Immunogenic compositions and uses thereof

Abstract

The present disclosure relates to compositions comprising a group B streptococcus (GBS) polysaccharide-protein conjugate and a respiratory syncytial virus (RSV) F protein and / or RNA polynucleotide encoding an antigen derived from RSV The present disclosure further relates to methods and uses of the compositions comprising a GBS polysaccharide-protein conjugate and RSV F protein and / or RNA polynucleotide encoding an antigen derived from RSV for the prevention of a disease or infection associated with GBS and / or RSV.

Description

[0001] PC072986A IMMUNOGENIC COMPOSITIONS AND USES THEREOF REFERENCE TO SEQUENCE LISTING 5 This application includes an electronically submitted sequence listing in .xml format. The .xml file contains a sequence listing entitled “PC072986A Sequence Listing.xml” created on July 23, 2015, and having a size of 66 KB. The sequence listing contained in this .xml file is part of the specification and is hereby incorporated by reference herein in its entirety. 10 FIELD The present disclosure relates to a composition comprising a capsular polysaccharide(CP) from group B streptococcus (GBS) conjugated to a carrier protein, and antigens derived from respiratory syncytial virus (RSV) polypeptides and / or polynucleotides encoding the RSV antigens for administration to a human. The present disclosure relates to the co-administration15 of a GBS vaccine comprising a capsular polysaccharide (CP) from GBS conjugated to a carrierprotein, and an RSV vaccine comprising antigens derived from RSV polypeptides and / orpolynucleotides encoding the RSV antigens.BACKGROUND20 Streptococcus agalactiae, also known as group B streptococcus (GBS), is anencapsulated, gram-positive bacterial pathogen that is a leading cause of invasive bacterial infections in young infants and pregnant women and a significant cause of infant morbidity and mortality globally. There are serotypes of GBS (Ia, Ib, II, III, IV, V, VI, VII, VIII, and IX)differentiated by the polysaccharide composition of their capsule. All GBS serotypes may25 cause disease, but there is variability in their global prevalence and virulence. GBS diseasemost frequently occurs in the very young—newborns and infants younger than 3 months ofage—and the elderly, especially older adults with comorbid conditions. However, GBS disease has been reported in individuals of all ages, and pregnant women may be particularly susceptible to it. 30 GBS colonizes the rectovaginal tracts in women, and colonization during pregnancy can expose the neonate to GBS before delivery due to ascending infection. GBS can also be passed to the infant during labor with passage through the birth canal. These events can result in serious, invasive disease in neonates and postneonatal infants, including sepsis, meningitis, and pneumonia resulting in high rates of mortality and for some survivors long-term morbidity.35 Among neonates and infants, GBS disease is categorized into 2 timeframes with early-onsetdisease (EOD) occurring between 0-6 days of life and late-onset disease (LOD) between 7-90 days of life. Infants with EOD generally present with respiratory distress, apnea, or other

[0002] signs of sepsis within the first 24 to 48 hours of life. LOD more commonly presents asmeningitis. Beyond the immediate dangers of acute illness, GBS in infancy has also beenassociated with long-term NDI in survivors of invasive disease. In addition to the poor neonatal outcomes described above, colonized pregnant women can also develop significant GBS 5 disease themselves, including urinary tract infections, chorioamnionitis (which may result in stillbirth, preterm delivery), postpartum endometritis, and sepsis. There is an unmet medical need for the prevention of GBS disease in pregnant women and their infants that could be addressed by an effective vaccine. To date, there are no vaccines licensed for the prevention of GBS in any country. Globally, there are various 10 prevention guidelines, which primarily include either universal microbiological screening or risk-based approaches to identify colonized pregnant women for whom intrapartum antibiotic prophylaxis (IAP) should be administered. Many countries around the world do not have the resources to implement IAP. Importantly, even in settings with optimal execution of these prevention policies, a significant burden of EOD remains and there has been no reduction in 15 LOD cases. In addition, IAP leads to unnecessary exposure to antibiotics. Reduction in antibiotic use has been identified as a global public health priority due to the potential for selection of resistance among bacterial pathogens. Maternal immunization with an efficacious GBS vaccine would address the shortcomings of current prophylactic antibiotic administration, provide significant benefit20 beyond the currently existing approaches, and importantly could prevent a significantproportion of the global burden of both EOD and LOD GBS in infants. A prophylactic GBSvaccine has the potential to limit the emergence of antibiotic resistance through avoidance ofantibiotic use and may therefore potentially have additional indirect impact on public healthGBS6 is an investigational hexavalent (serotypes Ia, Ib, II, III, IV and V) GBS conjugate 25 vaccine being developed for the prevention of GBS invasive disease in infants due to the 6 dominant serotypes worldwide by active immunization of pregnant women in their second or third trimester . The vaccine is intended for use in pregnant women to protect their neonates and infants through transfer of maternal antibodies. See Buurman et al. A Novel HexavalentCapsular Polysaccharide Conjugate Vaccine (GBS6) for the Prevention of Neonatal Group B 30 Streptococcal Infections by Maternal Immunization. J Infect Dis.2019 Jun 5;220(1):105-115. See Absalon et al. Safety and immunogenicity of a novel hexavalent group B streptococcusconjugate vaccine in healthy, non-pregnant adults: a phase 1 / 2, randomised, placebo- controlled, observer-blinded, dose-escalation trial. Lancet Infect Dis.2021 Feb;21(2):263-274. See Madhi et al. Potential for Maternally Administered Vaccine for Infant Group B35 Streptococcus. N. Engl. J. Med. 2023, 389, 215–227.The current standard of care, even when optimally implemented, has not been able tocompletely prevent all disease, including some cases of EOD, all cases of LOD, and all cases2

[0003] of preterm birth and stillbirth. By providing the GBS6 vaccination during the second or earlythird trimester of pregnancy, immunization could prevent GBS disease–associated outcomesthat may arise prepartum, such as preterm births and stillbirths, invasive disease through the first 90 days of life, as well as the long term neurodevelopmental sequelae frequently observed5 in survivors of invasive disease. GBS6 may also protect against adverse fetal outcomes by reducing GBS colonizationin pregnant women. It is anticipated pregnant women may also benefit directly from a GBSvaccine by the prevention of peripartum GBS disease. A vaccine to prevent invasive GBSdisease would be an important public health tool and is considered feasible given the10 longstanding recognition of the relationship between maternally transferred anticapsularantibodies and infant immunity to invasive GBS disease. The GBS6 vaccination would providesignificant benefit as an alternative approach to IAP for prevention of GBS disease in younginfants and can be more practically implemented in HIC settings as well as LMIC regions thathave the highest burden of disease.15 Respiratory syncytial virus (RSV) is a respiratory virus that infects the lungs andbreathing passages. RSV is a major cause of respiratory infection in all ages, which can resultin severe illness in both infants and older adults. In infants, RSV is the leading cause ofbronchiolitis and viral pneumonia and can lead to fatal respiratory distress, especially in those with risk factors including premature infants with underlying cardiopulmonary disease, or in20 the absence of effective healthcare systems. However, most cases of RSV disease occur inhealthy, full-term infants without risk factors. A maternal immunization to protect infants earlyin life is an attractive strategy for infant disease prevention. Adults ≥60 years of age have anincreased risk of RSV infection, which can trigger exacerbations of underlying comorbidconditions, such as COPD and CHF. RSV disease management in adults is limited to25 supportive measures, such as hydration and oxygenation. Prevention of RSV disease viaactive immunization has the potential to make a significant impact in this population, thereforemaking vaccine development a high priority. Two RSV protein subunit vaccines were approved in the United States 2023, ABRYSVOTM (Pfizer) and AREXVYTM (GSK). ABRYSVOTM is an RSV vaccine supplied as a30 lyophilized antigen component containing recombinant RSV stabilized prefusion F proteins, RSV preF A and RSV preF B, which is reconstituted at the time of use with a sterile water diluent. ABRYSVOTM was approved for active immunization of pregnant individuals at 32through 36 weeks gestational age for the prevention of lower respiratory tract disease (LRTD) and severe LRTD caused by RSV in infants from birth through 6 months of age and active35 immunization for the prevention of LRTD caused by RSV in individuals 60 years of age and older. AREXVYTM is an RSV vaccine supplied as a lyophilized recombinant RSV glycoproteinF stabilized in pre-fusion conformation (RSVPreF3) as the antigen component, which is 3

[0004] reconstituted at the time of use with AS01Eadjuvant as the suspension component. AREXVYTMwas approved for active immunization for the prevention of LRTD caused by RSV in individuals 60 years of age and older and individuals 50 through 59 years of age who are at increased risk for LRTD caused by RSV. 5 Given the global burden of GBS and RSV disease, there is an unmet medical need that could be addressed by an effective vaccine against both GBS and RSV for pregnant individuals to prevent disease in infants and for older individuals. Accordingly, improvedimmunogenic compositions to protect against both GBS and RSV infection are needed.10 SUMMARY OF THE INVENTIONThe present invention provides for compositions comprising a group B streptococcus(GBS) polysaccharide-protein conjugate comprising a capsular polysaccharide and a carrier protein; and at least a first and second respiratory syncytial virus (RSV) F protein. In a preferred aspect, the compositions described herein are immunogenic compositions.15 In some aspects, the conjugate comprises GBS capsular polysaccharides selectedfrom serotypes la, Ib, II, III, IV, V, Vl, Vll, VIll, and IX, such as serotypes la, Ib, and III; serotypesla, Ib, III and V; serotypes la, Ib, II, III and V; serotypes la, Ib, II, III, V and VII; or serotypes la,Ib, II, III, IV, and V. In some aspects, the conjugates further comprise at least one GBS capsularpolysaccharide from serotypes Vl, Vll, VIll, and IX. In one aspect, the carrier protein is CRM197,20 tetanus toxoid, or SCP. In one aspect, composition comprises about 5 µg / mL to about 50 µg / mL of each GBS capsular polysaccharide. In some aspects, the first and second RSV F proteins comprise a pair of cysteinemutations selected from the group consisting of: (1) 55C and 188C; (2) 103C and 148C; (3) 142C and 371C; and (4) 155C and 290C, wherein amino acid positions are numbered 25 according to SEQ ID NO: 1. In some aspects, the first RSV F protein is a F protein of subtype A. In one aspect, thefirst RSV F protein comprises a combination of mutations selected from the group consisting of: (1) combination of 103C, 148C, 190I, and 486S; (2) combination of 54H, 55C, 188C, and 486S; (3) combination of 54H, 103C, 148C, 190I, 296I, and 486S; (4) combination of 54H, 30 55C, 142C, 188C, 296I, and 371C; (5) combination of 55C, 188C, and 486S; (6) combination of 54H, 55C, 188C, and 190I; (7) combination of 55C, 188C, 190I, and 486S; (8) combination of 54H, 55C, 188C, 190I, and 486S; (9) combination of 155C, 190I, 290C, and 486S; (10) combination of 54H, 55C, 142C, 188C, 296I, 371C, 486S, 487Q, and 489S; (11) combination of 54H, 155C, 190I, 290C, and 296I; and (12) combination of 155C, 190F, 290C, and 207L; 35 wherein the amino acid positions are numbered according to SEQ ID NO: 1. In one aspect, the first RSV F protein comprises a F2 polypeptide comprising the amino acid sequence of 4

[0005] SEQ ID NO: 17 and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 18or 19. In one aspect, the first RSV F protein comprises a trimerization domain.In some aspects, the second RSV F protein is a F protein of subtype B. In one aspect, the second RSV F protein comprises a combination of selected from the group consisting of: 5 (1) combination of 103C, 148C, 190I, and 486S.; (2) combination of 54H, 55C, 188C, 486S; (3) combination of 54H, 103C, 148C, 190I, 296I, and 486S; (4) combination of 54H, 55C, 142C, 188C, 296I, and 371C; (5) combination of 55C, 188C, and 486S; (6) combination of 54H, 55C, 188C, and 190I; (7) combination of 55C, 188C, 190I, and 486S; (8) combination of 54H, 55C, 188C, 190I, and 486S; (9) combination of 155C, 190I, 290C, and 486S; (10) 10 combination of 54H, 55C, 142C, 188C, 296I, 371C, 486S, 487Q, and 489S; (11) combination of 54H, 155C, 190I, 290C, and 296I; and (12) combination of 155C, 190F, 290C, and 207L;wherein the amino acid positions are numbered according to SEQ ID NO: 1. In oneaspect, the second RSV F protein comprises a F2 polypeptide comprising the amino acidsequence of SEQ ID NO: 20 and a F1 polypeptide comprising the amino acid sequence of15 SEQ ID NO: 21 or 22. In one aspect, the second RSV F protein comprises a trimerizationdomain. In some aspects, the first and second RSV F proteins are a trimer in a prefusionconformation. In one aspect, the composition comprises about 40 µg / mL to about 80 µg / mL ofeach RSV F protein. 20 In some aspects, the composition further comprises sodium chloride, a surfactant, a buffer and at least one of sucrose, mannitol and glycine. In one aspect, the pH of thecomposition is between about 6 and about 8. In one aspect, the composition further comprisesan adjuvant. In one aspect, the composition does not comprise an adjuvant. In one aspect,the RSV F proteins are lyophilized and reconstituted with the GBS polysaccharide-protein25 conjugate in a liquid form. In one aspect, the GBS polysaccharide-protein conjugate islyophilized and reconstituted with the RSV F proteins in a liquid form. In another aspect, theRSV F proteins are in a liquid form and combined with the GBS polysaccharide-protein conjugate in a liquid form. In another aspect, the GBS polysaccharide-protein conjugate and RSV F proteins are lyophilized and reconstituted with a diluent or buffer. 30 In some aspect, the RSV F proteins maintained stability for at least 4 hours in the composition when compared to the stability of the RSV F proteins alone in a composition (without GBS polysaccharide-protein conjugate). In some aspects, the GBS polysaccharide-protein conjugate maintained stability for at least 4 hours in the composition when comparedto the stability of the GBS polysaccharide-protein conjugates alone in a composition (without35 RSV F proteins). 5

[0006] The present invention further provides for methods of eliciting an immune responseagainst group B streptococcus (GBS) and / or respiratory syncytial virus (RSV) in a subject, themethod comprises administering to the subject any of the compositions described herein.In some aspects, the methods include eliciting an immune response against group B 5streptococcus (GBS) in a subject, the method comprises administering to the subject any ofthe compositions described herein. In one aspect, the immune response elicited against GBSis comparable to the immune response elicited against GBS after administering a compositioncomprising GBS polysaccharide-protein conjugate alone (without RSV F proteins). In oneaspect, the immune response elicited comprises anti-GBS capsular polysaccharide IgG10 antibodies. In one aspect, the immune response elicited comprises anti-GBS capsularpolysaccharide IgG antibodies specific to at least one of serotypes la, Ib, II, III, IV, V, Vl, Vll, VIll, and IX. In one aspect, the immune response elicited comprises geometric mean IgGconcentrations comparable to geometric mean IgG concentrations elicited after administering a composition comprising GBS polysaccharide-protein conjugate alone (without RSV F15 proteins). In one aspect, the immune response elicited comprises functional antibodies againstGBS as determined by an opsonophagocytic assay (OPA). In one aspect, the immuneresponse elicited comprises functional antibodies against at least one GBS capsular polysaccharide selected from serotypes la, Ib, II, III, IV, V, Vl, Vll, VIll, and IX as determinedby OPA. In one aspect, the immune response elicited comprises geometric mean OPA titers20 comparable to geometric mean OPA titers elicited after administering a composition comprising GBS polysaccharide-protein conjugate alone (without RSV F proteins). In some aspects, the methods include eliciting an immune response against respiratory syncytial virus (RSV) in a subject, the method comprising administering to the subject any composition described herein. In one aspect, the immune response elicited25 against RSV is comparable to the immune response elicited against RSV after administeringa composition comprising RSV F protein alone (without GBS polysaccharide-protein conjugates). In one aspect, the immune response elicited comprises anti-RSV IgG antibodies.In one aspect, the immune response elicited comprises anti-RSV IgG antibodies specific to at least one of RSV A and RSV B F proteins. In one aspect, the immune response elicited30 comprises geometric mean IgG concentrations comparable to geometric mean IgG concentrations elicited after administering a composition comprising RSV F proteins (without GBS polysaccharide-protein conjugate). In one aspect, the immune response elicitedcomprises functional antibodies against RSV as determined by a neutralization assay. In oneaspect, the immune response elicited comprises functional antibodies against RSV A and RSV35 B. In one aspect, the immune response elicited comprises geometric mean neutralization titerscomparable to geometric mean neutralization titers elicited after administering a composition comprising RSV F proteins alone (without GBS polysaccharide-protein conjugate). 6

[0007] The present invention further provides for methods of preventing or reducing a disease, infection or condition associated with GBS and / or RSV in a subject, the method comprisingadministering to the subject any composition described herein.The present invention further provides for method of eliciting an immune response5 against group B streptococcus (GBS) and respiratory syncytial virus (RSV) in a subject, themethod comprising administering to the subject a first composition comprising a GBS capsular polysaccharide and a carrier protein; and a second composition comprising at least a first andsecond RSV F protein. The present invention further provides for methods of preventing or reducing a disease,10 infection or condition associated with GBS and / or RSV in a subject, the method comprising administering to the subject a first composition comprising a GBS capsular polysaccharide and a carrier protein; and a second composition comprising at least a first and second RSV Fprotein. In one aspect, the first and second compositions are administered concurrently or15 concomitantly. In one aspect, the second composition is administered within at most 1 hour,2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 24 hours, 48 hours, 3 days, 7 days, 2 weeks, 30 days, 2 months, 4 months, 6 months or 12 months of the administration of the first composition. In one aspect, the second composition is administered simultaneously to the administration of the first composition. 20 In some aspects, the subject is less than about 1 year of age, about 5 years of age or older, about 10 years of age or older, about 18 years of age or older, about 20 years of age or older, about 25 years of age or older, about 30 years of age or older, about 35 years of age or older, about 40 years of age or older, about 45 years of age or older, about 50 years of age or older, about 55 years of age or older, about 60 years of age or older, about 65 years of age or 25 older, about 70 years of age or older, about 75 years of age or older, about 80 years of age or older, about 85 years of age or older. In one aspect, the subject is pregnant or of child-bearingage. In one aspect, the subject is 60 years of age or older. In one aspect, the composition isadministered by intramuscular injection. The present invention further provides kits comprising: (i) a lyophilized composition30 comprising a first and second respiratory syncytial virus (RSV) F protein described herein, and(ii) a liquid composition comprising a group B streptococcus (GBS) polysaccharide-protein conjugate described herein for reconstituting the lyophilized composition.In some aspects, reconstituting the lyophilized composition with the liquid composition results in any one of the compositions described herein.35 The present invention further provides for composition comprising a GBSpolysaccharide-protein conjugate comprising a capsular polysaccharide and a carrier protein; 7

[0008] and an RNA polynucleotide comprising at least one open reading frame encoding a respiratory syncytial virus (RSV) polypeptide or an immunogenic fragment thereof. In some aspects, the RSV polypeptide comprises any one of the amino acid sequenceset forth in SEQ ID NO: 1 to 6. In one aspect, the RNA polynucleotide comprises any one of5 the sequences set forth in SEQ ID NO: 11 to 16. In one aspect, the RSV polypeptide is derivedfrom RSV subtype A and / or RSV subtype B. In one aspect, each of the RNA polynucleotidescomprise a 5’ cap, 5’ UTR, 3’ UTR, and poly-A tail. In one aspect, any one of the RNApolynucleotide comprise at least one modified nucleotide selected from the group consisting of pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4′-10 thiouridine, 5-methylcytosine, 5-methyluridine, 2-thio-1-methyl-1-deaza-pseudouridine, 2-thio- 1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio- dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy- pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, dihydropseudouridine, 5-methoxyuridine OR 2′-O-methyl uridine. In one aspect, each of the15 RNA polynucleotides is encapsulated in a lipid nanoparticle (LNP). In one aspect, the LNPcomprises a cationic lipid, a polymer-lipid, a neutral lipid, and a steroid or steroid analog. In some aspects, the conjugate comprises a GBS capsular polysaccharide selectedfrom serotypes la, Ib, II, III, IV, V, Vl, Vll, VIll, and IX. In one aspect, the conjugates compriseGBS capsular polysaccharides from serotypes la, Ib, and III; serotypes la, Ib, III and V;20 serotypes la, Ib, II, III and V; serotypes la, Ib, II, III, V and VII; or serotypes la, Ib, II, III, IV, andV. In one aspect, the conjugates further comprise at least one GBS capsular polysaccharidefrom serotypes Vl, Vll, VIll, and IX. In one aspect, the carrier protein is CRM197, tetanus toxoid,or SCP. The present invention further provides for methods of eliciting an immune response25 against GBS and / or RSV in a subject, the method comprising administering to the subject a composition comprising a GBS polysaccharide-protein conjugate comprising a capsularpolysaccharide and a carrier protein; and an RNA polynucleotide comprising at least one open reading frame encoding a respiratory syncytial virus (RSV) polypeptide or an immunogenic fragment thereof.30 The present invention further provides for methods of preventing or reducing a disease,infection or condition associated with GBS and / or RSV in a subject, the method comprising administering to the subject a composition comprising a GBS polysaccharide-proteinconjugate comprising a capsular polysaccharide and a carrier protein; and an RNA polynucleotide comprising at least one open reading frame encoding a respiratory syncytial35 virus (RSV) polypeptide or an immunogenic fragment thereof. The present invention further provides for methods of eliciting an immune responseagainst GBS and / or RSV in a subject, the method comprising administering to the subject a 8

[0009] first composition comprising a GBS polysaccharide-protein conjugate comprising a capsular polysaccharide and a carrier protein; and a second composition comprising an RNApolynucleotide comprising at least one open reading frame encoding at least one RSV polypeptide or an immunogenic fragment thereof. 5The present invention further provides for methods of preventing or reducing a disease,infection or condition associated with GBS and / or RSV in a subject, the method comprising administering to the subject a first composition comprising a GBS polysaccharide-protein conjugate comprising a capsular polysaccharide and a carrier protein; and a secondcomposition comprising an RNA polynucleotide comprising at least one open reading frame10 encoding at least one RSV polypeptide or an immunogenic fragment thereof. In some aspects, the first and second compositions are administered concurrently orconcomitantly. In one aspect, the second composition is administered within at most 1 hour,2 hours, 3 hours, 4 hours, 5 hours, 6 hours, 7 hours, 8 hours, 9 hours, 10 hours, 11 hours, 12 hours, 24 hours, 48 hours, 3 days, 7 days, 2 weeks, 30 days, 2 months, 4 months, 6 months15 or 12 months of the administration of the first composition. In one aspect, the second composition is administered simultaneously to the administration of the first composition. In some aspects, the subject is less than about 1 year of age, about 5 years of age or older, about 10 years of age or older, about 18 years of age or older, about 20 years of age or older, about 25 years of age or older, about 30 years of age or older, about 35 years of age or 20 older, about 40 years of age or older, about 45 years of age or older, about 50 years of age or older, about 55 years of age or older, about 60 years of age or older, about 65 years of age or older, about 70 years of age or older, about 75 years of age or older, about 80 years of age or older, about 85 years of age or older. In one aspect, the subject is pregnant or of child-bearingage. In one aspect, the subject is 60 years of age or older. In one aspect, the composition is25 administered by intramuscular injection. The present invention further provides for kits comprising: (i) a lyophilized compositioncomprising an RNA polynucleotide comprising at least one open reading frame encoding anRSV polypeptide or an immunogenic fragment thereof, and (ii) a liquid composition comprisinga GBS polysaccharide-protein conjugate for reconstituting the lyophilized composition. In30 some aspects, reconstituting the lyophilized composition with the liquid composition results in any composition described herein.BRIEF DESCRIPTION OF THE DRAWINGS FIGS.1A-1F show anti-GBS CPS IgG concentrations specific to 1A) serotype Ia, 1B) serotype35 Ib, 1C) serotype II, 1D) serotype III, 1E) serotype IV, and 1F) serotype V, at week 0 and 8 forRSVPreF, GBS6 and GBS6 + RSV PreF treatment groups. LLOQ = lower limit of quantification.ns = not statistically significant by two-tailed unpaired t-test. Bars represent geometric mean9

[0010] concentration and 95% confidence interval (CI). Differences in anti-CPS IgG concentrationsbetween GBS6 and GBS6+RSV PreF groups were evaluated by two-tailed unpaired t-test andwere not significant.FIGS. 2A-2F show OPA titers for 1A) serotype Ia, 1B) serotype Ib, 1C) serotype II, 1D)5 serotype III, 1E) serotype IV, and 1F) serotype V, at week 0 and 8 for GBS6, RSVPreF andGBS6 + RSV PreF treatment groups. Log10 titers are shown. LOD = limit of detection. Barsrepresent geometric mean concentration. Differences in OPA titers between GBS6 andGBS6+RSV PreF groups were evaluated by two-tailed unpaired t-test and were not significant.FIGS.3A-3B show 3A) anti-RSVPreF A (M37) and 3B) anti-RSVPreF B (B18537) IgG titers at10 week 8 for GBS6, RSVPreF and GBS6 + RSV PreF treatment groups. LOD = limit of detection.Numbers in each bar represent mean neutralization titer. Bars represent geometric meanconcentration and 95% confidence interval (CI). Differences in anti-RSV PreF IgG titersbetween RSV PreF and GBS6+RSV PreF groups were evaluated by two-tailed unpaired t-testand were not significant.15 FIGS. 4A-4B show 4A) anti-RSVPreF A (M37) and 4B) anti-RSVPreF B (B18537)neutralization titers at week 8 for GBS6, RSVPreF and GBS6 + RSV PreF treatment groups.LOD = limit of detection. Numbers in each bar represent mean neutralization titer. Barsrepresent geometric mean concentration and 95% confidence interval (CI). Differences in RSVneutralization titers in RSV PreF and GBS6+RSV PreF groups were evaluated by two-tailed20 unpaired t-test and were not significant.DETAILED DESCRIPTIONThe inventors of the present invention have discovered that a group B Streptococcus(GBS) composition may be formulated in combination with (e.g., being in a mix and / or being25 in association with) an RSV composition. For example, a lyophilized RSV composition may bereconstituted with an aqueous or liquid GBS composition, resulting in a subject receiving two vaccines in a single administration. In particular, a lyophilized RSV F protein composition (such as RSVPreF described herein) may be reconstituted with a GBS polysaccharide-proteinconjugate composition (such as GBS6 described herein) in aqueous or liquid form (see Table30 1 of Example 1). Combining vaccine compositions, such as GBS and RSV compositions, can presentchallenges such as maintaining the immunogenicity of each vaccine, ensuring formulationstability and addressing potential immune interference between components. For example,when multiple antigens are combined in a single vaccine composition, the presence of one 35 antigen can interfere with the immune response of another causing immune interference. This can lead to reduced antibody titers or a weaker overall immune response to certain components of the vaccine. Further, combining multiple antigens with potentially different10

[0011] physical and chemical properties can create issues with formulation stability. This challengeis particularly relevant with RSV vaccine compositions that target the prefusion F protein. Theprefusion conformation of the RSV protein F is relatively unstable making it prone to refoldinginto the postfusion state, which is less immunogenic. Thus, maintaining a stabilized prefusion 5 conformation is important for generating a strong and durable immune response. The Examples described herein assess the compatibility and immunogenicity of thecombination of a GBS polysaccharide-protein conjugate composition and an RSV F proteincomposition (see Examples 2 and 3). The results show no difference in total antigenicityobserved between the GBS composition and the combined GBS and RSV composition, and 10 no difference in prefusion content was observed between the RSV composition and the combined GBS and RSV composition. Thus, the combined GBS and RSV composition wasshown to be physically compatible and stability of the respective antigens was maintained for at least 4 hours (see Example 2).Further, the combined GBS and RSV composition elicited a similar immune response 15 to the GBS and RSV compositions administered individually at the same dose. In particular, the combination of the GBS and RSV composition did not impact the immunogenicity of the GBS conjugates as measured by total anti-CPS IgG concentrations and GBS OPA titers, anddid not impact the immunogenicity of the RSV F proteins as measured by total anti-RSVprefusion F protein A and B IgG concentrations and RSV neutralization titers. Thus, the20 combined GBS and RSV composition was shown to retain functional immunogenicity of theGBS conjugates and RSV F proteins (see Example 3).The data generated supports the development of a single vaccine against GBS(bacterial) and RSV (viral) disease and / or infection in infants through maternal immunizationand against GBS and RSV disease and / or infection in older adults through preventive25 vaccination. Thus, the present invention provides for additional compositions and dosageregimens for the immunization of subjects with GBS and RSV vaccines. The present disclosure provides for a single or combined composition that comprisescomponents of both a GBS composition and RSV composition described herein. In one aspect, the present disclosure provides for the co-administration of a GBS composition and30 RSV composition described herein. The present disclosure further provides for methods ofeliciting or inducing an immune response against GBS, RSV and / or GSB and RSV in a subject with the administration of a composition described herein. The present disclosure further provides for methods of preventing or reducing a disease, infection or condition associated with GBS, RSV and / or GSB and RSV in a subject with the administration of a composition35 described herein. 11

[0012] The present disclosure provides for a composition including a GBS polysaccharide-protein conjugate described herein; and an RSV F protein. In a preferred aspect, thecomposition is an immunogenic composition. The present disclosure provides for a composition that includes the components of a5 GBS composition that includes at least one polysaccharide-protein conjugate. The conjugatescomprise a capsular polysaccharide selected from GBS serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX conjugated to a carrier protein. In an aspect, the carrier protein is selected from CRM197, Diphtheria toxoid (DT), tetanus toxoid (TT), and Streptococcal C5a peptidase (SCP). In a preferred aspect, the GBS composition comprises conjugates that include10 capsular polysaccharides from GBS serotypes Ia, Ib, II, III, IV, and V and a carrier protein. Ina preferred aspect, the composition comprises conjugates that include a CRM197 carrier protein. In one aspect, the GBS composition comprises a total conjugate dose of at least about60 µg / mL, such as at least about 120 µg / mL or at least about 240 µg / mL. In a preferred aspect,15 the composition comprises a total conjugate dose of about 240 µg / mL. In another preferredaspect, the composition comprises a total conjugate dose of about 120 µg per 0.5 mL dose.In one aspect, the GBS composition comprises is at least about 10 µg / mL, such as at least about 20 µg / mL or at least about 40 µg / mL of each GBS capsular polysaccharide. In apreferred aspect, the composition comprises about 40 µg / mL of each GBS capsular20 polysaccharide. In another preferred aspect, the composition comprises about 20 µg of each GBS capsular polysaccharide per 0.5 mL dose. In one aspect, the composition includes the components of a GBS compositioncomprising a polysaccharide-protein conjugate described herein and an excipient, a buffer, and a surfactant. 25 In a preferred aspect, the GBS composition comprises histidine as the buffer. In one aspect, the buffer is at a concentration of between about 10 mM to about 25 mM, and most preferably about 20 mM. In a preferred aspect, the GBS composition comprises histidine at aconcentration of about 20 mM. In one aspect, the GBS composition is buffered to a pH range of from about 6.0 to 30 about 7.5, such as about 6.5 to about 7.5 and about 6.5 or about 7.0, and most preferably 6.0 to 7.0. In a preferred aspect, the GBS composition is buffered to a pH range of about 6.5. In a preferred aspect, the GBS composition comprises polysorbate 80 as the surfactant.In one aspect, the surfactant is at a concentration of between about 0.01% to about 0.03%, and most preferably about 0.02%. In a preferred aspect, the GBS composition comprises35 polysorbate 80 at a concentration of about 0.02%.In a preferred aspect, the GBS composition comprises sodium chloride as theexcipient. In one aspect, the excipient at a concentration of between about 10 mM to about12

[0013] 350 mM, such as about 10 mM to about 250 mM, about 130 mM to about 170 mM, and about225 mM to about 265 mM. In a preferred aspect, the GBS composition comprises sodium chloride at a concentration of about 150 mM. In a preferred aspect, the composition includes components of a GBS composition5 comprising 240 µg / mL of six GBS polysaccharide-protein conjugates (serotypes Ia, Ib, II, III,IV and V conjugated to CRM197) in 20 mM histidine, 150 mM NaCl, 0.02% polysorbate 80, pH6.5 (GBS6, see Table 1). In a preferred aspect, the GBS composition is in an aqueous or liquidform. In a preferred aspect, the composition including components of a GBS composition10 described herein delivers a 20 µg dose of each polysaccharide-protein conjugate, equivalentto a 120 µg dose of total polysaccharide-protein conjugate per 0.5 mL injection.In some aspects, the composition of the present disclosure includes GBS conjugates and compositions as described in WO2016 / 178123, WO2018 / 087635, and WO2022 / 043855 which are hereby incorporated by reference in their entirety. 15 The present disclosure provides for a composition that includes the components of an RSV composition that includes at least one RSV F protein. In one aspect, the RSV compositioncomprises a first RSV F protein comprising a F1 polypeptide and F2 polypeptide from the F protein of RSV subtype A, and a second RSV F protein comprising a F1 polypeptide and F2polypeptide from the F protein of RSV subtype B. In some aspects, the RSV composition20 comprises a first and second RSV F comprising amino acid substitutions 103C, 148C, 190I,and 486S. In a preferred aspect, the RSV composition comprises a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of 25 SEQ ID NO: 18 or 19 and the second RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the RSV composition comprises about 200 µg / mL to about 300 µg / mLtotal RSV F protein. In a preferred aspect, the composition comprises a total RSV F protein 30 dose of 240 µg / mL. In another preferred aspect, the composition comprises total RSV F protein dose of 120 µg per 0.5 mL dose. In one aspect, the RSV composition comprises about 100 µg / mL to about 140 µg / mLof each of a first and a second RSV F protein. In a preferred aspect, the composition comprises about 120 µg / mL of a first RSV F protein and 120 µg / mL of a second RSV F protein. In another 35 aspect, the composition comprises 60 µg of a first RSV F protein and 60 µg of a second RSV F protein per 0.5 mL dose. 13

[0014] In one aspect, the composition includes the components of an RSV compositioncomprising a first and a second RSV F protein described herein and a sugar, a surfactant, abuffer and an excipient. In a preferred aspect, the RSV composition comprises between about 2% and about 5 3% of sucrose, between about 4% and about 5% of mannitol, between about 0.01% and about 0.02% of polysorbate 80, between about 10 mM to about 30 mM of Tris and between about40mM and about 60 mM of sodium chloride. In another preferred aspect the RSV composition comprises about 2.25% of sucrose,about 4.5% of mannitol, about 0.015% of polysorbate 80, about 15 mM of Tris and about 37.510 mM of sodium chloride. In a preferred aspect, the composition includes components of an RSV compositioncomprising 240 µg / mL of ExpiCHO produced RSV prefusion F proteins (RSVPreF 847A andRSVPreF 847B) in 15 mM Tris, 2.25% sucrose, 4.5% mannitol, 37.5 mM NaCl, 0.015%polysorbate 80, pH 7.4 (RSVPreF, see Table 1). In a preferred aspect, RSV composition is15 lyophilized. In a preferred aspect, composition delivers a 60 µg dose of each RSVPreF protein,equivalent to 120 µg dose of total RSV F protein in a 0.5 mL IM injection.In some aspects, the composition of the present disclosure includes RSV F proteins and compositions as described in WO2017 / 109629, WO2024 / 069420 WO2024 / 089633 which20 are hereby incorporated by reference in their entirety. The present disclosure provides for a composition that includes the components of an RSV composition that includes at least one RNA polynucleotide including at least one openreading frame encoding at least one respiratory syncytial virus (RSV) antigen or animmunogenic fragment thereof.25 In some aspects, the RSV antigen is an RSV polypeptide. In some aspects, the RSVpolypeptide is RSV F protein. In some aspects, the RSV F protein is a full-length, truncated,fragment or variant thereof. In some aspects, the RSV F protein comprises at least onemutation. The RSV polypeptide to be included in the composition disclosed herein may be anyRSV F protein, preferably any RSV F protein in the prefusion conformation. In some aspects,30 the RSV polypeptide has at least 90%, 95%, 96%, 97%, 98% or 99% identity to the amino acid sequence selected from SEQ ID NO: 1 to 6. In some aspects, the RNA polynucleotide includes at least one open reading frameencoding at least one respiratory syncytial virus (RSV) polypeptide that comprises a sequenceset forth in any one of SEQ ID NO: 11-16. In some aspects, the RSV polypeptide is derived35 from RSV subtype A and / or RSV subtype B. In some aspects, each of the RNA polynucleotideincludes a 5’ cap, 5’ UTR, 3’ UTR, and poly-A tail. In some aspects, any one of said RNA polynucleotide includes at least one modified nucleotide, such as N1-methylpseudouridine. 14

[0015] In some aspects, the RNA polynucleotide is encapsulated in a lipid nanoparticle(LNP). In some embodiments, the LNP includes a cationic lipid, a polymer-lipid, a neutral lipid,and a steroid or steroid analog. In some aspects, the present invention includes RSV RNA polynucleotides and5 compositions as described in WO2024 / 089633, which is hereby incorporated by reference inits entirety. In a preferred aspect, the present disclosure provides for a composition including a GBS polysaccharide-protein conjugate comprising a GBS capsular polysaccharide and a carrier protein; and a first and a second RSV F protein.10 In another preferred aspect, the present disclosure provides for a compositionincluding GBS polysaccharide-protein conjugates comprising capsular polysaccharides fromserotypes la, Ib, II, III, IV, and V and CRM197; and a first and a second RSV F protein.DEFINITIONS 15 Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure have the meanings that are commonly understood by those of ordinary skill in the art. As used herein, the term “about” when used to modify a numerically defined parameter means that the parameter may vary by as much as 10% below or above the stated numerical20 value for that parameter. For example, a dose of about 5 mg means 5 mg ± 10%, i.e., it mayvary between 4.5 mg and 5.5 mg. Recitation of ranges of values herein is merely intended to serve as a shorthand method of referring individually to each separate value falling within the range. Unless otherwise indicated herein, each individual value is incorporated into the specification as if it25 was individually recited herein. The use of the word “a” or “an” when used in conjunction with the term “comprising” may mean “one,” but it is also consistent with the meaning of “one or more,” “at least one,” and “one or more than one.” The phrase “and / or” means “and” or “or.” To illustrate, A, B, and / or C includes: A alone, 30 B alone, C alone, a combination of A and B, a combination of A and C, a combination of B and C, or a combination of A, B, and C. In other words, “and / or” operates as an inclusive or. The phrase “essentially all” is defined as “at least 95%”; if essentially all members of a group have a certain property, then at least 95% of members of the group have that property. In some aspects, essentially all means equal to any one of, at least any one of, or between35 any two of 95, 96, 97, 98, 99, or 100% of members of the group have that property. 15

[0016] The compositions and methods for their use may “comprise,” “consist essentially of,” or “consist of” any of the ingredients or steps disclosed throughout the specification. Throughout this specification, unless the context requires otherwise, the words “comprising” (and any form of comprising, such as “comprise” and “comprises”), “having” (and any form of 5 having, such as “have” and “has”), “including” (and any form of including, such as “includes” and “include”) or “containing” (and any form of containing, such as “contains” and “contain”) are inclusive or open-ended and will be understood to imply the inclusion of a stated step or element or group of steps or elements but not the exclusion of any other step or element or group of steps or elements. It is contemplated that aspects described herein in the context of 10 the term “comprising” may also be implemented in the context of the term “consisting of” or “consisting essentially of.” Compositions and methods “consisting essentially of” any of the ingredients or steps disclosed limits the scope of the claim to the specified materials or steps which do not materially affect the basic and novel characteristic of the claimed disclosure. The words “consisting of” (and any form of consisting of, such as “consist of” and “consists of”) 15 means including, and limited to, whatever follows the phrase “consisting of.” Thus, the phrase “consisting of” indicates that the listed elements are required or mandatory, and that no other elements may be present. Reference throughout this specification to “one aspect,” “another aspect,” “an aspect,”“a particular aspect,” “a related aspect,” “a certain aspect,” “an additional aspect,” or “a further 20 aspect” or combinations thereof means that a particular feature, structure or characteristic described in connection with the aspect is included in at least one aspect of the present disclosure. Thus, the appearances of the foregoing phrases in various places throughout this specification are not necessarily all referring to the same aspect. Furthermore, the particular features, structures, or characteristics may be combined in any suitable manner in one or more25 aspects. The terms “inhibiting,” “decreasing,” or “reducing” or any variation of these terms includes any measurable decrease (e.g., a 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% decrease) or complete inhibition to achieve a desired result. The terms “improve,” “promote,” or “increase” or any 30 variation of these terms includes any measurable increase (e.g., a 5%, 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or 99% increase) to achieve a desired result or production of a protein or molecule. As used herein, the terms “reference,” “standard,” or “control” describe a value relative to which a comparison is performed. For example, an agent, subject, population, sample, or35 value of interest is compared with a reference, standard, or control agent, subject, population,sample, or value of interest. A reference, standard, or control may be tested and / or determined substantially simultaneously and / or with the testing or determination of interest for an agent, 16

[0017] subject, population, sample, or value of interest and / or may be determined or characterized under comparable conditions or circumstances to the agent, subject, population, sample, or value of interest under assessment. The term “vaccine” refers to a pharmaceutical composition comprising an immunogen 5 that is capable of eliciting a prophylactic or therapeutic immune response in a subject. Typically, a vaccine elicits an antigen-specific immune response to an antigen of a pathogen, for example a viral pathogen. The term “immunogenic” refers to the ability of a substance to cause, elicit, stimulate, or induce an immune response against a particular antigen, in an animal, whether in the10 presence or absence of an adjuvant. Immunogenicity can be measured by any method orassay known in the art, such as for example animal vaccination models, serum bactericidal assays (SBA), flow cytometry, and in vitro potency assays. The term "immune response" refers to any detectable response of a cell or cells of the immune system of a host mammal to a stimulus (such as an immunogen), including, but 15 not limited to, innate immune responses (e.g., activation of Toll receptor signaling cascade), cell-mediated immune responses (e.g., responses mediated by T cells, such as antigen- specific T cells, and non-specific cells of the immune system), and humoral immune responses(e.g., responses mediated by B cells, such as generation and secretion of antibodies into the plasma, lymph, and / or tissue fluids). Examples of immune responses include an alteration 20 (e.g., increase) in Toll-like receptor activation, lymphokine (e.g., cytokine (e.g., Th1, Th2 or Th17 type cytokines) or chemokine) expression or secretion, macrophage activation, dendritic cell activation, T cell (e.g., CD4+ or CD8+ T cell) activation, NK cell activation, B cell activation (e.g., antibody generation and / or secretion), binding of an immunogen (e.g., antigen (e.g.,immunogenic polypeptide) to an MHC molecule, induction of a cytotoxic T lymphocyte ("CTL") 25 response, induction of a B cell response (e.g., antibody production), and, expansion (e.g., growth of a population of cells) of cells of the immune system (e.g., T cells and B cells), and increased processing and presentation of antigen by antigen presenting cells. The term “immune response” also encompasses any detectable response to a particular substance (such as an antigen or immunogen) by one or more components of the 30 immune system of a vertebrate animal in vitro. The term “immunogen” refers to a compound, composition, or substance that is immunogenic as defined herein below. The term ‘immunogenic composition” refers to a composition comprising an immunogen. As used herein, the term “pharmaceutical composition” refers to an active agent, 35 formulated together with one or more pharmaceutically acceptable carriers. Pharmaceutical compositions may be immunogenic compositions. In some aspects, active agent is present in unit dose amount appropriate for administration in a therapeutic regimen that shows a 17

[0018] statistically significant probability of achieving a predetermined therapeutic effect when administered to a relevant population. In some aspects, pharmaceutical compositions may be specially formulated for parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or 5 suspension, or sustained-release formulation. As used herein, the term “vaccination” refers to the administration of an immunogenic composition intended to generate an immune response, for example to a disease-associated (e.g., disease-causing) agent (e.g., a bacteria or a virus). In some aspects, vaccination may be administered before, during, and / or after exposure to a disease-associated agent, and in 10 certain aspects, before, during, and / or shortly after exposure to the agent. In some aspects, vaccination includes multiple administrations, appropriately spaced in time, of a vaccine composition. In some aspects, vaccination generates an immune response to an infectious agent. In some aspects, vaccination generates an immune response to a tumor; in some such aspects, vaccination is “personalized” in that it is partly or wholly directed to epitope(s) (e.g., 15 which may be or include one or more neoepitopes) determined to be present in a particular individual’s tumors. As used herein, the term “combination therapy” refers to those situations in which a subject is simultaneously exposed to two or more therapeutic regimens (e.g., two or more therapeutic agents). In some aspects, the two or more regimens may be administered 20 simultaneously; in some aspects, such regimens may be administered sequentially (e.g., all “doses” of a first regimen are administered prior to administration of any doses of a second regimen); in some aspects, such agents are administered in overlapping dosing regimens. In some aspects, “administration” of combination therapy may involve administration of one or more agent(s) or modality(ies) to a subject receiving the other agent(s) or modality(ies) in the 25 combination. For clarity, combination therapy does not require that individual agents be administered together in a single composition (or even necessarily at the same time), although in some aspects, two or more agents, or active moieties thereof, may be administered together in a combination composition, or even in a combination compound (e.g., as part of a single chemical complex or covalent entity). 30 Those skilled in the art will appreciate that the term “dosing regimen” may be used to refer to a set of unit doses (typically more than one) that are administered individually to a subject, typically separated by periods of time. In some aspects, a given therapeutic agent has a recommended dosing regimen, which may involve one or more doses. In some aspects, a dosing regimen comprises a plurality of doses each of which is separated in time from other 35 doses. In some aspects, individual doses are separated from one another by a time period of the same length; in some aspects, a dosing regimen comprises a plurality of doses and at least two different time periods separating individual doses. In some aspects, all doses within 18

[0019] a dosing regimen are of the same unit dose amount. In some aspects, different doses within a dosing regimen are of different amounts. In some aspects, a dosing regimen comprises a first dose in a first dose amount, followed by one or more additional doses in a second dose amount different from the first dose amount. In some aspects, a dosing regimen comprises a 5 first dose in a first dose amount, followed by one or more additional doses in a second dose amount same as the first dose amount. In some aspects, a dosing regimen is correlated with a desired or beneficial outcome when administered across a relevant population (e.g., is a therapeutic dosing regimen). 10 GROUP B STREPTOCOCCUS (GBS) The present disclosure relates to compositions comprising polysaccharide-proteinconjugate comprising a capsular polysaccharide from group B streptococcus (GBS) and a carrier protein. In one aspect, the capsular polysaccharide is selected from the groupconsisting of serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII, and IX. In a preferred aspect, the15 compositions comprise polysaccharide-protein conjugates comprising capsularpolysaccharides from GBS serotypes Ia, Ib, II, III, IV and V, and a carrier protein.In an additional aspect, the polysaccharide-protein conjugate comprises a capsular polysaccharide having about 1.0 mM sialic acid per mM of polysaccharide, such as at least about 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, or 0.95 mM sialic acid per mM of polysaccharide. 20 In an additional aspect, the polysaccharide-protein conjugate comprises a capsular polysaccharide having a sialic acid level of greater than about 60%, greater than about 95%, or about 100%. In another aspect, the capsular polysaccharide may be desialylated up to about 40% (sialylation level greater than about 60%). In another aspect of the invention, the conjugate comprises a capsular polysaccharide 25 having a molecular weight of between about 5 kDa and about 1,000 kDa, between about 25 kDa and about 750 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 200 kDa, or between about 100 kDa and about 400 kDa. In a further aspect, the conjugate of the invention has a molecular weight of between about 300 kDa and about 20,000 kDa, such as between about 1,000 kDa and about 15,00030 kDa or between about 1,000 kDa and about 10,000 kDa. In one aspect, the conjugate comprises a capsular polysaccharide having between about 0% and about 40% O-acetylation, such as less than about 5%, less than about 4%, less than about 3%, less than about 2%, or less than about 1% O-acetylation. In one aspect, the conjugate comprises a capsular polysaccharide having at least35 about 0.1, 0.2, 0.3, 0.35 or about 0.4 mM O-acetate per mM saccharide repeating unit. Inanother aspect, the conjugate comprises a capsular polysaccharide having less than about 0.01, 0.02, 0.03, 0.04, or 0.05 mM O-acetate per mM saccharide repeating unit. 19

[0020] In one aspect, the conjugate comprises CRM197or tetanus toxoid as the carrier protein. In a preferred aspect, the carrier protein is CRM197. Capsular Polysaccharides As used herein, the term "saccharide" refers to a single sugar moiety or 5 monosaccharide unit as well as combinations of two or more single sugar moieties or monosaccharide units covalently linked to form disaccharides, oligosaccharides, and polysaccharides. The term "saccharide" may be used interchangeably with the term "carbohydrate." The polysaccharide may be linear or branched.A “monosaccharide” as used herein refers to a single sugar residue in an 10 oligosaccharide. The term "disaccharide" as used herein refers to a polysaccharide composed of two monosaccharide units or moieties linked together by a glycosidic bond. In one aspect, the polysaccharide is an oligosaccharide (OS). An “oligosaccharide” as used herein refers to a compound containing two or more monosaccharide units or moieties. Within the context of an oligosaccharide, an individual monomer unit or moiety is a 15 monosaccharide which is, or can be, bound through a hydroxyl group to another monosaccharide unit or moiety. Oligosaccharides can be prepared by either chemical synthesis from protected single residue sugars or by chemical degradation of biologically produced polysaccharides. Alternatively, oligosaccharides may be prepared by in vitro enzymatic methods. 20 In a preferred aspect, the polysaccharide (PS) refers to a linear or branched polymer of at least 5 monosaccharide units or moieties. For clarity, larger number of repeating units, wherein n is greater than about 5, such as greater than about 10, will be referred to herein asa polysaccharide. In one aspect, the polysaccharide is a cell surface polysaccharide. A cell surface 25 polysaccharide refers to a polysaccharide having at least a portion located on the outermost bacterial cell membrane or bacterial cell surface, including the peptidoglycan layer, cell wall, and capsule. Typically, a cell surface polysaccharide is associated with inducing an immune response in vivo. A cell surface polysaccharide may be a “cell wall polysaccharide” or a “capsular polysaccharide.” A cell wall polysaccharide typically forms a discontinuous layer on30 the bacterial surface. In one aspect, the polysaccharide is a capsular polysaccharide. A capsular polysaccharide refers to a glycopolymer that includes repeating units of one or more monosaccharides joined by glycosidic linkages. A capsular polysaccharide typically forms a capsule-like layer around a bacterial cell. "Capsular polysaccharide" or "capsule35 polysaccharide" refers to the polysaccharide capsule that is external to the cell wall of most isolates of streptococci. For example, all GBS capsular polysaccharides have a branched repeat structure with a terminal α2–3-linked sialic acid that is required for bacterial virulence. 20

[0021] Capsule-associated sialic acid (quantified by HPLC assay) has been detected in > 94% of invasive neonatal isolates from T.E.S.T. cultured in vitro. In one aspect, the capsular polysaccharides comprise their natural sialic acid level, such as about 100% or greater than about 95%. In another aspect, the capsular5 polysaccharides may be desialylated up to about 40% (sialylation level greater than about 60%), such as up to about 35% (sialylation level greater than about 65%), up to about 30% (sialylation level greater than about 70%), up to about 25% (sialylation level greater than about 75%), up to about 20% (sialylation level greater than about 80%), up to about 15% (sialylationlevel greater than about 85%), up to about 10% (sialylation level greater than about 90%), and10 up to about 5% (sialylation level greater than about 95%). It should be noted that 100% sialic acid level corresponds to about 1.0 mM sialic acid per mM of polysaccharide. Therefore, the capsular polysaccharides may have about 1.0 mMsialic acid per mM of polysaccharide, such as at least about 0.95 mM sialic acid per mM of polysaccharide. In a further aspect, the capsular polysaccharide may have at least about 0.615 mM sialic acid per mM of polysaccharide, such as at least about 0.65 mM sialic acid per mM of polysaccharide, at least about 0.7 mM sialic acid per mM of polysaccharide, at least about 0.75 mM sialic acid per mM of polysaccharide, at least about 0.8 mM sialic acid per mM of polysaccharide, at least about 0.85 mM sialic acid per mM of polysaccharide, at least about 0.9 mM sialic acid per mM of polysaccharide, or at least about 0.95 mM sialic acid per mM of20 polysaccharide. The terminal sialic residues of some capsular polysaccharide (CP) serotypes are partially O-acetylated (OAc) (Lewis, A.L., et al., Proceedings of the National Academy of Sciences USA, 101(30):11123-8 (2004)). Serotypes Ib, III, IV, V, VI, and IX are partially O- acetylated (up to ~40%), whereas serotypes Ia, II, and VII have little or no O-acetylation (less25 than about 5%) (Lewis 2004). In one aspect of the invention, the capsular polysaccharidescomprise their natural O-acetylation level (about 0% to about 40%). In another aspect, thecapsular polysaccharides may be de-O-acetylated (less than about 5%). The degree of O-acetylation of the polysaccharide or oligosaccharide can be determined by any method known in the art, for example, by proton NMR (Lemercinier, X., et al., Carbohydrate Research, 30 296:83-96 (1996); Jones, C., et al., Journal of Pharmaceutical and Biomedical Analysis, 30:1233-1247 (2002); Int’l Patent Appl. Pub. Nos. WO 2005 / 033148 and WO 00 / 56357). Another commonly used method is described by Hestrin, S., J. Biol. Chem., 180:249-261 (1949). Also, 100% O-acetate corresponds to about 1.0 mM O-acetate per mM of saccharide35 repeating unit. Accordingly, partially O-acetylated polysaccharides comprise at least about0.1, 0.2, 0.3, 0.35 or about 0.4 mM O-acetate per mM saccharide repeating unit. A de-O-21

[0022] acetylated polysaccharide comprises less than about 0.01, 0.02, 0.03, 0.04, or 0.05 mM O- acetate per mM saccharide repeating unit. Streptococcal microorganisms capable of causing invasive disease generally also are capable of producing a CP that encapsulates the bacterium and enhances its resistance to 5 clearance by host innate immune system. The CP serves to cloak the bacterial cell in a protective capsule that renders the bacteria resistant to phagocytosis and intracellular killing. Bacteria lacking a capsule are more susceptible to phagocytosis. Capsular polysaccharides are frequently an important virulence factor for many bacterial pathogens, including Haemophilus influenzae, Streptococcus pneumoniae, Neisseria meningitidis, and10 Staphylococcus aureus. The capsule polysaccharide can be used to serotype a particular species of bacteria. Typing is usually accomplished by reaction with a specific antiserum or monoclonal antibody generated to a specific structure or unique epitope characteristic of the capsule polysaccharide. There are ten GBS serotypes: Ia, Ib, and II–IX (Ferrieri, P., et al., Emerg.15 Infect. Dis. [Internet], 19(4) (2013), available at http: / / wwwnc.cdc.gov / eid / article / 19 / 4 / 12-1572_article. In one aspect of the invention, the polysaccharide is isolated from Streptococcusagalactiae. The polysaccharide may be isolated from any encapsulated strain of S. agalactiae, such as 090, A909 (ATCC Accession No. BAA-1138), 515 (ATCC Accession No. BAA-1177),20 B523, CJB524, MB 4052 (ATCC Accession No.31574), H36B (ATCC Accession No.12401), S40, S42, MB 4053 (ATCC Accession No. 31575), M709, 133, 7357, PFEGBST0267, MB 4055 (ATCC Accession No.31576), 18RS21 (ATCC Accession No. BAA-1175), S16, S20, V8 (ATCC Accession No.12973), DK21, DK23, UAB, 5401, PFEGBST0708, MB 4082 (ATCC Accession No.31577), M132, 110, M781 (ATCC Accession No. BAA-22), D136C(3) (ATCC 25 Accession No. 12403), M782, S23, 120, MB 4316 (M-732; ATCC Accession No. 31475), M132, K79, COH1 (ATCC Accession No. BAA-1176), PFEGBST0563, 3139 (ATCC Accession No.49446), CZ-NI-016, PFEGBST0961, 1169-NT1, CJB111(ATCC Accession No. BAA-23), CJB112, 2603 V / R (ATCC Accession No. BAA-611), NCTC 10 / 81, CJ11, PFEGBST0837, 118754, 114852, 114862, 114866, 118775, B 4589, B 4645, SS1214, CZ-30 PW-119, 7271, CZ-PW-045, JM9130013, JM9130672, IT-NI-016, IT-PW-62, and IT-PW-64. The polysaccharides described herein may be isolated by methods known in the art, including, for example, methods described herein. As used herein, “isolated” refers to being obtained from and separated from a particular source. The term “isolated” further refers to notbeing in its respective naturally occurring form, state, and / or environment. For example, 35 “isolated from streptococcus” refers to a matter that was obtained from and separated from a streptococcus cell. The isolated polysaccharide is not naturally occurring. The term "isolated" means that the material is removed from its original environment (e.g., the natural environment 22

[0023] if it is naturally occurring or from its host organism if it is a recombinant entity, or taken from one environment to a different environment). For example, an "isolated" capsule polysaccharide, protein or peptide is substantially free of cellular material or other contaminating proteins from the cell or tissue source from which the protein is derived, or 5 substantially free of chemical precursors or other chemicals when chemically synthesized, or otherwise present in a mixture as part of a chemical reaction. In the present disclosure, the proteins or polysaccharides may be isolated from the bacterial cell or from cellular debris, so that they are provided in a form useful in the manufacture of an immunogenic composition. The term "isolated" or "isolating" may include purifying, or purification, including methods for 10 purifying an isolated polysaccharide known in the art and / or methods described herein. The language "substantially free of cellular material" includes preparations of a polypeptide / protein in which the polypeptide / protein is separated from cellular components of the cells from which it is isolated or recombinantly produced. Thus, a capsule polysaccharide, protein or peptide that is substantially free of cellular material includes preparations of the capsule 15 polysaccharide, protein or peptide having less than about 30%, 20%, 10%, 5%, 2.5%, or 1% (by dry weight) of contaminating protein or polysaccharide or other cellular material. When the polypeptide / protein is recombinantly produced, it is also preferably substantially free of culture medium, i.e., culture medium represents less than about 20%, 10%, or 5% of the volume of the protein preparation. When polypeptide / protein or polysaccharide is produced by chemical 20 synthesis, it is preferably substantially free of chemical precursors or other chemicals, i.e., it is separated from chemical precursors or other chemicals which are involved in the synthesis of the protein or polysaccharide. Accordingly, such preparations of the polypeptide / protein or polysaccharide have less than about 30%, 20%, 10%, 5% (by dry weight) of chemical precursors or compounds other than polypeptide / protein or polysaccharide fragment of25 interest. In one aspect of the invention, the polysaccharide is isolated from a bacterium. Inanother aspect of the invention, the polysaccharide is produced recombinantly. In furtheraspect, the polysaccharide is synthetic or chemically synthesized according to conventional methods. In yet another aspect of the invention, the polysaccharide is prepared by expression30 in a surrogate host after cloning and expressing a biosynthetic pathway to produce the polysaccharide. In one aspect, the polysaccharide is immunogenic. For example, the inventors discovered that each polysaccharide described herein is capable of inducing or eliciting an immune response. The term “immunogenic” refers to an ability to initiate, trigger, cause, enhance, improve, and / or augment a humoral and / or cell-mediated immune response in a35 mammal. In one aspect, the mammal is a human, primate, rabbit, pig, mouse, etc.The molecular weight of the capsular polysaccharide is a consideration for use in immunogenic compositions. High molecular weight capsular polysaccharides are able to23

[0024] induce certain antibody immune responses due to a higher valence of the epitopes present on the antigenic surface. The isolation and purification of high molecular weight capsularpolysaccharides is contemplated for use in the conjugates, compositions and methods of the present disclosure. 5 However, in one aspect, the polysaccharide may be sized to a molecular weight (MW) range that is lower than the molecular weight of the native capsular polysaccharide prior to conjugation to a carrier protein. The size of the purified capsular polysaccharide is reduced in order to generate conjugates with advantageous filterability characteristics and / or yields. In one such aspect, the size of the purified capsular polysaccharide is reduced by high 10 pressure homogenization. High pressure homogenization achieves high shear rates by pumping the process stream through a flow path with sufficiently small dimensions. The shear rate is increased by using a larger applied homogenization pressure, and exposure time can be increased by recirculating the feed stream through the homogenizer. In one aspect, the polysaccharide described herein is capable of inducing opsonic 15 activity. In another aspect, the polysaccharide described herein is capable of inducing opsonic and phagocytic activity (e.g., opsonophagocytic activity). Opsonic activity or opsonization refers to a process by which an opsonin (for example, an antibody or a complement factor) binds to an antigen (e.g., an isolated polysaccharide described herein), which facilitates attachment of the antigen to a phagocyte or phagocytic 20 cell (e.g., a macrophage, dendritic cell, and polymorphonuclear leukocyte (PMNL). Some bacteria, such as, for example, encapsulated bacteria that are not typically phagocytosed due to the presence of the capsule, become more likely to be recognized by phagocytes when coated with an opsonic antibody. In one aspect, the polysaccharide induces an immune response, such as, e.g., an antibody, that is opsonic. In one aspect, the opsonic activity is25 against a Gram positive coccus, preferably against a Streptococcus species, more preferablyagainst at least one strain of S. agalactiae. In yet another aspect, the polysaccharide described herein is capable of inducing a bactericidal immune response. In one aspect, the bactericidal activity is against a Gram positive coccus, preferably against a Streptococcus species, more preferably against at least 30 one strain of S. agalactiae. Methods for measuring opsonization, phagocytosis, and / or bactericidal activity are known in the art, such as, for example, by measuring reduction in bacterial load in vivo (e.g.,by measuring bacteremia levels in mammals challenged with a Streptococcus species) and / orby measuring bacterial cell killing in vitro (e.g., an in vitro opsonophagocytic assay). In one 35 aspect, the polysaccharide is capable of inducing opsonic, phagocytic, and / or bactericidal activity as compared to an appropriate control, such as, for example, as compared to antisera raised against a heat-killed Gram positive coccus. 24

[0025] Serotype Ia One aspect includes a serotype Ia GBS capsular polysaccharide. The structure ofserotype Ia can be depicted as follows: a) 5 conjugation are10 between about 5 kDa and about 1,000 kDa, such as between about 25 kDa and about 750kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, 15 between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa,20 between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa,25 between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between 25 about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between 5 about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. In one preferred aspect, the molecular weight of the capsular 10 polysaccharide prior to conjugation is between about 25 kDa and about 200 kDa. In another preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 100 kDa and about 400 kDa. Any whole number integer within any of the aboveranges is contemplated as an aspect of the disclosure.In a particular aspect, a high pressure homogenization process is used to reduce the 15 size of native GBS capsular polysaccharide serotype Ia while preserving the structural features, such as sialic acid, of the polysaccharide. Serotype Ia capsular polysaccharides are less than about 5% O-acetylated. Some exemplary strains of serotype Ia capsular polysaccharides of the invention include 090, A909 (ATCC Accession No. BAA-1138), 515 (ATCC Accession No. BAA-1177), B523, CJB524, and20 MB 4052 (ATCC Accession No.31574). Serotype Ib One aspect includes a serotype Ib GBS capsular polysaccharide. The structure ofserotype Ib can be depicted as follows: a) 25 n

[0026] The molecular weight of serotype Ib capsular polysaccharides prior to conjugation are between about 5 kDa and about 1,000 kDa, such as between about 25 kDa and about 750kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, 5 between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa,10 between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa,15 between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between 20 about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 25 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa 30 and about 500 kDa. In one preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 25 kDa and about 400 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype Ib capsular polysaccharides are between about 0% and about 40% O- acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., less35 than about 5% O-acetylated). Some exemplary strains of serotype Ib capsularpolysaccharides of the invention include H36B (ATCC Accession No.12401), S40, S42, MB 4053 (ATCC Accession No.31575), M709, 133, 7357, and PFEGBST0267. 27

[0027] Serotype II One aspect includes a serotype II GBS capsular polysaccharide. The structure ofserotype II can be depicted as follows: a) OH HO OH OHCOO-O5 n njugation are 10 between about 5 kDa and about 1,000 kDa, such as between about 25 kDa and about 750kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, 15 between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa,20 between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa,25 between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between 28

[0028] about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 5 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa 10 and about 500 kDa. In one preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 25 kDa and about 400 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype II capsular polysaccharides are less than about 5% O-acetylated. Some exemplary strains of serotype II capsular polysaccharides of the invention include MB 4055 15 (ATCC Accession No. 31576), 18RS21 (ATCC Accession No. BAA-1175), S16, S20, V8 (ATCC Accession No.12973), DK21, DK23, UAB, 5401, and PFEGBST0708. Serotype III One aspect includes a serotype III GBS capsular polysaccharide. The structure ofserotype III can be depicted as follows: 20 a) 25 conjugation are between about 5 kDa and about 1,000 kDa, such as between about 25 kDa and about 750kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, 29

[0029] between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa,5 between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa,10 between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between 15 about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 20 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa 25 and about 500 kDa. In one preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 25 kDa and about 200 kDa. In another preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 100 kDa and about 400 kDa. Any whole number integer within any of the aboveranges is contemplated as an aspect of the disclosure. In a particular aspect, a high pressure30 homogenization process is used to reduce the size of native GBS capsular polysaccharide serotype III while preserving the structural features, such as sialic acid, of the polysaccharide. Serotype III capsular polysaccharides are between about 0% and about 40% O- acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., lessthan about 5% O-acetylated). Some exemplary strains of serotype III capsular35 polysaccharides of the invention include MB 4082 (ATCC Accession No.31577), M132, 110, M781 (ATCC Accession No. BAA-22), D136C(3) (ATCC Accession No.12403), M782, S23, 30

[0030] 120, MB 4316 (M-732; ATCC Accession No.31475), M132, K79, COH1 (ATCC Accession No. BAA-1176), and PFEGBST0563. Serotype IV One aspect includes a serotype IV GBS capsular polysaccharide. The structure of5 serotype IV can be depicted as follows: a) b) 10 Th cular w of serotype IV capsular polysaccharides prior to conjugation are between about 5 kDa and about 1,000 kDa, such as between about 25 kDa and about 750kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, 15 between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa,20 between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa,31 between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, 5 between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between 10 about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa 15 and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. In one preferred aspect, the molecular weight of the capsular polysaccharide prior to conjugation is between about 25 kDa and about 400 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype IV capsular polysaccharides are between about 0% and about 40% O-20 acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., lessthan about 5% O-acetylated). Some exemplary strains of serotype IV capsularpolysaccharides of the invention include 3139 (ATCC Accession No.49446), CZ-NI-016, and PFEGBST0961. Serotype V25 One aspect includes a serotype V GBS capsular polysaccharide. The structure ofserotype V can be depicted as follows: a) 30

[0031] b) ccharides prior to conjugation are e ween a ou a an a ou , a, suc as e ween about 25 kDa and about 7505 kDa, between about 25 kDa and about 500 kDa, between about 25 kDa and about 450 kDa, between about 25 kDa and about 400 kDa, between about 25 kDa and about 350 kDa, between about 25 kDa and about 300 kDa, between about 25 kDa and about 250 kDa, between about 25 kDa and about 200 kDa, between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa,10 between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa,15 between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, 20 between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between 25 about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 30 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. In one preferred aspect, the molecular weight of the capsular 33

[0032] polysaccharide prior to conjugation is between about 25 kDa and about 400 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype V capsular polysaccharides are between about 0% and about 40% O- acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., less5 than about 5% O-acetylated). Some exemplary strains of serotype V capsular polysaccharides of the invention include 1169-NT1, CJB111 (ATCC Accession No. BAA-23), CJB112, 2603 V / R (ATCC Accession No. BAA-611), NCTC 10 / 81, CJ11, and PFEGBST0837. Serotype VI 10 GBS Serotype VI capsular polysaccharides are described by von Hunolstein, C., et al., Infection and Immunity, 6194):1272-1280 (1993), the disclosures of which are hereby incorporated by reference in their entirety. The structure of serotype VI can be depicted as follows: a) 15 conjugation are 20 between about 5 kDa and about 1,000 kDa, such as between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, 25 between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, 34

[0033] between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between 5 about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 10 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa 15 and about 500 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype VI capsular polysaccharides are between about 0% and about 40% O- acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., lessthan about 5% O-acetylated). 20 Some exemplary strains of serotype IIII capsular polysaccharides of the invention include 118754, 114852, 114862, 114866, 118775, B 4589, B 4645, SS1214, and CZ-PW- 119. Serotype VII GBS Serotype VII capsular polysaccharides are described by Kogan, G., et al., 25 Carbohydrate Research, 277(1):1-9 (1995), the disclosures of which are hereby incorporated by reference in their entirety. The repeating unit of serotype VII is as follows: accharides prior to conjugation are between about 5 kDa and about 1,000 kDa, such as between about 50 kDa and about 750 30 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, 35

[0034] between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa,5 between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between 10 about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 15 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa 20 and about 500 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure. Serotype VII capsular polysaccharides are less than about 5%O-acetylated. Some exemplary strains of serotype VII capsular polysaccharides of theinvention include 7271 and CZ-PW-045. Serotype VIII 25 B Serotype VIII capsular polysaccharides are described by Kogan, G., et al., The Journal of Biological Chemistry, 271(15):8786-8790 (1996), the disclosures of which are hereby incorporated by reference in their entirety. The repeating unit of serotype VIII is asfollows: 30 VIII capsular polysaccharides prior to conjugation are between about 5 kDa and about 1,000 kDa, such as between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, 36

[0035] between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa,5 between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, 10 between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between 15 about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 20 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure.Serotype VIII capsular polysaccharides are between about 0% and about 40% O-25 acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., lessthan about 5% O-acetylated). Some exemplary strains of serotype VIII capsularpolysaccharides of the invention include JM9130013 and JM9130672. Serotype IX GBS Serotype IX capsular polysaccharides have previously been described by Berti,30 F., et al., The Journal of Biological Chemistry, 289(34):23437-2348 (2014), and others. However, the configuration of the GlcpNAc in the backbone of GBS serotype IX polysaccharide is alpha (α), which is different from previously published structural details that have proposed this linkage to be in the β configuration. The structure of serotype IX can bemore accurately depicted as follows: 37

[0036] This corresponding structure for GBS serotype IX may also be represented as follows: . 5 o conjugation are between about 5 kDa and about 1,000 kDa, such as between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa,10 between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa, between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa,15 between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between 20 about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between 25 about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between 38

[0037] about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. Any whole number integer within any of the above ranges is contemplated 5as an aspect of the disclosure.Serotype IX capsular polysaccharides are between about 0% and about 40% O- acetylated. In one aspect of the invention, the polysaccharide is de-O-acetylated (i.e., lessthan about 5% O-acetylated). Some exemplary strains of serotype IX capsular polysaccharides of the invention include IT-NI-016, IT-PW-62, and IT-PW-64. 10 In one aspect, serotype Ia, Ib, II, III, IV, V, VI, VII, VIII and / or IX capsular polysaccharides comprise its natural sialic acid level, such as about 100% or greater than about 95%. In another aspect, the capsular polysaccharides may be desialylated up to about40% (sialylation level greater than about 60%), such as up to about 35% (sialylation level greater than about 65%), up to about 30% (sialylation level greater than about 70%), up to 15 about 25% (sialylation level greater than about 75%), up to about 20% (sialylation level greater than about 80%), up to about 15% (sialylation level greater than about 85%), up to about 10% (sialylation level greater than about 90%), or up to about 5% (sialylation level greater than about 95%) prior to conjugation. In another aspect, serotype Ia, Ib, II, III, IV, V, VI, VII, VIII and / or IX capsular 20 polysaccharide have about 1.0 mM sialic acid per mM of polysaccharide, such as at least about 0.95 mM sialic acid per mM of polysaccharide prior to conjugation. In a further aspect,the capsular polysaccharide may have at least about 0.6 mM sialic acid per mM of polysaccharide, such as at least about 0.65 mM sialic acid per mM of polysaccharide, at least about 0.7 mM sialic acid per mM of polysaccharide, at least about 0.75 mM sialic acid per mM 25 of polysaccharide, at least about 0.8 mM sialic acid per mM of polysaccharide, at least about 0.85 mM sialic acid per mM of polysaccharide, at least about 0.9 mM sialic acid per mM of polysaccharide, or at least about 0.95 mM sialic acid per mM of polysaccharide prior to conjugation. Polysaccharide-Protein Conjugates 30 As used herein, "conjugates" comprise a capsule polysaccharide usually of a desired range of molecular weight and a carrier protein, wherein the capsule polysaccharide is conjugated to the carrier protein. Conjugates may or may not contain some amount of free capsule polysaccharide. As used herein, "free capsule polysaccharide" refers to capsule polysaccharide that is non-covalently associated with (i.e., non-covalently bound to, adsorbed 35 to or entrapped in or with) the conjugated capsular polysaccharide-carrier protein. The terms "free capsule polysaccharide," "free polysaccharide" and "free saccharide" may be used interchangeably and are intended to convey the same meaning. Regardless of the nature of 39

[0038] the carrier molecule, it can be conjugated to the capsular polysaccharide either directly or through a linker. As used herein, "to conjugate", "conjugated" and "conjugating" refers to a process whereby a bacterial capsular polysaccharide is covalently attached to the carrier molecule. Conjugation enhances the immunogenicity of the bacterial capsular polysaccharide. 5 The conjugation can be performed according to the methods described below or by processes known in the art. A “conjugate immunogenic composition,” as used herein, refers to an immunogenic composition wherein the immunogenic material includes an antigenic polysaccharide that is covalently linked to a carrier protein to produce a polysaccharide-protein conjugate. In one 10 aspect, a polysaccharide-protein conjugate of the invention may be formulated as a multivalent immunogenic composition. As used herein, the term “molecular weight” of polysaccharide or of carrier protein- polysaccharide conjugate refers to molecular weight calculated by size exclusion chromatography (SEC) combined with multiangle laser light scattering detector (MALLS). 15 As used herein, a “polysaccharide-protein conjugate” refers to a polysaccharide molecule conjugated to a protein carrier molecule through one or more covalent bonds. It maybe desirable to conjugate the polysaccharide to a protein from another species known to be immunogenic in the target host. Accordingly, in one aspect, the carrier molecule is a carrier protein. As defined herein, such a foreign protein is referred to as a “carrier protein.” Carrier20 proteins serve to enhance the antigenicity and immunogenicity of the polysaccharide. As used herein, the term “carrier effect” refers to the process where the antigenicity and immunogenicity of a weakly immunogenic or non-immunogenic molecule is enhanced, by being attached to a more immunogenic molecule as carrier (e.g., a heterologous protein). In this case, the polysaccharide in the combined polysaccharide-protein conjugate becomes 25 more immunogenic than if it were presented alone. Carrier proteins contain T cell epitopes for stimulating T-cell help for producing antibody responses. "Carrier protein" or "protein carrier" as used herein, refers to any protein molecule that may be conjugated to an antigen (such as the capsular polysaccharides) against which an immune response is desired. Conjugation of an antigen such as a polysaccharide to a carrier 30 protein can render the antigen immunogenic. Carrier proteins are preferably proteins that are non-toxic and non-reactogenic and obtainable in sufficient amount and purity. Examples of carrier proteins are toxins, toxoids or any mutant cross-reactive material (CRM197) of the toxin from tetanus, diphtheria, pertussis, Pseudomonas species, E. coli, Staphylococcus species,and Streptococcus species. Carrier proteins should be amenable to standard conjugation35 procedures. In an aspect, the carrier protein is Streptococcal C5a peptidase (SCP). In anotheraspect of the present disclosure, CRM197 is used as the carrier protein.40

[0039] Cross-reacting materials or CRMs are especially useful for some aspects of the present disclosure. One may produce genetically altered proteins, which are antigenically similar to the certain bacterial toxins, yet non-toxic. These are called “cross reacting materials”, or CRMs. CRM197 (Wyeth / Pfizer Inc., Sanford, NC) is noteworthy since it has a single amino 5 acid change from the native diphtheria toxin and is immunologically indistinguishable from it. See Pappenheimer, A.M., et al., Immunochem., 9(9):891-906 (1972); U.S. Pat. No.5,614,382,the disclosures of which are hereby incorporated by reference in their entirety. CRM197is a non-toxic variant (i.e., toxoid) of diphtheria toxin isolated from cultures of Corynebacterium diphtheriae strain C7 (β197) grown in casamino acids and yeast extract-based medium.10 CRM197 is purified through ultra-filtration, ammonium sulfate precipitation, and ion-exchange chromatography. A culture of C. diphtheriae strain C7 (β197), which produces CRM197 protein,has been deposited with the American Type Culture Collection, Rockville, Maryland and has been assigned accession number ATCC 53281. Other diphtheria toxoids are also suitable for use as carrier proteins. CRM3201 is a genetically manipulated variant of pertussis toxin. See 15 Black, W.J., et al., Science, 240(4852):656-659 (1988), the disclosure of which is hereby incorporated by reference in its entirety. Streptococcal C5a peptidase (SCP) is a cell wall anchored virulence protein encoded by members of the beta hemolytic streptococcus genus that proteolytically inactivates the alpha fragment of complement component 5 (C5a) that is responsible for polymorphonuclear 20 cell recruitment to the site of infection (2005. PNAS.102(51):18391.) It is a target of protective antibodies, wherein IgG antibodies directed against SCP can mediate opsonophagocytosis. Additionally, SCP could serve as a carrier protein to enhance the immune response to conjugated GBS CPS polysaccharide haptens. In addition to a diphtheria toxoid (DT), CRM197, SCP, and a pertussis toxoid, further 25 examples of carrier proteins include a tetanus toxoid (TT), a cholera toxoid (e.g., as described in Int’l Patent Appl. Pub. No. WO 2004 / 083251), an E. coli heat labile toxoid (LT), an E. coli heat stable toxoid (ST), pneumolysin from S. pneumonia (wild-type or mutant with reduced toxicity), pneumococcal surface protein A (PspA), pneumococcal adhesin protein A (PsaA), a C5a peptidase from Streptococcus, hemolysin from Staphylococcal aureus , Nontypeable30 Haemophilus influenzae (NTHi) proteins, Haemophilus influenzae protein D, Clostridiumperfringens exotoxins / toxoid, hepatitis B surface antigen, hepatitis B core antigen, rotavirus VP 7 protein, and respiratory syncytial virus F and G protein, ovalbumin, keyhole limpet haemocyanin (KLH), bovine serum albumin (BSA), purified protein derivative of tuberculin (PPD), a Pseudomonas exotoxin or its derivatives, including a recombinantly-produced35 nontoxic mutant Pseudomonas aeruginosa Exotoxin A, and nontoxic Pseudomonasaeruginosa ExoProtein A (rEPA). Bacterial outer membrane proteins such as outer membraneprotein complex c (OMPC), porins, transferrin binding proteins, or C. difficile enterotoxin (toxin41

[0040] A) and cytotoxin (toxin B) can also be used. Other proteins, such as ovalbumin, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) or purified protein derivative of tuberculin (PPD) can also be used as carrier proteins. In a preferred aspect, the carrier protein is a diphtheria toxoid. More preferably, the carrier protein is CRM197. In another aspect of the5 invention, the carrier protein is tetanus toxoid. For the synthesis of a multivalent conjugate immunogenic composition, polysaccharide-protein conjugates may be produced by conjugating a mixture of polysaccharides purified from bacteria of two different species to a carrier protein. Alternatively, a multivalent conjugate immunogenic composition may be produced by 10 combining polysaccharides purified from bacteria of two or more different serotypes of the same bacteria and conjugating them as a mixture to a carrier protein. Alternatively, polysaccharide-protein conjugates produced by reacting a single type of polysaccharide with carrier protein in separate reactions using different polysaccharides, may be mixed. Thus, a multivalent immunogenic composition may include a carrier protein bearing a homogeneous15 or a heterogeneous population of linked polysaccharides. After conjugation of the capsular polysaccharide to the carrier protein, the polysaccharide-protein conjugates are purified (enriched with respect to the amount of polysaccharide-protein conjugate) by a variety of techniques. These techniques include, e.g., concentration / diafiltration operations, precipitation / elution, column chromatography, and 20 depth filtration. As described above, the present disclosure relates to compositions comprisingpolysaccharide-protein conjugates comprising GBS capsular polysaccharides conjugated to carrier proteins. In one aspect the conjugates comprise a GBS serotype Ia capsular polysaccharide conjugated to a carrier protein, a GBS serotype Ib capsular polysaccharide 25 conjugated to a carrier protein, a GBS serotype II capsular polysaccharide conjugated to a carrier protein, a GBS serotype IIII capsular polysaccharide conjugated to a carrier protein, a GBS serotype IV capsular polysaccharide conjugated to a carrier protein, a GBS serotype V capsular polysaccharide conjugated to a carrier protein, a GBS serotype VI capsular polysaccharide conjugated to a carrier protein, a GBS serotype VII capsular polysaccharide 30 conjugated to a carrier protein, a GBS serotype VIII capsular polysaccharide conjugated to a carrier protein, and / or a GBS serotype IX capsular polysaccharide conjugated to a carrier protein. In one aspect, the polysaccharides have a molecular weight of between about 5 kDaand 1,000 kDa; the conjugates have molecular weights of between about 300 kDa and about 20,000 kDa; and the conjugates comprise less than about 40% free polysaccharide relative to 35 total polysaccharide. In one aspect, the conjugates comprise less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% free polysaccharide relative to total polysaccharide. 42

[0041] In one aspect, the capsular polysaccharide has a molecular weight before conjugation of between about 5 kDa and about 1,000 kDa, such as between about 50 kDa and about 750 kDa, between about 50 kDa and about 500 kDa, between about 50 kDa and about 450 kDa, between about 50 kDa and about 400 kDa, between about 50 kDa and about 350 kDa, 5 between about 50 kDa and about 300 kDa, between about 50 kDa and about 250 kDa, between about 50 kDa and about 200 kDa, between about 75 kDa and about 750 kDa, between about 75 kDa and about 500 kDa, between about 75 kDa and about 450 kDa, between about 75 kDa and about 400 kDa, between about 75 kDa and about 350 kDa, between about 75 kDa and about 300 kDa, between about 75 kDa and about 250 kDa,10 between about 75 kDa and about 200 kDa, between about 100 kDa and about 750 kDa, between about 100 kDa and about 700 kDa, between about 100 kDa and about 650 kDa, between about 100 kDa and about 600 kDa, between about 100 kDa and about 550 kDa, between about 100 kDa and about 500 kDa, between about 100 kDa and about 450 kDa, between about 100 kDa and about 400 kDa, between about 100 kDa and about 350 kDa, 15 between about 100 kDa and about 300 kDa, between about 200 kDa and 750 kDa, between about 200 kDa and about 700 kDa, between about 200 kDa and about 650 kDa, between about 200 kDa and about 600 kDa, between about 200 kDa and about 550 kDa, between about 200 kDa and about 500 kDa, between about 200 kDa and about 450 kDa, between about 200 kDa and about 400 kDa, , between about 250 kDa and about 750 kDa, between 20 about 250 kDa and about 700 kDa, between about 250 kDa and about 650 kDa, between about 250 kDa and about 600 kDa, between about 250 kDa and about 550 kDa, between about 250 kDa and about 500 kDa, between about 250 kDa and about 450 kDa, between about 250 kDa and about 400 kDa, between about 300 kDa and 750 kDa, between about 300 kDa and about 700 kDa, between about 300 kDa and about 650 kDa, between about 300 kDa 25 and about 600 kDa, between about 300 kDa and about 550 kDa, or between about 300 kDa and about 500 kDa. Any whole number integer within any of the above ranges is contemplated as an aspect of the disclosure. In an aspect, a GBS serotype Ia, Ib, II, III, IV, V, VI, VII, VIII,and / or IX capsular polysaccharide has a molecular weight of any of the above ranges. In one aspect, the conjugate has a molecular weight of between about 300 kDa and 30 about 20,000 kDa, such as between about 300 kDa and about 15,000 kDa, between about 300 kDa and about 10,000 kDa, between about 300 kDa and about 9,000 kDa, between about 300 kDa and about 8,000 kDa, between about 300 kDa and about 7,000 kDa, between about 300 kDa and about 6,000 kDa, between about 300 kDa and about 5,000 kDa, between about 300 kDa and about 4,000 kDa, between about 300 kDa and about 3,000 kDa, between about 35 300 kDa and about 2,000 kDa, between about 300 kDa and about 1,000 kDa, between about 500 kDa and about 20,000 kDa, between about 500 kDa and about 15,000 kDa, between about 500 kDa and about 10,000 kDa, between about 500 kDa and about 9,000 kDa, between 43

[0042] about 500 kDa and about 8,000 kDa, between about 500 kDa and about 7,000 kDa, between about 500 kDa and about 6,000 kDa, between about 500 kDa and about 5,000 kDa, between about 500 kDa and about 4,000 kDa, between about 500 kDa and about 3,000 kDa, between about 500 kDa and about 2,000 kDa, between about 500 kDa and about 1,000 kDa, between 5 about 1,000 kDa and about 20,000 kDa, between about 1,000 kDa and about 15,000 kDa, between about 1,000 kDa and about 10,000 kDa, between about 1,000 kDa and about 9,000 kDa, between about 1,000 kDa and about 8,000 kDa, between about 1,000 kDa and about 7,000 kDa, between about 1,000 kDa and about 6,000 kDa, between about 1,000 kDa and about 5,000 kDa, between about 1,500 kDa and about 20,000 kDa, between about 1,500 kDa 10 and about 15,000 kDa, between about 1,500 kDa and about 10,000 kDa, between about 1,500 kDa and about 9,000 kDa, between about 1,500 kDa and about 8,000 kDa, between about 1,500 kDa and about 7,000 kDa, between about 1,500 kDa and about 6,000 kDa, between about 1,500 kDa and about 5,000 kDa, between about 2,000 kDa and about 20,000 kDa, between about 2,000 kDa and about 15,000 kDa, between about 2,000 kDa and about 10,000 15 kDa, between about 2,000 kDa and about 9,000 kDa, between about 2,000 kDa and about 8,000 kDa, between about 2,000 kDa and about 7,000 kDa, between about 2,000 kDa and about 6,000 kDa, between about 2,500 kDa and about 20,000 kDa, between about 2,500 kDa and about 15,000 kDa, between about 2,500 kDa and about 10,000 kDa, between about 2,500 kDa and about 9,000 kDa, between about 2,500 kDa and about 8,000 kDa, between about 20 2,500 kDa and about 7,000 kDa, between about 2,500 kDa and about 6,000 kDa, between about 3,000 kDa and about 20,000 kDa, between about 3,000 kDa and about 15,000 kDa, between about 3,000 kDa and about 10,000 kDa, between about 3,000 kDa and about 9,000 kDa, between about 3,000 kDa and about 8,000 kDa, between about 3,000 kDa and about 7,000 kDa, or between about 3,000 kDa and about 6,000 kDa. In an aspect, a GBS serotype25 Ia, Ib, II, III, IV, V, VI, VII, VIII, and / or IX capsular polysaccharide conjugate has a molecular weight of any of the above ranges. In one aspect, the conjugates of the invention have at least about 0.6, 0.65, 0.7, 0.75, 0.8, 0.85, 0.9, 0.95, 0.97 or 0.98 mM sialic acid per mM polysaccharide. In a preferred aspect, the conjugates have at least about 0.9 or 0.95 mM sialic acid per mM polysaccharide. In an30 aspect, a GBS serotype Ia, Ib, II, III, IV, V, VI, VII, VIII, and / or IX capsular polysaccharide conjugate has a sialic acid content of at least any of the above value. In an aspect, the conjugate of the invention comprises less than about 0.01, 0.02, 0.03, 0.04, or 0.05 mM O-acetate per mM saccharide repeating unit. In another aspect, the conjugate comprises at least about 0.1, 0.2, 0.3, 0.35 or about 0.4 mM O-acetate per mM35 saccharide repeating unit. In an aspect, a GBS serotype Ia, Ib, II, III, IV, V, VI, VII, VIII, and / orIX capsular polysaccharide conjugate has an O-acetate content of any of the above value. 44

[0043] In a further aspect, the conjugate comprises less than about 40%, less than about 35%, less than about 30%, less than about 25%, less than about 20%, less than about 15%, less than about 10%, or less than about 5% of free GBS capsular polysaccharide compared to the total amount of GBS capsular polysaccharide. In a preferred aspect the conjugate5 comprises less than about 5% of unreacted free saccharide compared to the total amount of GBS capsular polysaccharide. In yet another aspect, the ratio (weight by weight) of GBS capsular polysaccharide to carrier protein in the conjugate is between about 0.5 and about 3.0. In one aspect, the ratio of GBS capsular polysaccharide to carrier protein in the conjugate is between about 0.5 and 10 about 2.0, between about 0.5 and about 1.5, between about 0.5 and about 1.0, between about 1.0 and about 1.5, or between about 1.0 and about 2.0. In a preferred aspect, the ratio of GBScapsular polysaccharide to carrier protein in the conjugate is between about 0.8 and about 1.0. In another aspect, the degree of conjugation of the conjugate is between 2 and 15, 15 between 2 and 13, between 2 and 10, between 2 and 8, between 2 and 6, between 2 and 5, between 2 and 4, between 3 and 15, between 3 and 13, between 3 and 10, between 3 and 8, between 3 and 6, between 3 and 5, between 3 and 4, between 5 and 15, between 5 and 10, between 8 and 15, between 8 and 12, between 10 and 15, or between 10 and 12. In a preferredaspect, the degree of conjugation of the conjugate is between 2 and 5. 20 Conjugation Conjugation may be direct, where the atoms from the polysaccharide are covalently bonded to atoms from the protein surface. Alternatively, conjugation may be through a linker molecule, which reacts with both the polysaccharide and the protein and connects the two, tethering the carbohydrate to the protein. 25 Where a carrier and one or more antigens such as a polysaccharide are conjugated (i.e., covalently associated), conjugation may be by any chemical method, process or genetic technique known in the art. For example, a carrier polypeptide and one or more antigens selected from a group comprising a carbohydrate, an oligosaccharide, a lipid, a lipooligosaccharide, a polysaccharide, an oligosaccharide-protein conjugate, a 30 polysaccharide-protein conjugate, a peptide-protein conjugate, an oligosaccharide-peptide conjugate, a polysaccharide-peptide conjugate, a protein-protein conjugate, a lipooligosaccharide-protein conjugate, a polysaccharide-protein conjugate, or any combination thereof, may be conjugated by techniques, including, but not limited to: (1) direct coupling via protein functional groups (e.g., thiol-thiol linkage, amine-carboxyl linkage, amine- 35 aldehyde linkage; enzyme direct coupling); (2) homobifunctional coupling of amines (e.g., using bis-aldehydes); (3) homobifunctional coupling of thiols (e.g., using bis-maleimides); (4) homobifunctional coupling via photoactivated reagents (5) heterobifunctional coupling of 45

[0044] amines to thiols (e.g., using maleimides); (6) heterobifunctional coupling via photoactivated reagents (e.g., the β-carbonyidiazo family); (7) introducing amine-reactive groups into a poly- or oligosaccharide via cyanogen bromide activation or carboxymethylation; (8) introducing thiol-reactive groups into a poly- or oligosaccharide via a heterobifunctional compound such5 as maleimido-hydrazide; (9) protein-lipid conjugation via introducing a hydrophobic group into the protein and (10) protein-lipid conjugation via incorporating a reactive group into the lipid. Also, contemplated are heterobifunctional “non-covalent coupling” techniques such the Biotin- Avidin interaction. Other methods well known in the art for effecting conjugation of oligosaccharides and polysaccharides to immunogenic carrier proteins are also within the 10 scope of some aspects of the invention. In an aspect, the polysaccharide-protein conjugates are obtained by activating polysaccharide with 1-cyano-4-dimethylamino pyridinium tetrafluoroborate (CDAP) to form a cyanate ester. The activated polysaccharide may be coupled directly or via a spacer (linker) group to an amino group on the carrier protein. For example, the spacer could be cystamine 15 or cysteamine to give a thiolated polysaccharide which could be coupled to the carrier via a thioether linkage obtained after reaction with a maleimide-activated carrier protein (for example using GMBS) or a haloacetylated carrier protein (for example using iodoacetimide, SIB, SlAB, sulfo-SIAB, SIA, or SBAP). In one aspect, the cyanate ester (optionally made by CDAP chemistry) is coupled with 20 hexane diamine or adipic acid dihydrazide (ADH) and the amino-derivatised saccharide is conjugated to the carrier protein using carbodiimide (e.g., EDAC or EDC) chemistry via a carboxyl group on the protein carrier. Such conjugates are described for example in Int’l Patent Appl. Pub. Nos. WO 93 / 15760, WO 95 / 08348, and WO 96 / 29094. Other suitable techniques use carbodiimides, hydrazides, active esters, norborane, p- 25 nitrobenzoic acid, N-hydroxysuccinimide, S--NHS, EDC, and TSTU. Many are described in Int’l Patent Appl. Pub. No. WO 98 / 42721. Conjugation may involve a carbonyl linker which may be formed by reaction of a free hydroxyl group of the saccharide with 1,1 carbonyldiimidazole (CDI) or 1,1 carboyl di 1,2,4 triazole (CDT)_(See Bethell, et al., J. Biol.Chem., 254:2572-2574 (1979); Hearn, et al., J. Chromatogr., 218:509-518 (1981)) followed by 30 reaction with a protein to form a carbamate linkage. This may involve reduction of the anomeric terminus to a primary hydroxyl group, optional protection / deprotection of the primary hydroxyl group, reaction of the primary hydroxyl group with CDI / CDT to form a CDI / CDT carbamate intermediate, and coupling the CDI / CDT carbamate intermediate with an amino group on a protein. 35 In preferred aspects, the GBS capsular polysaccharide-protein conjugates of the invention are prepared using reductive amination. Reductive amination involves two steps: (1) oxidation of the polysaccharide to generate aldehyde functionalities from vicinal diols in 46

[0045] individual hexasaccharide unit and (2) reduction of the activated polysaccharide and a carrier protein to form a conjugate. In an aspect, GBS capsular polysaccharide is activated (oxidized) by a process comprising the steps of: 5 (a) reacting isolated GBS capsular polysaccharide with an oxidizing agent; and (b) quenching the oxidation reaction by addition of a quenching agent resulting in an activated GBS capsular polysaccharide. RESPIRATORY SYNCYTIAL VIRUS (RSV)10 RSV is a member of the Paramyxoviridae family. Its genome consists of a single-stranded, negative-sense RNA molecule that encodes 11 proteins, including nine structural proteins (three glycoproteins and six internal proteins) and two non-structural proteins. The structural proteins include three transmembrane surface glycoproteins: the attachment protein G, fusion protein F, and the small hydrophobic small hydrophobic (SH) protein. There are two 15 subtypes of RSV, A and B. They differ primarily in the G glycoprotein, while the sequence of the F glycoprotein is more conserved between the two subtypes. The mature F glycoprotein has three general domains: ectodomain (ED), transmembrane domain (TM), and a cytoplasmic tail (CT). CT contains a single palmitoylated cysteine residue. The F glycoproteinof human RSV is initially translated from the mRNA as a single 574-amino acid polypeptide 20 precursor (referred to “F0” or “F0 precursor”), which contains a signal peptide sequence (amino acids 1-25) at the N-terminus. Upon translation the signal peptide is removed by a signal peptidase in the endoplasmic reticulum. The remaining portion of the F0 precursor (i.e., residues 26-574) may be further cleaved at two polybasic sites (a.a.109 / 110 and 136 / 137) by cellular proteases (in particular furin), removing a 27-amino acid intervening sequence 25 designated pep27 (amino acids 110-136) and generating two linked fragments designated F1 (C-terminal portion; amino acids 137-574) and F2 (N-terminal portion; amino acids 26-109). F1 contains a hydrophobic fusion peptide at its N-terminus and two heptad-repeat regions (HRA and HRB). HRA is near the fusion peptide, and HRB is near the TM domain. The F1 and F2 fragments are linked together through two disulfide bonds. Either the uncleaved F0 protein30 without the signal peptide sequence or a F1-F2 heterodimer can form an RSV F protomer.Three such protomers assemble to form the final RSV F protein complex, which is a homotrimer of the three protomers. The F proteins of subtypes A and B are about 90 percent identical in amino acid sequence. An example sequence of the F0 precursor polypeptide forthe A subtype is provided in SEQ ID NO: 1 (A2 strain; GenBank GI: 138251; Swiss Prot 35 P03420), and for the B subtype is provided in SEQ ID NO: 2 (18537 strain; GenBank GI: 138250; Swiss Prot P13843). SEQ ID NO: 1 and SEQ ID NO: 2 are both 574 amino acid 47

[0046] sequences. The signal peptide sequence for SEQ ID NO: 1 and SEQ ID NO: 2 has also been reported as amino acids 1-25 (GenBank and UniProt). In both sequences the TM domain isfrom approximately amino acids 530 to 550 but has alternatively been reported as 525-548. The cytoplasmic tail begins at either amino acid 548 or 550 and ends at amino acid 574, with 5 the palmitoylated cysteine residue located at amino acid 550. RSV F protein is a primary antigen explored for RSV vaccines. The RSV F proteintrimer mediates fusion between the virion membrane and the host cellular membrane and also promotes the formation of syncytia. In the virion prior to fusion with the membrane of the host cell, the largest population of F molecules forms a lollipop-shaped structure, with the TM10 domain anchored in the viral envelope. This conformation is referred to as the pre-fusion conformation. Pre-fusion RSV F is recognized by monoclonal antibodies (mAbs) D25, AM22, and MPE8, without discrimination between oligomeric states. Pre-fusion F trimers are specifically recognized by mAb AM14. During RSV entry into cells, the F protein rearrangesfrom the pre-fusion state (which may be referred to herein as “pre-F”), through an intermediate15 extended structure, to a post-fusion state (“post-F”). During this rearrangement, the C-terminalcoiled-coil of the pre-fusion molecule dissociates into its three constituent strands, which then wrap around the globular head and join three additional helices to form the post-fusion six helix bundle. If a pre-fusion RSV F trimer is subjected to increasingly harsh chemical orphysical conditions, such as elevated temperature, it undergoes structural changes. Initially,20 there is loss of trimeric structure (at least locally within the molecule), and then rearrangement to the post-fusion form, and then denaturation of the domains. To prevent viral entry, F-specific neutralizing antibodies presumably must bind the pre-fusion conformation of F on the virion, or potentially the extended intermediate, before the viral envelope fuses with a cellular membrane. Thus, the pre-fusion form of the F protein is considered the preferred conformation 25 as the desired vaccine antigen. Mutants of the RSV F protein have been provided to increase prefusion stability (see for example PCT application No WO2017 / 109629). RSV vaccine clinical studies have shownthat some F protein subunit-based vaccines are safe and immunogenic. The term “F0 polypeptide” (F0) refers to the precursor polypeptide of the RSV F 30 protein, which is composed of a signal polypeptide sequence, a F1 polypeptide sequence, a pep27 polypeptide sequence, and a F2 polypeptide sequence. With rare exceptions the F0polypeptides of the known RSV strains consist of 574 amino acids. The term “F1 polypeptide” (F1) refers to a polypeptide chain of a mature RSV F protein.Native F1 includes approximately residues 137-574 of the RSV F0 precursor and is composed35 of (from N- to C-terminus) an extracellular region (approximately residues 137-524), atransmembrane domain (approximately residues 525-550), and a cytoplasmic domain(approximately residues 551-574). As used herein, the term encompasses both native F1 48

[0047] polypeptides and F1 polypeptides including modifications (e.g., amino acid substitutions, insertions, or deletions) from the native sequence, for example, modifications designed to stabilize an RSV F protein mutant or to enhance the immunogenicity of an RSV F protein mutant. 5 The term “F2 polypeptide” (F2) refers to the polypeptide chain of a mature RSV F protein. Native F2 includes approximately residues 26-109 of the RSV F0 precursor. As used herein, the term encompasses both native F2 polypeptides and F2 polypeptides including modifications (e.g., amino acid substitutions, insertions, or deletions) from the native sequence, for example, modifications designed to stabilize an RSV F protein mutant in a 10 prefusion conformation or to enhance the immunogenicity of an RSV F protein mutant. In native RSV F protein, the F2 polypeptide is linked to the F1 polypeptide by two disulfide bonds to form a F2-F1 heterodimer. The term “foldon” or “foldon domain” refers to an amino acid sequence that is capable of forming trimers. One example of such foldon domains is the peptide sequence derived from 15 bacteriophage T4 fibritin, which has the sequence of GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 23). The term “mutant” of a wild-type RSV F protein, “mutant” of an RSV F protein, “RSV Fprotein mutant,” or “modified RSV F protein” refers to a polypeptide that displays introducedmutations relative to a wild-type F protein and is immunogenic against the wild-type F protein. 20 The term “mutation” refers to deletion, addition, or substitution of amino acid residues in the amino acid sequence of a protein or polypeptide as compared to the amino acid sequence of a reference protein or polypeptide. Throughout the specification and claims, the substitution of an amino acid at one particular location in the protein sequence is referred to using a notation "(amino acid residue in wild type protein)(amino acid position)(amino acid 25 residue in engineered protein)". For example, a notation Y75A refers to a substitution of a tyrosine (Y) residue at the 75th position of the amino acid sequence of the reference protein by an alanine (A) residue (in a mutant of the reference protein). In cases where there isvariation in the amino acid residue at the same position among different wild-type sequences, the amino acid code preceding the position number may be omitted in the notation, such as 30 “75A.” The term “native” or “wild-type” protein, sequence, or polypeptide refers to a naturally existing protein, sequence, or polypeptide that has not been artificially modified by selective mutations. The term “pep27 polypeptide” or “pep27” refers to a 27-amino acid polypeptide that is 35 excised from the F0 precursor during maturation of the RSV F protein. The sequence of pep27 is flanked by two furin cleavage sites that are cleaved by a cellular protease during F protein maturation to generate the F1 and F2 polypeptides. 49

[0048] The term “AM14” refers to an antibody described in WO 2008 / 147196 A2, which ishereby incorporated by reference in its entirety. The term “AM22” refers to an antibody described in WO 2011 / 043643 A1, which is hereby incorporated by reference in its entirety. The term “D25” refers to an antibody described in WO 2008 / 147196 A2, which is hereby5 incorporated herein by reference in its entirety. The term “prefusion conformation” refers to a structural conformation adopted by an RSV F protein or mutant thereof that can be specifically bound by (i) antibody D25 or AM22 when the RSV F protein or mutant is in the form of a monomer or trimer, or (ii) by antibody AM14 when the RSV F protein mutant is in the form of a trimer. The prefusion trimer 10 conformation is a subset of prefusion conformations. As used herein, an RSV F protein or polypeptide or mutant thereof in prefusion conformation may be denoted as “RSV preF”. The term “postfusion conformation” refers to a structural conformation adopted by theRSV F protein that is not specifically bound by D25, AM22, or AM14. Native F protein adoptsthe postfusion conformation subsequent to the fusion of the virus envelope with the host 15 cellular membrane. RSV F protein may also assume the postfusion conformation outside the context of a fusion event, for example, under stress conditions such as heat and low osmolality, when extracted from a membrane, when expressed as an ectodomain, or upon storage. The term “mutation” refers to deletion, addition, or substitution of amino acid residues in the amino acid sequence of a protein or polypeptide as compared to the amino acid sequence of 20 a reference protein or polypeptide. Throughout the specification and claims, the substitution of an amino acid at one particular location in the protein sequence is referred to using a notation "(amino acid residue in wild type protein)(amino acid position)(amino acid residue in engineered protein)". For example, a notation Y75A refers to a substitution of a tyrosine (Y) residue at the 75th position of the amino acid sequence of the reference protein by an alanine25 (A) residue (in a mutant of the reference protein). In cases where there is variation in the aminoacid residue at the same position among different wild-type sequences, the amino acid code preceding the position number may be omitted in the notation, such as “75A.” The term “native” or “wild-type” protein, sequence, or polypeptide refers to a naturally existing protein, sequence, or polypeptide that has not been artificially modified by selective30 mutations. In some aspects, the RSV F protein is an RSV F protein of subtype A. In some aspects, the RSV F protein is an RSV F protein of subtype B. As used herein the terms “subtype” and“subgroup” are used interchangeably. As used herein the term “strain” refers to a specific isolate within each subtype or subgroup. 35 Antibodies to RSV F protein are prevalent after natural infection and following vaccination and have been shown to neutralize viral activity in vitro. As used herein, the term “respiratory syncytial virus” or “RSV” is not limited to any particular strain or variant.50

[0049] RSV F Protein The present disclosure provides for compositions that include a respiratory syncytialvirus (RSV) F protein. In some aspects, the RSV F protein is an RSV F protein of subtype A.In some aspects, the RSV F protein is an RSV F protein of subtype B. In some aspects, the 5 RSV F protein is a mutant of wild type RSV F protein. In some aspects, the RSV F protein is a mutant of wild type RSV F protein of subtype A. In some aspects, the RSV F protein is a mutant of wild type RSV F protein of subtype B. In some aspects, the composition includes atleast a first and second RSV F protein. In some aspects, the present disclosure provides for compositions that include an RSV10 F protein that is a mutant of wild type RSV F protein, wherein the mutants display introduced mutations in the amino acid sequence relative to the amino acid sequence of thecorresponding wild-type RSV F protein and are immunogenic against the wild-type RSV F protein in the prefusion conformation or against a virus comprising the wild-type F protein. In certain aspects, the RSV F mutants possess certain beneficial characteristics, such as 15 increased immunogenic properties or improved stability in the prefusion conformation of the mutants or prefusion trimeric conformation of the mutant, as compared to the corresponding wild-type F protein. In some aspects, the composition includes at least a first and second RSVF protein comprising mutations. The amino acid mutations in the RSV F protein mutantsinclude amino acid substitutions, deletions, or additions relative to a wild-type RSV F protein. 20 In some aspects, the RSV F protein is an RSV protein mutant as described in WO2017 / 109629, which is hereby incorporated by reference in its entirety.In view of the substantial conservation of RSV F sequences, a person of ordinary skill in the art can easily compare amino acid positions between different native RSV F sequences to identify corresponding RSV F amino acid positions between different RSV strains and 25 subtypes. For example, across nearly all identified native RSV F0 precursor proteins, the furin cleavage sites fall in the same amino acid positions. Thus, the conservation of native RSV F protein sequences across strains and subtypes allows use of a reference RSV F sequence for comparison of amino acids at particular positions in the RSV F protein. For the purposes of this disclosure (unless context indicates otherwise), the RSV F 30 protein amino acid positions are given with reference to the amino acid sequence of the full length native F precursor polypeptide of the RSV A2 strain; corresponding to GenInfo Identifier GI 138251 and Swiss Prot identifier P03420 (SEQ ID NO: 1). Structure of the RSV F Protein Mutants The present disclosure provides for compositions that include mutants that further35 comprise a trimerization domain. In some aspects, either the F1 polypeptide or the F2 polypeptide includes at least one introduced modification (e.g., amino acid substitution) as described herein. In some other aspects each of the F1 polypeptide and F2 polypeptide51

[0050] includes at least one introduced modification (e.g., amino acid substitution) as described herein. F1 Polypeptide and F2 Polypeptide of the RSV F Mutants In some aspects, the compositions comprise an RSV F mutant in the mature form of5 the RSV F protein, which comprises two separate polypeptide chains, namely the F1polypeptide and F2 polypeptide. In some other aspects, the F2 polypeptide is linked to the F1polypeptide by one or two disulfide bonds to form a F2 / F1 polypeptide heterodimer. In stillother aspects, the RSV F mutants are in the form a single chain protein, wherein the F2 polypeptide is linked to the F1 polypeptide by a peptide bond or peptide linker. Any suitable10 peptide linkers for joining two polypeptide chains together may be used. Examples of suchlinkers include G, GG, GGG, GS, and SAIG linker sequences. The linker may also be the full length pep27 sequence or a fragment thereof, which full length pep27 sequence correspondsto amino acids at positions 110-136 of SEQ ID NO: 1. The F1 polypeptide chain of the mutant may be of the same length as the full length 15 F1 polypeptide of the corresponding wild-type RSV F protein; however, it may also have deletions, such as deletions of 1 up to 60 amino acid residues from the C-terminus of the full- length F1 polypeptide. A full-length F1 polypeptide of the RSV F mutants corresponds to aminoacid positions 137-574 of the native RSV F0 precursor, and includes (from N- to C-terminus)an extracellular region (residues 137-524), a transmembrane domain (TM) (residues 525-20 550), and a cytoplasmic domain (CT) (residues 551-574). It should be noted that amino acidresidues 514 onwards in a native F1 polypeptide sequence are optional sequences in a F1 polypeptide of the RSV F mutants, and therefore may be absent from the F1 polypeptide of the mutant. In some aspects, the F1 polypeptide of the RSV F mutants lacks the entire cytoplasmic 25 domain. In other aspects, the F1 polypeptide lacks the cytoplasmic domain and a portion of or all entire transmembrane domain. In some specific aspects, the mutant comprises a F1 polypeptide wherein the amino acid residues from position 510, 511, 512, 513, 514, 515, 520, 525, or 530 through 574 are absent. Typically, for mutants that are linked to trimerizationdomain, such as a foldon, amino acids 514 through 574 can be absent. Thus, in some specific 30 aspect, amino acid residues 514 through 574 are absent from the F1 polypeptide of the mutant. In still other specific aspects, the F1 polypeptide of the RSV F mutants comprises orconsists of amino acid residues 137-513 of a native F0 polypeptide sequence, such as any alternative F0 precursor sequence such as those disclosed in SEQ ID NOS: 1, 2, 4, 6, and81-270 of WO2017109629, which is hereby incorporated by reference in its entirety.35 The F1 polypeptide of the RSV F mutant may include a C-terminal linkage to a trimerization domain, such as a foldon. Many of the sequences of the RSV F mutants disclosedherein include a sequence of protease cleavage site, such as thrombin cleavage site 52

[0051] (LVPRGS), protein tags, such as 6x His-tag (HHHHHH) and Streptag II (WSHPGFEK), or linker sequences (such as GG and GS) that are not essential for the function of the RSV F protein, such as for induction of an immune response. A person skilled in the art will recognizesuch sequences, and when appropriate, understand that these sequences are not included in 5 a disclosed RSV F mutant. The F1 polypeptide and F2 polypeptide of the RSV F protein mutants to which one or more mutations are introduced can be from any wild-type RSV F proteins known in the art or discovered in the future, including, without limitations, the F protein amino acid sequence ofRSV subtype A, and subtype B strains, including A2 Ontario and Buenos Aires, or any other 10 subtype. In some aspects, compositions of the present disclosure include RSV F mutantscomprising a F1 and / or a F2 polypeptide from an RSV A virus, for example, a F1 and / or F2polypeptide from an RSV F0 precursor protein set forth in any one of SEQ ID NOs: 1, 4 and81-210 of WO2017109629, which sequences are hereby incorporated by reference in their15 entireties, to which one or more mutations are introduced. In some other aspects, compositions of the present disclosure include RSV F mutants comprising a F1 and / or a F2 polypeptide from an RSV B virus, for example, a F1 and / or F2polypeptide from an RSV F0 precursor protein set forth in any one of SEQ ID NOs: 2, 6 and 211- 263 of WO2017 / 109629, which sequences are hereby incorporated by reference in their20 entireties, to which one or more mutations are introduced. In still other aspects, thecompositions include RSV F mutants comprising a F1 and / or a F2 polypeptide from an RSVbovine virus, for example, a F1 and / or F2 polypeptide from an RSV F0 precursor protein set forth in any one of SEQ ID NOs: 264-270 of WO2017109629, which sequences are hereby incorporated by reference in their entireties, to which one or more mutations are introduced.25 In some aspects, compositions of the present disclosure include RSV F proteinmutants disclosed in WO2009 / 079796, WO2010 / 149745, WO2011 / 008974,WO2014 / 160463, WO2014 / 174018, WO2014 / 202570, WO2015 / 013551, WO2015 / 177312, WO2017 / 005848, WO2017 / 174564, WO2017 / 005844 and WO2018 / 109220. The RSV F proteins disclosed in these references are hereby incorporated by reference in their entirety. 30 Trimerization Domains In some aspects, compositions of the present disclosure include RSV F mutantscomprising a trimerization domain. In some aspects, the trimerization domain promotes the formation of trimer of three F1 / F2 heterodimers. Several exogenous multimerization domains that promote formation of stable trimers 35 of soluble proteins are known in the art. Examples of such multimerization domains that can be linked to a mutant provided by the present disclosure include: (1) the GCN4 leucine zipper (Harbury et al. 1993 Science 262: 1401-1407); (2) the trimerization motif from the lung 53

[0052] surfactant protein (Hoppe et al.1994 FEB S Lett 344: 191-195); (3) collagen (McAlinden et al. 2003 Biol Chem 278:42200-42207); and (4) the phage T4 fibritin foldon (Miroshnikov et al. 1998 Protein Eng 11:329-414). In some aspects, a foldon domain is linked to a F mutant atthe C-terminus of F1 polypeptide. In specific aspects, the foldon domain is a T4 fibritin foldon 5 domain, such as the amino acid sequence GYIPEAPRDGQAYVRKDGEWVLLSTFL (SEQ ID NO: 23). Typically, the multimerization domain is positioned C-terminal to the F1 polypeptide. It may join directly to the F1 polypeptide chain. Optionally, the multimerization domain is connected to the F1 polypeptide via a linker, such as an amino acid linker, for example the10 sequence GG, GS, or SAIG. The linker can also be a longer linker (for example, including therepeat sequence GG). Numerous conformationally neutral linkers are known in the art that can be used in the mutants provided by the present disclosure. In some aspects, the RSV Fmutant comprising a foldon domain include a protease cleavage site for removing the foldon domain from the F1 polypeptide, such as a thrombin site between the F1 polypeptide and the15 foldon domain. In a preferred aspect, the RSV F protein to be included in the compositiondisclosed herein comprises a T4 fibritin foldon domain linked at the C-terminus of the F1 polypeptide by an SAIG linker. Introduced Mutations in the RSV F Protein Mutants The compositions including the RSV F protein mutants provided herein comprise a F1 20 polypeptide and a F2 polypeptide, wherein (1) either the F1 polypeptide or (2) the F2 polypeptide, or (3) both the F1 polypeptide and F2 polypeptide include one or more introduced amino acid mutations relative to the amino acid sequence of the corresponding native F protein. The introduction of such amino acid mutations in the RSV F mutants may confer abeneficial property to the mutants, such as enhanced immunogenicity, improved stability, or25 formation or improved stability of certain desired physical form or conformation of the mutants. Such introduced amino acid mutations are referred to as “engineered disulfide bond mutations,” “cavity filling mutations,” or “electrostatic mutations,” and are described in detailherein below. RSV F mutants that include any additional mutations are also encompassed bythe invention so long as the immunogenic property of the mutants is not substantially adversely30 affected by the additional mutations. Engineered Disulfide Bond Mutations In some aspects, the present disclosure provides for compositions comprising RSV Fmutants that include one or more engineered disulfide bond mutations. In some aspects, theRSV F protein is a mutant of a wild-type RSV F protein, wherein the introduced amino acid35 mutations are a pair of amino acid residues in a wild-type RSV F protein to a pair of cysteines (”engineered disulfide mutation”). The introduced pair of cysteine residues allows for formation 54

[0053] of a disulfide bond between the cysteine residues that stabilize the protein’s conformation or oligomeric state, such as the prefusion conformation. In some aspects, the compositions include RSV F protein mutants comprising only oneengineered disulfide mutation (“single engineered disulfide mutation”). In some other aspects,5 the compositions include RSV F protein mutants comprising at least two engineered disulfidemutations, wherein each pair of the cysteine residues of the engineered disulfide mutations are appropriately positioned when RSV F protein mutant is in pre-fusion conformation (“double engineered disulfide mutation”). Examples of specific pairs of such mutations include: 55C and 188C; 155C and 290C; 103C and 148C; and 142C and 371C, such as S55C and L188C;10 S155C and S290C; A103C and I148C; T103C and I148C and L142C and N371C. In some aspects, the present disclosure provides for compositions including RSV F protein mutants, wherein the amino acid mutations are mutation of a pair of amino acid residues in the HRB region (approximately amino acids 476 – 524) of an RSV F protein to apair of cysteines. The introduced pair of cysteine residues allows for formation of a disulfide15 bond between the cysteine residues from two adjacent F2-F1 mutant protomers of a trimer. The disulfide linking two protomers in a trimer serves to stabilize the mutant in a trimeric state. Examples of specific pairs of such mutations include: 508C and 509C; 515C and 516C; 522C and 523C, such as K508C and S509C, N515C and V516C, or T522C and T523C.Cavity Filling Mutations20 In some aspects, compositions of the present disclosure include RSV F proteinmutants comprising one or more cavity filling mutations. The term “cavity filling mutation” refersto the substitution of an amino acid residue in the wild-type RSV F protein by an amino acid that is expected to fill an internal cavity of the mature RSV F protein. In some aspects, suchcavity-filling mutations contribute to stabilizing the pre-fusion conformation of an RSV F protein25 mutant. Examples of amino acids that may be replaced with the goal of cavity filling includesmall aliphatic (e.g. Gly, Ala, and Val) or small polar amino acids (e.g. Ser and Thr) and aminoacids that are buried in the prefusion conformation, but exposed to solvent in the postfusion conformation. Examples of the replacement amino acids include large aliphatic amino acids(Ile, Leu and Met) or large aromatic amino acids (His, Phe, Tyr and Trp).30 In some aspects, the compositions include an RSV F protein mutant comprising acavity filling mutation selected from the group consisting of: (1) substitution of the amino acid at position 55, 62, 155, 190, or 290 with I, Y, L, H, orM; (2) substitution of the amino acid at position 54, 58, 189, 219, or 397 with I, Y, L, H, or 35 M; (3) substitution of the amino acid at position 151 with A or H; (4) substitution of the amino acid at position 147 or 298 with I, L, H, or M; 55

[0054] (5) substitution of the amino acid at position 164, 187, 192, 207, 220, 296, 300, or 495 with I, Y, or H; and (6) substitution of the amino acid at position 106 with W. In some particular aspects, the compositions include an RSV F protein comprising at5 least one cavity filling mutation selected from the group consisting of: T54H, S190I, and V296I. Electrostatic Mutations In some aspects, compositions of the present disclosure include RSV F protein mutants comprising one or more electrostatic mutations. The term “electrostatic mutation”refers to an amino acid mutation introduced to a wild-type RSV F protein that decreases ionic 10 repulsion or increase ionic attraction between residues in a protein that are proximate to each other in the folded structure. In some aspects, an electrostatic mutation may be introduced to improve trimer stability. In some aspects, the RSV F protein mutant includes an electrostaticsubstitution that reduces repulsive ionic interactions or increases attractive ionic interactions with acidic residues of Glu487 and Asp489 from another protomer of RSV F trimer.15 In some aspects, the compositions include an RSV F protein mutant comprising anelectrostatic mutation selected from the group consisting of: (1) substitution of the amino acid at position 82, 92, or 487 by D, F, Q, T, S, L, or H; (2) substitution of the amino acid at position 315, 394, or 399 by F, M, R, S, L, I, Q, or T; (3) substitution of the amino acid at position 392, 486, or 489 by H, S, N, T, or P; and20 (4) substitution of the amino acid at position 106 or 339 by F, Q, N, or W. In some particular aspects, the compositions comprise an RSV F protein comprising anelectrostatic mutation D486S.Combination of Mutations In some aspects, compositions of the present disclosure include RSV F protein25 mutants comprising a combination of two or more different types of mutations selected fromengineered disulfide mutations, cavity filling mutations, and electrostatic mutations. In some aspects, the compositions may include RSV protein F mutants comprising atleast one engineered disulfide bond mutation and at least one cavity filling mutation. For example, the compositions may include RSV F protein mutants comprising a combination of30 mutations as set forth in Table 4 of WO2017109629, which are hereby incorporated by reference in their entireties. In some further aspects, the compositions may include RSV F protein mutantscomprising at least one engineered disulfide mutation and at least one electrostatic mutation.For example, the compositions may include RSV F protein mutants comprising a combination35 of mutations as set forth in Table 5 of WO2017109629, which are hereby incorporated by reference in their entireties. 56

[0055] In still other aspects, the compositions may include RSV F protein mutants comprisingat least one engineered disulfide mutation, at least one cavity filling mutation, and at least one electrostatic mutation. For example, the compositions may include RSV F protein mutantscomprising a combination of mutations as set forth in Table 6 of WO2017109629, which are5 hereby incorporated by reference in their entireties. In some aspects, compositions of the present disclosure include RSV F protein mutants comprising a combination of mutations relative to the corresponding wild-type RSV Fprotein, wherein the combination of mutations is selected from the group consisting of: (1) combination of 103C, 148C, 190I, and 486S; 10 (2) combination of 54H 55C 188C 486S; (3) combination of 54H, 103C, 148C, 190I, V96I, and 486S; (4) combination of 54H, 55C, 142C, 188C, 296I, and 371C; (5) combination of 55C, 188C, and 486S; (6) combination of 54H, 55C, 188C, and 190I; 15 (7) combination of 55C, 188C, 190I, and 486S; (8) combination of 54H, 55C, 188C, 190I, and 486S; (9) combination of 155C, 190I, 290C, and 486S; (10) combination of 54H, 55C, 142C, 188C, 296I, 371C, 486S, 487Q, and 489S; (11) combination of 54H, 155C, 190I, 290C, and 296I, and, 20 (12) combination of 155C, 190F, 290C, and 207L. In some aspects, compositions of the present disclosure include RSV F proteinmutants comprising a combination of mutations relative to the corresponding wild-type RSV Fprotein, wherein the combination of mutations is selected from the group consisting of: (1) combination of 215P and 486N; 25 (2) combination of 66E, 215P, and 486N; (3) combination of 66E, 76V, 215P, and 486N; and (4) combination of 66E, 67I, 76V, 215P, and 486N. In some aspects, the compositions include an RSV F protein mutant of subtype Acomprising the mutations 103C, 148C, 190I, and 486S. In some aspects, the compositions30 comprise an RSV F protein mutant of subtype B comprising the mutations 103C, 148C, 190I,and 486S. In some aspects, the compositions include an RSV F protein mutant of subtype Acomprising the mutations 54H, 103C, 148C, 190I, and 486S. In some aspects, thecompositions include an RSV F protein mutant of subtype B comprising the mutations 54H,35 103C, 148C, 190I, and 486S. In some aspects, the compositions include an RSV F protein mutant of subtype Acomprising the mutations 54H, 55C, 188C, and 486S. In some aspects, the compositions57

[0056] include an RSV F protein mutant of subtype B comprising the mutations 54H, 55C, 188C, and486S. In some aspects, the compositions include an RSV F protein mutant of subtype Acomprising the mutations 155C, 190F, 290C, and 207L. In some aspects, the compositions5 include an RSV F protein mutant of subtype B comprising the mutations 155C, 190F, 290C,and 207L. In some particular aspects, the compositions include RSV F protein mutantscomprising a combination of mutations relative to the corresponding wild-type RSV F protein,wherein the combination of mutations is selected from the group consisting of:10 (1) combination of T103C, I148C, S190I, and D486S;(2) combination of A103C, I148C, S190I, and D486S;(3) combination of T54H, S55C, L188C D486S;(4) combination of T54H, T / A103C, I148C, S190I, V296I, and D486S; (5) combination of T54H, S55C, L142C, L188C, V296I, and N371C; 15 (6) combination of S55C, L188C, and D486S; (7) combination of T54H, S55C, L188C, and S190I; (8) combination of S55C, L188C, S190I, and D486S; (9) combination of T54H, S55C, L188C, S190I, and D486S; (10) combination of S155C, S190I, S290C, and D486S; 20 (11) combination of T54H, S55C, L142C, L188C, V296I, N371C, D486S, E487Q, and D489S; (12) combination of T54H, S155C, S190I, S290C, and V296I, and, (13) combination of S155C, S190F, S290C, and V207L. In some aspects, the compositions include RSV F protein mutants comprising a25 combination of mutations relative to the corresponding wild-type RSV F protein, wherein the combination of mutations is selected from the group consisting of: (1) combination of S215P and D486N; (2) combination of K66E, S215P, and D486N; (3) combination of K66E, I76V, S215P, and D486N; and 30 (4) combination of K66E, N67I, I76V, S215P, and D486N. In some aspects, compositions of the present disclosure include an RSV F proteinmutant comprising a F1 and / or a F2 polypeptide set forth in any one of SEQ ID NOS: 41-80,279-326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties. For example, the composition may include an RSV F mutant comprising a F135 polypeptide and a F2 polypeptide selected from the group consisting of: a F2 polypeptidecomprising the amino acid sequence of SEQ ID NO: 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 58

[0057] 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, or 325; and a F1 polypeptidecomprising the amino acid sequence of SEQ ID NO: 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62,64, 66, 68, 70, 72, 74, 76, 78, 80, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, or 326 of WO2017109629, which5 sequences are hereby incorporated by reference in their entireties. In some aspects, a foldon domain is linked to an RSV F mutant described herein,wherein the foldon domain is linked to the C-terminus of the F1 polypeptide and comprises the amino acid sequence of SEQ ID NO: 23. The present disclosure provides for a composition including a mutant of a wild-type10 RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the mutantcomprises at least one introduced amino acid mutation relative to the amino acid sequence of the wild-type RSV F protein, wherein the introduced amino acid mutation is a pair of cysteine mutations selected from the group consisting of: (1) 55C and 188C; (2) 103C and 148C; and(3) 142C and 371C, and wherein amino acid positions are numbered according to SEQ ID15 NO: 1. In some aspects, the mutant further comprises at least one cavity filling mutation, andat least one electrostatic mutation, wherein the cavity filling mutation is selected from the group consisting of: (1) substitution of the amino acid at position 62, 155, 190, or 290 with I, Y, L, H, or M; 20 (2) substitution of the amino acid at position 54, 58, 189, 219, or 397 with I, Y, L, H, or M; (3) substitution of the amino acid at position 151 with A or H; (4) substitution of the amino acid at position 147 or 298 with I, L, H, or M; and (5) substitution of the amino acid at position 164, 187, 192, 207, 220, 296, 300, or 49525 with I, Y, or H, and wherein the electrostatic mutation is selected from the group consisting of: (1) substitution of the amino acid at position 82, 92, or 487 by D, F, Q, T, S, L, or H; (2) substitution of the amino acid at position 315, 394, or 399 by F, M, R, S, L, I, Q, or T; 30 (3) substitution of the amino acid at position 392, 486, or 489 by H, S, N, T, or P; and (4) substitution of the amino acid at position 106 or 339 by F, Q, N, or W. In some aspects, the C-terminus of the F1 polypeptide is linked to a trimerization domain. In some aspects, the F2 polypeptide comprises RSV F amino acid positions 26-109and F1 polypeptide comprises RSV F amino acid positions 137-513. In some aspects, the35 wild-type RSV is subtype A, subtype B, strain A2, strain Ontario, or strain Buenos Aires. In some aspects, the pair of cysteine mutations is selected from the group consisting of: (1) 55C and 188C; and (2) 103C and 148C. 59

[0058] In some aspects, the cavity filling mutation is selected from the group consisting of: (1) substitution of the amino acid at position 190 with I, Y, or M;(2) substitution of the amino acid at position 54 with I or H; and (3) substitution of the amino acid at position 296 with I. 5 In some aspects, the electrostatic mutation is selected from the group consisting of: (1) substitution of the amino acid at position 487 with D, Q, or H; (2) substitution of the amino acid at position 489 with H, S, or N; and (3) substitution of the amino acid at position 486 with H, S, or T. In some aspects, (i) the pair of cysteine mutations is selected from the group consisting 10 of: (1) 55C and 188C and (2) 103C and 148C; (ii) the cavity filling mutation is selected from the group consisting of: (1) substitution of the amino acid at positions 190 with I, Y, or M; (2) substitution of the amino acid at position 54 with I or H; and (3) substitution of the amino acid at position 296 with I; and (iii) the electrostatic mutation is selected from the group consistingof: (1) substitution of the amino acid at position 487 with D, Q, or H; (2) substitution of the 15 amino acid at position 486 with H, S, or T; and (3) substitution of the amino acid at position 489 with H, S, or N. In some aspects, the mutant comprises a combination of introduced amino acidmutations selected from the group consisting of: (1) combination of 103C, 148C, 190I, and 486S; (2) combination of 54H, 55C, 188C, and 486S; (3) combination of 54H, 103C, 148C, 20 190I, 296I, and 486S; (4) combination of 54H, 55C, 142C, 188C, 296I, and 371C; (5) combination of 55C, 188C, and 486S; (6) combination of 54H, 55C, 188C, and 190I; (7) combination of 55C, 188C, 190I, and 486S; and (8) combination of 54H, 55C, 188C, 190I, and 486S. In a preferred aspect, the mutant comprises a cysteine (C) at position 103 (103C) and25 at position 148 (148C), an isoleucine (I) at position 190 (190I), and a serine (S) at position 486 (486S), wherein the mutant comprises a F1 polypeptide and a F2 polypeptide, and the F2polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptidecomprises the amino acid sequence of SEQ ID NO: 18. In another preferred aspect, the mutant comprises a cysteine (C) at position 103 (103C) and at position 148 (148C), an 30 isoleucine (I) at position 190 (190I), and a serine (S) at position 486 (486S), wherein the mutant comprises a F1 polypeptide and a F2 polypeptide, and the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain). In a preferred aspect,the mutant comprises a cysteine (C) at position 103 (103C) and at position 148 (148C), an35 isoleucine (I) at position 190 (190I), and a serine (S) at position 486 (486S), wherein the mutant comprises a F1 polypeptide and a F2 polypeptide, and the F2 polypeptide comprises the 60

[0059] amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acidsequence of SEQ ID NO: 21. In another preferred aspect, the mutant comprises a cysteine (C) at position 103(103C) and at position 148 (148C), an isoleucine (I) at position 190 (190I), and a serine (S) at 5position 486 (486S), wherein the mutant comprises a F1 polypeptide and a F2 polypeptide,and the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). In some aspects, the mutant further comprises at least one pair of introduced cysteine10 mutations in the HRB region. In some aspects, the at least one pair of cysteine mutations in the HRB region is selected from the group consisting of: (1) 508C and 509C; (2) 515C and 516C; and (3) 522C and 523C. The present disclosure provides for a composition including a mutant of a wild-type RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the mutant15 comprises at least one introduced amino acid mutation relative to the amino acid sequence of the wild-type RSV F protein, and wherein the introduced amino acid mutation comprises: (i) a pair of cysteine mutations 155C and 290C; and (ii) one or more cavity filling mutations selected from the group consisting of: (1) substitution of the amino acid at position 190 with I; 20 (2) substitution of the amino acid at position 54 with I, Y, L, H, or M; (3) substitution of the amino acid at position 296 with I, Y, H, and wherein amino acid positions are numbered according to SEQ ID NO: 1. In some aspects, the cavity filling mutation is selected from the group consisting of: (1) substitution of the amino acid at position 190 with I; 25 (2) substitution of the amino acid at position 54 with H; and (3) substitution of the amino acid at position 296 with I. In some aspects, the mutant is in the form of a trimer. In some aspects, the mutant hasincreased stability as compared with the corresponding wild-type RSV F protein, wherein the stability is measured by binding of the mutant with antibody AM14. In some aspects, the wild- 30 type RSV is subtype A or subtype B. In some aspects, the cavity filing mutation is selected from the group consisting of: 54H, 190I, and 296I. In some aspects, the mutant further comprises an electrostatic mutation. In someaspects, the electrostatic mutation is selected from the group consisting of: 35 (1) substitution of the amino acid at position 82, 92, or 487 by D, F, Q, T, S, L, or H; (2) substitution of the amino acid at position 315, 394, or 399 by F, M, R, S, L, I, Q, or T; 61

[0060] (3) substitution of the amino acid at position 392, 486, or 489 by H, S, N, T, or P; and (4) Substitution of the amino acid at position 106 or 339 by F, Q, N, or W. In some aspects, the mutant of a wild-type RSV F protein comprises a F1 polypeptideand a F2 polypeptide, wherein the mutant comprises at least one introduced amino acid5 mutation relative to the amino acid sequence of the wild-type RSV F protein, and wherein the introduced amino acid mutation comprises: (i) the pair of cysteine mutations 155C and 290C; (ii) a cavity filling mutation; and (iii) an electrostatic mutation, wherein the cavity filling mutationis selected from the group consisting of: (1) substitution of the amino acid at position 62 with I, Y, L, H, or M; 10 (2) substitution of the amino acid at position 190 with I; (3) substitution of the amino acid at position 54, 58, 189, 219, or 397 with I, Y, L, H, or M; (4) substitution of the amino acid at position 151 with A or H; (5) substitution of the amino acid at position 147 or 298 with I, L, H, or M; and 15 (6) substitution of the amino acid at position 164, 187, 192, 207, 220, 296, 300, or 495 with I, Y, H, wherein the electrostatic mutation is 486S, and wherein the amino acid positions are numbered according to SEQ ID NO: 1.In some aspects, the mutant comprises a combination of amino acid mutations20 selected from the group consisting of: (1) combination of 155C, 290C, and 54H; (2) combination of 155C, 290C, 296I; (3) combination of 155C, 290C, 54H, and 296Y; (4) combination of 155C, 290C, and 190I; 25 (5) combination of 155C, 290C, 54H, and 190I; (6) combination of 155C, 290C, 54H, 496S; (7) combination of 155C, 290C, 190I, and 486S; (7) combination of 155C, 290C, 296I; and 486S; (9) combination of 155C, 290C, 54H, 190I; and 486S; 30 (10) combination of 155C, 290C, 54H, 296I, and 486S (11) combination of 155C, 290C, 190I, 296I, and 485S; (12) combination of 155C, 290C, 54H, 190I, 296I, and 486S; (13) combination of 155C, 290C, 190I, and 296I; and (14) combination of 155C, 290C, 54H, 190I, and 296I.35 In some aspects, the mutant of a wild-type RSV F protein comprises a F1 polypeptideand a F2 polypeptide, wherein the mutant comprises at least one introduced amino acidmutation relative to the amino acid sequence of the wild-type RSV F protein, and wherein the 62

[0061] introduced amino acid mutation comprises: (i) the pair of cysteine mutations 155C and 290C; (ii) at least one cavity filling mutation; and (iii) at least one pair of cysteine mutations in the HRB region, wherein the cavity filling mutation is selected from the group consisting of: (1) substitution of the amino acid at position 62 with I, Y, L, H, or M; 5 (2) substitution of the amino acid at position 190 with I; (3) substitution of the amino acid at position 54, 58, 189, 219, or 397 with I, Y, L, H, or M; (4) substitution of the amino acid at position 151 with A or H; (5) substitution of the amino acid at position 147 or 298 with I, L, H, or M; and 10 (6) substitution of the amino acid at position 164, 187, 192, 207, 220, 296, 300, or 495 with I, Y, H, wherein the at least one pair of cysteine mutations in the HRB region is selected from the group consisting of: (1) 508C and 509C; (2) 515C and 516C; and 15 (3) 522C and 523C, and wherein the amino acid positions are numbered according to SEQ ID NO: 1. In some aspects, the composition includes an RSV F protein of subtype A comprisingthe mutations S155C, S190F, S290C, and V207L. In some aspects, the composition includes an RSV F protein of subtype B comprising the mutations S155C, S190F, S290C, and V207L.20 In some aspects, the composition includes an RSV F protein of subtype A comprisingthe mutations S155C, S190F, and S290C. In some aspects, the composition includes an RSVF protein of subtype B comprising the mutations S155C, S190F, and S290C.Nucleic Acids The present disclosure further provides a composition including nucleic acid molecules25 that encode an RSV F protein mutant described herein above. These nucleic acid moleculesinclude DNA, cDNA, and RNA sequences. Nucleic acid molecules that encode only a F2polypeptide or only a F1 polypeptide of an RSV F mutant are also encompassed by theinvention. The nucleic acid molecule can be incorporated into a vector, such as an expressionvector.30 In some particular aspects, the nucleic acid molecule encodes an RSV F mutant, wherein themutant comprises a F1 polypeptide and a F2 polypeptide, and the F2 polypeptide comprisesthe amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18. In another particular aspect, the nucleic acid molecule encodes an RSV F mutant, wherein the mutant comprises a F1 polypeptide and a F2 polypeptide, and35 the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker andtrimerization domain). In some particular aspects, the nucleic acid molecule encodes63

[0062] an RSV F mutant, wherein the mutant comprises a F1 polypeptide and a F2 polypeptide, andthe F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In another particular aspect, the nucleic acid molecule encodes an RSV F mutant, wherein the mutant comprises5 a F1 polypeptide and a F2 polypeptide, and the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). In some particular aspects, the nucleic acid molecule encodes an RSV F mutant,wherein the mutant comprises a F1 and / or a F2 polypeptide set forth in any one of SEQ ID 10 NOS: 41-80, 279-326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties. For example, the nucleic acid molecule encodes the RSV F mutant comprising a F1 polypeptide and a F2 polypeptide selected from the group consisting of: a F2 polypeptide comprising the amino acid sequence of SEQ ID NO: 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 71, 73, 75, 77, 79, 279, 281, 283, 285, 287, 289, 291, 293, 295,15 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, or 325; and a F1polypeptide comprising the amino acid sequence of SEQ ID NO: 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, or 326 ofWO2017109629, which sequences are hereby incorporated by reference in their entireties. 20 RSV RNA The present disclosure further provides for compositions including the components of an RSV composition comprising RNA polynucleotides. In one aspect, the present disclosureprovides for compositions comprising an immunogenic RNA polynucleotide encoding an25 amino acid sequence, e.g., an immunogenic antigen, comprising a respiratory syncytial virus (RSV) protein, an immunogenic variant thereof, or an immunogenic fragment of the RSV protein or the immunogenic variant thereof, e.g., an antigenic peptide or protein. Thus, the immunogenic antigen comprises an epitope of an RSV protein for inducing an immuneresponse against RSV, in the subject. RNA polynucleotide encoding an immunogenic antigen 30 is administered to provide (following expression of the polynucleotide by appropriate target cells) antigen for induction, e.g., stimulation, priming, and / or expansion, of an immune response, e.g., antibodies and / or immune effector cells. In one aspect, the immune response to be induced according to the present disclosure is both B cell-mediated immune response,e.g., an antibody-mediated immune response as well as T-cell-mediated immune response. 35 In one aspect, the immune response is an anti-RSV immune response. The RSV compositions provided herein comprise RNA polynucleotides comprising RNA (as the active principle) that may be translated into one or more proteins in a recipient’s 64

[0063] cells. In addition to wild type, codon-optimized or mutant sequences encoding the antigen sequence, the RNA polynucleotides may contain one or more structural elements optimized for maximal efficacy of the RNA with respect to stability and translational efficiency (5′ cap, 5′ UTR, subgenomic promoter, 3′ UTR, poly-A-tail). In one aspect, the RNA polynucleotides 5 contain all of these elements. The RNA polynucleotides described herein may be complexed with lipids and / or proteins to generate RNA-particles (e.g., lipid nanoparticles (LNPs)) for administration. In one aspect, the RNA polynucleotides described herein are complexed with lipids to generate RNA-lipid nanoparticles (e.g. RNA-LNPs) for administration. In one aspect,the RNA polynucleotides described herein are complexed with proteins for administration. In 10 one aspect, the RNA polynucleotides described herein are complexed with lipids and proteins for administration. If a combination of different RNA polynucleotides is used, the RNA polynucleotides may be complexed together or complexed separately with lipids and / or proteins to generate RNA-particles for administration. The present disclosure provides for RNA polynucleotides and RNA-LNPs that include15 at least one open reading frame (ORF) encoding an RSV antigen.The present disclosure provides for RNA polynucleotides and RNA-LNPs that include at least one ORF encoding an RSV polypeptide comprising an amino acid sequence selectedfrom SEQ ID NO: 1 to 6. In some aspects, the RSV polypeptide has, has at least, or has atmost 90%, 91%, 92%, 93%, 94%, 95, 96%, 97%, 98% or 99% or higher identity to any of the 20 amino acid sequences of SEQ ID NO: 1 to 6. In some aspects, the RSV polypeptide consists of any of the amino acid sequences of SEQ ID NO: 1 to 6. The present disclosure provides for RNA polynucleotides and RNA-LNPs comprising at least one ORF transcribed from a DNA nucleic acid sequence selected from SEQ ID NO: 7 to 10. In some aspects, the RNA polynucleotide comprises an ORF transcribed from a nucleic25 acid sequence that has, has at least, or has at most 90%, 91%, 92%, 93%, 94%, 95, 96%, 97%, 98% or 99% or higher identity to any of the nucleic acid sequences of SEQ ID NO: 7 to 10. In some aspects, the RNA polynucleotide comprises an ORF transcribed from a nucleicacid sequence that consists of any of the nucleic acid sequences of SEQ ID NO: 7 to 10. The present disclosure further provides for RNA polynucleotides and RNA-LNPs30 comprising at least one ORF comprising an RNA nucleic acid sequence selected from SEQID NO: 11 to 16. In some aspects, the RNA polynucleotide comprises a nucleic acid sequencethat has, has at least, or has at most 90%, 91%, 92%, 93%, 94%, 95, 96%, 97%, 98% or 99% identity to any of the nucleic acid sequences of SEQ ID NO: 11 to 16. In some aspects, the RNA polynucleotide comprises a nucleic acid sequence that consists of any of the nucleic acid35 sequences of SEQ ID NO: 11 to 16. In some aspects, each uridine of any of SEQ ID NO: 11 to 16 is replaced by N1-methylpseudouridine (Ψ) (e.g., modified RNA; modRNA).65

[0064] The present disclosure further provides for RNA polynucleotides and RNA-LNPs that include a 5’ untranslated region (5’-UTR) and / or a 3’ untranslated region (3’-UTR). In some aspects, the RNA polynucleotide includes a 5’ untranslated region (5’-UTR). In some aspects,the 5’ UTR comprises a sequence selected from SEQ ID NO: 24 and 25. In some aspects, the5 5′ UTR comprises a sequence having at least 90%, 91%, 92%, 93%, 94%, 95, 96%, 97%,98% or 99% or higher identity to SEQ ID NO: 24 or 25. In some aspects, the 5′ UTR comprisesa sequence selected from SEQ ID NO: 24 and 25. In some aspects, the 5′ UTR comprises asequence consisting of SEQ ID NO: 24 or 25. The present disclosure further provides for RNA polynucleotides that include at least10 one open reading frame that was generated from codon-optimized DNA. In some aspects, the open reading frame comprises a G / C content of at least, at most, exactly, or between (inclusive or exclusive) any two of 50%, 51%, 52%, 53%, 54%, 55%, 56%, 57%, 58%, 59%, 60%, 61%, 62%, 63%, 64%, 65%, 66%, 67%, 68%, 69%, 70%, 71%, 72%, 73%, 74%, or 75% or more.The present disclosure further provides for immunogenic compositions including the 15 RNA polynucleotides described herein. The RNA polynucleotides may be formulated in, encapsulated in, complex with, bound to or adsorbed on a lipid nanoparticle (LNP) (e.g., RSV RNA-LNPs) in such immunogenic compositions. In some aspects, the lipid nanoparticleincludes at least one of a cationic lipid, a polymer conjugated lipid (e.g., a PEGylated lipid), and at least one structural lipid (e.g., a neutral lipid and a steroid or steroid analog). In some20 aspects, 1, 2, 3, or more of the foregoing lipids can be excluded from the lipid nanoparticle. In some aspects, the lipid nanoparticle includes a cationic lipid. In some aspects, thecationic lipid is (4-hydroxybutyl)azanediyl)bis(hexane-6,1-diyl)bis(2-hexyldecanoate) (ALC- 0315). In some aspects, the lipid nanoparticle includes a polymer conjugated lipid. In some25 aspects, the lipid nanoparticle includes a PEGylated lipid, also referred to as a PEG-lipid. Insome aspects, the PEGylated lipid is PEG-modified phosphatidylethanolamine, PEG-modifiedphosphatidic acid, PEG-modified ceramides (e.g., PEG-CerC14 or PEG-CerC20), PEG- modified dialkylamines, PEG-modified diacylglycerols, PEG-modified dialkylglycerols, 2- [(polyethylene glycol)-2000]-N,N-ditetradecylacetamide, glycol-lipids including PEG-c-DOMG,30 PEG-c-DMA, PEG-s-DMG, N-[(methoxy polyethylene glycol)2000)carbamoyl]-1,2- dimyristyloxlpropyl-3-amine (PEG-c-DMA), and PEG-2000-DMG, PEGylated diacylglycerol (PEG-DAG) such as 1-(monomethoxy-polyethyleneglycol)-2,3-dimyristoylglycerol (PEG- DMG), a PEGylated phosphatidylethanolamine (PEG-PE), a PEG succinate diacylglycerol (PEG-S-DAG) such as 4-O-(2’,3′- di(tetradecanoyloxy)propyl-1-O-((o-35 methoxy(polyethoxy)ethyl)butanedioate (PEG-S-DMG), a PEGylated ceramide (PEG-cer), or a PEG dialkoxypropylcarbamate such as co-methoxy(polyethoxy)ethyl-N- (2,3di(tetradecanoxy)propyl)carbamate or 2,3-di(tetradecanoxy)propyl-N-(u>- 66

[0065] methoxy(polyethoxy)ethyl)carbamate. In some aspects, 1, 2, 3, 4, 5, or more of the foregoing PEGylated lipids can be excluded from the RNA polynucleotide. In some aspects, thePEGylated lipid is 2-[(polyethylene glycol)-2000]-N,N-ditetradecylacetamide (ALC-0159). In some aspects, the lipid nanoparticle includes at least one structural lipid, such as a5 neutral lipid. In some aspects, the neutral lipid is 1,2-distearoyl-sn-glycero-3-phosphocholine(DSPC), distearoylphosphatidylcholine (DSPC), dioleoylphosphatidylcholine (DOPC), dipalmitoylphosphatidylcholine (DPPC), dioleoylphosphatidylglycerol (DOPG), dipalmitoylphosphatidylglycerol (DPPG), dioleoyl-phosphatidylethanolamine (DOPE), palmitoyloleoylphosphatidylcholine (POPC), palmitoyl-oleoyl-phosphatidylethanolamine10 (POPE), dioleoyl- phosphatidylethanolamine 4-(N-maleimidomethyl)-cyclohexane-1-carboxylate (DOPE-mal), dipalmitoylphosphatidylethanolamine (DPPE), dimyristoylphosphoethanolamine (DMPE), distearoylphosphatidylethanolamine (DSPE), 16- O-monomethyl PE, 16-O-dimethyl PE, 18-1-trans PE, 1-stearoyl-2- oleoylphosphatidyethanolamine (SOPE), and / or 1,2-dielaidoyl-sn-glycero-3- 15 phosphoethanolamine (transDOPE). In some aspects, 1, 2, 3, 4, 5, or more of the foregoing structural lipids can be excluded from the RNA polynucleotide. In some aspects, the neutrallipid is 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC). In some aspects, the lipid nanoparticle includes a second structural lipid, such as asteroid or steroid analog. In some aspects, the steroid or steroid analog is cholesterol.20 In some aspects, the RNA polynucleotides and RNA-LNPs include a 3’ untranslated region (3’-UTR). In some aspects, the 3’ UTR comprises a sequence selected from any ofSEQ ID NO: 26 to 29. In some aspects, the 3′ UTR comprises a sequence having at least90%, 91%, 92%, 93%, 94%, 95, 96%, 97%, 98% or 99% or higher identity to any of SEQ ID NO: 26 to 29. In some aspects, the 3′ UTR comprises a sequence selected from any of SEQ25 ID NO: 26 to 29. In some aspects, the 3′ UTR comprises a sequence consisting of any of SEQID NO: 26 to 29.The present disclosure further provides RNA polynucleotides comprising stabilized RNA. The present disclosure further provides for RNA polynucleotides that include RNA having at least one modified nucleotide (e.g., modified RNA; modRNA). In some aspects, the modified 30 nucleotide is pseudouridine, N1-methylpseudouridine, N1-ethylpseudouridine, 2-thiouridine, 4′-thiouridine, 5-methylcytosine, 5-methyluridine, 2-thio-1-methyl-1-deaza-pseudouridine, 2- thio-1-methyl-pseudouridine, 2-thio-5-aza-uridine, 2-thio-dihydropseudouridine, 2-thio- dihydrouridine, 2-thio-pseudouridine, 4-methoxy-2-thio-pseudouridine, 4-methoxy- pseudouridine, 4-thio-1-methyl-pseudouridine, 4-thio-pseudouridine, 5-aza-uridine, 35 dihydropseudouridine, 5-methoxyuridine, or 2′-O-methyl uridine. In some aspects, the modified nucleotide is N1-methylpseudouridine (Ψ). In some aspects, 1, 2, 3, 4, 5, or more of the foregoing modified nucleotides can be excluded from the RNA polynucleotide. 67

[0066] The present disclosure further provides for RNA polynucleotides that are messenger- RNA (mRNA) or self-replicating RNA. In some aspects, the RNA is a mRNA.The present disclosure further provides for RNA polynucleotides and RNA-LNPs that include a 5’ cap moiety. In some aspects, the 5′ cap moiety is (3′OMe) - m27,3′-OGppp (m12′-5O)ApG. The present disclosure further provides for RNA polynucleotides and RNA-LNPs that include a 3’ poly-A tail. In some aspects, the poly-A tail comprises a sequence having SEQ IDNO: 30. In some aspects, the poly-A tail length may contain +1 / -1 A. In some aspects, the RNA polynucleotide includes a 5’ UTR and 3’ UTR. In someaspects, the RNA polynucleotide includes a 5’ cap, 5’ UTR, and 3’ UTR. In some aspects, the10 RNA polynucleotide includes a 5’ cap, 5’ UTR, 3’ UTR, and poly-A tail. In some aspects, theRNA polynucleotide includes a 5’ UTR, 3’ UTR, and poly-A tail. In some aspects, 1, 2, 3, ormore of the foregoing elements can be excluded from the RNA polynucleotide. In some aspects, each uridine of any of the 5′ UTR, 3′ UTR, and poly-A tail is replaced by N1-methylpseudouridine (Ψ) (e.g., modified RNA; modRNA).15 In some aspects, the present invention includes RSV RNA polynucleotides andcompositions as described in WO2024 / 089633, which is hereby incorporated by reference inits entirety. COMPOSITIONS20 The present disclosure provides for a composition that includes components of a GBScomposition and an RSV composition as described herein. In a preferred aspect, thecomposition comprises GBS6 (as described herein) and an RSV vaccine, such ABRYSVOTM(RSVPreF) or AREXVYTM. In a preferred aspect, a lyophilized RSV composition describedherein is reconstituted with an aqueous or liquid GBS composition described herein to .25 GBS Compositions The present disclosure provides for a composition that includes components of a GBS composition described herein. The amount of a particular conjugate in a composition is generally calculated basedon total polysaccharide, conjugated and non-conjugated, for that conjugate. For example, a 30 GBS capsular polysaccharide conjugate with 20% free polysaccharide will have about 80 µg / mL of conjugated GBS capsular polysaccharide and about 20 µg / mL of non-conjugatedGBS capsular polysaccharide in a 100 µg / mL GBS capsular polysaccharide dose. The proteincarrier contribution to the conjugate is usually not considered when calculating the dose of a conjugate. The amount of conjugate can vary depending upon the streptococcal serotype.35 Generally, each dose will comprise about 0.01 mg / mL to about 100 µg / mL of eachpolysaccharide, particularly about 1 µg / mL to about 70 µg / mL, and more particularly about 568

[0067] µg / mL to about 50 µg / mL. The "immunogenic amount" of the different polysaccharidecomponents in the immunogenic composition, may diverge and each may comprise about 0.01 µg / mL, about 0.1 µg / mL, about 0.25 µg / mL, about 0.5 µg / mL, about 1 µg / mL, about 2 µg / mL, about 3 µg / mL, about 4 µg / mL, about 5 µg / mL, about 6 µg / mL, about 7 µg / mL, about 5 8 µg / mL, about 9 µg / mL, about 10 µg / mL, about 15 µg / mL, about 20 µg / mL, about 25 µg / mL, about 30 µg / mL, about 40 µg / mL, about 50 µg / mL, about 60 µg / mL, about 70 µg / mL, about 80µg / mL, about 90 µg / mL, or about 100 µg / mL of any particular polysaccharide antigen. A doseor immunogenic amount of a multivalent immunogenic composition would indicate the dose of each polysaccharide unless indicated otherwise. For example, a 10 µg / mL dose of a10 hexavalent immunogenic composition would contain 10 µg / mL of each of the sixpolysaccharides for a total antigen or conjugate dose of 60 µg / mL. In one aspect, the GBS composition comprises about 5 µg / mL to 20 µg / mL of each polysaccharide. In a preferredaspect, the GBS composition comprises about 20 µg / mL of each polysaccharide.In one aspect of the invention, the total conjugate dose is at least about 60 µg / mL,15 such as at least about 120 µg / mL or at least about 240 µg / mL. In one aspect, the totalconjugate dose is about 60 µg / mL to about 600 µg / mL, such as about 60 µg / mL to about 540µg / mL, about 60 µg / mL to about 480 µg / mL, about 60 µg / mL to about 420 µg / mL, about 60µg / mL to about 360 µg / mL, about 60 µg / mL to about 300 µg / mL, about 60 µg / mL to about 240µg / mL, about 60 µg / mL to about 180 µg / mL, about 60 µg / mL to about 120 µg / mL, about 12020 µg / mL to about 600 µg / mL, about 120 µg / mL to about 540 µg / mL, about 120 µg / mL to about480 µg / mL, about 120 µg / mL to about 420 µg / mL, about 120 µg / mL to about 360 µg / mL, about120 µg / mL to about 300 µg / mL, about 120 µg / mL to about 240 µg / mL, about 120 µg / mL toabout 180 µg / mL, about 180 µg / mL to about 600 µg / mL, about 180 µg / mL to about 540 µg / mL,about 180 µg / mL to about 480 µg / mL, about 180 µg / mL to about 420 µg / mL, about 180 µg / mL25 to about 360 µg / mL, about 180 µg / mL to about 300 µg / mL, about 180 µg / mL to about 240µg / mL, about 240 µg / mL to about 600 µg / mL, about 240 µg / mL to about 540 µg / mL, about240 µg / mL to about 480 µg / mL, about 240 µg / mL to about 420 µg / mL, about 240 µg / mL toabout 360 µg / mL, or about 240 µg / mL to about 300 µg / mL. In one aspect, the total conjugatedose is about 60 µg / mL to about 360 µg / mL, and more preferably about 120 µg / mL to about30 240 µg / mL, and most preferably about 240 µg / mL. In one aspect of the invention, GSB composition comprises at least about 10 µg / mL,such as at least about 20 µg / mL or at least about 40 µg / mL of each polysaccharide. In apreferred aspect, the composition comprises about 40 µg / mL of each polysaccharide. In one aspect, the GBS composition includes at least one of any polysaccharide 35 described herein and a pharmaceutically acceptable excipient, buffer, stabilizer, adjuvant, a cryoprotectant, a salt, a divalent cation, a non-ionic detergent or surfactant, an inhibitor of free radical oxidation, a diluent or a carrier, or mixture thereof. 69

[0068] In one aspect, the buffer is selected from, but not limited to HEPES, PIPES, MES, Tris (trimethamine), phosphate, acetate, borate, citrate, glycine, histidine and succinate. In apreferred aspect, the buffer is histidine or phosphate. In one aspect, the GBS composition comprises a buffer at a concentration of from 5 about 5 mM to about 50 mM, about 5 mM to about 40 mM, about 5 mM to about 30 mM, about 5 mM to about 20 mM, about 5 mM to about 10 mM, about 10 mM to about 50 mM, about 10 mM to about 40 mM, about 10 mM to about 35 mM, about 10 mM to about 30 mM, about 10 mM to about 25 mM, about 10 mM to about 20 mM, about 10 mM to about 15 mM, about 15 mM to about 50 mM, about 15 mM to about 40 mM, about 15 mM to about 35 mM, about 1510 mM to about 30 mM, about 15 mM to about 25 mM, or about 15 mM to about 20 mM. In apreferred aspect, the GBS composition comprises a buffer at a concentration of about 10 mM to about 25 mM, such as about 15 mM to about 25 mM, and most preferably about 20 mM. Ina preferred aspect, the GBS composition comprises histidine at a concentration of about 20 mM. 15 In some aspect, the GBS composition is buffered to within a pH range of about 5.0 to about 7.5, such as 5.3 to about 7.5, about 5.5 to about 7.5, about 6.0 to about 7.5, about 6.5 to about 7.5, about 5.3 to about 7.1, about 5.5 to about 7.0, about 6.0 to about 7.0, about 6.0 to about 6.5, about 6.3 to about 7.0, or about 6.5 to about 7.0. In another aspect, theformulation is buffered to a pH of about 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, 7.0, 7.1,20 7.2, 7.3, 7.4, or 7.5. In a preferred aspect, the GBS composition is buffered to a pH range offrom about 6.0 to about 7.5, such as about 6.5 to about 7.5 and about 6.5 or about 7.0, and most preferably 6.0 to 7.0. In a preferred aspect, the GBS composition is buffered to a pH range of about 6.5. In some aspects, the GBS composition may comprise one or more non-ionic 25 surfactants, including but not limited to polyoxyethylene sorbitan fatty acid esters, polysorbate 80 (TWEEN 80), polysorbate 60 (TWEEN 60), polysorbate 40 (TWEEN 40), polysorbate 20 (TWEEN 20), and polyoxyethylene alkyl ethers, including but not limited to BRIJ 58, BRIJ 35, as well as others such as TRITON X-100; TRITON X- 114, NP40, SPAN 85 and thePLURONIC series of non-ionic surfactants (e.g., PLURONIC 121). In one aspect, the GBS 30 composition comprises polysorbate 80 or polysorbate 40, preferably polysorbate 80 (PS80). In one aspect, the GBS composition comprises a surfactant at a concentration of from about 0.001% to about 2% (v / w), about 0.001% to about 1%, about 0.001% to about 0.5%, about 0.001% to about 0.1%, about 0.001% to about 0.05%, about 0.001% to about 0.01%, about 0.001% to 0.005%, about 0.005% to about 2%, about 0.005% to about 1%, about 35 0.005% to about 0.5%, about 0.005% to about 0.1%, about 0.005% to about 0.05%, about 0.005% to about 0.01%, about 0.01% to about 2%, about 0.01% to about 1%, about 0.01% to about 0.5%, about 0.01% to about 0.1%, about 0.01% to about 0.05%, about 0.01% to about 70

[0069] 0.04%, about 0.01% to about 0.03%, about 0.015% to about 2%, about 0.015% to about 1%, about 0.015% to about 0.5%, about 0.015% to about 0.1%, about 0.015% to about 0.05%, about 0.015% to about 0.04%, about 0.015% to about 0.03%, about 0.02% to about 2%, about 0.02% to about 1%, about 0.02% to about 0.5%, about 0.02% to about 0.1%, about 0.02% to 5 about 0.05%, about 0.02% to about 0.04%, about 0.02% to about 0.03%, about 0.05% to about 2%, about 0.05% to about 1%, about 0.05% to about 0.5%, about 0.05% to about 0.1%, about 0.1% to about 2%, about 0.1% to about 1%, about 0.1% to about 0.5% or about 0.1% to about 0.25%. In a preferred aspect, the immunogenic composition comprises a surfactantat a concentration of about 0.01% to about 0.03%, and most preferably about 0.02%. In a 10 preferred aspect, the GBS composition comprises polysorbate 80 at a concentration from about 0.001% to about 2% (with up to about 0.25% being preferred) or polysorbate 40 at a concentration from about 0.001% to 1% (with up to about 0.5% being preferred). In a preferred aspect, the GBS composition comprises polysorbate 80 at a concentration of about 0.02%.Suitable pharmaceutical excipients include starch, glucose, lactose, sucrose, 15 trehalose, raffinose, stachyose, melezitose, dextran, mannitol, lactitol, palatinit, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, glycine, arginine, lysine, sodium chloride (NaCl), dried skim milk, glycerol, propylene glycol, water, ethanol and the like. In a preferred aspect, the excipient is sodium chloride. In one aspect, the GBS composition comprises an excipient at a concentration of from 20 about 10 mM to about 500 mM, about 10 mM to about 450 mM, about 10 mM to about 400 mM, about 10 mM to about 350 mM, about 10 mM to about 300 mM, about 10 mM to about 250 mM, about 10 mM to about 200 mM, about 10 mM to about 150 mM, about 10 mM to about 100 mM, about 10 mM to about 50 mM, about 10 mM to about 30 mM, about 10 mM to about 20 mM, 20 mM to about 500 mM, about 20 mM to about 450 mM, about 20 mM to about 25 400 mM, about 20 mM to about 350 mM, about 20 mM to about 300 mM, about 20 mM to about 250 mM, about 20 mM to about 200 mM, about 20 mM to about 150 mM, about 20 mM to about 100 mM, about 20 mM to about 50 mM, about 20 mM to about 30 mM, 50 mM to about 500 mM, about 50 mM to about 450 mM, about 50 mM to about 400 mM, about 50 mM to about 350 mM, about 50 mM to about 300 mM, about 50 mM to about 250 mM, about 50 30 mM to about 200 mM, about 50 mM to about 150 mM, about 50 mM to about 100 mM, about 100 mM to about 500 mM, about 100 mM to about 450 mM, about 100 mM to about 400 mM, about 100 mM to about 350 mM, about 100 mM to about 300 mM, about 100 mM to about 250 mM, about 100 mM to about 200 mM, about 100 mM to about 150 mM, about 120 mM to about 470 mM, about 120 mM to about 420 mM, about 120 mM to about 370 mM, about 120 mM to 35 about 320 mM, about 120 mM to about 270 mM, about 120 mM to about 220 mM, about 120 mM to about 170 mM, about 150 mM to about 500 mM, about 150 mM to about 450 mM, about 150 mM to about 400 mM, about 150 mM to about 350 mM, about 150 mM to about 300 mM, 71

[0070] about 150 mM to about 250 mM, about 150 mM to about 200 mM, about 200 mM to about 500 mM, about 200 mM to about 450 mM, about 200 mM to about 400 mM, about 200 mM to about 350 mM, about 200 mM to about 300 mM, about 200 mM to about 250 mM, about 225 mM to about 465 mM, about 225 mM to about 425 mM, about 225 mM to about 365 mM, about 225 5 mM to about 325 mM, about 225 mM to about 265 mM, about 250 mM to about 500 mM, about 250 mM to about 450 mM, about 250 mM to about 400 mM, about 250 mM to about 350 mM, about 250 mM to about 300 mM, about 300 mM to about 500 mM, about 300 mM to about 450 mM, about 300 mM to about 400 mM, about 300 mM to about 350 mM, about 350 mM to about 500 mM, about 350 mM to about 450 mM, about 350 mM to about 400 mM, about 400 mM to10 about 500 mM, about 400 mM to about 450 mM, or about 450 mM to about 500 mM. In apreferred aspect, the GBS composition comprises an excipient at a concentration of fromabout 10 mM to about 350 mM, such as about 10 mM to about 250 mM, about 130 mM toabout 170 mM, and about 225 mM to about 265 mM, and most preferably about 150 mM orabout 245 mM. In a preferred aspect, the excipient is sodium chloride at a concentration of 15 about 150 mM. In one aspect, the GBS composition may comprise one or more adjuvant. Non-limiting examples of adjuvants that can be used in the composition of the present disclosure includethe RIBI adjuvant system (Ribi Inc., Hamilton, Mont.); mineral gels, such as aluminum hydroxide gel; water-in-oil emulsions, such as Freund's complete and incomplete adjuvants; 20 Block copolymer (CytRx, Atlanta Ga.); SAF-M (Chiron, Emeryville, Calif.); AMPHIGEN® adjuvant; saponin; Quil A or other saponin fraction; monophosphoryl lipid A; and Avridine lipid- amine adjuvant. Non-limiting examples of oil-in-water emulsions useful as an adjuvant in the vaccine of the invention include MF59 (U.S. Patent No.6,299,884) (containing 5% Squalene, 0.5% polysorbate 80, and 0.5% Span 85 (optionally containing various amounts of MTP-PE)25 formulated into submicron particles using a microfluidizer such as Model 110Y microfluidizer (Microfluidics, Newton, MA)), and SAF (containing 10% Squalene, 0.4% polysorbate 80, 5% pluronic-blocked polymer L121, and thr-MDP, either microfluidized into a submicron emulsion or vortexed to generate a larger particle size emulsion); modified SEAM62 (containing 5% (v / v) squalene (Sigma), 1% (v / v) SPAN®85 detergent (ICI Surfactants), 0.7% (v / v) polysorbate30 80 detergent (ICI Surfactants), 2.5% (v / v) ethanol, 200 µg / mL Quil A, 100 µg / mL cholesterol,and 0.5% (v / v) lecithin); and modified SEAM 1 / 2 (containing 5% (v / v) squalene, 1% (v / v) SPAN® 85 detergent, 0.7% (v / v) polysorbate 80 detergent, 2.5% (v / v) ethanol, 100 µg / mL QuilA, and 50 µg / mL cholesterol).Suitable adjuvants used to enhance an immune response further include, without35 limitation, MPL™ (3-O-deacylated monophosphoryl lipid A, Corixa, Hamilton, MT), which isdescribed in U.S. Patent No.4,912,094. Also suitable for use as adjuvants are synthetic lipid A analogs or aminoalkyl glucosamine phosphate compounds (AGP), or derivatives or analogs 72

[0071] thereof, which are available from Corixa (Hamilton, MT), and which are described in U.S. Patent No. 6,113,918. One such AGP is 2-[(R)-3-Tetradecanoyloxy¬tetradecanoyl¬amino] ethyl 2-Deoxy-4-O-phosphono-3-O-[(R)-3-tetra¬decanoyoxy¬tetrade¬canoyl]-2-[(R)-3- tetradecanoyloxy¬tetradecanoyl-amino]-b-D-glucopyranoside, which is also known as 529 5 (formerly known as RC529). This 529 adjuvant is formulated as an aqueous form (AF) or as a stable emulsion (SE). Still other adjuvants include a cyclodextrin derivative (U.S. Patent No. 6,165,995); a polyanionic polymer (U.S. Patent No. 6,610,310); muramyl peptides, such as N-acetyl- muramyl-L-threonyl-D-isoglutamine (thr-MDP), and N-acetyl-normuramyl-L-alanine-2-(1'-2' 10 dipalmitoyl-sn-glycero-3-hydroxy¬phosphoryl¬oxy)-ethylamine (MTP-PE); Amphigen; Avridine; L121 / squalene; D-lactide-polylactide / glycoside; pluronic polyols; killed Bordetella; saponins, such as Stimulon™ QS-21 (Antigenics, Framingham, MA.), described in U.S. PatentNo. 5,057,540; Mycobacterium tuberculosis; bacterial lipopolysaccharides; synthetic polynucleotides such as oligonucleotides containing a CpG motif (e.g., U.S. Patent No. 15 6,207,646); IC-31 (Intercell AG, Vienna, Austria), described in European Patent Nos. 1,296,713 and 1,326,634; a pertussis toxin (PT) or mutant thereof, a cholera toxin or mutant thereof (e.g., U.S. Patent Nos.7,285,281, 7,332,174, 7,361,355 and 7,384,640); or an E. coli heat-labile toxin (LT) or mutant thereof, particularly LT-K63, LT-R72 (e.g., U.S. Patent Nos. 6,149,919, 7,115,730 and 7,291,588).20 In one aspect, the GBS composition comprises an adjuvant at a concentration of fromabout 0.1 mg / mL to about 1.0 mg / mL, 0.1 mg / mL to about 0.9 mg / mL, 0.1 mg / mL to about 0.8mg / mL, 0.1 mg / mL to about 0.7 mg / mL, 0.1 mg / mL to about 0.6 mg / mL, 0.1 mg / mL to about0.5 mg / mL, 0.1 mg / mL to about 0.4 mg / mL, 0.1 mg / mL to about 0.3 mg / mL, 0.1 mg / mL toabout 0.2 mg / mL, 0.25 mg / mL to about 0.95 mg / mL, 0.25 mg / mL to about 0.85 mg / mL, 0.2525 mg / mL to about 0.75 mg / mL, 0.25 mg / mL to about 0.65 mg / mL, 0.25 mg / mL to about 0.55mg / mL, 0.25 mg / mL to about 0.45 mg / mL, 0.25 mg / mL to about 0.35 mg / mL, 0.5 mg / mL toabout 1.0 mg / mL, 0.5 mg / mL to about 0.9 mg / mL, 0.5 mg / mL to about 0.8 mg / mL, 0.5 mg / mLto about 0.75 mg / mL, 0.5 mg / mL to about 0.7 mg / mL, 0.5 mg / mL to about 0.65 mg / mL, 0.5mg / mL to about 0.6 mg / mL, 0.75 mg / mL to about 1.0 mg / mL, 0.75 mg / mL to about 0.9530 mg / mL, 0.75 mg / mL to about 0.9 mg / mL, and 0.75 mg / mL to about 0.85 mg / mL. In a preferredaspect, the GBS composition comprises an adjuvant at a concentration of from about 0.25mg / mL to about 0.75 mg / mL, and most preferably about 0.5 mg / mL.In a preferred aspect, the adjuvant is an aluminum-based at a concentration of about 0.5 mg / mL. In one such aspect, the aluminum-based adjuvant is aluminum phosphate or 35 aluminum hydroxyl phosphate. In another aspect, the aluminum-based adjuvant is aluminum hydroxide. In a preferred aspect, the GBS composition is in an aqueous or liquid form.73

[0072] In another aspect, the GBS composition is lyophilized, optionally in the presence of at least one excipient. In a preferred aspect, the at least one excipient is selected from the group consisting of starch, glucose, lactose, sucrose, trehalose, raffinose, stachyose, melezitose, dextran, mannitol, lactitol, palatinit, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, 5glycerol monostearate, talc, glycine, arginine, lysine, sodium chloride (NaCl), dried skim milk,glycerol, propylene glycol, water, and ethanol. In a preferred aspect, the at least one excipient is selected from the group consisting of sucrose, mannitol, and glycine. In a particular aspect, the at least one excipient is sucrose. In another aspect, the lyophilized composition comprisesan additional excipient. In one such aspect, the additional excipient is mannitol or glycine.10 In another aspect, the lyophilized GBS composition comprises about 1% (w / v) to about 10% (w / v) of at least one saccharide, such as about 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5% or 10.0%. In a preferredaspect, the lyophilized composition comprises greater than about 5.5% (w / v) of at least one excipient, such as greater than about 7.0%(w / v). In a further aspect, the lyophilized 15 composition comprises about 1% (w / v) to about 10% (w / v) of an additional excipient, such as about 1.5%, 2.0%, 2.5%, 3.0%, 3.5%, 4.0%, 4.5%, 5.0%, 5.5%, 6.0%, 6.5%, 7.0%, 7.5%, 8.0%, 8.5%, 9.0%, 9.5% or 10.0%. In a preferred aspect, the lyophilized composition comprises about 1% (w / v) to about 10% (w / v) of the at least one excipient and about 1% (w / v) to about 10% (w / v) of the additional excipient. 20 In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising a GBS polysaccharide-protein conjugate as described herein, a buffer, a surfactant, and an excipient. In one aspect, the GBS composition comprises a polysaccharide-protein conjugate as described herein, a buffer, a surfactant, an excipient, and optionally an adjuvant. 25 In one such aspect, the composition includes the components of a GBS composition comprising a GBS polysaccharide-protein conjugate, a buffer, a surfactant, and an excipient, wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In another such aspect, the GBS composition comprises a GBS polysaccharide-protein conjugate, a buffer, a surfactant, an excipient, and optionally an adjuvant, wherein the capsular30 polysaccharide has a sialic acid level of greater than about 60%. In one aspect, the composition includes the components of a GBS composition comprising a GBS polysaccharide-protein conjugate, histidine, polysorbate 80, and sodium chloride. In one particular aspect, the GBS composition comprises a GBS polysaccharide- protein conjugate, histidine, polysorbate 80, and sodium chloride, wherein the capsular35 polysaccharide has a sialic acid level of greater than about 60%. In another aspect, the composition includes the components of a GBS composition comprising a GBS polysaccharide-protein conjugate, histidine, polysorbate 80, sodium 74

[0073] chloride, and optionally aluminum phosphate. In another aspect, the GBS composition comprises a GBS polysaccharide-protein conjugate, histidine, polysorbate 80, sodium chloride, and optionally aluminum phosphate, wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. 5 In an aspect, the GBS composition is buffered to a pH of about 6.0 to about 7.5. In another aspect, the GBS composition is buffered to a pH of about 6.0 to about 7. In a preferred aspect, the GBS composition is buffered to a pH of about 6.5. In a preferred aspect, a composition of the present disclosure includes the components of a GBS composition comprising about 5 µg / mL to about 50 µg / mL of each polysaccharide, 10 about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In a preferred aspect, the GBS composition comprises about 5µg / mL to about 50 µg / mL of each polysaccharide, about 10 mM to about 25 mM of histidine,about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of 15 sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In another aspect, the composition includes the components of a GBS composition comprises about 5 µg / mL to about 50 µg / mL of each polysaccharide, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, about 10 mM to about 20 250 mM of sodium chloride, and optionally about 0.25 mg / mL to about 0.75 mg / mL of aluminum as aluminum phosphate, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In another aspect, the GBS composition comprises about 5 µg / mL to about 50 µg / mL of each polysaccharide, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, about 10 mM to about 250 mM of sodium chloride, and 25 optionally about 0.25 mg / mL to about 0.75 mg / mL of aluminum as aluminum phosphate, wherein the composition is buffered to a pH of about 6.0 to about 7.5 and wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In a preferred aspect, a composition of the present disclosure includes the components of a GBS composition comprising about 40 µg / mL of each polysaccharide, about 20 mM of 30 histidine, about 0.02% (v / w) of polysorbate 80, and about 150 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In a preferred aspect, the GBS composition comprises about 40 µg / mL of each polysaccharide, about 20 mM of histidine, about 0.02% (v / w) of polysorbate 80, and about 150 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular35 polysaccharide has a sialic acid level of greater than about 60%. In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising at least one, 75

[0074] two, three, four, five, six, seven or more capsular polysaccharide selected from GBS serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII, and IX and a carrier protein, a buffer, a surfactant, and an excipient, wherein the composition is buffered to a pH of about 6.0 to about 7.5. The GBS composition optionally further comprises an adjuvant. 5 In one particular aspect, the composition includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising at least one, two, three, four, five, six, seven or more capsular polysaccharide selected from GBS serotypes Ia,Ib, II, III, IV, V, VI, VII, VIII, and IX and a carrier protein, histidine, polysorbate 80, and sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. The GBS10 composition optionally further comprises aluminum phosphate. In a preferred aspect, the composition includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising about 5 µg / mL to about 50 µg / mL of at least one, two, three, four, five, six, seven or more polysaccharide selected from GBS serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII, and IX and a carrier protein, about 10 mM to 15 about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. The GBS composition optionally further comprises about 0.25 mg / mL to about 0.75 mg / mL of aluminum as aluminum phosphate. In one aspect, a composition of the present disclosure includes the components of a 20 GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, and II and a carrier protein. In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, and III and a carrier protein. 25 In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II and III and a carrier protein. In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular30 polysaccharide from GBS serotypes Ia, Ib, II, III and V and a carrier protein. In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein. In one aspect, a composition of the present disclosure includes the components of a 35 GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II, III, V and VII and a carrier protein. 76

[0075] In one aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV, V and VI and a carrier protein. In one aspect, a composition of the present disclosure includes the components of a 5 GBS composition comprising GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, a buffer, a surfactant, and an excipient, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In one aspect, the GBS composition comprises GBS polysaccharide-protein conjugates comprising a capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier 10 protein, a buffer, a surfactant, and an excipient, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular polysaccharides have a sialic acid level of greater than about 60%. In one particular aspect, the composition includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising capsular 15 polysaccharides from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, histidine, polysorbate 80, and sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In one particular aspect, the GBS composition comprises GBS polysaccharide- protein conjugates comprising capsular polysaccharides from GBS serotypes Ia, Ib, II, III, IVand V and a carrier protein, histidine, polysorbate 80, and sodium chloride, wherein the 20 composition is buffered to a pH of about 6.0 to about 7.5 and wherein the capsular polysaccharides have a sialic acid level of greater than about 60%. In a preferred aspect, a composition of the present disclosure includes the components of a GBS composition comprising GBS polysaccharide-protein conjugates comprising about 5 µg / mL to about 50 µg / mL of each capsular polysaccharide from GBS serotypes Ia, Ib, II, III, 25 IV and V and a carrier protein, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In a preferred aspect, the GBS composition comprises GBS polysaccharide-protein conjugates comprising about 5 µg / mL to about 50 µg / mL of each capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V 30 and a carrier protein, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In a preferred aspect, the composition includes the components of a GBS composition 35 comprising GBS polysaccharide-protein conjugates comprising about 40 µg / mL of each capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, about 20 mM of histidine, about 0.02% (v / w) of polysorbate 80, and about 150 mM of sodium77

[0076] chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In another preferred aspect, the composition comprises about 20 µg of each GBS capsular polysaccharide per 0.5 mL dose. In a preferred aspect, the GBS composition comprises GBS polysaccharide-protein 5 conjugate comprising about 40 µg / mL of each capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, about 20 mM of histidine, about 0.02% (v / w) of polysorbate 80, and about 150 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In another preferred aspect, the composition comprises about 2010 µg of each GBS capsular polysaccharide per 0.5 mL dose. In a preferred aspect, a composition of the present disclosure includes the components of a GBS composition comprising about 60 µg / mL, about 120 µg / mL or about 240 µg / mL total GBS polysaccharide-protein conjugate comprising capsular polysaccharides from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, about 10 mM to about 25 mM of histidine, 15 about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In a preferred aspect, the GBS composition comprising about 60 µg / mL, about 120 µg / mL or about 240 µg / mL total GBS polysaccharide-protein conjugate comprising capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, about 10 mM to about 25 mM 20 of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular polysaccharide has a sialic acid level of greater than about 60%. In a preferred aspect, a composition of the present disclosure includes the components of a GBS composition comprising about 240 µg / mL total GBS polysaccharide-protein25 conjugate comprising capsular polysaccharides from GBS serotypes Ia, Ib, II, III, IV and V anda carrier protein, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5. In another preferred aspect, the composition comprises a total conjugate dose of about 120 µg per 0.5 mL dose. 30 In a preferred aspect, the GBS composition comprising about 240 µg / mL total GBS polysaccharide-protein conjugate comprising capsular polysaccharide from GBS serotypes Ia, Ib, II, III, IV and V and a carrier protein, about 10 mM to about 25 mM of histidine, about 0.01% to about 0.03% (v / w) of polysorbate 80, and about 10 mM to about 250 mM of sodium chloride, wherein the composition is buffered to a pH of about 6.0 to about 7.5, and wherein the capsular 35 polysaccharide has a sialic acid level of greater than about 60%. In another preferred aspect, the composition comprises a total conjugate dose of about 120 µg per 0.5 mL dose. 78

[0077] In some aspects, a composition of the present disclosure includes the components of an immunogenic GBS composition as described herein. In some aspects, a composition of the present disclosure includes the components of GBS6 as described in Table 1 herein.5 In a preferred aspect, the GBS composition is in an aqueous or liquid form. In another aspect, the GBS composition is lyophilized. RSV Compositions The present disclosure provides for a composition that includes components of an RSV composition described herein. 10 In one aspect, a composition of the present disclosure includes the components of an RSV composition comprising a F1 polypeptide and F2 polypeptide derived from the F proteinof RSV subtype B. In some aspects, the F1 polypeptide and F2 polypeptide in the composition are derived from the F protein of RSV subtype A. In some aspects, the composition comprises a first RSV F protein comprising a F1 polypeptide and F2 polypeptide from the F protein of 15 RSV subtype A, and a second RSV F protein comprising a F1 polypeptide and F2 polypeptide from the F protein of RSV subtype B. In some aspects, the composition is a vaccine. In some particular aspects, the composition comprises an RSV F protein mutant,wherein the mutant comprises a cysteine (C) at position 103 (103C) and at position 148 (148C), an isoleucine (I) at position 190 (190I), and a serine (S) at position 486 (486S).20 In another particular aspect, the composition comprises an RSV F mutant, wherein themutant comprises a histidine (H) at position 54, a cysteine (C) at positions 103 and 148, an isoleucine (I) at positions 190 and 296, and a serine (S) at position 486. In another particular aspect, the composition comprises an RSV F mutant, wherein themutant comprises a histidine (H) at position 54, a cysteine (C) at positions 55 and 188, and a25 serine (S) at position 486. In another particular aspect, the composition comprises an RSV F mutant, wherein themutant comprises a histidine (H) at position 54, a cysteine (C) at positions 55 and 188, an isoleucine (I) at position 190 (190I), and a serine (S) at position 486. In another particular aspect, the composition comprises an RSV F mutant, wherein30 each mutant comprises a cysteine (C) at positions 155 (155C) and 290 (290C), a phenylalanine (F) at position 190 (190F) and a leucine (L) at position 207 (207L). In some aspects, the composition comprises an RSV F protein mutant comprises a F1and / or a F2 polypeptide set forth in any one of SEQ ID NOS: 41-80, 279-326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties.35 For example, the composition may comprises an RSV F mutant comprising a F1 polypeptideand a F2 polypeptide selected from the group consisting of: a F2 polypeptide comprising the amino acid sequence of SEQ ID NO: 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61, 63, 65, 67, 69, 79

[0078] 71, 73, 75, 77, 79, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, or 325; and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72,74, 76, 78, 80, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 5 310, 312, 314, 316, 318, 320, 322, 324, or 326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties. In some particular aspects, the composition comprises an RSV F protein mutantcomprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid10 sequence of SEQ ID NO: 18. In another particular aspect, the composition comprises an RSV Fprotein mutant comprising a F1 polypeptide and a F2 polypeptide, wherein the F2polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain).15 In some particular aspects, the composition comprises an RSV F protein mutantcomprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In another particular aspect, the composition comprises an RSV Fprotein mutant comprising a F1 polypeptide and a F2 polypeptide, wherein the F220 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). In some aspects, the composition comprises the RSV F mutant linked to a foldon domain, wherein the foldon domain is linked to the C-terminus of the F1 polypeptide and 25 comprises the amino acid sequence of SEQ ID NO: 23. In some aspects, a composition of the present disclosure includes the components of an RSV composition comprising two or more different RSV F mutants. The two or moredifferent RSV F mutants may comprise the same introduced amino acid mutations butcomprise a F1 polypeptide and F2 polypeptide from different RSV strains or subtypes, such 30 as RSV subtype A and RSV subtype B. The two or more different RSV F mutants may comprise different introduced amino acid mutations. In some aspects, the composition includes the components of an RSV compositioncomprising two different mutants comprising the same introduced amino acid mutations, wherein one of the mutants comprises a F1 polypeptide and F2 polypeptide from RSV subtype35 A and wherein the other mutant comprises a F1 polypeptide and F2 polypeptide from RSV subtype B. In some specific aspects, the two different mutants comprise the same combination of amino acid substitutions selected from the group consisting of: 80

[0079] (1) a combination of amino acid substitutions 103C, 148C, 190I, and 486S; (2) a combination of amino acid substitutions 54H, 55C, 188C, and 486S; (3) a combination of amino acid substitutions 54H, 103C, 148C, 190I, 296I, and 486S; (4) a combination of amino acid substitutions 54H, 55C, 142C, 188C, 296I, and 371C; 5 (5) a combination of amino acid substitutions 55C, 188C, and 486S; (6) a combination of amino acid substitutions 54H, 55C, 188C, and 190I ; (7) a combination of amino acid substitutions 55C, 188C, 190I, and 486S;(8) a combination of amino acid substitutions 54H, 55C, 188C, 190I, and 486S;(9) a combination of amino acid substitutions 155C, 190I, 290C, and 486S; 10 (10) a combination of amino acid substitutions 54H, 55C, 142C, 188C, 296I, 371C, 486S, 487Q, and 489S; (11) a combination of amino acid substitutions 54H, 155C, 190I, 290C, and 296I; and(12) combination of 155C, 190F, 290C, and 207L. In some particular aspects, the composition includes the components of an RSV 15 composition comprising a first and second RSV F protein mutant, wherein each mutant comprises a cysteine (C) at position 103 (103C) and at position 148 (148C), an isoleucine (I) at position 190 (190I), and a serine (S) at position 486 (486S). In another particular aspect, the composition includes the components of an RSV composition comprising a first and second RSV F mutant, wherein each mutant comprises a20 histidine (H) at position 54, a cysteine (C) at positions 103 and 148, an isoleucine (I) atpositions 190 and 296, and a serine (S) at position 486. In another particular aspect, the composition includes the components of an RSV composition comprising a first and second RSV F mutant, wherein each mutant comprises a histidine (H) at position 54, a cysteine (C) at positions 55 and 188, and a serine (S) at position25 486. In another particular aspect, the composition includes the components of an RSV composition comprising a first and second RSV F mutant, wherein each mutant comprises a histidine (H) at position 54, a cysteine (C) at positions 55 and 188, an isoleucine (I) at position 190 (190I), and a serine (S) at position 486. 30 In another particular aspect, the composition includes the components of an RSV composition comprising a first and second RSV F mutant, wherein each mutant comprises a cysteine (C) at positions 155 (155C) and 290 (290C), a phenylalanine (F) at position 190 (190F) and a leucine (L) at position 207 (207L). In some aspects, the composition includes the components of an RSV composition35 comprising a first and second RSV F protein mutant each comprising a F1 and a F2polypeptide set forth in any one of SEQ ID NOS: 41-80, 279-326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties. For example, the 81

[0080] composition may comprises a first and second RSV F mutant each comprising a F1 polypeptide and a F2 polypeptide selected from the group consisting of: a F2 polypeptide comprising the amino acid sequence of SEQ ID NO: 41, 43, 45, 47, 49, 51, 53, 55, 57, 59, 61,63, 65, 67, 69, 71, 73, 75, 77, 79, 279, 281, 283, 285, 287, 289, 291, 293, 295, 297, 299, 301, 5 303, 305, 307, 309, 311, 313, 315, 317, 319, 321, 323, or 325; and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 42, 44, 46, 48, 50, 52, 54, 56, 58, 60, 62, 64, 66, 68, 70, 72, 74, 76, 78, 80, 280, 282, 284, 286, 288, 290, 292, 294, 296, 298, 300, 302, 304, 306, 308, 310, 312, 314, 316, 318, 320, 322, 324, or 326 of WO2017109629, which sequences are hereby incorporated by reference in their entireties. 10 In some particular aspects, a composition of the present disclosure includes the components of an RSV composition comprising a first RSV F protein mutant comprising a F1polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18. In another particular aspect, the composition comprises a first RSV F protein15 mutant comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptidecomprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain). In some particular aspects, a composition of the present disclosure includes the components of an RSV composition comprising a second RSV F protein mutant comprising a20 F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In another particular aspect, the composition comprises a second RSV F protein mutant comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptidecomprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the 25 amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). In a preferred aspect, a composition of the present disclosure includes the components of an RSV composition comprising a first RSV F protein mutant comprising a F1 polypeptideand a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18, and30 a second RSV F protein mutant comprising a F1 polypeptide and a F2 polypeptide, whereinthe F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some aspects, the composition comprises a first and a second RSV F mutant linked to a foldon domain, wherein the foldon domain is linked to the C-terminus of the F1 polypeptide and comprises the amino 35 acid sequence of SEQ ID NO: 23. In another preferred aspect, a composition of the present disclosure includes the components of an RSV composition comprising a first RSV F protein mutant comprising a F182

[0081] polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain), and a second RSV F protein mutant comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide5 comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). In a preferred aspect, a composition of the present disclosure includes the components of an RSV composition comprising a first RSV F protein mutant comprising a F2 polypeptideand F1 polypeptide from the F protein of RSV subtype A comprising the amino acid sequence 10 of SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 18, respectively, and a second RSV F protein mutant comprising a F2 polypeptide and F1 polypeptide from the F protein of RSV subtype B comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 21, respectively, wherein the C-terminus of the F1 polypeptide of the first and second RSV F protein mutant is linked to a trimerization domain, wherein the 15 trimerization domain is a phage T4 foldon domain, such as the amino acid sequence of SEQ ID NO: 23. In another preferred aspect, a composition of the present disclosure includes the components of an RSV composition comprising a first RSV F protein mutant comprising a F2polypeptide and F1 polypeptide from the F protein of RSV subtype A comprising the amino 20 acid sequence of SEQ ID NO: 17 and the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain), respectively, and a second RSV F protein mutant comprising a F2 polypeptide and F1 polypeptide from the F protein of RSV subtype B comprising the amino acid sequence of SEQ ID NO: 20 and the amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). 25 The present disclosure provides for compositions including the components of an RSV composition comprising any RSV F protein as described herein, a sugar, a surfactant, a buffer and an excipient. In one aspect, the sugar is at least one of sucrose, mannitol and glycine at a concentration of between about 5 mg / mL and about 100 mg / mL. 30 In one aspect, the composition comprises sucrose at a concentration of between about 10 mg / mL and about 100 mg / mL, such as about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mg / mL; between about 10 mg / mL and about 90 mg / mL; between about 10 mg / mL and about 50 mg / mL; about 22.5 mg / mL; about 30 mg / mL; about 60 mg / mL; or about 90 mg / mL. In a preferred aspect, the composition comprises about 22.5 mg / mL of sucrose. In another35 preferred aspect, the composition comprises about 11.3 mg of sucrose per 0.5 mL. 83

[0082] In one aspect, the composition comprises between about 1% and about 9% sucrose; between about 1% and about 5% sucrose; about 2%; about 3%; or between about 2% and about 3% sucrose;. In a preferred aspect, the composition comprises about 2.25% sucrose. In one aspect, the composition comprises glycine at a concentration of between about 5 10 mg / mL and about 100 mg / mL, such as about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 100 mg / mL; between about 10 mg / mL and about 90 mg / mL; between about 10 mg / mL and about 50 mg / mL; about 22.5 mg / mL; about 30 mg / mL; about 60 mg / mL; or about 90 mg / mL. In one aspect, the composition comprises mannitol at a concentration of between about 10 mg / mL and about 100 mg / mL, such as about 5, 10, 20, 30, 40, 50, 60, 70, 80, 90 or 10 100 mg / mL; between about 10 mg / mL and about 90 mg / mL; between about 10 mg / mL and about 50 mg / mL; about 22.5 mg / mL; about 30 mg / mL; about 45 mg / mL; about 60 mg / mL; or about 90 mg / mL. In a preferred aspect, the composition comprises about 45 mg / mL of mannitol. In another preferred aspect, the composition comprises about 22.5 mg of mannitol per 0.5 mL. 15 In one aspect, the composition comprises between about 1% and about 9% mannitol; between about 1% and about 5% of mannitol; about 2%; about 3%; about 4%; about 5%; or between about 4% and 5% of mannitol. In a preferred aspect, the composition comprises about 4.5% mannitol. In one aspect, sucrose is at a concentration of between about 10 mg / mL and about 70 20 mg / mL and mannitol is at a concentration of between about 10 mg / mL and about 70 mg / mL. In one aspect, sucrose is at a concentration of about 30 mg / mL and mannitol is at aconcentration of about 60 mg / mL. In a preferred aspect, sucrose is at a concentration of about 22.5 mg / mL and mannitol is at a concentration of about 45 mg / mL. In another preferred aspect, sucrose is at a concentration of about 11.3 mg and mannitol is at a concentration of about 22.525 mg per 0.5 mL. In another aspect, sucrose is at a concentration of between about 10 mg / mL and about 70 mg / mL and glycine is at a concentration of between about 10 mg / mL and about 70 mg / mL.In one aspect, sucrose is at a concentration of about 30 mg / mL and glycine is at a concentration of about 60 mg / mL. In one aspect, sucrose is at a concentration of about 22.530 mg / mL and glycine is at a concentration of about 45 mg / mL. In another aspect, mannitol is at a concentration of between about 10 mg / mL and about 70 mg / mL and glycine is at a concentration of between about 10 mg / mL and 70 mg / mL. In one aspect, mannitol is at a concentration of about 30 mg / mL and glycine is at a concentration of about 60 mg / mL. In one aspect, mannitol is at a concentration of about 22.5 mg / mL and glycine35 is at a concentration of about 45 mg / mL. In one aspect, the surfactant is at least one of polysorbate 20 (TweenTM20), polysorbate 40 (TweenTM40), polysorbate 60 (TweenTM60), polysorbate 65 (TweenTM65), 84

[0083] polysorbate 80 (TweenTM80), polysorbate 85 (TweenTM85), TritonTMN-101, TritonTMX-100, oxtoxynol 40, nonoxynol-9, triethanolamine, triethanolamine polypeptide oleate, polyoxyethylene-660 hydroxystearate (PEG-15, SolutolTMH15), polyoxyethylene-35- ricinoleate (Cremophor ELTM), soy lecithin, poloxamer, hexadecylamine, octadecylamine, 5 octadecyl amino acid esters, lysolecithin, dimethyl-dioctadecylammonium bromide, methoxyhexadecylgylcerol, pluronic polyols, polyamines (e.g., pyran, dextransulfate, poly IC, carbopol), peptides (e.g., muramyl peptide and dipeptide, dimethylglycine, tuftsin), oil emulsions, mineral gels (e.g., aluminum phosphate) and immune stimulating complexes (ISCOMS). In one aspect, the surfactant is a polysorbate. In one aspect, the surfactant is 10 polysorbate 20 or polysorbate 80. In a preferred aspect, the composition comprises polysorbate 80. In one aspect, the concentration of the surfactant is from about 0.01 mg / ml to about 10 mg / ml, from about 0.01 mg / ml to about 5.0 mg / ml, from about 0.01 mg / ml to about 2.0 mg / ml, from about 0.01 mg / ml to about 1.0 mg / ml, from about 0.1 mg / ml to about 1.0 mg / ml, 15 from about 0.1 mg / ml to about 0.5 mg / ml, from about 0.1 mg / ml to about 0.3 mg / ml or from about 0.1 mg / ml to about 0.25 mg / ml. In one aspect, the concentration of the surfactant is about 0.05, 0.06, 0.07, 0.08, 0.09, 0.10, 0.11, 0.12, 0.13, 0.14, 0.15, 0.16, 0.17, 0.18, 0.19, 0.20, 0.21, 0.22, 0.23, 0.24, 0.25, 0.26, 0.27, 0.28, 0.29 or 0.30 mg / mL. In one aspect, the concentration of the surfactant is about 0.15 mg / mL. In another aspect, the concentration of20 the surfactant is about 0.20 mg / mL. In a preferred aspect, the composition comprises between about 0.15 mg / mL and 0.20 mg / mL polysorbate 80. In an aspect, the composition comprises about 0.15 mg / mL polysorbate 80. In another aspect, the composition comprises about 0.20 mg / mL polysorbate 80. In another preferred aspect, the composition comprises about 0.08 mg of polysorbate 8025 per 0.5 mL. In one aspect, the composition comprises between about 0.01% and about 1% polysorbate 80; between about 0.01% and about 0.05% of polysorbate 80; about 0.015%; about 0.02%; about 0.025%; about 0.03%; about 0.04%; about 0.05% sucrose; or between about between about 0.01% and about 0.02% of polysorbate 80. In a preferred aspect, the30 composition comprises about 0.015% of polysorbate 80. In one aspect, the buffer is selected from histidine, phosphate, phosphoric acid, ascorbate, maleic acid, glycine, ascorbic acid, bicarbonate and carbonic acid, gluconate, edetate, malate, imidazole, Tris, phosphate, and mixtures thereof. In an aspect, the buffer is histidine or Tris. In a preferred aspect, the buffer is Tris (tris(hydroxymethyl) aminomethane). 35 In one aspect, the concentration of the buffer is between about 0.5 mM and about 50 mM, preferably about 5 mM to about 40 mM, more preferably about 10 mM to about 30 mM, increasingly preferably about 15 to about 25 mM. Preferably, the concentration of the buffer is 85

[0084] about 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM or 30 mM. In one aspect, the concentration of the buffer is about 15 mM. In another aspect, the concentration of the buffer is about 20 mM. In a preferred aspect, the composition comprises between about 10 mM to about 305 mM Tris. In an aspect, the composition comprises about 20 mM Tris. In a preferred aspect, the composition comprises about 15 mM Tris. In another preferred aspect, the compositioncomprises about 0.11 mg tromethamine and about 1.04 mg tromethamine hydrochloride per0.5 mL. In one aspect, the pH of the RSV composition is about 7.0, 7.1, 7.2, 7.3, 7.4, 7.5, 7.6, 10 7.7, 7.8, 7.9 or 8.0 or is between about 7.1 and about 7.7, or is about 7.1, 7.2, 7.3, 7.4, 7.5, 7.6 or 7.7, more preferably 7.4. In one aspect, the excipient is sodium chloride at a concentration of between about 20mM and about 250 mM. In one aspect, the sodium chloride is at a concentration of about 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, 120, 130, 140, 150, 160, 170, 180, 190, 200, 210, 220, 15 230, 240 or 250 mM. In another aspect, the sodium chloride is at a concentration of between about 20 mM and about 100 mM. In another aspect, the sodium chloride is at a concentration of between about 40mM and about 60 mM. In a preferred aspect, RSV composition comprises about 37.5 mM sodium chloride. In one aspect, the composition comprises between about 1 mg / mL and about 5 mg / mL 20 sodium chloride; between about 1 mg / mL and about 4 mg / mL sodium chloride; about 1.5 mg / mL; about 2 mg / mL; about 2.1 mg / mL; about 2.2 mg / mL; about 2.3 mg / mL; about 2.4 mg / mL; about 2.5 mg / mL; or about 3 mg / mL sodium chloride. In a preferred aspect, the composition comprises about 2.2 mg / mL sodium chloride. In another preferred aspect, the composition comprises about 1.1 mg / mL sodium chloride per 0.5 mL. 25 In the present disclosure, the RSV composition may further comprise an immunomodulatory agent, such as an adjuvant. Examples of suitable adjuvants include aluminum salts such as aluminum hydroxide and / or aluminum phosphate; oil-emulsion compositions (or oil-in-water compositions), including squalene-water emulsions, such as MF59 (see e.g., WO 90 / 14837); saponin formulations, such as, for example, QS21 and 30 Immunostimulating Complexes (ISCOMS) (see e.g., U.S. Pat. No.5,057,540; WO 90 / 03184, WO 96 / 11711, WO 2004 / 004762, WO 2005 / 002620); bacterial or microbial derivatives, examples of which are monophosphoryl lipid A (MPL), 3-O-deacylated MPL (3dMPL), CpG- motif containing oligonucleotides, ADP-ribosylating bacterial toxins or mutants thereof, such as E. coli heat labile enterotoxin LT, cholera toxin CT, and the like. It is also possible to use35 vector-encoded adjuvant, e.g., by using heterologous nucleic acid that encodes a fusion of the oligomerization domain of C4-binding protein (C4 bp) to the antigen of interest (e.g., Solabomi et al., 2008, Infect Immun 76: 3817-23). In certain aspects the compositions herein comprise 86

[0085] aluminum as an adjuvant, e.g., in the form of aluminum hydroxide, aluminum phosphate, aluminum potassium phosphate, or combinations thereof, in concentrations of 0.05-5 mg, e.g., from 0.075-1.0 mg, of aluminum content per dose. The present disclosure provides for compositions including the components of an RSV 5 composition comprising a first and a second RSV F protein as described herein, a sugar, a surfactant, a buffer and an excipient. The present disclosure provides for compositions including the components of an RSV composition comprising a first and a second RSV F protein as described herein, between about 2% and about 3% of sucrose, between about 4% and about 5% of mannitol, between10 about 0.01% and about 0.02% of polysorbate 80, between about 10 mM to about 30 mM ofTris and between about 40mM and about 60 mM of sodium chloride. In a preferred aspect, the RSV composition comprises a first and a second RSV F protein as described herein, about 2.25% of sucrose, about 4.5% of mannitol, about 0.015% of polysorbate 80, about 15 mM of Tris and about 37.5 mM of sodium chloride.15 The present disclosure provides for compositions including the components of an RSV composition comprising a first and a second RSV F protein as described herein, between about 10 mg / mL and about 50 mg / mL of sucrose, between about 10 mg / mL and about 50 mg / mL of mannitol, between about 0.15 mg / mL and 0.20 mg / mL of polysorbate 80, betweenabout 0.2 mg / mL to about 0.3 mg / mL of tromethamine, between about 2 mg / mL to about 320 mg / mL of tromethamine, and between about 2 mg / mL to about 3 mg / mL of sodium chloride.In a preferred aspect, the RSV composition comprises a first and a second RSV F protein as described herein, about 22.5 mg / mL of sucrose, about 45 mg / mL of mannitol, about 0.16 mg / mL of polysorbate 80, about 0.22 mg / mL of tromethamine, about 2.08 mg / mL oftromethamine, and about 2.2 mg / mL of sodium chloride.25 In a preferred aspect, the RSV composition comprises a first and a second RSV F protein as described herein, about 11.3 mg / mL of sucrose, about 22.5 mg / mL of mannitol, about 0.08 mg / mL of polysorbate 80, about 0.11 mg / mL of tromethamine, about 1.04 mg / mLof tromethamine, and about 1.1 mg / mL of sodium chloride per 0.5 mL dose.The present disclosure provides for compositions including the components of an RSV 30 composition comprising about 100 µg / mL to about 140 µg / mL of each of a first and a second RSV F protein as described herein. In a preferred aspect, the composition comprises 120 µg / mL of a first RSV F protein and 120 µg / mL of a second RSV F protein. In some aspects, the composition comprises 60 µg of a first RSV F protein and 60 µg of a second RSV F protein per 0.5 mL dose. 35 In some aspects, the composition comprises about 200 µg / mL to about 300 µg / mL total RSV F protein as described herein. In a preferred aspect, the composition comprises 240 87

[0086] µg / mL total RSV F protein as described herein. In some aspects, the composition comprises 120 µg total RSV F protein as described herein per 0.5 mL dose. The present disclosure provides for compositions including the components of an RSV composition comprising about 100 µg / mL to about 140 µg / mL, preferably about 120 µg / mL, of 5 each of a first and a second RSV F protein as described herein, a sugar, a surfactant, a buffer and an excipient. The present disclosure provides for compositions including the components of an RSV composition comprising about 200 µg / mL to about 300 µg / mL, preferably about 240 µg / mL, of total RSV F protein as described herein, a sugar, a surfactant, a buffer and an excipient.10 The present disclosure provides for compositions including the components of an RSV composition comprising about 100 µg / mL to about 140 µg / mL, preferably about 120 µg / mL, of each of a first and a second RSV F protein as described herein, between about 2% and about 3% of sucrose, between about 4% and about 5% of mannitol, between about 0.01% and about 0.02% of polysorbate 80, between about 10 mM to about 30 mM of Tris and between about15 40 mM and about 60 mM of sodium chloride. The present disclosure provides for compositions including the components of an RSV composition comprising about 200 µg / mL to about 300 µg / mL, preferably about 240 µg / mL, total RSV F protein as described herein, between about 2% and about 3% of sucrose, between about 4% and about 5% of mannitol, between about 0.01% and about 0.02% of polysorbate20 80, between about 10 mM to about 30 mM of Tris and between about 40mM and about 60mM of sodium chloride. The present disclosure provides for compositions including the components of an RSV composition comprising about 100 µg / mL to about 140 µg / mL, preferably about 120 µg / mL, of each of a first and a second RSV F protein as described herein, about 2.25% of sucrose, about25 4.5% of mannitol, about 0.015% of polysorbate 80, about 15 mM of Tris and about 37.5 mMof sodium chloride. The present disclosure provides for compositions including the components of an RSV composition comprising about 200 µg / mL to about 300 µg / mL, preferably about 240 µg / mL, total RSV F protein as described herein, about 2.25% of sucrose, about 4.5% of mannitol,30 about 0.015% of polysorbate 80, about 15 mM of Tris and about 37.5 mM of sodium chloride.The present disclosure provides for compositions including the components of an RSV composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV F protein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F protein comprises a F1 polypeptide and F2 polypeptide from the F protein of RSV subtype A, and the35 second RSV F protein comprises a F1 polypeptide and F2 polypeptide from the F protein ofRSV subtype B, wherein each RSV F protein comprises amino acid substitutions 103C, 148C, 190I, and 486S. 88

[0087] The present disclosure provides for compositions including the components of an RSV composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV Fprotein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F proteincomprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the 5 amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 and the second RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21. In some aspects, the first and a second RSV F proteins are linked to a foldon domain, wherein 10 the foldon domain is linked to the C-terminus of the F1 polypeptide and comprises the amino acid sequence of SEQ ID NO: 23. The present disclosure provides for compositions including the components of an RSV composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV F protein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F protein 15 comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 19 (including linker and trimerization domain) and the second RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the 20 amino acid sequence of SEQ ID NO: 22 (including linker and trimerization domain). The present disclosure provides for compositions including the components of an RSV composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV F protein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the 25 amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22, a sugar, a surfactant, a buffer and an excipient. 30 The present disclosure provides for compositions including the components of an RSV composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV Fprotein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F proteincomprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid 35 sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 89

[0088] 22, between about 2% and about 3% of sucrose, between about 4% and about 5% of mannitol, between about 0.01% and about 0.02% of polysorbate 80, between about 10 mM toabout 30 mM of Tris and between about 40mM and about 60 mM of sodium chloride. The present disclosure provides for compositions including the components of an RSV 5 composition comprising about 240 µg / mL total RSV F protein (about 120 µg / mL of a first RSV F protein and about 120 µg / mL of a second RSV F protein), wherein the first RSV F protein comprises a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprises a F1 polypeptide 10 and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22, about 2.25% of sucrose, about 4.5% of mannitol, about 0.015% of polysorbate 80, about 15 mM of Tris and about 37.5 mM of sodium chloride. In another preferred aspect, the composition comprises about 240 µg / mL total RSV F 15 protein (about 120 µg / mL of a first RSV F protein and about 120 µg / mL of a second RSV F protein),per 0.5 mL dose. In some aspects, a composition of the present disclosure includes the components of an immunogenic RSV composition as described herein. In some aspects, a composition of the present disclosure includes the components of20 RSVPreF as described in Table 1 herein.In some aspects, a composition of the present disclosure includes the components of ABRYSVOTMor AREXVYTM. The present disclosure provides for an RSV composition that is lyophilized or in aaqueous or liquid form. In a preferred aspect, the RSV composition is lyophilized. Lyophilized 25 RSV compositions are described in WO2024 / 069420, which are hereby incorporated by reference in their entirety. Such lyophilized compositions can be obtained by lyophilization of the aqueous compositions using lyophilization methods known to the skilled person or preferably using the lyophilization methods disclosed in WO2024 / 069420. Acomposition of the present disclosure includes the components of a lyophilized RSV30 composition that may be reconstituted with any suitable diluent. In certain aspects, the diluent is water for injection. In certain aspects, the diluent comprises sodium chloride. In a preferred aspect, the composition comprises a lyophilized RSV composition reconstituted with a GBS composition described herein. 35 METHODS OF USE The term "subject" refers to a mammal, bird, fish, reptile, or any other animal. The term "subject" also includes humans. The term "subject" also includes household pets. Non limiting 90

[0089] examples of household pets include: dogs, cats, pigs, rabbits, rats, mice, gerbils, hamsters, guinea pigs, ferrets, birds, snakes, lizards, fish, turtles, and frogs. The term "subject" also includes livestock animals. Non limiting examples of livestock animals include: alpaca, bison, camel, cattle, deer, pigs, horses, llamas, mules, donkeys, sheep, goats, rabbits, reindeer, yak, 5 chickens, geese, and turkeys. As used herein, "treatment" (including variations thereof, for example, "treat" or "treated") refers to any one or more of the following: (i) the prevention of infection or reinfection, as in a traditional vaccine, (ii) the reduction in the severity of or the elimination of symptoms, and (iii) the substantial or complete elimination of the pathogen or disorder in question. Hence, 10 treatment may be effected prophylactically (prior to infection) or therapeutically (following infection). In the present disclosure, prophylactic or therapeutic treatments can be used. According to a particular aspect of the present disclosure, compositions and methods areprovided which treat, including prophylactically and / or therapeutically immunize, a host animal against a microbial infection (e.g., a bacterium such as S. agalactiae and or virus such as15 RSV). The methods and uses of the present disclosure are useful for conferring prophylacticand / or therapeutic immunity to a subject. The methods of the present disclosure can also bepracticed on subjects for biomedical research applications. In an aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS in a subject by administering to the subject an effective amount of an 20 immunogenic composition described herein. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising a GBS polysaccharide-protein conjugate, and an RSV F protein.In one aspect, the invention relates to a method of eliciting and / or inducing an immune 25 response against GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and acarrier protein, and at least a first and second RSV F protein.In one aspect, the invention relates to a method of eliciting and / or inducing an immune 30 response against GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and afirst and second RSV F protein comprising amino acid substitutions 103C, 148C, 190I, and486S relative to the corresponding wild-type RSV F protein. 35 In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising 91

[0090] capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV Fprotein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptidecomprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a 5 F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS in a subject by administering to the subject an effective amount of an 10 immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM. 15 In an aspect, the present disclosure relates to a method of preventing, treating, ameliorating or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition described herein. In an aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an20 effective amount of an immunogenic composition described herein. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein. 25 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V,VI, VII, VIII and IX and a carrier protein, and at least a first and second RSV F protein.30 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and acarrier protein, and a first and second RSV F protein comprising amino acid substitutions35 described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to thecorresponding wild-type RSV F protein. 92

[0091] In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and 5 CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the 10 amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS in a subject by administering to the subject aneffective amount of an immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or15 condition associated with GBS in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM. In another aspect, the invention relates to a method of eliciting and / or inducing an immune response against RSV in a subject by administering to the subject an effective amount 20 of an immunogenic composition described herein. In one aspect, the invention relates to a method of inducing an immune response against RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein.In one aspect, the invention relates to a method of eliciting and / or inducing an immune 25 response against RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and acarrier protein, and at least a first and second RSV F protein. In one aspect, the invention relates to a method of eliciting and / or inducing an immune 30 response against RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and afirst and second RSV F protein comprising amino acid substitutions described herein, such asamino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type35 RSV F protein. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against RSV in a subject by administering to the subject an effective amount of an 93

[0092] immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV Fprotein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptidecomprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the 5 amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of eliciting and / or inducing an immune 10 response against RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM. 15 In an aspect, the present disclosure relates to a method of preventing, treating, ameliorating or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject an effective amount of an immunogenic composition described herein. In an aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject an20 effective amount of an immunogenic composition described herein. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or conditionassociated with RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein. 25 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject aneffective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first and second RSV F protein.30 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject aneffective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and acarrier protein, and a first and second RSV F protein comprising amino acid substitutions 35 described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type RSV F protein. 94

[0093] In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and 5 CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the 10 amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with RSV in a subject by administering to the subject aneffective amount of an immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or15 condition associated with RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM. In an aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS and RSV in a subject by administering to the subject an effective 20 amount of an immunogenic composition described herein. In one aspect, the invention relates to a method of inducing an immune response against GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising aGBS polysaccharide-protein conjugate and an RSV F protein.In one aspect, the invention relates to a method of eliciting and / or inducing an immune 25 response against GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first and second RSV F protein.In one aspect, the invention relates to a method of eliciting and / or inducing an immune 30 response against GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrierprotein, and a first and second RSV F protein comprising amino acid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the35 corresponding wild-type RSV F protein. In one aspect, the invention relates to a method of eliciting and / or inducing an immuneresponse against GBS and RSV in a subject by administering to the subject an effective 95

[0094] amount of an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide 5 comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of eliciting and / or inducing an immune 10 response against GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of eliciting and / or inducing an immune response against GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS6 and an RSV composition, such ABRYSVOTMor AREXVYTM. 15 In an aspect, the present disclosure relates to a method of preventing, treating, ameliorating or reducing a disease, infection or condition associated with GBS and / or RSV in a subject by administering to the subject an effective amount of an immunogenic composition described herein. In an aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject by administering to20 the subject an effective amount of an immunogenic composition described herein. In one aspect, the invention relates to a method of preventing or reducing a disease, infection orcondition associated with GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein.25 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject by administering to thesubject an effective amount of an immunogenic composition comprising GBS polysaccharide- protein conjugates comprising at least one capsular polysaccharide from serotypes Ia, Ib, II,III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first and second RSV F protein.30 In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject by administering to thesubject an effective amount of an immunogenic composition comprising GBS polysaccharide- protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising amino acid substitutions35 described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to thecorresponding wild-type RSV F protein. 96

[0095] In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject by administering to the subject an effective amount of an immunogenic composition comprising GBS polysaccharide- protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V 5 and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the 10 amino acid sequence of SEQ ID NO: 21 or 22. In one aspect, the invention relates to a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject by administering to thesubject an effective amount of an immunogenic composition comprising GBS6 and RSVPreF. In one aspect, the invention relates to a method of preventing or reducing a disease, infection15 or condition associated with GBS and RSV in a subject by administering to the subject aneffective amount of an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM. In an aspect, the invention relates to an immunogenic composition described hereinfor use as a medicament. In one aspect, the invention relates to an immunogenic composition20 comprising a GBS polysaccharide-protein conjugate and an RSV F protein for use as amedicament. In one aspect, the invention relates to the immunogenic composition described hereinfor use in a method of eliciting and / or inducing an immune response against GBS in a subject. In one aspect, the invention relates to the immunogenic composition described herein for use 25 in a method of eliciting and / or inducing an immune response against RSV in a subject. In another aspect, the invention relates to the immunogenic composition described herein for use in a method of eliciting and / or inducing an immune response against GBS and RSV in asubject. In one aspect, the invention relates to an immunogenic composition comprising a GBS30 polysaccharide-protein conjugate and an RSV F protein for use in a method of eliciting and / orinducing an immune response against GBS in a subject. In one aspect, the invention relatesto an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein for use in a method of eliciting and / or inducing an immune response againstRSV in a subject. In another aspect, the invention relates to an immunogenic composition35 comprising a GBS polysaccharide-protein conjugate and an RSV F protein for use in a methodof eliciting and / or inducing an immune response against GBS and RSV in a subject.97

[0096] In one aspect, the invention relates to an immunogenic composition comprising a GBS polysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first andsecond RSV F protein for use in a method of eliciting and / or inducing an immune response 5against GBS in a subject. In one aspect, the invention relates to an immunogenic compositioncomprising a GBS polysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, andat least a first and second RSV F protein for use in a method of eliciting and / or inducing an immune response against RSV in a subject. In another aspect, the invention relates to an 10 immunogenic composition comprising a GBS polysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and acarrier protein, and at least a first and second RSV F protein for use in a method of eliciting and / or inducing an immune response against GBS and RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS 15 polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising aminoacid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type RSV F protein for use in a method of eliciting and / or inducing an immune response against GBS in a subject. 20 In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and 25 the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of eliciting and / or inducing an immune response against GBS in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS 30 polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising aminoacid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type RSV F protein for use in a method of eliciting and / or inducing an immune response against RSV in a subject. 35 In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 98

[0097] polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide 5 comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of eliciting and / or inducing an immune response against RSV in a subject. In another aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising10 amino acid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type RSV F protein for use in a method of eliciting and / or inducing an immune response against GBS and RSV in a subject. In another aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes 15 Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide 20 comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of eliciting and / or inducing an immune response against GBS and RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS6 and RSVPreF for use in a method of eliciting and / or inducing an immune response against GBS in a subject. In one aspect, the invention relates to an immunogenic composition25 comprising GBS6 and RSVPreF for use in a method of eliciting and / or inducing an immune response against RSV in a subject. In another aspect, the invention relates to an immunogenic composition GBS6 and RSVPreF for use in a method of eliciting and / or inducing an immune response against GBS and RSV in a subject.n one aspect, the invention relates to an immunogenic composition comprising GBS6 30 and an RSV vaccine, such ABRYSVOTMor AREXVYTMfor use in a method of eliciting and / or inducing an immune response against GBS in a subject. In one aspect, the invention relatesto an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTMfor use in a method of eliciting and / or inducing an immune response against RSV in a subject. In another aspect, the invention relates to an immunogenic composition GBS6 35 and an RSV vaccine, such ABRYSVOTMor AREXVYTMfor use in a method of eliciting and / or inducing an immune response against GBS and RSV in a subject. 99

[0098] In one aspect, the invention relates to the immunogenic composition described herein for use in a method of preventing or reducing a disease, infection or condition associated withGBS in a subject. In one aspect, the invention relates to the immunogenic compositiondescribed herein for use in a method of preventing or reducing a disease, infection or condition5 associated with RSV in a subject. In another aspect, the invention relates to the immunogenic composition described herein for use in a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject.In one aspect, the invention relates to an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein for use in a method of preventing or10 reducing a disease, infection or condition associated with GBS in a subject. In one aspect, theinvention relates to an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an RSV F protein for use in a method of preventing or reducing a disease,infection or condition associated with RSV in a subject. In another aspect, the invention relates to an immunogenic composition comprising a GBS polysaccharide-protein conjugate and an15 RSV F protein for use in a method of preventing or reducing a disease, infection or conditionassociated with GBS and RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising a GBS polysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first and20 second RSV F protein for use in a method of preventing or reducing a disease, infection or condition associated with GBS in a subject. In one aspect, the invention relates to animmunogenic composition comprising a GBS polysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and acarrier protein, and at least a first and second RSV F protein for use in a method of preventing 25 or reducing a disease, infection or condition associated with RSV in a subject. In another aspect, the invention relates to an immunogenic composition comprising a GBSpolysaccharide-protein conjugate comprising at least one capsular polysaccharide from serotypes Ia, Ib, II, III, IV, V, VI, VII, VIII and IX and a carrier protein, and at least a first andsecond RSV F protein for use in a method of preventing or reducing a disease, infection or30 condition associated with GBS and RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising aminoacid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 35 486S relative to the corresponding wild-type RSV F protein for use in a method of preventing or reducing a disease, infection or condition associated with GBS in a subject.100

[0099] In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 5 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of preventing or reducing a disease, infection or condition associated with GBS in a subject. 10 In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprising aminoacid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I, and 486S relative to the corresponding wild-type RSV F protein for use in a method of preventing 15 or reducing a disease, infection or condition associated with RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of preventingor reducing a disease, infection or condition associated with RSV in a subject. 25 In another aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and a carrier protein, and a first and second RSV F protein comprisingamino acid substitutions described herein, such as amino acid substitutions 103C, 148C, 190I,and 486S relative to the corresponding wild-type RSV F protein for use in a method of 30 preventing or reducing a disease, infection or condition associated with against GBS and RSV in a subject. In another aspect, the invention relates to an immunogenic composition comprising GBS polysaccharide-protein conjugates comprising capsular polysaccharides from serotypes Ia, Ib, II, III, IV and V and CRM197, and a first RSV F protein comprising a F1 polypeptide and35 a F2 polypeptide, wherein the F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 17 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 18 or 19 and the second RSV F protein comprising a F1 polypeptide and a F2 polypeptide, wherein the 101

[0100] F2 polypeptide comprises the amino acid sequence of SEQ ID NO: 20 and the F1 polypeptide comprises the amino acid sequence of SEQ ID NO: 21 or 22 for use in a method of preventingor reducing a disease, infection or condition associated with against GBS and RSV in a subject. 5 In one aspect, the invention relates to an immunogenic composition comprising GBS6 and RSVPreF for use in a method of preventing or reducing a disease, infection or condition associated with GBS in a subject. In one aspect, the invention relates to an immunogeniccomposition comprising GBS6 and RSVPreF for use in a method of preventing or reducing a disease, infection or condition associated with RSV in a subject. In another aspect, the 10 invention relates to an immunogenic composition GBS6 and RSVPreF for use in a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject. In one aspect, the invention relates to an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTMfor use in a method of preventing or15 reducing a disease, infection or condition associated with GBS in a subject. In one aspect, theinvention relates to an immunogenic composition comprising GBS6 and an RSV vaccine, such ABRYSVOTM or AREXVYTM for use in a m method of preventing or reducing a disease,infection or condition associated with RSV in a subject. In another aspect, the invention relates to an immunogenic composition GBS6 and an RSV vaccine, such ABRYSVOTMor AREXVYTM20 for use in a method of preventing or reducing a disease, infection or condition associated with GBS and RSV in a subject. In one aspect, the immune response elicited against GBS is comparable to the immuneresponse elicited against GBS after administering an immunogenic composition comprising GBS polysaccharide-protein conjugate alone (without RSV F proteins). 25 In one aspect, the immune response elicited comprises anti-GBS capsular polysaccharide IgG antibodies. In a particular aspect of the methods provided herein, the subject is a human.In a particular aspect of the methods and uses provided herein, the subject is a femaleplanning to become pregnant or a pregnant female. In one such aspect, the pregnant female30 is in her second half of pregnancy, such as at least 20 weeks, at least 27 weeks or at least 32weeks gestation. In a preferred aspect, the pregnant female is at 27 weeks to 36 weeks gestation. In another preferred aspect, the pregnant female is at 20 weeks to 36 weeksgestation. In a preferred aspect, the pregnant female is at 32 weeks to 36 weeks gestation.In some particular aspects, the subject is a child, such as an infant.35 In another aspect, the subject is an older adult, such as an adult 50 years of age or older, 65 years of age or older, and 85 years of age or older. In a further aspect, the subject is immunocompromised. In one aspect, the subject may have a medical condition selected102

[0101] from the group consisting of obesity, diabetes, HIV infection, cancer, cardiovascular disease, or liver disease. Subjects that may be selected for treatment include those that are at risk for developingan GBS and / or RSV infection because of exposure or the possibility of exposure to GBS 5and / or RSV. The entire birth cohort is included as a relevant population for immunization,which could be done by beginning an immunization regimen anytime from birth to 6 months ofage, from 6 months of age to 5 years of age, in pregnant women (or women of child-bearing age) to protect their infants by passive transfer of antibody, family members of newborn infants or those still in utero, and subjects greater than 50 years of age.10 An immunogenic or effective amount of the composition administered to the subject isan amount that is sufficient to elicit an immune response against GBS and / or RSV. Animmunogenic or effective amount of a composition can be determined by doing a doseresponse study in which subjects are immunized with gradually increasing amounts of the immunogenic composition and the immune response analyzed to determine the optimal 15 dosage. Starting points for the study can be inferred from immunization data in animal models. The dosage amount can vary depending upon specific conditions of the individual. The amount can be determined in routine trials by means known to those skilled in the art. An immunologically effective amount of the composition in an appropriate number of doses is administered to the subject to elicit an immune response. The dosage amount can 20 vary depending upon specific conditions of the individual, such as age and weight. This amount can be determined in routine trials by means known to those skilled in the art. Administration of the compositions provided by the present disclosure, such as pharmaceutical compositions, can be carried out using standard routes of administration. Non- limiting aspects include parenteral administration, such as intradermal, intramuscular, 25 subcutaneous, transcutaneous, mucosal, or oral administration. The total dose of the composition provided to a subject during one administration can be varied as is known to the skilled practitioner. It is also possible to provide one or more booster administrations of one or more of the vaccine compositions. If a boosting vaccination is performed, typically, such a boosting 30 vaccination will be administered to the same subject at a moment between one week and 10 years, preferably between two weeks and six months, after administering the composition to the subject for the first time (which is in such cases referred to as "priming vaccination"). In certain aspects, the administration comprises a priming administration and at least one booster administration. In certain other aspects, the administration is provided annually.35 In a preferred aspect, the compositions described herein are a single pharmaceuticaldosage formulation comprising components of a GBS composition and an RSV composition,e.g., co-administration. In another aspect the GBS composition and the RSV composition are 103

[0102] separate pharmaceutical dosage formulations. When separate dosage formulations are used,the GBS and RSV compositions can be administered at essentially the same time, e.g., concurrently, or at separately staggered times, e.g., sequentially. The present disclosure further provides for methods of administering a composition 5 comprising a lyophilized RSV F protein or lyophilized RNA polynucleotide encoding a RSV protein composition described herein reconstituted with a liquid GBS polysaccharide-protein conjugate composition described. In another aspect, the present disclosure provides formethods of administering a composition comprising a lyophilized RSV F protein or lyophilized RNA polynucleotide encoding a RSV protein composition described herein reconstituted with10 a diluent or buffer and combined or mixed with a liquid GBS polysaccharide-protein conjugatecomposition described herein. In a further aspect, the present disclosure provides for methods of administering a composition comprising a lyophilized GBS polysaccharide-protein conjugate composition described herein reconstituted with a liquid RSV F protein or liquid RNA polynucleotide15 encoding a RSV protein composition described herein. In another aspect, the presentdisclosure provides for methods of administering a composition comprising a lyophilized GBS polysaccharide-protein conjugate composition described herein reconstituted with a diluent or buffer and combined or mixed with a liquid RSV F protein or liquid RNA polynucleotide encoding a RSV protein composition described herein. 20 In another aspect, the present disclosure provides for methods of administering a composition comprising a liquid RSV F protein or RNA polynucleotide encoding a RSV proteincomposition described herein and combined or mixed with a liquid GBS polysaccharide-protein conjugate composition described herein in a liquid.In yet another aspect, the present disclosure provides for methods of administering a25 composition comprising a lyophilized RSV F protein or lyophilized RNA polynucleotideencoding a RSV protein composition described herein reconstituted with a diluent or bufferand combined or mixed with a lyophilized GBS polysaccharide-protein conjugate compositiondescribed herein reconstituted with a diluent or buffer . The present disclosure further provides for a method of eliciting an immune response30 against GBS and / or RSV in a subject, the method comprising administering to the subject acomposition comprising a lyophilized RSV F protein or lyophilized RNA polynucleotide encoding a RSV protein composition described herein reconstituted with a liquid GBS polysaccharide-protein conjugate composition described herein. In another aspect, the present disclosure provides for a method of eliciting an immune response against GBS and / or35 RSV in a subject, the method comprising administering to the subject a compositioncomprising a lyophilized RSV F protein or lyoph...

Claims

WHAT IS CLAIMED IS:

1. A composition comprising a group B streptococcus (GBS) polysaccharide-proteinconjugate comprising a capsular polysaccharide and a carrier protein; and at least a first 5 and second respiratory syncytial virus (RSV) F protein.

2. The composition of claim 1, wherein the conjugate comprises a GBS capsularpolysaccharide selected from serotypes la, Ib, II, III, IV, V, Vl, Vll, VIll, and IX.

3. The composition of claims 1 or 2, wherein the conjugates comprise GBS capsularpolysaccharides from serotypes la, Ib, and III; serotypes la, Ib, III and V; serotypes la, Ib,10 II, III and V; serotypes la, Ib, II, III, V and VII; or serotypes la, Ib, II, III, IV, and V, andoptionally wherein the conjugates further comprise at least one GBS capsular polysaccharide from serotypes VI, VII, VIII and IX.

4. The composition of any one of claims 1-3, wherein the carrier protein is CRM197, tetanustoxoid, or SCP.15 5. The composition of any one of claims 1-4, wherein the composition comprises about 5µg / mL to about 50 µg / mL of each GBS capsular polysaccharide, optionally wherein the composition comprises at least about 40 µg / mL of each GBS capsular polysaccharide, and further optionally wherein the composition comprises at least about 20 µg of each GBScapsular polysaccharide per 0.5 mL dose.20 6. The composition of any one of claims 1-5, wherein the composition comprises about 60µg / mL to about 360 µg / mL total GBS polysaccharide-protein conjugate, optionally wherein the composition comprises at least about 240 µg / mL total GBS polysaccharide-proteinconjugate, and further optionally wherein the composition comprises at least about 120 µg total GBS polysaccharide-protein conjugate per 0.5 mL dose.25 7. The composition of any one of claims 1-6, wherein the first and second RSV F proteinscomprise a pair of cysteine mutations selected from the group consisting of: (1) 55C and 188C; (2) 103C and 148C; (3) 142C and 371C; and (4) 155C and 290C, wherein amino acid positions are numbered according to SEQ ID NO:

1.

8. The composition of any one of claims 1-7, wherein the first RSV F protein is a F protein of30 subtype A, and / or the second RSV F protein is a F protein of subtype B.1429. The composition of any one of claims 1-8, wherein the first RSV F protein comprises a F2polypeptide comprising the amino acid sequence of SEQ ID NO: 17 and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 18 or 19.

10. The composition of any one of claims 1-9, wherein the second RSV F protein comprises5 a F2 polypeptide comprising the amino acid sequence of SEQ ID NO: 20 and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 21 or 22.

11. The composition of any one of claims 1-10, wherein the composition comprises about 100µg / mL to about 140 µg / mL of each RSV F protein, optionally wherein the composition comprises about 120 µg / mL of each RSV F protein, and further optionally wherein the10 composition comprises about 60 µg of each RSV F protein per 0.5 mL dose.

12. The composition of any one of claims 1-11, wherein the composition comprises about 200µg / mL to about 300 µg / mL total RSV F protein, optionally wherein the composition comprises about 240 µg / mL total RSV F protein, and further optionally wherein the composition comprises about 120 µg total RSV F protein per 0.5 mL dose.15 13. The composition of any one of claims 1-12, wherein the RSV F proteins are lyophilizedand reconstituted with the GBS polysaccharide-protein conjugate in a liquid form 14. A composition comprising: (i) a liquid composition comprising group B streptococcus(GBS) polysaccharide-protein conjugates comprising capsular polysaccharides fromserotypes la, Ib, II, III, IV, and V and a carrier protein; and (ii) a lyophilized composition20 comprising a first respiratory syncytial virus (RSV) F protein comprising a F2 polypeptidecomprising the amino acid sequence of SEQ ID NO: 17 and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 18 or 19; and second RSV F protein comprisinga F2 polypeptide comprising the amino acid sequence of SEQ ID NO: 20 and a F1 polypeptide comprising the amino acid sequence of SEQ ID NO: 21 or 22; wherein the25 lyophilized composition is reconstituted with the liquid composition.

15. The composition of any one of claims 1-14, wherein the composition further comprisessodium chloride, a surfactant, a buffer and at least one of sucrose, mannitol and glycine, and optionally wherein the composition further comprises sodium chloride, polysorbate 80,histidine, Tris, sucrose and mannitol.30 16. The composition of any one of claims 1-15, wherein the RSV F proteins maintain stabilityfor at least 4 hours in the composition when compared to the stability of the RSV F proteins alone in a composition (without GBS polysaccharide-protein conjugate) and / or the GBS143polysaccharide-protein conjugate maintains stability for at least 4 hours in the compositionwhen compared to the stability of the GBS polysaccharide-protein conjugates alone in a composition (without RSV F proteins).

17. A method of eliciting an immune response against group B streptococcus (GBS) and / or5 respiratory syncytial virus (RSV) in a subject, the method comprising administering to the subject a composition of any one of claims 1-16.

18. A method of eliciting an immune response against group B streptococcus (GBS) in asubject, the method comprising administering to the subject a composition of any one of claims 1-16.10 19. The method of claim 17 or 18, wherein the immune response elicited against GBS iscomparable to the immune response elicited against GBS after administering acomposition comprising GBS polysaccharide-protein conjugate alone (without RSV F proteins).

20. The method of any one of claims 17-19, wherein the immune response elicited comprises15 geometric mean IgG concentrations comparable to geometric mean IgG concentrations elicited after administering a composition comprising GBS polysaccharide-protein conjugate alone (without RSV F proteins) and / or the immune response elicited comprisesgeometric mean OPA titers comparable to geometric mean OPA titers elicited after administering a composition comprising GBS polysaccharide-protein conjugate alone20 (without RSV F proteins).

21. A method of eliciting an immune response against respiratory syncytial virus (RSV) in asubject, the method comprising administering to the subject a composition of any one of claims 1-16.

22. The method of claim 17 or 21, wherein the immune response elicited against RSV is25 comparable to the immune response elicited against RSV after administering a composition comprising RSV F protein alone (without GBS polysaccharide-protein conjugates).

23. The method of any one of claims 17 or 21-22, wherein the immune response elicitedcomprises geometric mean IgG concentrations comparable to geometric mean IgG 30 concentrations elicited after administering a composition comprising RSV F proteins (without GBS polysaccharide-protein conjugate) and / or the immune response elicitedcomprises geometric mean neutralization titers comparable to geometric mean 144neutralization titers elicited after administering a composition comprising RSV F proteinsalone (without GBS polysaccharide-protein conjugate).

24. A method of preventing or reducing a disease, infection or condition associated with GBSand / or RSV in a subject, the method comprising administering to the subject the 5 composition according to any one of claims 1-16.

25. The method of any one of claims 17-24, wherein the subject is less than about 1 year ofage, about 5 years of age or older, about 10 years of age or older, about 18 years of age or older, about 20 years of age or older, about 25 years of age or older, about 30 years of age or older, about 35 years of age or older, about 40 years of age or older, about 45 years 10 of age or older, about 50 years of age or older, about 55 years of age or older, about 60 years of age or older, about 65 years of age or older, about 70 years of age or older, about 75 years of age or older, about 80 years of age or older, about 85 years of age or older.

26. The method of any one of claims 17-25, wherein the subject is pregnant or of child-bearingage.15 27. The method of any one of claims 17-25, wherein the subject is 60 years of age or older.

28. A kit comprising:(i) a first composition comprising a first and second respiratory syncytial virus (RSV) Fprotein, and (ii) a second composition comprising a group B streptococcus (GBS) polysaccharide-20 protein conjugate wherein combining the first composition with the second composition results in a composition of any one of claims 1-16.

29. A kit comprising:(i) a lyophilized composition comprising a first and second respiratory syncytial virus25 (RSV) F protein, and (ii) a liquid composition comprising a group B streptococcus (GBS) polysaccharide- protein conjugate for reconstituting the lyophilized composition; wherein reconstituting the lyophilized composition with the liquid composition results in a composition of any one of claims 1-16.30 30. A composition comprising a GBS polysaccharide-protein conjugate comprising a capsularpolysaccharide and a carrier protein; and an RNA polynucleotide comprising at least oneopen reading frame encoding a respiratory syncytial virus (RSV) polypeptide or an145immunogenic fragment thereof, wherein each of the RNA polynucleotides is encapsulated in a lipid nanoparticle (LNP).

31. A method of eliciting an immune response against GBS and / or RSV in a subject, themethod comprising administering to the subject a composition of claim 30.5 146

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