 50 mM and about 100 mM, between about 60 mM and about 100 mM, between about 70 mM and about 100 mM, between about 80 mM and about 100 mM, between about 5 mM and about 80 mM, between about 10 mM and about 80 mM, between about 20 mM and about 80 mM, between about 30 mM and about 80 mM, between about 40 mM and about 80 mM, between about 50 mM and about 80 mM, between about 60 mM and about 80 mM, between about 70 mM and about 80 mM, between about 5 mM and about 70 mM, between about 10 mM and about 70 mM, between about 20 mM and about 70 mM, between about 30 mM and about 70 mM, between about 40 mM and about 70 mM, between about 50 mM and about 70 mM, between about 60 mM and about 70 mM, between about 5 mM and about 60 mM, between about 10 mM and about 60 mM, between about 20 mM and about 60 mM, between about 30 mM and about 60 mM, between about 40 mM and about 60 mM, between about 50 mM and about 60 mM, between about 5 mM and about 50 mM, between about 10 mM and about 50 mM, between about 20 mM and about 50 mM, between about 30 mM and about 50 mM, between about 40 mM and about 50 mM, between about 5 mM and about 40 mM, between about 10 mM and about 40 mM, between about 20 mM and about 40 mM, between about 30 mM and about 40 mM, between about 5 mM and about 30mM, between about 10 mM and about 30 mM, between about 20 mM and about 30 mM, between about 5 mM and about 20 mM, between about 10 mM and about 20 mM, or between about 5 mM and about 10 mM. In certain embodiments, the liquid arenavirus formulation comprises a polyvalent anion as disclosed herein at a concentration of about 150 mM, about 125 mM, about 100 mM, about 80 mM, about 70 mM, about 60 mM, about 50 mM, about 40 mM, about 30 mM, about 20 mM, or about 10 mM.

[0109] In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of between about 10 mM and about 60 mM. In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of between about 40 mM and about 60 mM. In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of between about 20 mM and about 30 mM. In certain embodiments, the liquid arenavirus formulation comprises the polyvalent anion at a concentration of about 25 mM.

[0110] In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of up to about 150 mM, up to about 125 mM, up to about 100 mM, up to about 80 mM, up to about 70 mM, up to about 60 mM, up to about 50 mM, up to about 40 mM, up to about 30 mM, up to about 20 mM, or up to about 10 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of at least about 5 mM, at least about 10 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, at least about 50 mM, at least about 60 mM, at least about 70 mM, at least about 80 mM, at least about 100 mM, or at least about 125 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of between about 5 mM and about 150 mM, between about 10 mM and about 150 mM, between about 20 mM and about 150 mM, between about 30 mM and about 150 mM, between about 40 mM and about 150 mM, between about 50 mM and about 150 mM, between about 60 mM and about 150 mM, between about 70 mM and about 150 mM, between about 80 mM and about 150 mM, between about 100 mM and about 150 mM, between about 125 mM and about 150 mM, between about 5 mM and about 125 mM, between about 10 mM and about 125 mM, between about 20 mM and about 125 mM, between about 30 mM and about 125 mM, between about 40 mM and about 125 mM, between about 50 mM and about 125 mM, between about 60 mM and about 125 mM, between about 70 mM and about 125 mM, between about 80 mM and about 125 mM, between about 100 mM and about 125 mM, between about 5 mM and about 100 mM, between about 10 mM and about 100 mM, between about 20 mM and about 100mM, between about 30 mM and about 100 mM, between about 40 mM and about 100 mM, between about 50 mM and about 100 mM, between about 60 mM and about 100 mM, between about 70 mM and about 100 mM, between about 80 mM and about 100 mM, between about 5 mM and about 80 mM, between about 10 mM and about 80 mM, between about 20 mM and about 80 mM, between about 30 mM and about 80 mM, between about 40 mM and about 80 mM, between about 50 mM and about 80 mM, between about 60 mM and about 80 mM, between about 70 mM and about 80 mM, between about 5 mM and about 70 mM, between about 10 mM and about 70 mM, between about 20 mM and about 70 mM, between about 30 mM and about 70 mM, between about 40 mM and about 70 mM, between about 50 mM and about 70 mM, between about 60 mM and about 70 mM, between about 5 mM and about 60 mM, between about 10 mM and about 60 mM, between about 20 mM and about 60 mM, between about 30 mM and about 60 mM, between about 40 mM and about 60 mM, between about 50 mM and about 60 mM, between about 5 mM and about 50 mM, between about 10 mM and about 50 mM, between about 20 mM and about 50 mM, between about 30 mM and about 50 mM, between about 40 mM and about 50 mM, between about 5 mM and about 40 mM, between about 10 mM and about 40 mM, between about 20 mM and about 40 mM, between about 30 mM and about 40 mM, between about 5 mM and about 30 mM, between about 10 mM and about 30 mM, between about 20 mM and about 30 mM, between about 5 mM and about 20 mM, between about 10 mM and about 20 mM, , or between about 5 mM and about 10 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate as disclosed herein at a concentration of about 150 mM, about 125 mM, about 100 mM, about 80 mM, about 70 mM, about 60 mM, about 50 mM, about 40 mM, about 30 mM, about 20 mM, or about 10 mM.

[0111] In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of between about 10 mM and about 60 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of between about 40 mM and about 60 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of between about 20 mM and about 30 mM. In certain embodiments, the liquid arenavirus formulation comprises citrate at a concentration of about 25 mM.

[0112] In certain embodiments, the liquid arenavirus formulation comprises a polyvalent anion disclosed herein, wherein the polyvalent anion (e.g., citrate) is at a concentration that (i) reduces the formation of subvisible particles during manufacturing, and / or (ii) reduces theloss of viral titer during filtration. In certain embodiments, the filtration comprises bioburden reduction filtration (e.g., 0.45 pm filtration), sterile filtration (e.g., 0.2 pm or 0.22 pm filtration), or a combination thereof. In certain embodiments, the manufacturing comprises a purification process. In certain embodiments, the manufacturing comprises at least one chromatography step.

[0113] Subvisible particles are particles that are too large for analysis by size exclusion chromatography (SEC) (e.g., at least about 0.3 pm), but too small to be visible to the unaided eye (e.g., no more than about 100 pm). In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the formation of subvisible particles during manufacturing as compared to a control formulation without the polyvalent anion. In certain embodiments, the subvisible particles have a size of at least about 0.3 pm, at least about 0.5 pm, at least about 1 pm, at least about 5 pm, at least about 10 pm, at least about 15 pm, at least about 20 pm, at least about 25 pm, at least about 30 pm, at least about 35 pm, at least about 40 pm, at least about 45 pm, at least about 50 pm, at least about 55 pm, at least about 60 pm, at least about 65 pm, at least about 70 pm, at least about 75 pm, at least about 80 pm, at least about 85 pm, at least about 90 pm, or at least about 95 pm. In certain embodiments, the subvisible particles have a size of less than 100 pm. In certain embodiments, the subvisible particles have a size of at least about 1 pm. In certain embodiments, the subvisible particles have a size of at least about 10 pm. In certain embodiments, the subvisible particles have a size of at least about 25 pm.

[0114] Size distribution, shape, and concentration of the subvisible particles can be measured by any suitable methods known in the art, for example, light obscuration particle count techniques and microscopic particle count techniques. In certain embodiments, the size distribution, shape, and concentration of the subvisible particles can be measured by flow imaging microscopy (FIM). For example, flow imaging microscopy (FIM) can be used to measure the size distribution, shape and concentration of particles that are about 1 pm to several hundred pm in size. Sample analysis can be performed on a FlowCam 8100 (Fluid Imaging Technologies, Inc.) equipped with a 10x objective, a FOV80 flow cell and a precision syringe of 1 mL. Flow is set to 0.15 mL / min with an autoimage rate of 19 images per second, resulting in an efficiency of about 71%. The equivalent spherical diameter (ESD) is measured, which is a calculated value based on the volume of the particles. MODDE Pro (Satorius Stedim Data Analytics AB) can be used for evaluation. Data are fitted by multiple linear regression (MLR) model.

[0115] In certain embodiments, the polyvalent anion e.g., citrate) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is up to about 5%, up to about 10%, up to about 15%, up to about 20%, up to about 25%, up to about 30%, up to about 35%, up to about 40%, up to about 45%, up to about 50%, up to about 60%, up to about 70%, or up to about 80% of the concentration of subvisible particles in a control formulation without the polyvalent anion as measured by FIM. In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is about 5%, about 10%, about 15%, about 20%, about 25%, about 30%, about 35%, about 40%, about 45%, about 50%, about 60%, about 70%, or about 80% of the concentration of subvisible particles in a control formulation without the polyvalent anion, as measured by FIM. In certain embodiments, the control formulation is otherwise identical to the liquid arenavirus formulation disclosed herein except not comprising the polyvalent anion or comprising a different polyvalent anion.

[0116] Viral titer can be measured by any methods known in the art. In certain embodiments, the viral titer can be measured by replication competent virus focus forming units (RCV-FFU) assay. In certain embodiments, the viral titer can be measured by focus forming units (FFU) assay.

[0117] Any assay known to the skilled artisan can be used for measuring the infectivity of an arenavirus vector formulation. For example, the virus / vector titer can be determined by a focus forming unit assay (FFU assay). Briefly, complementing cells, e.g., HEK293-TVL cells are plated and inoculated with different dilutions of a virus vector sample. After a period of incubation, the cells form a monolayer and the virus attach to the cells. The monolayer is then covered with methylcellulose. After further incubation, the originally infected cells release viral progeny. Due to the methylcellulose overlay, the spread of the new viruses is restricted to neighboring cells. Consequently, each infectious particle produces a circular zone of infected cells called a focus. Such foci can be made visible and thus countable using antibodies against LCMV- NP or PICV-NP or another protein expressed by the arenavirus particle or the tri-segmented arenavirus particle and a HRP -based color reaction. The titer of a virus / vector can be calculated in focus-forming units per milliliter (FFU / mL). Similarly, the proportion of tri-segmented, replication competent virus particles can be determined using non-complementing cells are used, e.g., HEK293. This allows onlytri-segmented virus particles to infect neighboring cells. The titer of the replication competent virus / vector (RCV) can be calculated in focus-forming units per milliliter (RCV FFU / mL).

[0118] In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer prior to the filtration as measured by RCV-FFU or FFU assay. In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% of the viral titer prior to the filtration as measured by RCV-FFU or FFU assay.

[0119] In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 400%, or at least about 500% more than the viral titer in a control formulation without the polyvalent anion after the filtration as measured by RCV-FFU or FFU assay. In certain embodiments, the polyvalent anion (e.g., citrate) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 400%, about 500% more than the viral titer in a control formulation without the polyvalent anion after the filtration as measured by RCV-FFU or FFU assay. In certain embodiments, the control formulation is otherwise identical to the liquid arenavirus formulation disclosed herein except not comprising the polyvalent anion or comprising a different polyvalent anion.

[0120] In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of up to about 50 mM, up to about 40 mM, up to about 30 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, between about 20 mM and about 50 mM, between about 20mM and about 40 mM, between about 20 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 40 mM, or between about 40 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of about 20 mM, about 30 mM, about 40 mM, or about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of up to about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of between about 20 mM and about 50 mM.

[0121] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of up to about 50 mM, up to about 40 mM, up to about 30 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, between about 20 mM and about 50 mM, between about 20 mM and about 40 mM, between about 20 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 40 mM, or between about 40 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of about 20 mM, about 30 mM, about 40 mM, or about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of up to about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of between about 20 mM and about 50 mM.5.1.2 Carbohydrates

[0122] In certain embodiments, the liquid arenavirus formulation disclosed herein further comprises a carbohydrate. Without being bound by theory, addition of one or more carbohydrates to the liquid arenavirus formulation disclosed herein can increase the stability of the arenavirus particles during freezing and thawing. Without being bound by any particular theory, the addition of one or more carbohydrates to the formulation described herein increases the stability of arenavirus particles during the freeze / thaw process by reducing the aggregation and / or inactivation of the arenavirus particles.

[0123] In certain embodiments, the carbohydrate for use in the formulations and methods disclosed herein is a sugar or a polyol. Suitable sugars that can be used with the present disclosure include, but are not limited to, monosaccharides (e.g., glucose, galactose, ribose, mannose, rhamnose, talose, xylose, allose, and arabinose), disaccharides (e.g., trehalose, sucrose, maltose, isomaltose, cellibiose, gentiobiose, laminaribose, xylobiose, mannobiose,lactose, and fructose), trisaccharides (e.g., acarbose, raffinose, melizitose, panose, and cellotriose), and sugar polymers (e.g., dextran, xanthan, pullulan, cyclodextrins, amylose, amylopectin, starch, celloologosaccharides, cellulose, maltooligosaccharides, glycogen, chitosan, and chitin). Polyols, also known as sugar alcohols, are a class of low molecular weight polyols commonly obtained by hydrogenation of sugars. Suitable polyols that can be used with the present disclosure include, but are not limited to, mannitol, sorbitol, arabitol, erythritol, maltitol, xylitol, glycitol, glycol, polyglycitol, polyethylene glycol, polypropylene glycol, isomalt, hydrogenated starch hydrosylates, and glycerol.

[0124] In certain embodiments, the liquid arenavirus formulation disclosed herein further comprises a carbohydrate selected from the group consisting of sucrose, trehalose, mannitol, sorbitol, dextran, and combinations thereof. In certain embodiments, the carbohydrate is sucrose, trehalose, or a combination thereof. In certain embodiments, the carbohydrate is sucrose. In certain embodiments, the carbohydrate is trehalose.

[0125] In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of up to about 20% (w / v), up to about 15% (w / v), up to about 10% (w / v), up to about 5% (w / v), up to about 1% (w / v), up to about 0.5% (w / v), or up to about 0.1% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of at least about 0.1% (w / v), at least about 0.5% (w / v), at least about 1% (w / v), at least about 5% (w / v), at least about 10% (w / v), at least about 15% (w / v), or at least about 20% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of between about 0.1% (w / v) and about 20% (w / v), between about 0.5% (w / v) and about 20% (w / v), between about 1% (w / v) and about 20% (w / v), between about 5% (w / v) and about 20% (w / v), between about 10% (w / v) and about 20% (w / v), between about 15% (w / v) and about 20% (w / v), between about 0.1% (w / v) and about 15% (w / v), between about 0.5% (w / v) and about 15% (w / v), between about 1% (w / v) and about 15% (w / v), between about 5% (w / v) and about 15% (w / v), between about 10% (w / v) and about 15% (w / v), between about 0.1% (w / v) and about 10% (w / v), between about 0.5% (w / v) and about 10% (w / v), between about 1% (w / v) and about 10% (w / v), between about 5% (w / v) and about 10% (w / v), between about 0.1% (w / v) and about 5% (w / v), between about 0.5% (w / v) and about 5% (w / v), between about 1% (w / v) and about 5% (w / v), between about 0.1% (w / v) and about 1% (w / v), or between about 0.5% (w / v) and about 1% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of about 20% (w / v), about 15% (w / v), about 10% (w / v), about 5% (w / v), about 1% (w / v), about 0.5% (w / v), or about 0.1% (w / v).

[0126] In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of between about 3% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of between about 4% (w / v) and about 6% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the carbohydrate at a concentration of about 5% (w / v).

[0127] In certain embodiments, the liquid arenavirus formulation comprises sucrose at a concentration of up to about 20% (w / v), up to about 15% (w / v), up to about 10% (w / v), up to about 5% (w / v), up to about 1% (w / v), up to about 0.5% (w / v), or up to about 0.1% (w / v). In certain embodiments, the liquid arenavirus formulation disclosed herein comprises sucrose at a concentration of at least about 0.1% (w / v), at least about 0.5% (w / v), at least about 1% (w / v), at least about 5% (w / v), at least about 10% (w / v), at least about 15% (w / v), or at least about 20% (w / v). In certain embodiments, the liquid arenavirus formulation comprises sucrose at a concentration of between about 0.1% (w / v) and about 20% (w / v), between about 0.5% (w / v) and about 20% (w / v), between about 1% (w / v) and about 20% (w / v), between about 5% (w / v) and about 20% (w / v), between about 10% (w / v) and about 20% (w / v), between about 15% (w / v) and about 20% (w / v), between about 0.1% (w / v) and about 15% (w / v), between about 0.5% (w / v) and about 15% (w / v), between about 1% (w / v) and about 15% (w / v), between about 5% (w / v) and about 15% (w / v), between about 10% (w / v) and about 15% (w / v), between about 0.1% (w / v) and about 10% (w / v), between about 0.5% (w / v) and about 10% (w / v), between about 1% (w / v) and about 10% (w / v), between about 5% (w / v) and about 10% (w / v), between about 0.1% (w / v) and about 5% (w / v), between about 0.5% (w / v) and about 5% (w / v), between about 1% (w / v) and about 5% (w / v), between about 0.1% (w / v) and about 1% (w / v), or between about 0.5% (w / v) and about 1% (w / v). In certain embodiments, the liquid arenavirus formulation comprises sucrose at a concentration of about 20% (w / v), about 15% (w / v), about 10% (w / v), about 5% (w / v), about 1% (w / v), about 0.5% (w / v), or about 0.1% (w / v).

[0128] In certain embodiments, the liquid arenavirus formulation comprises sucrose at a concentration of between about 3% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation comprises sucrose at a concentration of between about 4% (w / v) and about 6% (w / v). In certain embodiments, a liquid arenavirus formulation disclosed herein comprises sucrose at a concentration of about 5% (w / v).

[0129] In certain embodiments, the liquid arenavirus formulation comprises trehalose at a concentration of up to about 20% (w / v), up to about 15% (w / v), up to about 10% (w / v), up toabout 5% (w / v), up to about 1% (w / v), up to about 0.5% (w / v), or up to about 0.1% (w / v). In certain embodiments, the liquid arenavirus formulation disclosed herein comprises trehalose at a concentration of at least about 0.1% (w / v), at least about 0.5% (w / v), at least about 1% (w / v), at least about 5% (w / v), at least about 10% (w / v), at least about 15% (w / v), or at least about 20% (w / v). In certain embodiments, the liquid arenavirus formulation comprises trehalose at a concentration of between about 0.1% (w / v) and about 20% (w / v), between about 0.5% (w / v) and about 20% (w / v), between about 1% (w / v) and about 20% (w / v), between about 5% (w / v) and about 20% (w / v), between about 10% (w / v) and about 20% (w / v), between about 15% (w / v) and about 20% (w / v), between about 0.1% (w / v) and about 15% (w / v), between about 0.5% (w / v) and about 15% (w / v), between about 1% (w / v) and about 15% (w / v), between about 5% (w / v) and about 15% (w / v), between about 10% (w / v) and about 15% (w / v), between about 0.1% (w / v) and about 10% (w / v), between about 0.5% (w / v) and about 10% (w / v), between about 1% (w / v) and about 10% (w / v), between about 5% (w / v) and about 10% (w / v), between about 0.1% (w / v) and about 5% (w / v), between about 0.5% (w / v) and about 5% (w / v), between about 1% (w / v) and about 5% (w / v), between about 0.1% (w / v) and about 1% (w / v), or between about 0.5% (w / v) and about 1% (w / v). In certain embodiments, the liquid arenavirus formulation comprises trehalose at a concentration of about 20% (w / v), about 15% (w / v), about 10% (w / v), about 5% (w / v), about 1% (w / v), about 0.5% (w / v), or about 0.1% (w / v).

[0130] In certain embodiments, the liquid arenavirus formulation comprises trehalose at a concentration of between about 3% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation comprises trehalose at a concentration of between about 4% (w / v) and about 6% (w / v). In certain embodiments, a liquid arenavirus formulation disclosed herein comprises trehalose at a concentration of about 5% (w / v).

[0131] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during freezing and thawing. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during freezing and thawing.

[0132] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during freezing and thawing, wherein the concentration of subvisible particles in the formulation is up to about 10%, up to about 20%, up to about 30%, up to about 40%, up to about 50%, up to about 60%, up to about 70%, or up to about 80% of the concentration of subvisible particles in a control formulation without the carbohydrate after the freezing and thawing as measured by FIM. Incertain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during freezing and thawing, wherein the concentration of subvisible particles in the formulation is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% of the concentration of subvisible particles in a control formulation without the carbohydrate after the freezing and thawing as measured by FIM. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the carbohydrate or comprising a different carbohydrate.

[0133] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during freezing and thawing, wherein the viral titer in the formulation after the freezing and thawing is at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer in the formulation before the freezing and thawing as measured by RCV-FFU or FFU assay. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during freezing and thawing, wherein the viral titer in the formulation after the freezing and thawing is about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% of the viral titer in the formulation before the freezing and thawing as measured by RCV-FFU or FFU assay.

[0134] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during freezing and thawing, wherein the viral titer in the formulation after the freezing and thawing is at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 400%, or at least about 500% more than the viral titer in a control formulation without the carbohydrate after the freezing and thawing as measured by RCV-FFU or FFU assay. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during freezing and thawing, wherein the viral titer in the formulation after the freezing and thawing is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 400%, about 500% more than the viral titer in a control formulation without the carbohydrate after the freezing and thawing as measured by RCV-FFU or FFU assay. In certainembodiments, the control formulation is otherwise identical to the liquid arenavirus formulation disclosed herein except not comprising the carbohydrate or comprising a different carbohydrate.

[0135] In certain embodiments, the freezing and thawing comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 7, at least 8, at least 9, at least 10, or more freezing and thawing (F / T) cycles. In certain embodiments, the freezing and thawing comprises 3 F / T cycles. In certain embodiments, the freezing and thawing comprises 5 F / T cycles. In certain embodiments, the temperature of the freezing condition is about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.

[0136] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during storage. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during storage. In certain embodiments, the arenavirus formulation is subject to bioburden reduction filtration and / or sterile filtration before storage.

[0137] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during storage, wherein the concentration of subvisible particles in the formulation after the storage is up to about 10%, up to about 20%, up to about 30%, up to about 40%, up to about 50%, up to about 60%, up to about 70%, or up to about 80% of the concentration of subvisible particles in a control formulation without the carbohydrate after the storage, as measured by FIM. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the formation of subvisible particles during storage, wherein the concentration of subvisible particles in the formulation after the storage is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% of the concentration of subvisible particles in a control formulation without the carbohydrate after the storage, as measured by FIM. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the carbohydrate or comprising a different carbohydrate.

[0138] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during storage, wherein the viral titer in the formulation after the storage is at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer in the formulation before the storage, as measured by RCV-FFU or FFU assay. In certain embodiments, the carbohydrate(e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during storage, wherein the viral titer in the formulation after the storage is about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% of the viral titer in the formulation before the storage, as measured by RCV-FFU or FFU assay.

[0139] In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during storage, wherein the viral titer in the formulation after the storage is at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 400%, or at least about 500% more than the viral titer in a control formulation without the carbohydrate after the storage, as measured by RCV-FFU or FFU assay. In certain embodiments, the carbohydrate (e.g., sucrose, trehalose) is at a concentration that reduces the loss of viral titer during storage, wherein the viral titer in the formulation after the storage is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 400%, about 500% more than the viral titer in a control formulation without the carbohydrate after the storage, as measured by RCV-FFU or FFU assay. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the carbohydrate or comprising a different carbohydrate.

[0140] In certain embodiments, the temperature of the storage is above about 2°C, room temperature (e.g., 20°C - 25°C), between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C. In certain embodiments, the temperature of the storage is about -20°C. In certain embodiments, the temperature of the storage is about -65°C. In certain embodiments, the temperature of the storage is about -80°C. In certain embodiments, the storage lasts at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 3 years, at least about 4 years, at least about 5 years, or longer. In certain embodiments, the storage lasts at least about 6 months. In certain embodiments, the storage lasts at least about 9 months. In certain embodiments, the storage lasts at least about 18 months.

[0141] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of up to about 10% (w / v), up to about 8% (w / v), up to about 6% (w / v), up to about 4% (w / v), at least about 3% (w / v), at least about 6% (w / v), at least about 8% (w / v), between about 3% and about 10% (w / v), between about 3% (w / v) and about 8% (w / v), between about 3% and about 6% (w / v), between about 4% and about 10% (w / v), between about 4% and about 8% (w / v), between about 4% and about 6% (w / v), between about 6% and about 10% (w / v), between about 6% and about 8% (w / v), or between about 8% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of about 5% (w / v).

[0142] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of up to about 10% (w / v), up to about 8% (w / v), up to about 6% (w / v), up to about 4% (w / v), at least about 3% (w / v), at least about 6% (w / v), at least about 8% (w / v), between about 3% and about 10% (w / v), between about 3% (w / v) and about 8% (w / v), between about 3% and about 6% (w / v), between about 4% and about 10% (w / v), between about 4% and about 8% (w / v), between about 4% and about 6% (w / v), between about 6% and about 10% (w / v), between about 6% and about 8% (w / v), or between about 8% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of about 3%, about 4%, about 5%, about 6%, about 7%, about 8%, about 9%, or about 10%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of about 5% (w / v).5.1.3 Salts

[0143] In certain embodiments, the liquid arenavirus formulation comprises a salt. Without being bound by any theory, addition of one or more salts to the liquid arenavirus formulations disclosed herein can increase the stability of the arenavirus particles during freezing and thawing. Without being bound by any particular theory, the addition of salts to the formulations described herein increases the ionic strength for enhanced stability of the virus particles and prevents aggregate formation of the virus particles for improved filterability during the process of manufacturing the arenavirus formulations disclosed herein.

[0144] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises a salt selected from the group consisting of sodium chloride (NaCl), sodium phosphate (NasPCh), potassium phosphate (K3PO4), potassium chloride (KC1), magnesium chloride (MgCh), sodium sulfate (Na2SO4), potassium sulfate (K2SO4), ammonium sulfate ((NEU^SC ), and combinations thereof. In certain embodiments, the liquid arenavirus formulation disclosed herein comprises sodium chloride.

[0145] In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of up to about 450 mM, up to about 400 mM, up to about 350 mM, up to about 300 mM, up to about 250 mM, up to about 225 mM, up to about 200 mM, up to about 175 mM, up to about 150 mM, up to about 125 mM, up to about 100 mM, up to about 75 mM, or up to about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of at least about 50 mM, at least about 75 mM, at least about 100 mM, at least about 125 mM, at least about 150 mM, at least about 175 mM, at least about 200 mM, at least about 225 mM, at least about 250 mM, at least about 300 mM, at least about 350 mM, at least about 400 mM, or at least about 450 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of between about 50 mM and about 450 mM, between about 50 mM and about 400 mM, between about 50 mM and about 350 mM, between about 50 mM and about 300 mM, between about 50 mM and about 250 mM, between about 50 mM and about 225 mM, between about 50 mM and about 200 mM, between about 50 mM and about 175 mM, between about 50 mM and about 150 mM, between about 50 mM and about 125 mM, between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 450 mM, between about 75 mM and about 400 mM, between about 75 mM and about 350 mM, between about 75 mM and about 300 mM, between about 75 mM and about 250 mM, between about 75 mM and about 225 mM, between about 75 mM and about 200 mM, between about 75 mM and about 175 mM, between about 75 mM and about 150 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, between about 100 mM and about 450 mM, between about 100 mM and about 400 mM, between about 100 mM and about 350 mM, between about 100 mM and about 300 mM, between about 100 mM and about 250 mM, between about 100 mM and about 225 mM, between about 100 mM and about 200 mM, between about 100 mM and about 175 mM, between about 100 mM and about 150 mM, between about 100 mM and about 125 mM, between about 125 mM and about 450 mM, between about 125 mM and about 400 mM, between about 125 mM and about 350 mM, between about 125 mM and about 300 mM, between about 125 mM and about 250 mM,between about 125 mM and about 225 mM, between about 125 mM and about 200 mM, between about 125 mM and about 175 mM, between about 125 mM and about 150 mM, between about 150 mM and about 450 mM, between about 150 mM and about 400 mM, between about 150 mM and about 350 mM, between about 150 mM and about 300 mM, between about 150 mM and about 250 mM, between about 150 mM and about 225 mM, between about 150 mM and about 200 mM, between about 150 mM and about 175 mM, between about 175 mM and about 450 mM, between about 175 mM and about 400 mM, between about 175 mM and about 350 mM, between about 175 mM and about 300 mM, between about 175 mM and about 250 mM, between about 175 mM and about 225 mM, between about 175 mM and about 200 mM, between about 200 mM and about 450 mM, between about 200 mM and about 400 mM, between about 200 mM and about 350 mM, between about 200 mM and about 300 mM, between about 200 mM and about 250 mM, between about 200 mM and about 225 mM, between about 225 mM and about 450 mM, between about 225 mM and about 400 mM, between about 225 mM and about 350 mM, between about 225 mM and about 300 mM, between about 225 mM and about 250 mM, between about 250 mM and about 450 mM, between about 250 mM and about 400 mM, between about 250 mM and about 350 mM, between about 250 mM and about 300 mM, between about 300 mM and about 450 mM, between about 300 mM and about 400 mM, between about 300 mM and about 350 mM, between about 350 mM and about 450 mM, between about 350 mM and about 400 mM, or between about 400 mM and about 450 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of about 450 mM, about 400 mM, about 350 mM, about 300 mM, about 250 mM, about 225 mM, about 200 mM, about 175 mM, about 150 mM, about 125 mM, about 100 mM, about 75 mM, about 50 mM, or 0 mM.

[0146] In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of between about 50 mM and about 250 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of between about 100 mM and about 150 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of about 125 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of between about 50 mM and about 100 mM. In certain embodiments, the liquid arenavirus formulation comprises the salt at a concentration of about 75 mM.

[0147] In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of up to about 450 mM, up to about 400 mM, up to about 350mM, up to about 300 mM, up to about 250 mM, up to about 225 mM, up to about 200 mM, up to about 175 mM, up to about 150 mM, up to about 125 mM, up to about 100 mM, up to about 75 mM, or up to about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of at least about 50 mM, at least about 75 mM, at least about 100 mM, at least about 125 mM, at least about 150 mM, at least about 175 mM, at least about 200 mM, at least about 225 mM, at least about 250 mM, at least about 300 mM, at least about 350 mM, at least about 400 mM, or at least about 450 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of between about 50 mM and about 450 mM, between about 50 mM and about 400 mM, between about 50 mM and about 350 mM, between about 50 mM and about 300 mM, between about 50 mM and about 250 mM, between about 50 mM and about 225 mM, between about 50 mM and about 200 mM, between about 50 mM and about 175 mM, between about 50 mM and about 150 mM, between about 50 mM and about 125 mM, between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 450 mM, between about 75 mM and about 400 mM, between about75 mM and about 350 mM, between about 75 mM and about 300 mM, between about 75 mM and about 250 mM, between about 75 mM and about 225 mM, between about 75 mM and about 200 mM, between about 75 mM and about 175 mM, between about 75 mM and about 150 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, between about 100 mM and about 450 mM, between about 100 mM and about 400 mM, between about 100 mM and about 350 mM, between about 100 mM and about 300 mM, between about 100 mM and about 250 mM, between about 100 mM and about 225 mM, between about 100 mM and about 200 mM, between about 100 mM and about 175 mM, between about 100 mM and about 150 mM, between about 100 mM and about 125 mM, between about 125 mM and about 450 mM, between about 125 mM and about 400 mM, between about 125 mM and about 350 mM, between about 125 mM and about 300 mM, between about 125 mM and about 250 mM, between about 125 mM and about 225 mM, between about 125 mM and about 200 mM, between about 125 mM and about 175 mM, between about 125 mM and about 150 mM, between about 150 mM and about 450 mM, between about 150 mM and about 400 mM, between about 150 mM and about 350 mM, between about 150 mM and about 300 mM, between about 150 mM and about 250 mM, between about 150 mM and about 225 mM, between about 150 mM and about 200 mM, between about 150 mM and about 175 mM, between about 175 mM and about 450 mM, between about 175 mM and about 400 mM, between about 175 mM and about 350 mM,between about 175 mM and about 300 mM, between about 175 mM and about 250 mM, between about 175 mM and about 225 mM, between about 175 mM and about 200 mM, between about 200 mM and about 450 mM, between about 200 mM and about 400 mM, between about 200 mM and about 350 mM, between about 200 mM and about 300 mM, between about 200 mM and about 250 mM, between about 200 mM and about 225 mM, between about 225 mM and about 450 mM, between about 225 mM and about 400 mM, between about 225 mM and about 350 mM, between about 225 mM and about 300 mM, between about 225 mM and about 250 mM, between about 250 mM and about 450 mM, between about 250 mM and about 400 mM, between about 250 mM and about 350 mM, between about 250 mM and about 300 mM, between about 300 mM and about 450 mM, between about 300 mM and about 400 mM, between about 300 mM and about 350 mM, between about 350 mM and about 450 mM, between about 350 mM and about 400 mM, or between about 400 mM and about 450 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of about 450 mM, about 400 mM, about 350 mM, about 300 mM, about 250 mM, about 225 mM, about 200 mM, about 175 mM, about 150 mM, about 125 mM, about 100 mM, about 75 mM, about 50 mM, or 0 mM.

[0148] In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of between about 50 mM and about 250 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of between about 100 mM and about 150 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of about 125 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of between about 50 mM and about 100 mM. In certain embodiments, the liquid arenavirus formulation comprises sodium chloride at a concentration of about 75 mM.

[0149] In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the formation of subvisible particles during manufacturing. In certain embodiments, the manufacturing comprises a purification process. In certain embodiments, the manufacturing comprises at least one chromatography step. In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the loss of viral titer during filtration. In certain embodiments, the filtration comprises bioburden reduction filtration, sterile filtration, or a combination thereof.

[0150] In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is up to about 10%, up to about 20%, up to about30%, up to about 40%, up to about 50%, up to about 60%, up to about 70%, or up to about 80% of the concentration of subvisible particles in a control formulation without the salt, as measured by FIM. In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% of the concentration of subvisible particles in a control formulation without the salt, as measured by FIM. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the salt or comprising a different salt.

[0151] In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer in the formulation prior to the filtration, as measured by RCV-FFU or FFU assay. In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% of the viral titer in the formulation prior to the filtration, as measured by RCV-FFU or FFU assay.

[0152] In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 400%, or at least about 500% more than the viral titer in a control formulation without the salt after the filtration, as measured by RCV-FFU or FFU assay. In certain embodiments, the salt (e.g., sodium chloride) is at a concentration that reduces the loss of viral titer during filtration, wherein the viral titer in the formulation after the filtration is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 400%, about 500% more than the viral titer in a control formulation without the salt after the filtration, as measured by RCV-FFU or FFU assay. In certain embodiments, the control formulation isotherwise identical to the presently disclosed formulation except not comprising the salt or comprising a different salt.

[0153] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of up to about 125 mM, up to about 100 mM, up to about 75 mM, at least about 50 mM, at least about 75 mM, at least about 100 mM, between about 50 mM and about 125 mM, between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, or between about 100 mM and about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of about 50 mM, about 75 mM, about 100 mM, or about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of up to about 125 mM or between about 50 mM and about 125 mM.

[0154] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of up to about 125 mM, up to about 100 mM, up to about 75 mM, at least about 50 mM, at least about 75 mM, at least about 100 mM, between about 50 mM and about 125 mM, between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, or between about 100 mM and about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of about 50 mM, about 75 mM, about 100 mM, or about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM.5.1.4 Buffers

[0155] In certain embodiments, the liquid arenavirus formulation comprises a buffer. A buffer refers to a buffered solution that resists changes in pH by the action of its acid-base conjugate components.

[0156] The pH of the buffer used herein can generally be in the range of a physiological pH (e.g., between about 6 to about 8); however, one of skill in the art can appreciate that a wider range of pH may be necessary for stability of the arenavirus particles and / or other components present in a particular arenavirus formulation disclosed herein. In certain embodiments, a buffer disclosed herein can have a pH up to about 6.5, up to about 7.0, up toabout 7.5, up to about 8.0, up to about 8.5, up to about 9.0, at least about 6.5, at least about 7.0, at least about 7.5, at least about 8.0, at least about 8.5, at least about 9.0, between about 6.5 and about 9.0, between about 6.5 and about 8.5, between about 6.5 and about 8.0, between about 6.5 and about 7.5, between about 6.5 and about 7.0, between about 7.0 and about 7.5, between about 7.5 and about 8.0, between about 8.0 and about 8.5, or between about 8.5 and about 9.0. In certain embodiments, a buffer disclosed herein can have a pH of between about 7.0 and about 9.0. In certain embodiments, a buffer disclosed herein can have a pH of about 6.5, about 6.6, about 6.7, about 6.8, about 6.9, about 7.0, about 7.1, about 7.2, about 7.3, about 7.4, about 7.5, about 7.6, about 7.7, about 7.8, about 7.9, about 8.0, about 8.1, about 8.2, about 8.3, about 8.4, about 8.5, about 8.6, about 8.7, about 8.8, about 8.9, or about 9.0. In certain embodiments, a buffer disclosed herein can have a pH of about 8.0.

[0157] Examples of suitable buffers that can be used with the present disclosure include, but are not limited to, phosphate (e.g., potassium phosphate, sodium phosphate), citrate (e.g., sodium citrate), sodium acetate, sodium succinate, histidine, imidazole, bicarbonate, carbonate, HEPES, TRIS, and the like. In certain embodiments, a liquid arenavirus formulation disclosed herein can comprise a buffer selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof. A HEPES (N-2-Hydroxyethylpiperazine-N'-2-ethanesulfonic Acid (CsHislShChS)) buffer as used herein refers to a zwitterionic N-substituted aminosulfonic acid buffer that can have a pKa of 7.5 at 25°C and may be useful in the pH 6.8-8.2 range. A TRIS (C4H11NO3) buffer as used herein can have a pKa of 8.1 at 25°C and may be useful in the pH range of 7.0-9.0. In certain embodiments, the liquid arenavirus formulation disclosed herein comprises a HEPES buffer. In certain embodiments, the liquid arenavirus formulation disclosed herein comprises a TRIS buffer.

[0158] In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of up to about 50 mM, up to about 45 mM, up to about 40 mM, up to about 35 mM, up to about 30 mM, up to about 25 mM, up to about 20 mM, up to about 15 mM, up to about 10 mM, up to about 5 mM, or up to about 1 mM. In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of at least about 1 mM, at least about 5 mM, at least about 10 mM, at least about 15 mM, at least about 20 mM, at least about 25 mM, at least about 30 mM, at least about 35 mM, at least about 40 mM, at least about 45 mM, or at least about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of between about 1 mM and about 50 mM, between about 1 mM and about 45 mM, between about 1 mM and about 40 mM, between about 1 mMand about 35 mM, between about 1 mM and about 30 mM, between about 1 mM and about 25 mM, between about 1 mM and about 20 mM, between about 1 mM and about 15 mM, between about 1 mM and about 10 mM, between about 1 mM and about 5 mM, between about 5 mM and about 50 mM, between about 5 mM and about 45 mM, between about 5 mM and about 40 mM, between about 5 mM and about 35 mM, between about 5 mM and about 30 mM, between about 5 mM and about 25 mM, between about 5 mM and about 20 mM, between about 5 mM and about 15 mM, between about 5 mM and about 10 mM, between about 10 mM and about 50 mM, between about 10 mM and about 45 mM, between about 10 mM and about 40 mM, between about 10 mM and about 35 mM, between about 10 mM and about 30 mM, between about 10 mM and about 25 mM, between about 10 mM and about 20 mM, between about 10 mM and about 15 mM, between about 15 mM and about 50 mM, between about 15 mM and about 45 mM, between about 15 mM and about 40 mM, between about 15 mM and about 35 mM, between about 15 mM and about 30 mM, between about 15 mM and about 25 mM, between about 15 mM and about 20 mM, between about 20 mM and about 50 mM, between about 20 mM and about 45 mM, between about 20 mM and about 40 mM, between about 20 mM and about 35 mM, between about 20 mM and about 30 mM, between about 20 mM and about 25 mM, between about 25 mM and about 50 mM, between about 25 mM and about 45 mM, between about 25 mM and about 40 mM, between about 25 mM and about 35 mM, between about 25 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 45 mM, between about 30 mM and about 40 mM, between about 30 mM and about 35 mM, between about 35 mM and about 50 mM, between about 35 mM and about 45 mM, between about 35 mM and about 40 mM, between about 40 mM and about 50 mM, between about 40 mM and about 45 mM, or between about 45 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of about 50 mM, about 45 mM, about 40 mM, about 35 mM, about 30 mM, about 25 mM, about 20 mM, about 15 mM, about 10 mM, about 5 mM, or about 1 mM.

[0159] In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of between about 1 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of between about 10 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation comprises the buffer at a concentration of between about 5 mM and about 15 mM. In certain embodiments, the liquid arenavirus formulation disclosed herein comprises the buffer at a concentration of about 10 mM.

[0160] In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of up to about 50 mM, up to about 45 mM, up to about 40 mM, up to about 35 mM, up to about 30 mM, up to about 25 mM, up to about 20 mM, up to about 15 mM, up to about 10 mM, up to about 5 mM, or up to about 1 mM. In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of at least about 1 mM, at least about 5 mM, at least about 10 mM, at least about 15 mM, at least about 20 mM, at least about 25 mM, at least about 30 mM, at least about 35 mM, at least about 40 mM, at least about 45 mM, or at least about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of between about 1 mM and about 50 mM, between about 1 mM and about 45 mM, between about 1 mM and about 40 mM, between about 1 mM and about 35 mM, between about 1 mM and about 30 mM, between about 1 mM and about 25 mM, between about 1 mM and about 20 mM, between about 1 mM and about 15 mM, between about 1 mM and about 10 mM, between about 1 mM and about 5 mM, between about 5 mM and about 50 mM, between about 5 mM and about 45 mM, between about 5 mM and about 40 mM, between about 5 mM and about 35 mM, between about 5 mM and about 30 mM, between about 5 mM and about 25 mM, between about 5 mM and about 20 mM, between about 5 mM and about 15 mM, between about 5 mM and about 10 mM, between about 10 mM and about 50 mM, between about 10 mM and about 45 mM, between about 10 mM and about 40 mM, between about 10 mM and about 35 mM, between about 10 mM and about 30 mM, between about 10 mM and about 25 mM, between about 10 mM and about 20 mM, between about 10 mM and about 15 mM, between about 15 mM and about 50 mM, between about 15 mM and about 45 mM, between about 15 mM and about 40 mM, between about 15 mM and about 35 mM, between about 15 mM and about 30 mM, between about 15 mM and about 25 mM, between about 15 mM and about 20 mM, between about 20 mM and about 50 mM, between about 20 mM and about 45 mM, between about 20 mM and about 40 mM, between about 20 mM and about 35 mM, between about 20 mM and about 30 mM, between about 20 mM and about 25 mM, between about 25 mM and about 50 mM, between about 25 mM and about 45 mM, between about 25 mM and about 40 mM, between about 25 mM and about 35 mM, between about 25 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 45 mM, between about 30 mM and about 40 mM, between about 30 mM and about 35 mM, between about 35 mM and about 50 mM, between about 35 mM and about 45 mM, between about 35 mM and about 40 mM, between about 40 mM and about 50 mM, between about 40 mM and about 45 mM, or between about 45 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulationcomprises TRIS at a concentration of about 50 mM, about 45 mM, about 40 mM, about 35 mM, about 30 mM, about 25 mM, about 20 mM, about 15 mM, about 10 mM, about 5 mM, or about 1 mM.

[0161] In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of between about 1 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of between about 10 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation comprises TRIS at a concentration of between about 5 mM and about 15 mM. In certain embodiments, the liquid arenavirus formulation disclosed herein comprises TRIS at a concentration of about 10 mM.

[0162] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration of up to about 20 mM, up to about 18 mM, up to about 15 mM, at least about 10 mM, at least about 15 mM, at least about 18 mM, between about 10 mM and about 20 mM, between about 10 mM and about 18 mM, between about 10 mM and about 15 mM, between about 15 mM and about 20 mM, between about 15 mM and about 18 mM, or between about 18 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration of about 10 mM, about 12 mM, about 14 mM, about 16 mM, about 18 mM, or about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration of about 10 mM.

[0163] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration of up to about 20 mM, up to about 18 mM, up to about 15 mM, at least about 10 mM, at least about 15 mM, at least about 18 mM, between about 10 mM and about 20 mM, between about 10 mM and about 18 mM, between about 10 mM and about 15 mM, between about 15 mM and about 20 mM, between about 15 mM and about 18 mM, or between about 18 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration of about 10 mM, about 12 mM, about 14 mM, about 16 mM, about 18 mM, or about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration of about 10 mM.5.1.5 Surfactants

[0164] In certain embodiments, the liquid arenavirus formulation comprises a surfactant. Without being bound by any theory, addition of one or more surfactants to the liquid arenavirus formulations disclosed herein can prevent or reduce the formation of subvisible particles in the arenavirus formulations.

[0165] Suitable surfactants that can be used with the present disclosure include, but are not limited to, alkylphenyl alkoxylates, alcohol alkoxylates, fatty amine alkoxylates, polyoxyethylene glycerol fatty acid esters, castor oil alkoxylates, fatty acid alkoxylates, fatty acid amide alkoxylates, fatty acid polydiethanolamides, lanolin ethoxylates, fatty acid polyglycol esters, isotridecyl alcohol, fatty acid amides, methylcellulose, fatty acid esters, silicone oils, alkyl polyglycosides, glycerol fatty acid esters, polyethylene glycol, polypropylene glycol, polyethylene glycol / polypropylene glycol block copolymers, polyethylene glycol alkyl ethers, polypropylene glycol alkyl ethers, polyethylene glycol / polypropylene glycol ether block copolymers, polyacrylates, acrylic acid graft copolymers, alkylaryl sulfonates, phenyl sulfonates, alkyl sulfates, alkyl sulfonates, alkyl ether sulfates, alkyl aryl ether sulfates, alkyl polyglycol ether phosphates, polyaryl phenyl ether phosphates, alkyl sulfosuccinates, olefin sulfonates, paraffin sulfonates, petroleum sulfonates, taurides, sarcosides, fatty acids, alkylnaphthalenesulfonic acids, naphthalenesulfonic acids, lignosulfonic acids, condensates of sulfonated naphthalenes, lignin-sulfite waste liquor, alkyl phosphates, quaternary ammonium compounds, amine oxides, betaines, and the like. In certain embodiments, the surfactant is a nonionic surfactant, an ionic surfactant, or a combination thereof. In certain embodiments, the surfactant is a polysorbate, a polyethylene glycol sorbitan monolaurate (Tween®20), a polyoxy ethylenesorbitan monooleate (Tween®80), a block copolymer of polyethylene and polypropylene glycol e.g., a pol oxamer or Pluronic®), or a combination thereof.

[0166] In certain embodiments, the surfactant is a poloxamer. A poloxamer is a nonionic triblock copolymer composed of a central hydrophobic chain of polyoxypropylene (polypropylene oxide)) flanked by two hydrophilic chains of polyoxyethylene (poly(ethylene oxide)). Because the lengths of the polymer blocks can be customized, many different pol oxamers that have slightly different properties are available for use herein. For the generic term, a poloxamer is commonly named with the letter “P” (for poloxamer) followed by three digits: the first two digits multiplied by 100 give the approximate molecular mass of the poly oxypropylene core, and the last digit multiplied by 10 gives the percentage polyoxyethylene content (e.g., P407 = poloxamer with a polyoxypropylene molecular massof 4000 g / mol and a 70% polyoxyethylene content). In certain embodiments, the surfactant for use with the present disclosure is selected from the group consisting of pol oxamer 188, poloxamer 407, and combinations thereof.

[0167] In certain embodiments, the surfactant for use with the present disclosure is a polysorbate. Polysorbates are oily liquids derived from ethoxylated sorbitan (a derivative of sorbitol) esterified with fatty acids. Many examples of polysorbates are available, wherein the number following “polysorbate” relates to the type of major fatty acid associated with the molecule. For example, monolaurate is indicated by “20” in “polysorbate 20” and monooleate by “80” in “polysorbate 80.” In certain embodiments, the surfactant for use with the present disclosure is selected from the group consisting of polysorbate 20, polysorbate 80, and combinations thereof.

[0168] In certain embodiments, the surfactant is a poloxamer, a polysorbate, or a combination thereof. In certain embodiments, the surfactant is selected from the group consisting of poloxamer 188, poloxamer 407, polysorbate 20, polysorbate 80, and combinations thereof. In certain embodiments, the surfactant is poloxamer 188.

[0169] In certain embodiments, the liquid arenavirus formulation comprises the surfactant at a concentration of up to about 0.9% (w / v), up to about 0.8% (w / v), up to about 0.7% (w / v), up to about 0.6% (w / v), up to about 0.5% (w / v), up to about 0.4% (w / v), up to about 0.3% (w / v), up to about 0.2% (w / v), up to about 0.1% (w / v), up to about 0.09% (w / v), up to about 0.08% (w / v), up to about 0.07% (w / v), up to about 0.06% (w / v), up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), up to about 0.01% (w / v), up to about 0.009% (w / v), up to about 0.008% (w / v), up to about 0.007% (w / v), up to about 0.006% (w / v), up to about 0.005% (w / v), up to about 0.004% (w / v), up to about 0.003% (w / v), up to about 0.002% (w / v), or up to about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the surfactant at a concentration of at least about 0.001% (w / v), at least about 0.002% (w / v), at least about 0.003% (w / v), at least about 0.004% (w / v), at least about 0.005% (w / v), at least about 0.006% (w / v), at least about 0.007% (w / v), at least about 0.008% (w / v), at least about 0.009% (w / v), at least about 0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), at least about 0.05% (w / v), at least about 0.06% (w / v), at least about 0.07% (w / v), at least about 0.08% (w / v), at least about 0.09% (w / v), at least about 0.1% (w / v), at least about 0.2% (w / v), at least about 0.3% (w / v), at least about 0.4% (w / v), at least about 0.5% (w / v), at least about 0.6% (w / v), at least about 0.7% (w / v), at least about 0.8% (w / v), or at least about 0.9% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the surfactant at aconcentration of between about 0.001% (w / v) and about 0.9% (w / v), between about 0.01% (w / v) and about 0.9% (w / v), between about 0.05% (w / v) and about 0.9% (w / v), between about 0.1% (w / v) and about 0.9% (w / v), between about 0.5% (w / v) and about 0.9% (w / v), between about 0.001% (w / v) and about 0.5% (w / v), between about 0.01% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.5% (w / v), between about 0.1% (w / v) and about 0.5% (w / v), between about 0.001% (w / v) and about 0.1% (w / v), between about 0.01% (w / v) and about 0.1% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), between about 0.001% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04% (w / v), between about 0.02% (w / v) and about 0.04% (w / v), or between about 0.001% (w / v) and about 0.01% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the surfactant at a concentration of about 0.9% (w / v), about 0.8% (w / v), about 0.7% (w / v), about 0.6% (w / v), about 0.5% (w / v), about 0.4% (w / v), about 0.3% (w / v), about 0.2% (w / v), about 0.1% (w / v), about 0.09% (w / v), about 0.08% (w / v), about 0.07% (w / v), about 0.06% (w / v), about 0.05% (w / v), about 0.04% (w / v), about 0.03% (w / v), about 0.02% (w / v), about 0.01% (w / v), about 0.009% (w / v), about 0.008% (w / v), about 0.007% (w / v), about 0.006% (w / v), about 0.005% (w / v), about 0.004% (w / v), about 0.003% (w / v), about 0.002% (w / v), or about 0.001% (w / v).

[0170] In certain embodiments, the liquid arenavirus formulation comprises the surfactant at a concentration of between about 0.01% (w / v) and about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the surfactant at a concentration of about 0.03% (w / v).

[0171] In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of up to about 0.9% (w / v), up to about 0.8% (w / v), up to about 0.7% (w / v), up to about 0.6% (w / v), up to about 0.5% (w / v), up to about 0.4% (w / v), up to about 0.3% (w / v), up to about 0.2% (w / v), up to about 0.1% (w / v), up to about 0.09% (w / v), up to about 0.08% (w / v), up to about 0.07% (w / v), up to about 0.06% (w / v), up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), up to about 0.01% (w / v), up to about 0.009% (w / v), up to about 0.008% (w / v), up to about 0.007% (w / v), up to about 0.006% (w / v), up to about 0.005% (w / v), up to about 0.004% (w / v), up to about 0.003% (w / v), up to about 0.002% (w / v), or up to about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of at least about 0.001% (w / v), at least about 0.002% (w / v), at least about 0.003% (w / v), at least about 0.004% (w / v), at least about 0.005% (w / v), at least about 0.006% (w / v), at least about 0.007% (w / v), at least about 0.008% (w / v), at least about 0.009% (w / v), at least about0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), at least about 0.05% (w / v), at least about 0.06% (w / v), at least about 0.07% (w / v), at least about 0.08% (w / v), at least about 0.09% (w / v), at least about 0.1% (w / v), at least about 0.2% (w / v), at least about 0.3% (w / v), at least about 0.4% (w / v), at least about 0.5% (w / v), at least about 0.6% (w / v), at least about 0.7% (w / v), at least about 0.8% (w / v), or at least about 0.9% (w / v). In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of between about 0.001% (w / v) and about 0.9% (w / v), between about 0.01% (w / v) and about 0.9% (w / v), between about 0.05% (w / v) and about 0.9% (w / v), between about 0.1% (w / v) and about 0.9% (w / v), between about 0.5% (w / v) and about 0.9% (w / v), between about 0.001% (w / v) and about 0.5% (w / v), between about 0.01% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.5% (w / v), between about 0.1% (w / v) and about 0.5% (w / v), between about 0.001% (w / v) and about 0.1% (w / v), between about 0.01% (w / v) and about 0.1% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), between about 0.001% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04% (w / v), between about 0.02% (w / v) and about 0.04% (w / v), or between about 0.001% (w / v) and about 0.01% (w / v). In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of about 0.9% (w / v), about 0.8% (w / v), about 0.7% (w / v), about 0.6% (w / v), about 0.5% (w / v), about 0.4% (w / v), about 0.3% (w / v), about 0.2% (w / v), about 0.1% (w / v), about 0.09% (w / v), about 0.08% (w / v), about 0.07% (w / v), about 0.06% (w / v), about 0.05% (w / v), about 0.04% (w / v), about 0.03% (w / v), about 0.02% (w / v), about 0.01% (w / v), about 0.009% (w / v), about 0.008% (w / v), about 0.007% (w / v), about 0.006% (w / v), about 0.005% (w / v), about 0.004% (w / v), about 0.003% (w / v), about 0.002% (w / v), or about 0.001% (w / v).

[0172] In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of between about 0.01% (w / v) and about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation comprises poloxamer 188 at a concentration of about 0.03% (w / v).

[0173] In certain embodiments, the surfactant (e.g., poloxamer 188) is at a concentration that reduces the formation of subvisible particles during manufacturing. In certain embodiments, the manufacturing comprises a purification process. In certain embodiments, the manufacturing comprises at least one chromatography step.

[0174] In certain embodiments, the surfactant (e.g., poloxamer 188) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is up to about 10%, up to about 20%,up to about 30%, up to about 40%, up to about 50%, up to about 60%, up to about 70%, or up to about 80% of the concentration of subvisible particles in a control formulation without the surfactant, as measured by FIM. In certain embodiments, the surfactant (e.g., pol oxamer 188) is at a concentration that reduces the formation of subvisible particles during manufacturing, wherein the concentration of subvisible particles in the formulation is about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, or about 80% of the concentration of subvisible particles in a control formulation without the surfactant, as measured by FIM. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the surfactant or comprising a different surfactant.

[0175] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), at least about 0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), between about 0.01% and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04% (w / v), between about 0.01% and about 0.03% (w / v), between about 0.01% and about 0.02% (w / v), between about 0.02% and about 0.05% (w / v), between about 0.02% and about 0.04% (w / v), between about 0.02% and about 0.03% (w / v), between about 0.03% and about 0.05% (w / v), between about 0.03% and about 0.04% (w / v), or between about 0.04% and about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of about 0.01%, about 0.02%, about 0.03%, about 0.04%, or about 0.05%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of about 0.03% (w / v).

[0176] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises pol oxamer 188 at a concentration of up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), at least about 0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), between about 0.01% and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04%(w / v), between about 0.01% and about 0.03% (w / v), between about 0.01% and about 0.02%(w / v), between about 0.02% and about 0.05% (w / v), between about 0.02% and about 0.04%(w / v), between about 0.02% and about 0.03% (w / v), between about 0.03% and about 0.05%(w / v), between about 0.03% and about 0.04% (w / v), or between about 0.04% and about0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises poloxamer 188 at a concentration of about 0.01%, about 0.02%, about 0.03%, about 0.04%, or about 0.05%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises poloxamer 188 at a concentration of about 0.03% (w / v).5.1.6 Stabilizers

[0177] In certain embodiments, the liquid arenavirus formulation comprises a stabilizer. Without being bound by any theory, addition of the stabilizer to the liquid arenavirus formulations disclosed herein inhibits non-specific adsorption of virus particles throughout manufacture, formulation, and / or storage, and / or stabilizes the arenavirus formulations during the freezing, thawing and storage process.

[0178] In certain embodiments, the stabilizer is a protein. In certain embodiments, the protein is selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof. In certain embodiments, the stabilizer is an albumin. In certain embodiments, the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof. In certain embodiments, the stabilizer is rHSA.

[0179] In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of up to about 3% (w / v), up to about 2% (w / v), up to about 1% (w / v), up to about 0.5% (w / v), up to about 0.1% (w / v), up to about 0.05% (w / v), up to about 0.01% (w / v), up to about 0.005% (w / v), or up to about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of at least about 3% (w / v), at least about 2% (w / v), at least about 1% (w / v), at least about 0.5% (w / v), at least about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.01% (w / v), at least about 0.005% (w / v), or at least about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of between about 0.001% (w / v) and about 3% (w / v), between about 0.005% (w / v) and about 3% (w / v), between about 0.01% (w / v) and about 3% (w / v), between about 0.05% (w / v) and about 3% (w / v), between about 0.1% (w / v) and about 3% (w / v), between about 0.5% (w / v) and about 3% (w / v), between about 1% (w / v) and about 3% (w / v), between about 2% (w / v) and about 3% (w / v), between about 0.001% (w / v) and about 2% (w / v), between about 0.005% (w / v) and about 2% (w / v), between about 0.01% (w / v) and about 2% (w / v), between about 0.05% (w / v) and about 2% (w / v), between about 0.1% (w / v) and about 2% (w / v), between about 0.5% (w / v) and about 2% (w / v), between about 1% (w / v) and about 2% (w / v), between about 0.001% (w / v) andabout 1% (w / v), between about 0.005% (w / v) and about 1% (w / v), between about 0.01% (w / v) and about 1% (w / v), between about 0.05% (w / v) and about 1% (w / v), between about 0.1% (w / v) and about 1% (w / v), between about 0.5% (w / v) and about 1% (w / v), between about 0.001% (w / v) and about 0.5% (w / v), between about 0.005% (w / v) and about 0.5% (w / v), between about 0.01% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.5% (w / v), between about 0.1% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.3% (w / v), between about 0.05% (w / v) and about 0.2% (w / v), between about 0.05% (w / v) and about 0.15% (w / v), between about 0.001% (w / v) and about 0.1% (w / v), between about 0.005% (w / v) and about 0.1% (w / v), between about 0.01% (w / v) and about 0.1% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), between about 0.001% (w / v) and about 0.05% (w / v), between about 0.005% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.05% (w / v), between about 0.001% (w / v) and about 0.01% (w / v), between about 0.005% (w / v) and about 0.01% (w / v), or between about 0.001% (w / v) and about 0.005% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of about 3% (w / v), about 2% (w / v), about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v), about 0.01% (w / v), about 0.005% (w / v), or about 0.001% (w / v).

[0180] In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of between about 0.05% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation comprises the stabilizer at a concentration of about 0.1% (w / v).

[0181] In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of up to about 3% (w / v), up to about 2% (w / v), up to about 1% (w / v), up to about 0.5% (w / v), up to about 0.1% (w / v), up to about 0.05% (w / v), up to about 0.01% (w / v), up to about 0.005% (w / v), or up to about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of at least about 3% (w / v), at least about 2% (w / v), at least about 1% (w / v), at least about 0.5% (w / v), at least about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.01% (w / v), at least about 0.005% (w / v), or at least about 0.001% (w / v). In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of between about 0.001% (w / v) and about 3% (w / v), between about 0.005% (w / v) and about 3% (w / v), between about 0.01% (w / v) and about 3% (w / v), between about 0.05% (w / v) and about 3% (w / v), between about 0.1% (w / v) and about 3% (w / v), between about 0.5% (w / v) and about 3% (w / v), between about 1% (w / v) and about 3% (w / v), between about 2% (w / v) and about 3% (w / v), between about 0.001% (w / v) and about 2%(w / v), between about 0.005% (w / v) and about 2% (w / v), between about 0.01% (w / v) and about 2% (w / v), between about 0.05% (w / v) and about 2% (w / v), between about 0.1% (w / v) and about 2% (w / v), between about 0.5% (w / v) and about 2% (w / v), between about 1% (w / v) and about 2% (w / v), between about 0.001% (w / v) and about 1% (w / v), between about 0.005% (w / v) and about 1% (w / v), between about 0.01% (w / v) and about 1% (w / v), between about 0.05% (w / v) and about 1% (w / v), between about 0.1% (w / v) and about 1% (w / v), between about 0.5% (w / v) and about 1% (w / v), between about 0.001% (w / v) and about 0.5% (w / v), between about 0.005% (w / v) and about 0.5% (w / v), between about 0.01% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.5% (w / v), between about 0.1% (w / v) and about 0.5% (w / v), between about 0.05% (w / v) and about 0.3% (w / v), between about 0.05% (w / v) and about 0.2% (w / v), between about 0.05% (w / v) and about 0.15% (w / v), between about 0.001% (w / v) and about 0.1% (w / v), between about 0.005% (w / v) and about 0.1% (w / v), between about 0.01% (w / v) and about 0.1% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), between about 0.001% (w / v) and about 0.05% (w / v), between about 0.005% (w / v) and about 0.05% (w / v), between about 0.01% (w / v) and about 0.05% (w / v), between about 0.001% (w / v) and about 0.01% (w / v), between about 0.005% (w / v) and about 0.01% (w / v), or between about 0.001% (w / v) and about 0.005% (w / v). In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of about 3% (w / v), about 2% (w / v), about 1% (w / v), about 0.5% (w / v), about 0.1% (w / v), about 0.05% (w / v), about 0.01% (w / v), about 0.005% (w / v), or about 0.001% (w / v).

[0182] In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of between about 0.05% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation comprises rHSA at a concentration of about 0.1% (w / v).

[0183] In certain embodiments, the stabilizer (e.g., rHSA) is at a concentration that reduces the loss of viral titer during the freezing / thawing process and / or storage.

[0184] In certain embodiments, the stabilizer (e.g., rHSA) is at a concentration that reduces the loss of viral titer during the freezing / thawing process and / or storage, wherein the viral titer in the formulation after the freezing / thawing process and / or storage is at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer in the formulation before the freezing / thawing process and / or storage as measured by RCV-FFU or FFU assay. In certain embodiments, the stabilizer (e.g., rHSA) is at a concentration that reduces the loss of viral titer during the freezing / thawing process and / or storage, wherein the viral titer in the formulation after the freezing / thawing process and / orstorage is about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or about 100% of the viral titer in the formulation before the freezing / thawing process and / or storage as measured by RCV-FFU or FFU assay.

[0185] In certain embodiments, the stabilizer (e.g., rHSA) is at a concentration that reduces the loss of viral titer during the freezing / thawing process and / or storage, wherein the viral titer in the formulation after the freezing / thawing process and / or storage is at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 90%, at least about 100%, at least about 150%, at least about 200%, at least about 250%, at least about 300%, at least about 400%, or at least about 500% more than the viral titer in a control formulation without the stabilizer after the freezing / thawing process and / or storage, as measured by RCV-FFU or FFU assay. In certain embodiments, the stabilizer (e.g., rHSA) is at a concentration that reduces the loss of viral titer during the freezing / thawing process and / or storage, wherein the viral titer in the formulation after the freezing / thawing process and / or storage is about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 100%, about 150%, about 200%, about 250%, about 300%, about 400%, about 500% more than the viral titer in a control formulation without the stabilizer after the freezing / thawing process and / or storage, as measured by RCV-FFU or FFU assay. In certain embodiments, the control formulation is otherwise identical to the presently disclosed formulation except not comprising the stabilizer or comprising a different stabilizer.

[0186] In certain embodiments, the temperature of the storage is above about 2°C, room temperature (e.g., 20°C - 25°C), between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C. In certain embodiments, the temperature of the storage is between about 2°C and about 8°C. In certain embodiments, the storage lasts at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 3 years, at least about 4 years, at least about 5 years, or longer. In certain embodiments, the storage lasts at least about 1 month.

[0187] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of up to about0.15% (w / v), up to about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.1% (w / v), between about 0.05% and about 0.15% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), or between about 0.1% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of about 0.05%, about 0.08%, about 0.1%, about 0.12%, or about 0.15%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of about 0.1% (w / v).

[0188] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of up to about 0.15% (w / v), up to about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.1% (w / v), between about 0.05% and about 0.15% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), or between about 0.1% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of about 0.05%, about 0.08%, about 0.1%, about 0.12%, or about 0.15%. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of about 0.1% (w / v).5.1.7 Osmolality and pH

[0189] In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of at least about 200 mOsm / kg, at least about 300 mOsm / kg, at least about 400 mOsm / kg, at least about 500 mOsm / kg, at least about 600 mOsm / kg, at least about 700 mOsm / kg, at least about 800 mOsm / kg, at least about 900 mOsm / kg, at least about 1000 mOsm / kg, up to about 200 mOsm / kg, up to about 300 mOsm / kg, up to about 400 mOsm / kg, up to about 500 mOsm / kg, up to about 600 mOsm / kg, up to about 700 mOsm / kg, up to about 800 mOsm / kg, up to about 900 mOsm / kg, up to about 1000 mOsm / kg, between about 200 mOsm / kg and about 1000 mOsm / kg, between about 300 mOsm / kg and about 1000 mOsm / kg, between about 400 mOsm / kg and about 1000 mOsm / kg, between about 500 mOsm / kg and about 1000 mOsm / kg, between about 600 mOsm / kg and about 1000 mOsm / kg, between about 700 mOsm / kg and about 1000 mOsm / kg, between about 800 mOsm / kg and about 1000 mOsm / kg, between about 900 mOsm / kg and about 1000 mOsm / kg, between about 200 mOsm / kg and about 900 mOsm / kg, between about 300 mOsm / kg and about 900 mOsm / kg, between about 400 mOsm / kg and about 900 mOsm / kg, between about 500 mOsm / kg and about 900 mOsm / kg, between about 600 mOsm / kg and about 900 mOsm / kg, between about 700 mOsm / kg and about 900 mOsm / kg, between about800 mOsm / kg and about 900 mOsm / kg, between about 200 mOsm / kg and about 800 mOsm / kg, between about 300 mOsm / kg and about 800 mOsm / kg, between about 400 mOsm / kg and about 800 mOsm / kg, between about 500 mOsm / kg and about 800 mOsm / kg, between about 600 mOsm / kg and about 800 mOsm / kg, between about 700 mOsm / kg and about 800 mOsm / kg, between about 200 mOsm / kg and about 700 mOsm / kg, between about 300 mOsm / kg and about 700 mOsm / kg, between about 400 mOsm / kg and about 700 mOsm / kg, between about 500 mOsm / kg and about 700 mOsm / kg, between about 600 mOsm / kg and about 700 mOsm / kg, between about 200 mOsm / kg and about 600 mOsm / kg, between about 300 mOsm / kg and about 600 mOsm / kg, between about 400 mOsm / kg and about 600 mOsm / kg, between about 500 mOsm / kg and about 600 mOsm / kg, between about 200 mOsm / kg and about 500 mOsm / kg, between about 300 mOsm / kg and about 500 mOsm / kg, between about 400 mOsm / kg and about 500 mOsm / kg, between about 200 mOsm / kg and about 400 mOsm / kg, between about 300 mOsm / kg and about 400 mOsm / kg, or between about 200 mOsm / kg and about 300 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of about 200 mOsm / kg, about 300 mOsm / kg, about 400 mOsm / kg, about 500 mOsm / kg, about 600 mOsm / kg, about 700 mOsm / kg, about 800 mOsm / kg, about 900 mOsm / kg, or about 1000 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of between about 500 mOsm / kg and about 600 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of about 530 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein has an osmolality of about 400 mOsm / kg.

[0190] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0191] In certain embodiments, the liquid arenavirus formulation disclosed herein has a pH of between about 3.0 and about 9.0, between about 4.0 and about 9.0, between about 5.0 and about 9.0, between about 6.0 and about 9.0, between about 7.0 and about 9.0, between about 8.0 and about 9.0, between about 3.0 and about 8.0, between about 4.0 and about 8.0, between about 5.0 and about 8.0, between about 6.0 and about 8.0, between about 7.0 and about 8.0, between about 3.0 and about 7.0, between about 4.0 and about 7.0, between about5.0 and about 7.0, between about 6.0 and about 7.0, between about 3.0 and about 6.0, between about 4.0 and about 6.0, between about 5.0 and about 6.0, between about 3.0 and about 5.0, between about 4.0 and about 5.0, or between about 3.0 and about 4.0. In certain embodiments, the liquid arenavirus formulation disclosed herein has a pH of about 3.0, about 4.0, about 5.0, about 6.0, or about 7.0.

[0192] In certain embodiments, the liquid arenavirus formulation disclosed herein has a pH of between about 7.0 and about 9.0. In certain embodiments, the liquid arenavirus formulation disclosed herein has a pH of about 8.0.5.1.8 Exemplary Liquid Arenavirus Formulations

[0193] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus (see Section 5.3); (b) a polyvalent anion (see Section 5.1.1) at a concentration of between about 10 mM and about 60 mM; (c) a carbohydrate (see Section5.1.2) at a concentration of between about 3% and about 10% (w / v); (d) a salt (see Section5.1.3) at a concentration of between about 50 mM and about 250 mM; (e) a buffer (see Section 5.1.4) at a concentration of between about 10 mM and about 20 mM; and (f) a surfactant (see Section 5.1.5) at a concentration of between about 0.01% and about 0.05% (w / v), and (g) a stabilizer (see Section 5.1.6) at a concentration of between about 0.05 (w / v) and 0.15% (w / v); wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 600 mOsm / kg. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg. In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0194] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus (see Section 5.3); (b) a polyvalent anion (see Section 5.1.1) at a concentration of about 50 mM; (c) a carbohydrate (see Section 5.1.2) at a concentration of about 5% (w / v); (d) a salt (see Section 5.1.3) at a concentration of about 125 mM; (e) a buffer (see Section 5.1.4) at a concentration of about 10 mM; (f) a surfactant (see Section 5.1.5) at a concentration of about 0.03% (w / v), and (g) a stabilizer (see Section 5.1.6) at a concentration of about 0.1% (w / v); wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

[0195] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus (see Section 5.3); (b) a polyvalent anion (see Section 5.1.1) at a concentration of about 25 mM; (c) a carbohydrate (see Section 5.1.2) at a concentration of about 5% (w / v); (d) a salt (see Section 5.1.3) at a concentration of about 75 mM; (e) a buffer (see Section 5.1.4) at a concentration of about 10 mM; (f) a surfactant (see Section 5.1.5) at a concentration of about 0.03% (w / v), and (g) a stabilizer (see Section 5.1.6) at a concentration of about 0.1% (w / v); wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

[0196] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus (see Section 5.3); (b) citrate at a concentration of between about 10 mM and about 60 mM; (c) sucrose at a concentration of between about 3% and about 10% (w / v); (d) sodium chloride at a concentration of between about 50 mM and about 250 mM;(e) TRIS at a concentration of between about 10 mM and about 20 mM; (f) pol oxamer 188 at a concentration of between about 0.01% and about 0.05% (w / v); and (g) rHSA at a concentration of between about 0.05 (w / v) and 0.15% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 600 mOsm / kg.

[0197] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus; (b) citrate at a concentration of about 50 mM; (c) sucrose at a concentration of about 5% (w / v); (d) sodium chloride at a concentration of about 125 mM;(e) TRIS at a concentration of about 10 mM; (f) pol oxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

[0198] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus (see Section 5.3); (b) citrate at a concentration of between about 10 mM and about 60 mM; (c) trehalose at a concentration of between about 3% and about 10% (w / v); (d) sodium chloride at a concentration of between about 50 mM and about 250 mM; (e) TRIS at a concentration of between about 10 mM and about 20 mM; (f) pol oxamer 188 at a concentration of between about 0.01% and about 0.05% (w / v); and (g) rHSA at a concentration of between about 0.05 (w / v) and 0.15% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg.

[0199] In certain embodiments, the liquid arenavirus formulation disclosed herein comprises (a) an arenavirus; (b) citrate at a concentration of about 25 mM; (c) trehalose at a concentration of about 5% (w / v); (d) sodium chloride at a concentration of about 75 mM; (e) TRIS at a concentration of about 10 mM; (f) pol oxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

[0200] In certain embodiments, the liquid arenavirus formulation disclosed herein is suitable for lyophilization and comprises (a) an arenavirus (see Section 5.3); (b) a polyvalent anion (see Section 5.1.1) at a concentration of up to about 50 mM or between about 20 mM and 50 mM; (c) a carbohydrate (see Section 5.1.2) at a concentration of about 5% (w / v); (d) a salt (see Section 5.1.3) at a concentration of up to about 125 mM or between 50 mM and about 125 mM; (e) a buffer (see Section 5.1.4) at a concentration of about 10 mM; (f) a surfactant (see Section 5.1.5) at a concentration of about 0.03% (w / v), and (g) a stabilizer (see Section 5.1.6) at a concentration of about 0.1% (w / v); wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

[0201] In certain embodiments, the liquid arenavirus formulation suitable for lyophilization disclosed herein comprises (a) an arenavirus; (b) citrate at a concentration of up to about 50 mM or between about 20 mM and 50 mM; (c) trehalose at a concentration of about 5% (w / v); (d) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM; (e) TRIS at a concentration of about 10 mM; (f) poloxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.5.2 Lyophilized Arenavirus Formulations

[0202] The present disclosure further provides lyophilized arenavirus formulations. In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation disclosed herein (see Section 5.1). In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation disclosed herein (see Section 5.1), wherein the liquid arenavirus formulation is suitable for lyophilization.

[0203] In certain embodiments, the lyophilized arenavirus formulation comprises an arenavirus (see Section 5.3), and a polyvalent anion (see Section 5.1.1). In certain embodiments, the lyophilized arenavirus formulation comprises at least one of a carbohydrate (see Section 5.1.2), a salt (see Section 5.1.3), a buffer (see Section 5.1.4), a surfactant (see Section 5.1.5), and a stabilizer (see Section 5.1.6).

[0204] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of up to about 50 mM, up to about 40 mM, up to about 30 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, between about 20 mM and about 50 mM, between about 20 mM and about 40 mM, between about 20 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 40 mM, or between about 40 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of about 20 mM, about 30 mM, about 40 mM, or about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of up to about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the polyvalent anion disclosed herein at a concentration of between about 20 mM and about 50 mM.

[0205] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of up to about 50 mM, up to about 40 mM, up to about 30 mM, at least about 20 mM, at least about 30 mM, at least about 40 mM, between about 20 mM and about 50 mM, between about 20 mM and about 40 mM, between about 20 mM and about 30 mM, between about 30 mM and about 50 mM, between about 30 mM and about 40 mM, or between about 40 mM and about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of about 20 mM, about 30 mM, about 40 mM, or about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of up to about 50 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises citrate at a concentration of between about 20 mM and about 50 mM.

[0206] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of up to about 10% (w / v), up to about 8% (w / v), up to about 6% (w / v), up to about 4% (w / v), at least about 3% (w / v), at least about 6% (w / v), at least about 8% (w / v), between about 3% and about 10% (w / v), between about 3% (w / v) and about 8% (w / v), between about 3% and about 6% (w / v), between about 4% and about 10% (w / v), between about 4% and about 8% (w / v), between about 4% and about 6% (w / v), between about 6% and about 10% (w / v), between about 6% and about 8% (w / v), or between about 8% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of about 3% (w / v), about 4% (w / v), about 5% (w / v), about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), or about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the carbohydrate disclosed herein at a concentration of about 5% (w / v).

[0207] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of up to about 10% (w / v), up to about 8% (w / v), up to about 6% (w / v), up to about 4% (w / v), at least about 3% (w / v), at least about 6% (w / v), at least about 8% (w / v), between about 3% and about 10% (w / v), between about 3% (w / v) and about 8% (w / v), between about 3% and about 6% (w / v), between about 4% and about 10% (w / v), between about 4% and about 8% (w / v), between about 4% and about 6% (w / v), between about 6% and about 10% (w / v), between about 6% and about 8% (w / v), or between about 8% and about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of about 3% (w / v), about 4% (w / v), about 5% (w / v), about 6% (w / v), about 7% (w / v), about 8% (w / v), about 9% (w / v), or about 10% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises trehalose at a concentration of about 5% (w / v).

[0208] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of up to about 125 mM, up to about 100 mM, up to about 75 mM, at least about 50 mM, at least about 75 mM, at least about 100 mM, between about 50 mM and about 125 mM,between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, or between about 100 mM and about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of about 50 mM, about 75 mM, about 100 mM, or about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of up to about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the salt disclosed herein at a concentration of between about 50 mM and about 125 mM.

[0209] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of up to about 125 mM, up to about 100 mM, up to about 75 mM, at least about 50 mM, at least about 75 mM, at least about 100 mM, between about 50 mM and about 125 mM, between about 50 mM and about 100 mM, between about 50 mM and about 75 mM, between about 75 mM and about 125 mM, between about 75 mM and about 100 mM, or between about 100 mM and about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of about 50 mM, about 75 mM, about 100 mM, or about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of up to about 125 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises sodium chloride at a concentration of between about 50 mM and about 125 mM.

[0210] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration of up to about 20 mM, up to about 18 mM, up to about 15 mM, at least about 10 mM, at least about 15 mM, at least about 18 mM, between about 10 mM and about 20 mM, between about 10 mM and about 18 mM, between about 10 mM and about 15 mM, between about 15 mM and about 20 mM, between about 15 mM and about 18 mM, between about 18 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration of about 10 mM, about 12 mM, about 14 mM, about 16 mM, about 18 mM, or about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the buffer disclosed herein at a concentration about 10 mM.

[0211] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration of up to about 20 mM, up to about 18 mM, up to about 15 mM, at least about 10 mM, at least about 15 mM, at least about 18 mM, or between about 10 mM and about 20 mM, or between about 10 mM and about 18 mM, between about 10 mM and about 15 mM, between about 15 mM and about 20 mM, between about 15 mM and about 18 mM, between about 18 mM and about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration of about 10 mM, about 12 mM, about 14 mM, about 16 mM, about 18 mM, or about 20 mM. In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises TRIS at a concentration about 10 mM.

[0212] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), at least about 0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), between about 0.01% and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04% (w / v), between about 0.01% and about 0.03%(w / v), between about 0.01% and about 0.02% (w / v), between about 0.02% and about 0.05%(w / v), between about 0.02% and about 0.04% (w / v), between about 0.02% and about 0.03%(w / v), between about 0.03% and about 0.05% (w / v), between about 0.03% and about 0.04%(w / v), or between about 0.04% and about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of about 0.01% (w / v), about 0.02% (w / v), about 0.03% (w / v), about 0.04% (w / v), or about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the surfactant disclosed herein at a concentration of about 0.03% (w / v).

[0213] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises pol oxamer 188 at a concentration of up to about 0.05% (w / v), up to about 0.04% (w / v), up to about 0.03% (w / v), up to about 0.02% (w / v), at least about 0.01% (w / v), at least about 0.02% (w / v), at least about 0.03% (w / v), at least about 0.04% (w / v), between about 0.01% and about 0.05% (w / v), between about 0.01% (w / v) and about 0.04% (w / v), between about 0.01% and about 0.03% (w / v), between about0.01% and about 0.02% (w / v), between about 0.02% and about 0.05% (w / v), between about0.02% and about 0.04% (w / v), between about 0.02% and about 0.03% (w / v), between about0.03% and about 0.05% (w / v), between about 0.03% and about 0.04% (w / v), or between about 0.04% and about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises pol oxamer 188 at a concentration of about 0.01% (w / v), about 0.02% (w / v), about 0.03% (w / v), about 0.04% (w / v), or about 0.05% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises poloxamer 188 at a concentration of about 0.03% (w / v).

[0214] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of up to about 0.15% (w / v), up to about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.1% (w / v), between about 0.05% and about 0.15% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), or between about 0.1% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of about 0.05% (w / v), about 0.08% (w / v), about 0.1% (w / v), about 0.12% (w / v), or about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises the stabilizer disclosed herein at a concentration of about 0.1% (w / v).

[0215] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of up to about 0.15% (w / v), up to about 0.1% (w / v), at least about 0.05% (w / v), at least about 0.1% (w / v), between about 0.05% and about 0.15% (w / v), between about 0.05% (w / v) and about 0.1% (w / v), or between about 0.1% and about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of about 0.05% (w / v), about 0.08% (w / v), about 0.1% (w / v), about 0.12% (w / v), or about 0.15% (w / v). In certain embodiments, the liquid arenavirus formulation suitable for lyophilization comprises rHSA at a concentration of about 0.1% (w / v).

[0216] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises (a) an arenavirus (see Section 5.3); (b) a polyvalent anion (see Section 5.1.1) at a concentration of up to about 50 mM or between about 20 mM and 50 mM; (c) a carbohydrate (see Section 5.1.2) at a concentration of about5% (w / v); (d) a salt (see Section 5.1.3) at a concentration of up to about 125 mM or between 50 mM and about 125 mM; (e) a buffer (see Section 5.1.4) at a concentration of about 10 mM; (f) a surfactant (see Section 5.1.5) at a concentration of about 0.03% (w / v), and (g) a stabilizer (see Section 5.1.6) at a concentration of about 0.1% (w / v); wherein the liquid arenavirus formulation has a pH of about 8.0.

[0217] In certain embodiments, the lyophilized arenavirus formulation is prepared from a liquid arenavirus formulation suitable for lyophilization, wherein the liquid arenavirus formulation suitable for lyophilization comprises (a) an arenavirus; (b) citrate at a concentration of up to about 50 mM or between about 20 mM and 50 mM; (c) trehalose at a concentration of about 5% (w / v); (d) sodium chloride at a concentration of up to about 125 mM or between 50 mM and about 125 mM; (e) TRIS at a concentration of about 10 mM; (f) pol oxamer 188 at a concentration of about 0.03% (w / v); and (g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0.

[0218] Methods for preparing a lyophilized formulation from a liquid formulation are well known in the art. Examples of methods for preparing a lyophilized formulation from a liquid formulation suitable for use herein are described, for example, in Hansen et al., Freeze- drying of live virus vaccines: a review, T.R.M.; Vaccine 33, 2015 (5507-5519). Viruses (especially if live attenuated) are often instable in aqueous media if not frozen. This instability is primarily water-mediated and lyophilization aims at the removal of the bulk water present in the formulation. An exemplary method is provided herein. After 0.22 pm sterile filtration, the drug substance bulk solution is filled in glass vials and half stoppered with stoppers (both, vials and stoppers need to be suitable for lyophilization). The applied lyophilization cycle comprises the freezing step, the primary drying to remove most of the water by sublimation and the secondary drying to remove the bound water. The freezing rate is done at the fastest possible rate to avoid crystallization of formulation components and thereby damaging the virus. Primary drying (between -10 and -40°C) and secondary drying (between +10 and +40°C) is performed at the highest possible temperature and pressure (>0.03 mbar) without risking collapse of the cake or destabilization of the viral structure or infectivity. The duration of the respective drying steps will finally lead to a residual moisture content in the dried material between 0.5 to 2%. Lyophilization and the low residual moisture content facilitate storage at elevated temperatures (refrigerated or room temperature) without loss of infectivity over an extended period of time.

[0219] In certain embodiments, the lyophilized arenavirus formulation disclosed herein is reconstituted prior to the administration to a subject (see Section 5.6).

[0220] In certain embodiments, the lyophilized arenavirus formulation provided herein retains the viral titer after a period of storage. In certain embodiments, the lyophilized arenavirus formulation provided herein retains at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, or about 100% of the viral titer after a period of storage. In certain embodiments, the period of storage is up to about 3 months, up to about 4 months, up to about 5 months, up to about 6 months, up to about 7 months, up to about 8 months, up to about 9 months, up to about 10 months, up to about 11 months, up to about 12 months, up to about 18 months, up to about 24 months, up to about 3 years, up to about 4 years, up to about 5 years, or longer. In certain embodiments, the period of storage is about 1 month, about 2 months, about 3 months, about 4 months, about 5 months, about 6 months, about 7 months, about 8 months, about 9 months, about 10 months, about 11 months, about 12 months, about 18 months, about 24 months, about 3 years, about 4 years, about 5 years, or longer. In certain embodiments, the temperature of the storage is above about 2°C, room temperature (e.g., 20°C - 25°C), between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C. In certain embodiments, the viral titer is measured by RCV-FFU or FFU assay.5.3 Arenaviruses

[0221] Any arenaviruses known in the art can be included in the presently disclosed liquid and lyophilized formulations. In certain embodiments, the arenavirus is a genetically engineered arenavirus.

[0222] In certain embodiments, the arenavirus is lymphocytic choriomeningitis virus (LCMV). In certain embodiments, the arenavirus is Pichinde virus. In certain embodiments, the arenavirus is Tamiami virus. In certain embodiments, the arenavirus is Tacaribe virus.

[0223] In certain embodiments, the arenavirus is a non-replicating arenavirus. In certain embodiments, the arenavirus is a non-replicating arenavirus, wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles. In certain embodiments, the arenavirus is a nonreplicating arenavirus, and the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen. In certain embodiments, the arenavirus is a replicating arenavirus. In certain embodiments, the arenavirus is a replicating tri-segmented arenavirus. In certain embodiments, the replicating tri-segmented arenavirus is engineered such that the arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF. In certain embodiments, the replicating tri-segmentedarenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3’ UTR. In certain embodiments, the arenavirus is a replicating tri-segmented arenavirus, engineered such that an arenaviral ORF is separated over two or more mRNA transcripts. Non-limiting examples of non-replicating and replicating arenaviruses are described in International Publication Nos WO 2009 / 083210, WO 2016 / 075250, and WO 2021 / 089853, and US Publication Nos: US20100297172A1, US20170319673 Al, and US20220380805A1, the contents of each of which are incorporated by reference herein.

[0224] In certain embodiments, the arenavirus expresses an antigen. In certain embodiments, the arenavirus expresses at least one antigen selected from the group consisting of a tumor antigen, a tumor associated antigen, an antigen of a pathogen that causes an infectious disease, or an antigenic fragment of any of the foregoing. In certain embodiments, the arenavirus expresses at least one antigen selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.

[0225] In certain embodiments, the arenavirus expresses at least one cytomegalovirus (CMV) antigen. In certain embodiments, the at least one CMV antigen is selected from the group consisting of a CMV glycoprotein gB or an antigenic fragment thereof, a CMV tegument protein pp65 or an antigenic fragment thereof, and combinations thereof. In certain embodiments, the at least one CMV antigen is selected from the group consisting of a CMV glycoprotein gB with a truncation of the carboxy-terminus (i.e., gB(dCt)), pp65, and combinations thereof. In certain embodiments, the arenavirus expressing the at least one CMV antigen is a non-replicating arenavirus. Non-limiting examples of CMV-antigen expressing arenaviruses are described in International Publication No. WO 2015 / 082570 and US Publication No. US20160296619A1, the content of each of which is incorporated by reference herein.

[0226] In certain embodiments, the arenavirus expresses at least one human papilloma virus (HPV) antigen. In certain embodiments, the arenavirus expresses two HPV antigens which are fused together. In certain embodiments, the arenavirus expresses an HPV E7ZE6 fusion protein. In certain embodiments, the arenavirus expressing the at least one HPV antigen is a non-replicating arenavirus or a replicating tri-segmented arenavirus. In certain embodiments, the arenavirus expressing the at least one HPV antigen is LCMV. In certain embodiments, the arenavirus expressing the at least one HPV antigen is Pichinde virus. Nonlimiting examples of the HPV-antigen expressing arenaviruses are described in InternationalPublication Nos. WO 2016 / 198531 and WO2021 / 239471, and US Publication Nos. US20180179257A1 and US20230346906A1, the content of each of which is incorporated by reference herein.

[0227] In certain embodiments, the arenavirus expresses at least one prostate cancer antigen. In certain embodiments, the at least one prostate cancer antigen is selected from the group consisting of prostate specific antigen (PSA), prostate-specific membrane antigen (PSMA), prostatic acid phosphatase (PAP), and combinations thereof. In certain embodiments, the arenavirus expressing the at least one prostate cancer antigen is a replicating tri-segmented arenavirus. In certain embodiments, the arenavirus expressing the at least one prostate cancer antigen is LCMV. In certain embodiments, the arenavirus expressing the at least one prostate cancer antigen is Pichinde virus. Non-limiting examples of the prostate cancer antigen expressing arenaviruses are described in International Publication No. W02022 / 200373 and US Publication No. US20240174724 Al, the content of each of which is incorporated by reference herein.

[0228] In certain embodiments, the arenavirus expresses at least one hepatitis B virus (HBV) antigen. In certain embodiments, the at least one HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide, and combinations thereof. In certain embodiments, the arenavirus expressing the at least one HBV antigen is a non-replicating arenavirus or a replicating tri-segmented arenavirus. In certain embodiments, the arenavirus expressing the at least one HBV antigen is LCMV. In certain embodiments, the arenavirus expressing the at least one HBV antigen is Pichinde virus. Non-limiting examples of the HBV-antigen expressing arenaviruses are described in International Publications No. WO 2017 / 076988 and WO 2021 / 067181, and US Publication Nos. US20180319845A1 and US20210093712A1, the content of each of which is incorporated by reference herein.

[0229] In certain embodiments, the arenavirus expresses at least one human immunodeficiency virus (HIV) antigen. In certain embodiments, the at least one HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef. In certain embodiments, the arenavirus expressing the at least one HIV antigen is a non-replicating arenavirus or a replicating tri-segmented arenavirus. Incertain embodiments, the arenavirus expressing the at least one HIV antigen is LCMV. In certain embodiments, the arenavirus expressing the at least one HIV antigen is Pichinde virus. Non-limiting examples of the HIV-antigen expressing arenaviruses are described in International Publication No. WO 2022 / 155258 and US Publication No. US20220218813A1, the content of each of which is incorporated by reference herein.

[0230] In certain embodiments, the arenavirus expresses at least one antigenic fragment of a mutant KRAS. In certain embodiments, the arenavirus encodes from N- to C-terminus antigenic fragments of mutant KRAS comprising the mutations G13D, G12V, G12C, G12D, and G12R, respectively. In certain embodiments, the arenavirus expressing the antigenic fragments of mutant KRAS is a non-replicating arenavirus or a replicating tri-segmented arenavirus. In certain embodiments, the arenavirus expressing the antigenic fragment(s) of mutant KRAS is a “split” arenavirus particle. In certain embodiments, the arenavirus expressing the antigenic fragment(s) of mutant KRAS is LCMV. In certain embodiments, the arenavirus expressing the antigenic fragment(s) of mutant KRAS is Pichinde virus. Nonlimiting examples of the arenaviruses expressing the antigenic fragment(s) of mutant KRAS are described in International Patent Publication No. WO2023079153, the content of which is incorporated by reference herein.

[0231] In certain embodiments, the arenavirus expresses two or more CTA fragments (e.g., antigenic fragments or epitopes). In certain embodiments, the arenavirus expresses at least one fusion protein comprising two or more CTA fragments (e.g., antigenic fragments or epitopes). In certain embodiments, the arenavirus encodes a first fusion protein comprising two or more CTA fragments and a second fusion protein comprising two or more CTA fragments. In certain embodiments, the two or more CTA fragments of the at least one fusion protein, the first fusion protein, and / or the second fusion protein are independently selected from the group consisting of epitopes or antigenic fragments of PRAME, MAGE-A3, MAGE-A4, and MAGE-A6. In certain embodiments, the two or more CTA fragments of the first fusion protein comprise at least one PRAME fragment, at least one MAGE-A3 fragment, at least one MAGE-A4 fragment, and at least one MAGE-A6 fragment. In certain embodiments, the at least one PRAME fragment of the first fusion protein is selected from the group consisting of PRAME3-13, PRAME7-23, PRAME73-82, PRAME84-95, PRAME123-141, PRAME150-161, PRAME166-178, PRAME210-230, PRAME224-232, PRAME237-250, PRAME248- 256, PRAME267-277, PRAME278-292, PRAME315-327, PRAME325-334, PRAME425-433, PRAME435-443, PRAME445-458, PRAME472-491, PRAME485-509, and combinations thereof. In certain embodiments, the at least one MAGE- A3 fragment of the first fusion proteincomprises MAGE-A3135-149. In certain embodiments, the at least one MAGE-A4 fragment of the first fusion protein comprises MAGE- A4112-121 and / or MAGE-A4286-305. In certain embodiments, the at least one MAGE-A6 fragment of the first fusion protein comprises MAGE-A6167-184. In certain embodiments, the two or more CTA fragments of the second fusion protein comprise at least one MAGE-A3 fragment, at least one MAGE-A4 fragment, and at least one MAGE-A6 fragment. In certain embodiments, the at least one MAGE-A3 fragment of the second fusion protein is selected from the group consisting of MAGE- A3243- 255, MAGE- A3171-184, MAGE-A3284-296, MAGE-A3167-176, MAGE-A3192-202, MAGE-A3302- 311, MAGE-A370-81, MAGE-A3160-171, MAGE-A3143-151, and combinations thereof. In certain embodiments, the at least one MAGE-A4 fragment of the second fusion protein is selected from the group consisting of MAGE-A4136-155, MAGE-A4168-179, MAGE-A4190-199, MAGE-A477-89, MAGE-A4300-308, MAGE-A4175-183, MAGE-A4262-272, MAGE-A4222-246, MAGE-A4109-117, MAGE-A4205-218, MAGE-A4205-217, MAGE-A4155-168, or MAGE- A4266- 274, MAGE-A473-83, and combinations thereof. In certain embodiments, the at least one MAGE-A6 fragment of the second fusion protein is selected from the group consisting of MAGE-A6 196-204, MAGE-A6194-202, MAGE-A6231-241, MAGE-A6243-255, and combinations thereof. Sequences of exemplary CTA fragments are disclosed in Section 6 herein, and set forth in SEQ ID NOs: 1-51. In certain embodiments, the arenavirus expressing the first and second fusion proteins comprising two or more CTA fragments is a non-replicating arenavirus or a replicating tri-segmented arenavirus. In certain embodiments, the arenavirus expressing the first and second fusion proteins comprising two or more CTA fragments is a “split” arenavirus particle. In certain embodiments, the arenavirus expressing the first and second fusion proteins comprising two or more CTA fragments is LCMV. In certain embodiments, the arenavirus expressing the first and second fusion proteins comprising two or more CTA fragments is Pichinde virus. Non-limiting examples of the arenaviruses expressing the first and second fusion proteins comprising two or more CTA fragments are described in U.S. Provisional Application No 63 / 565,812 filed March 15, 2024, the content of which is incorporated by reference herein

[0232] In certain embodiments, the concentration of the arenavirus in the liquid arenavirus formulation disclosed herein (see Section 5.1) is between about 102RCV-FFU / ml and about 1010RCV-FFU / ml. In certain embodiments, the concentration of the arenavirus in the liquid arenavirus formulation disclosed herein (see Section 5.1) is between about 102RCV-FFU / ml and about 109RCV-FFU / ml, between about 102RCV-FFU / ml and about 108RCV-FFU / ml, between about 102RCV-FFU / ml and about 107RCV-FFU / ml, between about102RCV-FFU / ml and about 106RCV-FFU / ml, between about 102RCV-FFU / ml and about 105RCV-FFU / ml, between about 102RCV-FFU / ml and about 104RCV-FFU / ml, between about 102RCV-FFU / ml and about 103RCV-FFU / ml, between about 103RCV-FFU / ml and about IO10RCV-FFU / ml, between about 103RCV-FFU / ml and about 109RCV-FFU / ml, between about 103RCV-FFU / ml and about 108RCV-FFU / ml, between about 103RCV- FFU / ml and about 107RCV-FFU / ml, between about 103RCV-FFU / ml and about 106RCV- FFU / ml, between about 103RCV-FFU / ml and about 105RCV-FFU / ml, between about 103RCV-FFU / ml and about 104RCV-FFU / ml, between about 104RCV-FFU / ml and about IO10RCV-FFU / ml, between about 104RCV-FFU / ml and about 109RCV-FFU / ml, between about 104RCV-FFU / ml and about 108RCV-FFU / ml, between about 104RCV-FFU / ml and about 107RCV-FFU / ml, between about 104RCV-FFU / ml and about 106RCV-FFU / ml, between about 104RCV-FFU / ml and about 105RCV-FFU / ml, between about 105RCV-FFU / ml and about IO10RCV-FFU / ml, between about 105RCV-FFU / ml and about 109RCV-FFU / ml, between about 105RCV-FFU / ml and about 108RCV-FFU / ml, between about 105RCV- FFU / ml and about 107RCV-FFU / ml, between about 105RCV-FFU / ml and about 106RCV- FFU / ml, between about 106RCV-FFU / ml and about IO10RCV-FFU / ml, between about 106RCV-FFU / ml and about 109RCV-FFU / ml, between about 106RCV-FFU / ml and about 108RCV-FFU / ml, between about 106RCV-FFU / ml and about 107RCV-FFU / ml, between about 107RCV-FFU / ml and about IO10RCV-FFU / ml, between about 107RCV-FFU / ml and about 109RCV-FFU / ml, between about 107RCV-FFU / ml and about 108RCV-FFU / ml, between about 108RCV-FFU / ml and about IO10RCV-FFU / ml, between about 108RCV-FFU / ml and about 109RCV-FFU / ml, or between about 109RCV-FFU / ml and about IO10RCV-FFU / ml. In certain embodiments, the concentration of the arenavirus in the liquid arenavirus formulation disclosed herein (see Section 5.1) is between about 105RCV-FFU / ml to about 109RCV-FFU / ml, between about 105RCV-FFU / ml to about 8* 108RCV-FFU / ml, between about 105RCV-FFU / ml to about 6* 108RCV-FFU / ml, between about 105RCV-FFU / ml to about 5 * 108RCV-FFU / ml, between about 2* 105RCV-FFU / ml to about 109RCV-FFU / ml, between about 2* 105RCV-FFU / ml to about 8* 108RCV-FFU / ml, or between about 2* 105RCV-FFU / ml to about 6* 108RCV-FFU / ml. In certain embodiments, the concentration of the arenavirus in the liquid arenavirus formulation disclosed herein (see Section 5.1) is between about 2x l05RCV-FFU / ml to about 5x l08RCV-FFU / ml.5.4 Pharmaceutical Compositions

[0233] Provided herein are pharmaceutical compositions comprising a presently disclosed liquid arenavirus formulation (see Section 5.1). In certain embodiments, the pharmaceuticalcomposition comprises a liquid arenavirus formulation diluted from a presently disclosed liquid arenavirus formulation (see Section 5.1). As such, the concentrations of certain components in the pharmaceutical composition can be different from the concentrations of these components in the liquid arenavirus formulation from which a dilution is made.

[0234] In certain embodiments, the pharmaceutical composition comprises a presently disclosed lyophilized formulation (see Section 5.2). In certain embodiments, the pharmaceutical composition comprises a liquid arenavirus formulation reconstituted from a presently disclosed lyophilized formulation (see Section 5.2).

[0235] In certain embodiments, the pharmaceutical composition further comprises an adjuvant. In certain embodiments, the adjuvant is administered before, concomitantly with, or after the administration of the pharmaceutical composition. An adjuvant is a compound that, when administered in conjunction with or as part of a pharmaceutical composition described herein, augments, enhances, and / or boosts the immune response induced by the arenavirus; but when the compound is administered alone does not generate an immune response to the arenavirus or the antigen expressed by the arenavirus. Adjuvants that can be used with the present disclosure include, but are not limited to, mineral salt adjuvants or mineral salt gel adjuvants, particulate adjuvants, microparticulate adjuvants, mucosal adjuvants, and immunostimulatory adjuvants. Examples of adjuvants suitable for use herein are described in Rushit N et al., Overview of vaccine adjuvants, eds. Kolhe and Ohtake, PRACTICA ASPECTS OF VACCINE DEVELOPMENT, Academic Press, 2022 (9-25). Additional examples of adjuvants include, but are not limited to, aluminum salts (alum) (such as aluminum hydroxide, aluminum phosphate, and aluminum sulfate), 3 de-O-acylated monophosphoryl lipid A (MPL) (see GB 2220211), MF59® (Novartis), AS03 (GlaxoSmithKline), AS04 (GlaxoSmithKline), VacciGrade™ (AS03 -like adjuvant, InvivoGen), Montanide ISA™ VG (Seppic), AF03 (Sanofi), GLA-SE (Infectious Disease Research Institute (IDRI)), polysorbate 80 (Tween®80; ICL Americas, Inc.), imidazopyridine compounds (see International Application No. PCT / US2007 / 064857, published as International Publication No. W02007 / 109812), imidazoquinoxaline compounds (see International Application No. PCT / US2007 / 064858, published as International Publication No. W02007 / 109813) and saponins, such as QS21 (see Kensil et al., in VACCINE DESIGN: THE SUBUNIT AND ADJUVANT APPROACH, eds. Powell & Newman, Plenum Press, NY, 1995); U.S. Pat. No. 5,057,540). In certain embodiments, the adjuvant is Freund’s adjuvant (complete or incomplete). Other adjuvants are oil-in-water emulsions (such as squalene orpeanut oil), optionally in combination with immune stimulants, such as monophosphoryl lipid A (see Stoute et al., N. Engl. J. Med. 336, 86-91 (1997)).

[0236] In certain embodiments, the pharmaceutical composition disclosed herein can further comprise a pharmaceutically acceptable carrier. Pharmaceutically acceptable carrier, excipients, or diluents includes a carrier, excipient, or diluent approved by a regulatory agency of the Federal or a state government or listed in the U.S. Pharmacopeia or other generally recognized pharmacopeia for use in animals, and more particularly in humans. The term “carrier” refers to a diluent, excipient, or vehicle with which a therapeutic agent is administered. Pharmaceutically acceptable carrier, excipients or diluents includes any and all solvents, dispersion media, coatings, antibacterial agents, antifungal agents, isotonic and absorption delaying agents, and the like. Acceptable carriers, excipients, and / or diluents for therapeutic use are well known in the pharmaceutical art, and are described, for example, in REMINGTON'S PHARMACEUTICAL SCIENCES, Mack Publishing Co. (A. R. Gennaro edit. 1995). As one of skill in the art can appreciate, the choice of a pharmaceutically acceptable carrier, excipient, and / or diluent can be selected in regard to the intended route of administration and standard pharmaceutical practice.

[0237] In certain embodiments, the pharmaceutical composition comprises between about 102RCV-FFU / ml and about IO10RCV-FFU / ml of arenaviruses. In certain embodiments, the pharmaceutical composition comprises between about 102RCV-FFU / ml and about 109RCV- FFU / ml, between about 102RCV-FFU / ml and about 108RCV-FFU / ml, between about 102RCV-FFU / ml and about 107RCV-FFU / ml, between about 102RCV-FFU / ml and about 106RCV-FFU / ml, between about 102RCV-FFU / ml and about 105RCV-FFU / ml, between about102RCV-FFU / ml and about 104RCV-FFU / ml, between about 102RCV-FFU / ml and about103RCV-FFU / ml, between about 103RCV-FFU / ml and about IO10RCV-FFU / ml, between about 103RCV-FFU / ml and about 109RCV-FFU / ml, between about 103RCV-FFU / ml and about 108RCV-FFU / ml, between about 103RCV-FFU / ml and about 107RCV-FFU / ml, between about 103RCV-FFU / ml and about 106RCV-FFU / ml, between about 103RCV- FFU / ml and about 105RCV-FFU / ml, between about 103RCV-FFU / ml and about 104RCV- FFU / ml, between about 104RCV-FFU / ml and about IO10RCV-FFU / ml, between about 104RCV-FFU / ml and about 109RCV-FFU / ml, between about 104RCV-FFU / ml and about 108RCV-FFU / ml, between about 104RCV-FFU / ml and about 107RCV-FFU / ml, between about104RCV-FFU / ml and about 106RCV-FFU / ml, between about 104RCV-FFU / ml and about105RCV-FFU / ml, between about 105RCV-FFU / ml and about IO10RCV-FFU / ml, between about 105RCV-FFU / ml and about 109RCV-FFU / ml, between about 105RCV-FFU / ml andabout 108RCV-FFU / ml, between about 105RCV-FFU / ml and about 107RCV-FFU / ml, between about 105RCV-FFU / ml and about 106RCV-FFU / ml, between about 106RCV- FFU / ml and about IO10RCV-FFU / ml, between about 106RCV-FFU / ml and about 109RCV- FFU / ml, between about 106RCV-FFU / ml and about 108RCV-FFU / ml, between about 106RCV-FFU / ml and about 107RCV-FFU / ml, between about 107RCV-FFU / ml and about IO10RCV-FFU / ml, between about 107RCV-FFU / ml and about 109RCV-FFU / ml, between about 107RCV-FFU / ml and about 108RCV-FFU / ml, between about 108RCV-FFU / ml and about IO10RCV-FFU / ml, between about 108RCV-FFU / ml and about 109RCV-FFU / ml, or between about 109RCV-FFU / ml and about IO10RCV-FFU / ml of arenaviruses. In certain embodiments, the pharmaceutical composition comprises between about 105RCV-FFU / ml to about 109RCV-FFU / ml, between about 105RCV-FFU / ml to about 8* 108RCV-FFU / ml, between about 105RCV-FFU / ml to about 6* 108RCV-FFU / ml, between about 105RCV- FFU / ml to about 5 * 108RCV-FFU / ml, between about 2* 105RCV-FFU / ml to about 109RCV- FFU / ml, between about 2* 105RCV-FFU / ml to about 8* 108RCV-FFU / ml, or between about 2* 105RCV-FFU / ml to about 6* 108RCV-FFU / ml of arenaviruses. In certain embodiments, the pharmaceutical composition comprises between about 2* 105RCV-FFU / ml to about 5 / I O8RCV-FFU / ml of arenaviruses.5.5 Methods of Reducing Formation of Subvisible Particles or Loss of Viral Titer

[0238] The present disclosure provides methods for reducing the formation of subvisible particles in an arenavirus formulation. The present disclosure further provides methods for reducing the loss of viral titer (e.g., during manufacturing, filtration, freezing / thawing process(es) and / or storage of an arenavirus formulation). In certain embodiments, the method comprises combining an arenavirus with a solution to form a liquid arenavirus formulation disclosed in Section 5.1. In certain embodiments, the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (e.g., citrate) (see Section 5.1.1). In certain embodiments, the liquid arenavirus formulation further comprises at least one of a carbohydrate (e.g., sucrose or trehalose) (see Section 5.1.2), a salt (e.g., sodium chloride) (see Section 5.1.3), a buffer (e.g., TRIS) (see Section 5.1.4), a surfactant (e.g., poloxamer 188) (see Section 5.1.5), and a stabilizer (e.g., rHSA) (see Section 5.1.6). In certain embodiments, the liquid arenavirus formulation is suitable for lyophilization.

[0239] In certain embodiments, the method disclosed herein reduces the formation of subvisible particles during manufacturing. In certain embodiments, the manufacturing comprises a purification process. In certain embodiments, the manufacturing comprises at least one chromatography step. In certain embodiments, the method comprises combining anarenavirus with a solution to form a liquid arenavirus formulation disclosed in Section 5.1, wherein the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (see Section 5.1.1). In certain embodiments, the liquid arenavirus formulation further comprises a salt (see Section 5.1.3) and / or a surfactant (see Section 5.1.5). In certain embodiments, the polyvalent anion, salt, and / or the surfactant in the liquid arenavirus formulation is at a concentration that reduces the formation of subvisible particles during manufacturing. In certain embodiments, the liquid arenavirus formulation further comprises a carbohydrate (see Section 5.1.2), a buffer (see Section 5.1.4), and / or a stabilizer (see Section 5.1.6).

[0240] In certain embodiments, the method disclosed herein reduces the loss of viral titer during filtration. In certain embodiments, the filtration comprises bioburden reduction filtration, sterile filtration, or a combination thereof. In certain embodiments, the method comprises combining an arenavirus with a solution to form a liquid arenavirus formulation disclosed in Section 5.1, wherein the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (see Section 5.1.1). In certain embodiments, the liquid arenavirus formulation further comprises a salt (see Section 5.1.3). In certain embodiments, the polyvalent anion and / or the salt in the liquid arenavirus formulation is at a concentration that reduces the loss of viral titer during filtration. In certain embodiments, the liquid arenavirus formulation further comprises a carbohydrate (see Section 5.1.2), a buffer (see Section 5.1.4), a surfactant (see Section 5.1.5), and / or a stabilizer (see Section 5.1.6).

[0241] In certain embodiments, the subvisible particles reduced by the presently disclosed methods have a size of at least about 0.1 pm, at least about 0.5 pm, at least about 1 pm, at least about 5 pm, at least about 10 pm, at least about 15 pm, at least about 20 pm, at least about 25 pm, at least about 30 pm, at least about 35 pm, at least about 40 pm, at least about 45 pm, at least about 50 pm, at least about 55 pm, at least about 60 pm, at least about 65 pm, at least about 70 pm, at least about 75 pm, at least about 80 pm, at least about 85 pm, at least about 90 pm, at least about 95 pm, or at least about 100 pm. In certain embodiments, the subvisible particles reduced by the presently disclosed methods have a size of at least about 1 pm. In certain embodiments, the subvisible particles reduced by the presently disclosed method have a size of at least about 10 pm. In certain embodiments, the subvisible particles reduced by the presently disclosed method have a size of at least about 25 pm.

[0242] In certain embodiments, the method disclosed herein reduces the formation of subvisible particles during storage. In certain embodiments, the method disclosed herein reduces the loss of viral titer during storage. In certain embodiments, the method comprisescombining an arenavirus with a solution to form a liquid arenavirus formulation disclosed in Section 5.1, wherein the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (see Section 5.1.1), and further comprises a carbohydrate (see Section 5.1.2) and / or a stabilizer (see Section 5.1.6). In certain embodiments, the carbohydrate and / or the stabilizer in the liquid arenavirus formulation is at a concentration that reduces the formation of subvisible particles during storage. In certain embodiments, the liquid arenavirus formulation further comprises a salt (see Section 5.1.3), a buffer (see Section 5.1.4), and / or a surfactant (see Section 5.1.5).

[0243] In certain embodiments, the temperature of the storage is above about 2°C, room temperature (e.g., 20°C - 25°C), between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C. In certain embodiments, the temperature of the storage is about -20°C. In certain embodiments, the temperature of the storage is about -80°C. In certain embodiments, the storage lasts at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 12 months, at least about 18 months, at least about 24 months, at least about 3 years, at least about 4 years, at least about 5 years, or longer. In certain embodiments, the storage lasts at least about 6 months. In certain embodiments, the storage lasts at least about 9 months. In certain embodiments, the storage lasts at least about 18 months.

[0244] In certain embodiments, the method disclosed herein reduces the formation of subvisible particles after freezing and thawing. In certain embodiments, the method disclosed herein reduces the loss of viral titer after freezing and thawing. In certain embodiments, the method comprises combining an arenavirus with a solution to form a liquid arenavirus formulation disclosed in Section 5.1, wherein the liquid arenavirus formulation comprises an arenavirus (see Section 5.3) and a polyvalent anion (see Section 5.1.1), and further comprises a carbohydrate (see Section 5.1.2) and / or a salt (see Section 5.1.3). In certain embodiments, the carbohydrate and / or the salt in the liquid arenavirus formulation is at a concentration that reduces the formation of subvisible particles after freezing and thawing. In certain embodiments, the liquid arenavirus formulation further comprises a buffer (see Section 5.1.4), a surfactant (see Section 5.1.5), and / or a stabilizer (see Section 5.1.6).

[0245] In certain embodiments, the freezing and thawing comprises at least 1, at least 2, at least 3, at least 4, at least 5, at least 6, at least 6, at least 7, at least 8, at least 9, at least 10, or more freezing and thawing (F / T) cycles. In certain embodiments, the freezing and thawingcomprises 3 F / T cycles. In certain embodiments, the freezing and thawing comprises 5 F / T cycles. In certain embodiments, the temperate of the freezing condition is about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.5.6 Methods of Treatment

[0246] Provided herein are methods for treating or preventing a disease using the presently disclosed arenavirus formulations (see Section 5.1 and Section 5.2) and pharmaceutical compositions thereof (see Section 5.4).

[0247] In certain embodiments, the method comprises administering a liquid arenavirus formulation disclosed herein (see Section 5.1) to a subject in need thereof. In certain embodiments, the method comprises administering a liquid arenavirus formulation diluted from a liquid arenavirus formulation disclosed herein (see Section 5.1) to a subject in need thereof.

[0248] In certain embodiments, the method comprises diluting the liquid arenavirus formulation disclosed herein (see Section 5.1) prior to the administering to the subject. In certain embodiments, The diluting comprises adding a known volume of a buffer (see Section 5.1.4) or an appropriate diluent (e.g., sterile water, water for injection, or saline) to the liquid arenavirus formulation disclosed herein (see Section 5.1) to obtain a final arenavirus concentration suitable for administration to the subject. The diluting can be performed in accordance to and as required by a pharmacy manual, applicable regulation, guidelines, good clinical practices (GCP), aseptic preparation techniques, and / or standard operating procedures (SOPs).

[0249] In certain embodiments, the liquid arenavirus formulation comprises a therapeutically effective amount of the arenavirus.

[0250] In certain embodiments, the method comprises administering a lyophilized arenavirus formulation disclosed herein (see Section 5.2) to a subject in need thereof. In certain embodiments, the method comprises administering a liquid arenavirus formulation reconstituted from a lyophilized arenavirus formulation disclosed herein (see Section 5.2) to a subject in need thereof. In certain embodiments, the method comprises reconstituting the lyophilized arenavirus formulation prior to the administering to the subject.

[0251] Reconstitution of a lyophilized formulation is a skill commonly held in the art. In certain embodiments, the reconstitution comprises adding a known volume of liquid (e.g., sterile water, water for injection, buffer, or saline) to the lyophilized formulation. In certain embodiments, the reconstitution further comprises rotating or agitating (e.g., gently) the combined components. In certain embodiments, the reconstitution further comprisesconfirming substantially complete dissolution or suspension of all solids with the liquid component, e.g., by visual inspection. In certain embodiments, a reconstituted composition can be administered immediately or can be stored and / or incubated for a period of time before use. In certain embodiments, a reconstituted composition disclosed herein can be stored and / or incubated for a period of time at room temperature (e.g., about 20°C-25°C) or under refrigeration (e.g., about 4°C). In certain embodiments, a reconstituted composition disclosed herein can be stored for up to about 48 hours, up to about 36 hours, up to about 24 hours, up to about 12 hours, up to about 6 hours, up to about 2 hours, up to about 1 hour, or up to about 30 minutes prior to administration to the subject.

[0252] In certain embodiments, the method comprises administering the liquid arenavirus formulation or lyophilized formulation to the subject through intratumoral injection, oral, intradermal, intramuscular, intraperitoneal, intravenous, topical, subcutaneous, percutaneous, intranasal or inhalation routes, or via scarification (scratching through the top layers of skin, e.g., using a bifurcated needle. Specifically, subcutaneous, intramuscular or intravenous routes can be used.

[0253] In certain embodiments, the liquid arenavirus formulation or lyophilized formulation is administered to the subject intravenously, intramuscularly, intratumorally, or subcutaneously.

[0254] In certain embodiments, the method comprises administering a pharmaceutical composition disclosed herein (see Section 5.4) to a subject in need thereof.

[0255] Diseases that can be treated or prevented by the methods disclosed herein include neoplastic diseases and infectious diseases.

[0256] Neoplastic diseases that can be treated or prevented by the methods disclosed herein include acute lymphoblastic leukemia; acute lymphoblastic lymphoma; acute lymphocytic leukemia; acute myelogenous leukemia; acute myeloid leukemia (adult / childhood); adrenocortical carcinoma; AIDS-related cancers; AIDS-related lymphoma; anal cancer; appendix cancer; astrocytoma; atypical teratoid / rhabdoid tumor; basal-cell carcinoma; bile duct cancer, extrahepatic (cholangiocarcinoma); bladder cancer; bone osteosarcoma / malignant fibrous histiocytoma; brain cancer (adult / childhood); brain tumor, cerebellar astrocytoma (adult / childhood); brain tumor, cerebral astrocytoma / malignant glioma brain tumor; brain tumor, ependymoma; brain tumor, medulloblastoma; brain tumor, supratentorial primitive neuroectodermal tumors; brain tumor, visual pathway and hypothalamic glioma; brainstem glioma; breast cancer; bronchial adenomas / carcinoids;bronchial tumor; Burkitt lymphoma; cancer of childhood; carcinoid gastrointestinal tumor; carcinoid tumor; carcinoma of adult, unknown primary site; carcinoma of unknown primary; central nervous system embryonal tumor; central nervous system lymphoma, primary; cervical cancer; childhood adrenocortical carcinoma; childhood cancers; childhood cerebral astrocytoma; chordoma, childhood; chronic lymphocytic leukemia; chronic myelogenous leukemia; chronic myeloid leukemia; chronic myeloproliferative disorders; colon cancer; colorectal cancer; craniopharyngioma; cutaneous T-cell lymphoma; desmoplastic small round cell tumor; emphysema; endometrial cancer; ependymoblastoma; ependymoma; esophageal cancer; Ewing’s sarcoma in the Ewing family of tumors; extracranial germ cell tumor; extragonadal germ cell tumor; extrahepatic bile duct cancer; gallbladder cancer; gastric (stomach) cancer; gastric carcinoid; gastrointestinal carcinoid tumor; gastrointestinal stromal tumor; germ cell tumor: extracranial, extragonadal, or ovarian gestational trophoblastic tumor; gestational trophoblastic tumor, unknown primary site; glioma; glioma of the brain stem; glioma, childhood visual pathway and hypothalamic; hairy cell leukemia; head and neck cancer; heart cancer; hepatocellular (liver) cancer; Hodgkin lymphoma; hypopharyngeal cancer; hypothalamic and visual pathway glioma; intraocular melanoma; islet cell carcinoma (endocrine pancreas); Kaposi Sarcoma; kidney cancer (renal cell cancer); Langerhans cell histiocytosis; laryngeal cancer; lip and oral cavity cancer; liposarcoma; liver cancer (primary); lung cancer, non-small cell; lung cancer, small cell; lymphoma, primary central nervous system; macroglobulinemia, Waldenstrom; male breast cancer; malignant fibrous histiocytoma of bone / osteosarcoma; medulloblastoma; medulloepithelioma; melanoma; melanoma, intraocular (eye); Merkel cell cancer; Merkel cell skin carcinoma; mesothelioma; mesothelioma, adult malignant; metastatic squamous neck cancer with occult primary; mouth cancer; multiple endocrine neoplasia syndrome; multiple myeloma / plasma cell neoplasm; mycosis fungoides, myelodysplastic syndromes; myelodysplastic / myeloproliferative diseases; myelogenous leukemia, chronic; myeloid leukemia, adult acute; myeloid leukemia, childhood acute; myeloma, multiple (cancer of the bone-marrow); myeloproliferative disorders, chronic; nasal cavity and paranasal sinus cancer; nasopharyngeal carcinoma; neuroblastoma, non- small cell lung cancer; non-Hodgkin lymphoma; oligodendroglioma; oral cancer; oral cavity cancer; oropharyngeal cancer; osteosarcoma / malignant fibrous histiocytoma of bone; ovarian cancer; ovarian epithelial cancer (surface epithelial-stromal tumor); ovarian germ cell tumor; ovarian low malignant potential tumor; pancreatic cancer; pancreatic cancer, islet cell; papillomatosis; paranasal sinus and nasal cavity cancer; parathyroid cancer; penile cancer; pharyngeal cancer; pheochromocytoma; pineal astrocytoma; pineal germinoma; pinealparenchymal tumors of intermediate differentiation; pineoblastoma and supratentorial primitive neuroectodermal tumors; pituitary tumor; pituitary adenoma; plasma cell neoplasia / multiple myeloma; pleuropulmonary blastoma; primary central nervous system lymphoma; prostate cancer; rectal cancer; renal cell carcinoma (kidney cancer); renal pelvis and ureter, transitional cell cancer; respiratory tract carcinoma involving the NUT gene on chromosome 15; retinoblastoma; rhabdomyosarcoma, childhood; salivary gland cancer; sarcoma, Ewing family of tumors; Sezary syndrome; skin cancer (melanoma); skin cancer (non-melanoma); small cell lung cancer; small intestine cancer soft tissue sarcoma; soft tissue sarcoma; spinal cord tumor; squamous cell carcinoma; squamous neck cancer with occult primary, metastatic; stomach (gastric) cancer; supratentorial primitive neuroectodermal tumor; T-cell lymphoma, cutaneous (Mycosis Fungoides and Sezary syndrome); testicular cancer; throat cancer; thymoma; thymoma and thymic carcinoma; thyroid cancer; thyroid cancer, childhood; transitional cell cancer of the renal pelvis and ureter; urethral cancer; uterine cancer, endometrial; uterine sarcoma; vaginal cancer; vulvar cancer; and Wilms Tumor.

[0257] Infectious diseases that can be treated or prevented by the methods disclosed herein include acute flaccid myelitis, anaplasmosis, anthrax, babesiosis, brucellosis, campylobacteriosis, carbapenem-resistant infection, chancroid, chikungunya virus infection, chlamydia, Clostridium difficile infection, coccidioidomycosis fungal infection, Covid- 19, cryptosporidiosis, cyclosporiasis, dengue fever, diphtheria, E. coli infection, eastern equine encephalitis, Ebola hemorrhagic fever, ehrlichiosis, arboviral encephalitis, parainfectious encephalitis, enterovirus infection, giardiasis, glanders, gonococcal infection, granuloma inguinale, type b haemophilus influenza disease, hantavirus pulmonary syndrome, hemolytic uremic syndrome, hepatitis A, hepatitis B, hepatitis C, hepatitis D, hepatitis E, herpes, herpes zoster, histoplasmosis infection, acquired immunodeficiency syndrome, human papillomavirus infection, influenza, legionellosis, leprosy, leptospirosis, listeriosis, Lyme disease, lymphogranuloma venereum infection, malaria, measles, melioidosis, viral meningitis, viral meningitis, middle east respiratory syndrome, multisystem inflammatory syndrome, mumps, norovirus infection, pediculosis, pelvic inflammatory disease, pertussis, plague, pneumococcal disease, poliomyelitis, Powassan virus infection, psittacosis, pthiriasis, pustular rash disease, Q-fever, rabies, rickettsiosis, rubella, salmonellosis gastroenteritis, scabies infestation, sepsis, severe acute respiratory syndrome, shigellosis gastroenteritis, smallpox, staphyloccal infection, staphylococcal infection, streptococcal disease, streptococcal toxic-shock syndrome, syphilis, tetanus infection, trichomoniasis, trichonosisinfection, tuberculosis, tularemia, typhoid fever, typhus, varicella, vibrio cholera, vibriosis, viral hemorrhagic fever, west nile virus infection, coronavirus infection, yeast infection, yellow fever, yersenia, and zika virus infection.

[0258] Also provided herein are kits that can be used to perform the methods described herein. In certain embodiments, the kit provided herein comprises one or more containers. These containers can hold for storage the formulations or compositions disclosed herein (e.g., liquid arenavirus formulations, lyophilized formulations, pharmaceutical compositions). In certain embodiments, the kit further comprises a liquid (e.g., sterile water, water for injection, buffer, or saline) for diluting the liquid arenavirus formulation or reconstituting the lyophilized arenavirus formulation. Also included in the kit are instructions for use. These instructions describe, in sufficient detail, a treatment protocol for using the formulation and composition contained therein. For example, the instructions can include dosing and administration instructions as provided herein for the methods of treating a neoplastic disease or an infectious disease.6. SEQUENCES7. EXAMPLES

[0259] The following is a description of various methods and materials used in the studies. They are put forth so as to provide those of ordinary skill in the art with a complete disclosure and description of how to make and use the present invention, and are not intended to limit the scope of what the inventors regard as their invention, nor are they intended to represent that the experiments below were performed and are all of the experiments that may be performed. It is to be understood that exemplary descriptions written in the present tense were not necessarily performed, but rather that the descriptions can be performed to generate the data and the like associated with the teachings of the present invention. Efforts have been made to ensure accuracy with respect to numbers used (e.g., amounts, percentages, etc.), but some experimental errors and deviations should be accounted for.7.1 Example 1: Minimizing Loss of Viral Titer During Filtration

[0260] Sterility is required for the therapeutic use of virus formulations (e.g., pharmaceutical compositions) by parenteral administration. This is often achieved by subjecting the formulations to sterile filtration. Therefore, a formulation is needed, which allows adequate filterability (e.g., sterile filtration using 0.2 pm or 0.22 pm pore size). Careful selection and optimization of the excipients of the formulation is required in order to minimize aggregation of viral particles and formation of subvisible particles during manufacturing, purification and storage. Furthermore, the infectivity of the viral vectors in the formulation needs to be retained during storage. Thus, an ideal arenavirus formulation has the following features:(a) minimized loss of viral titer during purification process,(b) minimized formation of subvisible particles,(c) maintained biophysical stability of viral particles during freeze / thaw process, and(d) maintained biophysical stability of viral particles during storage.

[0261] Arenaviruses are enveloped, bi-segmented, ambisense, single stranded RNA (ssRNA) viruses. They have significant poly dispersity (pleomorphic / often spherical) with an approximate diameter of 50 nm to 300 nm. The virus particles tend to aggregate and / or form subvisible particles during manufacturing and storage. The formation of subvisible particles can result in significant loss of virus particles during sterile filtration (z.e., through a 0.2 pm or 0.22 pm pore size membrane). Therefore, the main challenge for formulations comprising arenavirus particles is the selection of a formulation that allows for sterile filtration through a filter with 0.2 pm or 0.22 pm pore size, avoiding a substantial loss of viral titer, and maintenance of the biophysical stability of the viral particles during subsequent storage.

[0262] Furthermore, the arenavirus formulations should allow for storage at a temperature of -20°C, or ideally 2°C - 8°C, to facilitate handling. In addition, the arenavirus formulations can also allow for storage at room temperature (e.g., 20°C - 25°C) for a defined short period of time.

[0263] A closed system, which is described in U.S. Patent No. 10,662,412, has been developed to allow virus preparation under full aseptic conditions, and thus dispenses the step of sterile filtration and reduces the loss of virus and improves virus recovery during purification. However, such closed system has the disadvantages of higher production costs and complex operating conditions, as compared to a manufacturing process using a terminal 0.2 pm or 0.22 pm sterile filtration.7.1.1 Presence of polyvalent anions in the arenavirus formulation reduced viral particle loss during sterile filtration

[0264] The effects of polyvalent anions (e.g., TSPP, citrate, malate, succinate and tartrate) on the sterile filtration recovery was evaluated. Respective viral titers were analyzed by replication competent virus focus forming units (RCV-FFU) assay prior to filtration and after filtration. The percentage of viral vector particles recovered after filtration was determined. The effects of polyanions on filtration recovery were calculated (see FIG. 1). Evaluation was performed by MODDE Pro (Satorius Stedim Data Analytics AB) software. A D-Optimal design was applied, and data were fitted by multiple linear regression (MLR) model. The effect plot showed the effects calculated as twice the MLR coefficient and sorted descending in absolute value (confidence level 95%). An excipient that had a positive effect on filterability was indicated by a positive value.

[0265] As demonstrated in FIG. 1, all tested polyanions showed a positive effect on filtration recovery, with TSPP and citrate having the strongest positive effects.7.1.2 Presence of NaCl and citrate in the arenavirus formulation reduced viral particle loss during sterile filtration

[0266] The recovery of virus particles after sterile filtration was analyzed as a function of increasing sodium chloride (NaCl) and citrate concentrations in the arenavirus formulation. Viral titers were determined by focus forming units (FFU) and RCV-FFU assays prior to and after sterile filtration. Evaluation was performed by MODDE Pro (Satorius Stedim Data Analytics AB). A D-Optimal design was applied, and data were fitted by multiple linear regression (MLR) model.

[0267] The results showed that increasing concentrations of NaCl and citrate in the arenavirus formulation significantly improved recoveries of virus particles after sterile filtration (see FIG. 2).7.1.3 TRIS / Citrate based formulation reduced the virus titer loss during reduction filtration

[0268] Small scale purification runs were performed to compare the virus particle recoveries during the manufacturing of the arenavirus formulations using a HEPES based formulation (10 mM HEPES, 150 mM NaCl, 20 mM Glycine, 1% rHSA, pH 7.4) or a TRIS- based formulation (10 mM Tris, 50 mM Citrate, 125 mM NaCl, 5% (w / v) Sucrose, 0.03% Poloxamer 188, 0.1% rHSA; pH 8.0). The influence of different formulations (HEPES-based vs. TRIS / Citrate-based) on virus titers and filtration recovery during the manufacturing process was compared.

[0269] Three batches of arenavirus formulations were processed with the HEPES-based formulation (see FIGS. 3A & 3B). Five batches of arenavirus formulations were processed with the TRIS / Citrate-based formulation (see FIGS. 4A & 4B). Virus titers and filtration recoveries were analyzed by quantification of RCV-FFU and FFU after each manufacturing steps (PS1-PS6). Table 1 shows the RCV-FFU and FFU recoveries of the final bioburden reduction filtration step.Table 1: Overview of RCV-FFU and FFU recoveries of the final bioburden reduction filtration step.

[0270] The 0.45 pm filtration recoveries for the HEPES-based batches were relatively low, ranging from 19% - 38% (RCV FFU) and 18 - 52% (FFU). By contrast, the filtration recoveries were considerably higher for the TRIS-based batches, ranging from 64 - 78% (RCV-FFU) and 64 - 107% (FFU) (see Table 1 and FIGS. 3A & 3B and FIGS. 4A & 4B).

[0271] The data demonstrated that the filtration recoveries following 0.45 pm bioburden reduction filtration were improved using the TRIS / Citrate-based formulation for the manufacture of the arenavirus formulation.7.2 Example 2: Minimizing formation of subvisible particles7.2.1 The addition of polyvalent anions to the arenavirus formulation reduced the formation of subvisible particles

[0272] Concentrations of subvisible particles in arenavirus formulations (10 mM Tris, 5% Sucrose, 50 mM NaCl; 0.03% Poloxamer 188; ~1% rHSA, pH 8.0) comprising 25 mM or 50 mM polyanions (TSPP, succinate, tartrate, and malate) were analyzed, after the second chromatography step.

[0273] Flow imaging microscopy (FIM) was used to measure the size distribution, shape and concentration of particles that are about 1 pm to several hundred pm in size. Sampleanalysis was performed on a FlowCam 8100 (Fluid Imaging Technologies, Inc.) equipped with a 10x objective, a FOV80 flow cell and a precision syringe of 1 mL. Flow was set to 0.15 mL / min with an autoimage rate of 19 images per second, resulting in an efficiency of about 71%. The equivalent spherical diameter (ESD) was measured, which is a calculated value based on the volume of the particles.

[0274] Arenavirus formulations comprising a polyanion at a concentration of 50 mM had significantly lower concentrations of subvisible particles as compared to arenavirus formulations comprising 25 mM polyanion (see FIG. 5). The observed effect was independent of the specific polyvalent anion used, indicating that a higher polyanion concentration reduced the interaction of viral particles (beside an increase of ionic strength) and therefore reduced the formation of subvisible particles during the purification process.7.2.2 NaCl, citrate and the surfactant poloxamer 188 decreased the formation of subvisible particles in the viral particle formulation

[0275] The effects of different concentrations of NaCl, citrate and poloxamer 188 on the formation of subvisible particles were determined for a TRIS / Citrate based arenavirus formulation (10 mM Tris, 5% (w / v) Sucrose, 0.1% rHSA, pH 8.0). Subvisible particles were analyzed by FIM. MODDE Pro (Satorius Stedim Data Analytics AB) was used for evaluation. Data were fitted by multiple linear regression (MLR) model.

[0276] Increasing concentrations of NaCl (up to 125 mM), citrate (up to 50 mM) as well as the surfactant poloxamer 188 (up to 0.05%) reduced the formation of subvisible particles in the arenavirus formulation (see FIG. 6). However, 125 mM NaCl and 35 to 50 mM citrate sufficiently suppressed the formation of subvisible particles without the presence of the surfactant poloxamer 188.7.2.3 Subvisible particle formation is reduced by the use of a TRIS / Citrate based formulation

[0277] The effect of the buffer used to prepare arenavirus formulations on formation of subvisible particles (>1 pm) during the manufacturing process was analyzed. Three batches of arenavirus formulations were processed with HEPES-based formulation (10 mM HEPES, 150 mM NaCl, 20 mM glycine, 1% rHSA; pH 7.4) and five batches of arenavirus formulations were processed with a TRIS / Citrate-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 5% (w / v) sucrose, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). Subvisible particles were analyzed by FIM during the manufacturing process. Specifically, the concentration of total subvisible particles (i.e., particles >1 pm in diameter) (FIG. 7A), particles >10 pm in diameter (FIG. 7B), and particles >25 pm in diameter (FIG. 7C) wasmeasured in the eluate resulting from each individual step during the manufacturing process). All samples underwent one freeze / thaw cycle before subvisible particle measurement.

[0278] The results showed that the formation of subvisible particles during the manufacture of arenavirus formulations was significantly reduced when using a TRIS / Citrate- based formulation compared to formulations prepared using an HEPES-based formulation (FIGS. 7A-7C).7.3 Example 3: Maintenance of Biophysical Stability of Viral Particles During Freeze / Thaw Process7.3.1 Addition of carbohydrates to the formulation stabilized viral particles during repeated freeze / thaw cycles and facilitated subsequent sterile filtration

[0279] To evaluate the influence of carbohydrates on the stability of arenavirus during freezing and thawing, arenavirus formulations were prepared using different combinations of polyanions (20 mM TSPP, 50 mM succinate, or 50 mM citrate) and carbohydrates (sucrose, trehalose, or sorbitol at a concentration of 5% (w / v) each) in a TRIS / Citrate-based formulation (10 mM Tris, 125 mM NaCl; 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). An arenavirus formulation without carbohydrates was prepared in a HEPES-based formulation (10 mM HEPES, 20 mM glycine, 150 mM NaCl, 1% rHSA; pH 7.4) and used as a reference. Viral titer was determined by RCV-FFU and FFU assays for each formulation prior to freezing, after three (3) freeze / thaw cycles, and after subsequent sterile filtration (0.22 pm). One (1) freeze / thaw cycle consisted of freezing the arenavirus formulation for at least one hour at -20°C in 2R Type I glass vials followed by thawing the formulation at room temperature until no ice crystals were visible (but not less than 20 minutes).

[0280] As shown in FIG. 8, the presence of carbohydrates (especially sucrose and trehalose) in the arenavirus formulations stabilized viral particles during the repeated freeze / thaw cycles. The differences in stabilization after three freeze / thaw cycles became more obvious after subsequent sterile filtration. Compared to the reference arenavirus formulation without a carbohydrate formulated in HEPES buffer, the TRIS / Citrate-based formulations supplemented with carbohydrates showed improved recovery after sterile filtration.7.3.2 Increasing concentrations of sucrose increased viral particle recovery after repeated freeze / thaw cycles

[0281] To further evaluate the influence of sucrose on the stability of arenavirus formulations during repeated freeze / thaw cycles, increasing concentrations of sucrose ranging from 0 to 10% (w / v) were added to arenavirus formulations prepared in a TRIS-basedformulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). The formulations were sterile filtered and subjected to five (5) freeze / thaw cycles. One (1) freeze / thaw cycle consisted of freezing the arenavirus formulation for at least one hour at -20°C followed by thawing the formulation at room temperature for no less than 20 minutes. The viral titer was determined by RCV-FFU assay for each of the formulations prior to sterile filtration, after sterile filtration, and after five freeze / thaw cycles (FIG. 9). RCV-FFU measurements were also used to assess viral titer recovery for formulations that were sterile filtered and subjected to five freeze / thaw cycles (FIG. 10).

[0282] As demonstrated in FIG. 9, viral titers in the high-sucrose formulations remained relatively constant after five freeze / thaw cycles and subsequent sterile filtration, whereas there was a considerable decrease in viral titers of formulations comprising no or only low concentrations of sucrose after five freeze / thaw cycles and, particularly, after sterile filtration. FIG. 10 shows that after sterile filtration and five freeze / thaw cycles, arenavirus formulations comprising sucrose concentrations of 3% or more had an increased percentage of virus titer recovery as compared to arenavirus formulations comprising no or only low concentrations of sucrose.

[0283] Data showed that the presence of sucrose at concentrations of > 3% in arenavirus formulations exerted a stabilizing effect on virus particles during repeated freezing and thawing while not influencing the filterability of the formulations.7.3.3 Presence of carbohydrates (e.g., sucrose) in arenavirus formulations exhibit no increase of subvisible particles during repeated freeze / thaw cycles at -20°C

[0284] The effect of carbohydrates (e.g., sucrose) on the formation of subvisible particles in arenavirus formulations subjected to repeated freeze / thaw cycles was assessed. Briefly, arenavirus formulations having a sucrose concentration of 5% were prepared in a TRIS / Citrate-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 5% sucrose, 0.03% poloxamer 188, 0.1% rHSA; pH 8.0). Subvisible particles were analyzed by FIM to determine the concentration of total subvisible particles (i.e., particles >1 pm in diameter), particles >10 pm in diameter, and particles >25 pm in diameter in the formulations prior to and after each of three (3) consecutive freeze / thaw cycles. One (1) freeze / thaw cycle consisted of freezing 0.7 mL of a solution comprising the arenavirus formulation in 2R Type I glass vials for at least one hour at -20°C followed by thawing the formulation at room temperature for no less than 20 minutes. The concentrations of subvisible particles were used to evaluate the stability of the arenavirus formulation during repeated freeze / thaw cycles.

[0285] As demonstrated in FIG. 11, no increase of subvisible particles during repeated freeze / thaw cycles at -20°C in the presence of carbohydrates (e.g., sucrose) could be observed.7.3.4 Presence of carbohydrates (e.g., sucrose) in arenavirus formulations prevented the increase of subvisible particles during repeated freeze / thaw cycles at - 80°C

[0286] The effect of carbohydrates (e.g., sucrose) on the formation of subvisible particles in arenavirus formulations subjected to repeated freeze / thaw cycles was assessed. In brief, arenavirus formulations were prepared in a TRIS-based formulation (10 mM Tris, 20 mM TSPP, 250 mM NaCl, 0.03% poloxamer 188; pH 8.0) with and without 5% sucrose. Subvisible particles were analyzed by FIM to determine the concentration of total subvisible particles (i.e., particles >1 pm in diameter) in the formulations prior to and after each of five (5) consecutive freeze / thaw cycles. One (1) freeze / thaw cycle consisted of freezing 15 mL of a solution comprising the arenavirus formulation in 50 mL polypropylene (PP) tubes for at least one hour at -80°C followed by thawing the formulation at room temperature until no ice crystals were visible.

[0287] FIG. 12 shows a direct comparison of subvisible particle concentrations in arenavirus formulations with and without 5% sucrose after each of the five consecutive freeze / thaw cycles. While the number of subvisible particles was significantly increased for the arenavirus formulation without sucrose, it was constant in the formulation comprising 5% sucrose. Similar to the data provided in Example 7.3.3, these data showed a significant stabilizing effect resulting from the use of sucrose in the formulation of the arenavirus formulations.7.4 Example 4: Maintaining Biophysical Stability of Viral Particles During Storage7.4.1 Recombinant human serum albumin (rHSA) increased viral particle stability during storage at 2-8°C

[0288] Stability studies were performed to evaluate the influence of rHSA on the stability of the viral particles during storage at 2°C - 8°C. In brief, rHSA at concentrations ranging from 0 to 1% were added to arenavirus formulations prepared in a TRIS-based formulation (10 mM Tris, 20 mM TSPP, 125 mM NaCl, 5% (w / v) sucrose, 0.015% poloxamer 188; pH 8.0). The arenavirus formulations were stored in PP tubes for a period of five weeks at 2- 8°C. Samples were drawn from the tubes in one-week intervals and viral titer was determined by quantification of RCV-FFU.

[0289] As can be seen in FIG. 13, increasing rHSA concentrations in the arenavirus formulation resulted in reduced loss of viral titer during storage at 2-8°C. A concentration of 0.1% rHSA was sufficient to stabilize the virus particles during storage at 2-8°C.7.4.2 Presence of carbohydrates in arenavirus formulations minimized the decline of infectious titer during storage at -20°C

[0290] To evaluate the influence of carbohydrates on viral titer stability during storage at -20°C, arenavirus formulations comprising different carbohydrates (sucrose, trehalose, or sorbitol) were prepared in a TRIS / Citrate-based formulation (10 mM Tris, 50 mM citrate, 125 mM NaCl, 0.03% (w / v) poloxamer 188, 0.1% (w / v) rHSA; pH 8.0). All carbohydrates used in the formulations had a concentration of 5% (w / v). The formulations were stored in glass vials for 18 months at -20°C. Viral titer was determined by quantification of RCV-FFU (FIG. 14A) and FFU (FIG. 14B) to evaluate virus stability during storage.

[0291] As shown in FIGS. 14A-14B, increased stability was demonstrated in formulations containing a carbohydrate compared to the formulations without carbohydrate. While the infectious titer decreased significantly over time in the formulation without a carbohydrate, the loss of infectivity was considerably less over time in the formulations containing carbohydrates. Trehalose was more effective at stabilizing viral particles during storage at -20°C than the other carbohydrates tested (i.e., sucrose and sorbitol).7.4.3 Presence of carbohydrates in arenavirus formulations minimized formation of subvisible particles during storage at -20°C

[0292] To evaluate the influence of carbohydrates on formation of subvisible particles during storage at -20°C, arenavirus compositions comprising either no carbohydrate or a concentration of 5% (w / v) of either trehalose, sucrose, or sorbitol were prepared. The arenavirus formulations were stored at -20°C and the formation of subvisible particles in the formulations was monitored over a storage period of 18 months at -20°C. Subvisible particles were analyzed by FIM prior to freezing as well as after 1, 2, 3, 6, 9, 12, and 18 months of storage to measure the concentration of total subvisible particles (i.e., particles >1 pm in diameter) in samples collected from the arenavirus formulations.

[0293] Data shown in FIG. 15 demonstrated that in the absence of a carbohydrate, the concentration of subvisible particles (>1 pm) in the arenavirus formulations significantly increased during storage at -20°C. The respective increase of particles was substantially reduced in formulations comprising 5% (w / v) of a carbohydrate (i.e., trehalose, sucrose, or sorbitol). Only limited formation of particles >10 pm or >25 pm in diameter was observed inthe formulations comprising carbohydrates during the 18 months storage period (data not shown).

[0294] These results showed that the addition of carbohydrates to arenavirus formulations did not only preserve viral infectious titers (as demonstrated in Example 7.4.2), but also reduced the formation of subvisible particles in the formulations during storage over a period of 18 months at -20°C.7.4.4 Presence of carbohydrates in arenavirus formulations stabilized the size of viral particles during storage at -20°C

[0295] To evaluate the influence of carbohydrates on the size of viral particles in the formulations during storage at -20°C, arenavirus formulations comprising either no carbohydrate or a concentration of 5% (w / v) of either trehalose, sucrose or sorbitol were prepared. The arenavirus formulations were stored at -20°C and the mean diameter of the virus particles was analyzed by Nanoparticle Tracking Analysis (NTA) prior to freezing as well as after 1, 2, and 3 months of storage. In brief, NTA measurements were performed using a NanoSight NS300 instrument (Malvern Instruments Ltd., Worcestershire, UK) equipped with a violet laser module (405 nm), a low volume flow cell, and a syringe pump. For each sample, five movies of 60 seconds were recorded at a constant syringe pump flow. NanoSight software version NTA 3.4 was used for both capturing and analysis of the movies.

[0296] Results shown in FIG. 16 indicated that the mean viral particle size remained relatively constant over time during storage at -20°C in the formulations containing 5% of a carbohydrate. In contrast, a slight increase of the average particle size was observed in the arenavirus formulation that did not comprise a carbohydrate. Results presented in FIGS. 17A-17B further show that an arenavirus formulation without carbohydrates had a slight increase in the number of larger particles (z.e., particles over 300 nm in diameter) after 3 months storage at -20°C (FIG. 17A; dashed line indicates 300 nm) compared to the number of larger particles in an arenavirus formulation containing 5% trehalose after 3 months storage at -20°C (FIG. 17B; dashed line indicates 300 nm).8. EQUIVALENTS

[0297] All patents and publications mentioned in this specification are incorporated herein by reference in their entireties. From the foregoing description, it will be apparent that variations and modifications can be made to the invention described herein to adopt it tovarious uses and conditions. Such embodiments are also within the scope of the following claims.

Claims

WHAT IS CLAIMED IS:

1. A liquid arenavirus formulation comprising:(a) an arenavirus; and(b) a polyvalent anion at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

2. The liquid arenavirus formulation of claim 1, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.

3. The liquid arenavirus formulation of claim 1, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

4. A liquid arenavirus formulation comprising:(a) an arenavirus; and(b) a polyvalent anion, wherein the polyvalent anion is at a concentration that (i) reduces the formation of subvisible particles during manufacturing, and / or (ii) reduces the loss of viral titer during filtration, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process.

5. The liquid arenavirus formulation of claim 4, wherein the polyvalent anion is at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

6. The liquid arenavirus formulation of claim 5, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.

7. The liquid arenavirus formulation of claim 5, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

8. The liquid arenavirus formulation of any one of claims 1-7, wherein the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.

9. The liquid arenavirus formulation of claim 8, wherein the polyvalent anion is citrate.

10. The liquid arenavirus formulation of claim 9, wherein the citrate is at a concentration of about 50 mM or about 25 mM.

11. The liquid arenavirus formulation of claim 9, wherein the liquid arenavirus formulation is suitable for lyophilization, and the citrate is at a concentration of up to about 50 mM or between about 20 mM and about 50 mM.

12. The liquid arenavirus formulation of any one of claims 1-11, further comprising at least one of:(c) a carbohydrate;(d) a salt;(e) a buffer;(f) a surfactant; and(g) a stabilizer.

13. The liquid arenavirus formulation of claim 12, wherein:(c) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v);(d) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM;(e) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM;(f) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or(g) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v), optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).

14. The liquid arenavirus formulation of claim 12 or 13, wherein(c) the carbohydrate is at a concentration of about 5% (w / v);(d) the salt is at a concentration of about 125 mM or about 75 mM;(e) the buffer is at a concentration of about 10 mM;(f) the surfactant is at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is at a concentration of about 0.1% (w / v).

15. The liquid arenavirus formulation of claim 12 or 13, wherein the liquid arenavirus formulation is suitable for lyophilization, and(c) the carbohydrate is at a concentration of about 5% (w / v);(d) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) the buffer is at a concentration of about 10 mM;(f) the surfactant is at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is at a concentration of about 0.1% (w / v).

16. The liquid arenavirus formulation of any one of claims 12-15, wherein:(c) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or a combination thereof;(d) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride;(e) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS;(f) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is pol oxamer 188; and / or(g) the stabilizer is selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albumin is lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the stabilizer is rHSA.

17. The liquid arenavirus formulation of claim 16, wherein:(c) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v);(d) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM;(e) the buffer is TRIS at a concentration of about 10 mM;(f) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is rHSA at a concentration of about 0.1%.

18. The liquid arenavirus formulation of claim 16, wherein the liquid arenavirus formulation is suitable for lyophilization, and:(c) the carbohydrate is trehalose at a concentration of about 5% (w / v);(d) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) the buffer is TRIS at a concentration of about 10 mM;(f) the surfactant is poloxamer 188 at a concentration of about 0.03% (w / v); and / or(g) the stabilizer is rHSA at a concentration of about 0.1%.

19. The liquid arenavirus formulation of any one of claims 1-18, wherein the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0.

20. The liquid arenavirus formulation of claim 19, wherein the pH is about 8.0.

21. The liquid arenavirus formulation of any one of claims 1-20, wherein the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg.

22. The liquid arenavirus formulation of claim 21, wherein the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

23. The liquid arenavirus formulation of claim 21, wherein the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

24. The liquid arenavirus formulation of any one of claims 21, wherein the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

25. The liquid arenavirus formulation of any one of claims 12-24, wherein (c)-(g) are present at concentrations to reduce the loss of viral titer during filtration, freezing and thawing, and / or storage; and / or reduce the formation of subvisible particles during manufacturing, freezing and thawing, and / or storage.

26. The liquid arenavirus formulation of claim 25, wherein the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C and about 8°C, above aboutl5°C, about -20°C or less than about -20°C, about -65°C or less than about - 65°C, or about -80°C.

27. A liquid arenavirus formulation comprising:(a) an arenavirus;(b) citrate at a concentration of about 50 mM;(c) sucrose at a concentration of about 5% (w / v);(d) NaCl at a concentration of about 125 mM;(e) TRIS at a concentration of about 10 mM; and(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

28. A liquid arenavirus formulation comprising:(a) an arenavirus;(b) citrate at a concentration of about 25 mM;(c) trehalose at a concentration of about 5% (w / v);(d) NaCl at a concentration of about 75 mM;(e) TRIS at a concentration of about 10 mM; and(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

29. A liquid arenavirus formulation suitable for lyophilization comprising:(a) an arenavirus;(b) citrate at a concentration of between about 20 mM and about 50 mM;(c) trehalose at a concentration of about 5% (w / v);(d) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(e) TRIS at a concentration of about 10 mM;(f) pol oxamer 188 at a concentration of about 0.03% (w / v); and(g) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

30. The liquid arenavirus formulation of any one of claims 1-29, wherein the concentration of the arenavirus is between about 102RCV-FFU / ml and about 1010RCV-FFU / ml, or between about 2* 105RCV-FFU / ml and about 5 108RCV-FFU / ml.

31. The liquid arenavirus formulation of any one of claims 1-30, wherein the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.

32. The liquid arenavirus formulation of any one of claims 1-31, wherein the arenavirus is a genetically engineered arenavirus.

33. The liquid arenavirus formulation of any one of claims 1-32, wherein the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.

34. The liquid arenavirus formulation of claim 33, wherein the antigen comprises a cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of gB(dCt), pp65, and combinations thereof.

35. The liquid arenavirus formulation of claim 33, wherein the antigen comprises a human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7 / E6 fusion protein.

36. The liquid arenavirus formulation of claim 33, wherein the antigen comprises a prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof.

37. The liquid arenavirus formulation of claim 33, wherein the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reversetranscriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide and combinations thereof.

38. The liquid arenavirus formulation of claim 33, wherein the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef.

39. The liquid arenavirus formulation of claim 33, wherein the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the at least one antigenic fragment from N- to C-terminus comprises the mutations G13D, G12V, G12C, G12D, and G12R respectively.

40. The liquid arenavirus formulation of claim 33, wherein the antigen comprises two or more cancer testis antigen (CTA) fragments, optionally wherein the two or more CTA fragments are selected from the group consisting of epitopes or antigenic fragments of preferentially expressed antigen in melanoma (PRAME), melanoma-associated antigen 3 (MAGE-A3), melanoma-associated antigen 4 (MAGE-A4), and melanoma-associated antigen 6 (MAGE-A6).

41. The liquid arenavirus formulation of any one of claims 1-40, wherein the arenavirus is a non-replicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles, further optionally wherein the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen.

42. The liquid arenavirus formulation of any one of claims 1-40, wherein the arenavirus is a replicating tri-segmented arenavirus.

43. The liquid arenavirus formulation of claim 42, wherein the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF.

44. The liquid arenavirus formulation of claim 43, wherein the tri-segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an Ssegment, wherein the ORF encoding the arenaviral glycoprotein (GP) is under control of an arenavirus 3 ’ UTR.

45. The liquid arenavirus formulation of claim 42, wherein the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.

46. A lyophilized arenavirus formulation prepared from the liquid arenavirus formulation of any one of claims 1-45.

47. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at a concentration of between about 5 mM and about 150 mM, optionally wherein the polyvalent anion is at a concentration of between about 10 mM and about 60 mM.

48. The method of claim 47, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.

49. The method of claim 47, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.

50. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises a polyvalent anion at a concentration that reduces the formation of subvisible particles during manufacturing, and / or reduces the loss of viral titer during filtration.

51. The method of claim 50, wherein the polyvalent anion is at a concentration of between about 5 mM and about 150 mM.

52. The method of claim 51, wherein the polyvalent anion is at a concentration of about 50 mM or about 25 mM.I l l53. The method of claim 51, wherein the liquid arenavirus formulation is suitable for lyophilization, and the polyvalent anion is at a concentration of between about 20 mM and about 50 mM.

54. The method of any one of claims 47-53, wherein the polyvalent anion is selected from the group consisting of citrate, tetrasodium pyrophosphate (TSPP), malate, succinate, tartrate, fumarate, maleate, and combinations thereof.

55. The method of claim 54, wherein the polyvalent anion is citrate.

56. The method of claim 55, wherein the citrate is at a concentration of about 50 mM or about 25 mM.

57. The method of claim 55, wherein the citrate is at a concentration of between about 10 mM and about 60 mM.

58. The method of any one of claims 47-57, wherein the liquid arenavirus formulation further comprises at least one of:(b) a carbohydrate;(c) a salt;(d) a buffer;(e) a surfactant; and(f) a stabilizer.

59. The method of claim 58, wherein:(b) the carbohydrate is at a concentration of between about 0.1% and about 20% (w / v), optionally wherein the carbohydrate is at a concentration of between about 3% and about 10% (w / v);(c) the salt is at a concentration of up to about 450 mM, optionally wherein the salt is at a concentration of between about 50 mM and about 250 mM;(d) the buffer is at a concentration of between about 1 mM and about 50 mM, optionally wherein the buffer is at a concentration of between about 10 mM and about 20 mM;(e) the surfactant is at a concentration of between about 0.001% and about 1% (w / v), optionally wherein the surfactant is at a concentration of between about 0.01% and about 0.05% (w / v); and / or(f) the stabilizer is at a concentration of between about 0.001% (w / v) and about 3% (w / v), optionally wherein the stabilizer is at a concentration of between about 0.05 (w / v) and 0.15% (w / v).

60. The method of claim 58 or 59, wherein(b) the carbohydrate is at a concentration of about 5% (w / v);(c) the salt is at a concentration of about 125 mM or about 75 mM;(d) the buffer is at a concentration of about 10 mM;(e) the surfactant is at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is at a concentration of about 0.1% (w / v).

61. The method of claim 58 or 59, wherein the liquid arenavirus formulation is suitable for lyophilization, and(b) the carbohydrate is at a concentration of about 5% (w / v);(c) the salt is at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) the buffer is at a concentration of about 10 mM;(e) the surfactant is at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is at a concentration of about 0.1% (w / v).

62. The method of any one of claims 58-61, wherein:(b) the carbohydrate is a sugar or a polyol; optionally wherein the sugar or polyol is selected from the group consisting of sucrose, trehalose, sorbitol, mannitol, dextran, and combinations thereof; further optionally wherein the carbohydrate is sucrose, trehalose, or a combination thereof; (c) the salt is selected from the group consisting of sodium chloride, sodium phosphate, potassium phosphate, potassium chloride, magnesium chloride, sodium sulfate, potassium sulfate, ammonium sulfate, and combinations thereof; optionally wherein the salt is sodium chloride;(d) the buffer is selected from the group consisting of phosphate, citrate, TRIS, HEPES, bicarbonate, histidine, and combinations thereof; optionally wherein the buffer is TRIS;(e) the surfactant is selected from the group consisting of pol oxamer 188, pol oxamer 407, polysorbate 20, polysorbate 80, and combinations thereof; optionally wherein the surfactant is pol oxamer 188; and / or(f) the stabilizer is a protein selected from the group consisting of albumins, recombinant proteins, plasma proteins, gelatin, and combinations thereof; optionally wherein the albuminis lactalbumin, human serum albumin, recombinant human serum albumin (rHSA), or a combination thereof; further optionally wherein the protein is rHSA.

63. The method of claim 62, wherein:(b) the carbohydrate is sucrose or trehalose at a concentration of about 5% (w / v);(c) the salt is sodium chloride at a concentration of about 125 mM or about 75 mM;(d) the buffer is TRIS at a concentration of about 10 mM;(e) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is rHSA at a concentration of about 0.1%.

64. The method of claim 62, wherein the liquid arenavirus formulation is suitable for lyophilization, and:(b) the carbohydrate is trehalose at a concentration of about 5% (w / v);(c) the salt is sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) the buffer is TRIS at a concentration of about 10 mM;(e) the surfactant is pol oxamer 188 at a concentration of about 0.03% (w / v); and / or(f) the stabilizer is rHSA at a concentration of about 0.1%.

65. The method of any one of claims 47-64, wherein the liquid arenavirus formulation has a pH of between about 7.0 and about 9.0, optionally wherein the pH is about 8.0.

66. The method of any one of claims 47-65, wherein the liquid arenavirus formulation has an osmolality of between about 200 mOsm / kg and about 1000 mOsm / kg.

67. The method of claim 66, wherein the liquid arenavirus formulation has an osmolality of between about 500 mOsm / kg and about 550 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

68. The method of claim 66, wherein the liquid arenavirus formulation has an osmolality of between about 350 mOsm / kg and about 450 mOsm / kg, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg.

69. The method of claims 66, wherein the liquid arenavirus formulation is suitable for lyophilization and has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

70. The method of any one of claims 58-69, wherein (b)-(f) are present at concentrations to reduce the loss of viral titer during filtration, reduce the formation of subvisible particles, maintain biophysical stability of the arenavirus during freeze and thaw process, and / or maintain biophysical stability of the arenavirus during storage.

71. The method of claim 70, wherein the temperature of the storage is above about 2°C, between about 20°C and about 25°C, between about 2°C and about 8°C, above about 15°C, about -20°C or less than about -20°C, about -65°C or less than about -65°C, or about -80°C.

72. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises:(a) citrate at a concentration of about 50 mM;(b) sucrose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of about 125 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0 optionally wherein the liquid arenavirus formulation has an osmolality of about 530 mOsm / kg.

73. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation comprises:(a) citrate at a concentration of about 25 mM;(b) trehalose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of about 75 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1%, wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the liquid arenavirus formulation has an osmolality of about 400 mOsm / kg74. A method for (i) reducing the formation of subvisible particles during manufacturing, and / or (ii) reducing the loss of viral titer during filtration, comprising combining an arenavirus with a solution to form a liquid arenavirus formulation, wherein the liquid arenavirus formulation is suitable for lyophilization and comprises:(a) citrate at a concentration of between 10 mM and about 60 mM;(b) trehalose at a concentration of about 5% (w / v);(c) sodium chloride at a concentration of up to about 125 mM or between about 50 mM and about 125 mM;(d) TRIS at a concentration of about 10 mM;(e) poloxamer 188 at a concentration of about 0.03% (w / v); and(f) rHSA at a concentration of about 0.1% (w / v), wherein the liquid arenavirus formulation has a pH of about 8.0, optionally wherein the filtration comprises bioburden reduction filtration or sterile filtration, and / or the manufacturing comprises a purification process, optionally wherein the liquid arenavirus formulation suitable for lyophilization has an osmolality of between about 300 mOsm / kg and about 600 mOsm / kg.

75. The method of any one of claims 47-74, wherein the concentration of the arenavirus is between about 102RCV-FFU / ml and about 1010RCV-FFU / ml, or between about 2* 105RCV-FFU / ml and about 5x l08RCV-FFU / ml.

76. The method of claims 47-75, wherein the arenavirus is lymphocytic choriomeningitis virus (LCMV), Pichinde virus, Tamiami virus, or Tacaribe virus.

77. The method of any one of claims 47-76, wherein the arenavirus is a genetically engineered arenavirus.

78. The method of any one of claims 47-77, wherein the arenavirus expresses an antigen, optionally wherein the antigen is selected from the group consisting of tumor antigens, tumor associated antigens, antigens of pathogens that can cause infectious diseases, cancer testis antigens (CTAs), and antigenic fragments of any of the foregoing.

79. The method of claim 78, wherein the antigen comprises at least one cytomegalovirus (CMV) antigen, optionally wherein the CMV antigen is selected from the group consisting of gB(dCt), pp65, and combinations thereof.

80. The method of claim 78, wherein the antigen comprises at least one human papilloma virus (HPV) antigen, optionally wherein the HPV antigen is a HPV E7ZE6 fusion protein.

81. The method of claim 78, wherein the antigen comprises at least one prostate cancer antigen, optionally wherein the prostate cancer antigen is selected from the group consisting of PSA, PSMA, PAP, and combinations thereof.

82. The method of claim 78, wherein the antigen comprises a hepatitis B virus (HBV) antigen, optionally wherein the HBV antigen is selected from the group consisting of a truncated HBV polymerase polypeptide comprising an inactivated reverse transcriptase domain and an inactivated RNase H, a fusion protein comprising an HBV core polypeptide and an HBV small surface antigen (sAg) polypeptide, and combinations thereof.

83. The method of claim 78, wherein the antigen comprises a human immunodeficiency virus (HIV) antigen, optionally wherein the HIV antigen is selected from the group consisting of gag, pol, nef, and combinations thereof, further optionally wherein the HIV antigen is a fusion polypeptide comprising a plurality of polypeptide segments of one or more HIV-1 proteins encoded by two or more HIV genes selected from gag, pol and nef.

84. The method of claim 78, wherein the antigen comprises at least one antigenic fragment of a mutant KRAS, optionally wherein the antigenic fragment of the mutant KRAS from N- to C-terminus comprises the mutations G13D, G12V, G12C, G12D, and G12R, respectively.

85. The method of claim 78, wherein the antigen comprises two or more cancer testis antigen (CTA) fragments, optionally wherein the two or more CTA fragments are selected from the group consisting of epitopes or antigenic fragments of preferentially expressed antigen in melanoma (PRAME), melanoma-associated antigen 3 (MAGE-A3), melanoma-associated antigen 4 (MAGE-A4), and melanoma-associated antigen 6 (MAGE-A6)86. The method of any one of claims 47-85, wherein the arenavirus is a non-replicating arenavirus, optionally wherein an arenavirus ORF has been removed or functionally inactivated such that the resulting arenavirus cannot produce further infectious progeny virus particles, further optionally wherein the ORF encoding the GP has been removed and replaced with a nucleotide sequence encoding an antigen.

87. The method of any one of claims 47-85, wherein the arenavirus is a replicating tri- segmented arenavirus.

88. The method of claim 87, wherein the tri-segmented arenavirus comprises an arenavirus ORF located in a position other than the wild-type position of said ORF.

89. The method of claim 88, wherein the tri-segmented arenavirus comprises one L segment and two S segments, wherein one of the two S segments is an S segment, wherein the ORF encoding the GP is under control of an arenavirus 3’ UTR.

90. The method of claim 87, wherein the tri-segmented arenavirus is engineered such that an arenaviral ORF is separated over two or more mRNA transcripts.

91. The method of any one of claims 47-90, further comprising lyophilizing the liquid arenavirus formulation.

92. A method of producing a lyophilized arenavirus formulation, comprising lyophilizing the liquid arenavirus formulation of any one of claims 1-45.

93. A method of treating or preventing a disease, comprising administering to a subject in need thereof the liquid arenavirus formation of any one of claims 1-45, optionally wherein the liquid arenavirus formulation is further diluted prior to the administration.

94. A method of treating or preventing a disease, comprising administering to a subject in need thereof a liquid arenavirus formulation reconstituted from the lyophilized formulation of claim 46, wherein the lyophilized formulation is reconstituted prior to the administration.

95. The method of claim 93 or 94, wherein the liquid arenavirus formulation is administered parenterally, optionally wherein the liquid arenavirus formulation is administered intravenously, intramuscularly, intratumorally, or subcutaneously.

96. The method of claims 93-95, wherein the disease is a neoplastic disease or an infectious disease.

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