Development of PNAG-based vaccines

Abstract

Provided herein are vaccine composition comprising poly-β-(l-6)-7N-acetylglucosamine (PNAG) antigen conjugated to capsid protein carrier, wherein said capsid comprises wild type or native sequence, or at least one mutation, and wherein the PNAG antigen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 glucosamine monomers, wherein at least one glucosamine monomer is acetylated.

Description

DEVELOPMENT OF PNAG-BASED VACCINESCross-Reference to Related ApplicationsThis application claims the benefit of U.S. Provisional Application No. 63 / 613,436 filed December 21, 2023, which is hereby incorporated by reference in its entirety.Government SupportThis invention was made with government support under grant Al 146210 awarded by the National Institutes of Health. The government has certain rights in the invention.Background of the InventionMicrobial infections continue to threaten global health, and this situation is further exacerbated by the emergence of antimicrobial resistant strains including those resistant to multiple drugs. In 2019, the Center for Disease Control and Prevention (CDC) estimated that 3 million antimicrobial-resistant infections occurred in the USA annually.[1]Antibiotic resistance is rising to dangerously high levels in all parts of the world with resistance observed to nearly all antibiotics that have been developed.[2]New strategies to prevent and treat infections are urgently needed.Parallel to the development of new antibiotics, vaccination is an important approach for combating pathogens.[3]Multiple antimicrobial vaccines have been implemented against infections such as those by Clostridium tetani, Bordetella pertussis and Streptococcus pneumoniae. However, despite these successes, there are no approved vaccines against many other deadly pathogens including Staphylococcus aureus (S. aureus), which caused over 300,000 infections and 20,000 deaths through bloodstream infection in 2017 in the USJ41 The prevalence of methicillin-resistant .S'. aureus (MRS A) further highlights the need for a vaccine to stem the rise of anti-microbial resistance.[5]Poly- -( l-6)-A-acetylglucosamine (PNAG) is an important carbohydrate antigen target as it is expressed on the surface of many types of pathogens and is often a virulence factor. Since naturally existing PNAG can bear varying numbers of free amines rather than A-acetylation in its glucosamine monomer units, it is not clear how the number and the position of the amines (amine / acetylation pattern) impacts its antigenicity.A carrier system is essential to deliver carbohydrate antigens to the immune system and elicit strong anti-carbohydrate antigens antibody responses. However, a potential drawback of protein carrier is that high anti-carrier antibody responses can be induced by the glyco-conjugate. As an example, GD3-KLH generated an-antiGD3 IgG titer of 300, while that for KLH were1,800,000. The high anti-carrier antibodies can significantly suppress the generation of anti- glycan antibodies. This phenomenon has been reported for carbohydrate based anti-microbial disease vaccines. It has been suggested that an ideal carrier should induce high levels of anti- glycan antibodies without strong anti-self antibodies.Accordingly, there is a great need in the art to identify potential therapeutic strategies and compositions that activate immune responses in the treatment and prevention of disease and infection. The development of an effective vaccine is a highly attractive strategy to combat microbial infection, especially with the rise of antimicrobial resistant strains. The cell wall of numerous bacteria, fungi, and protozoan parasites contain the polysaccharide PNAG. Provided herein are strategies and vaccines for targeting PNAG, which can be effective against microbial infection.SummaryAspects of the invention include vaccine compositions comprising a poly-P-(l-6)-N- acetylglucosamine (PNAG) antigen. In some embodiments, said PNAG antigen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 glucosamine monomers. In some such embodiments, at least one glucosamine monomer is acetylated.In some aspects of the invention, the vaccine composition comprises a poly-P-(l-6)-A- acetylglucosamine (PNAG) antigen, wherein the PNAG antigen comprises at least 5, 6, 7, 8, 9, or 10 glucosamine monomers comprising the following Formula I:wherein Rl, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof.In some embodiments of the disclosed invention, the PNAG antigen is at least a disaccharide, e.g., comprising at least two glucosamine monomers. Thus, the PNAG antigen may comprise 2-10 glucosamine monomers. In certain embodiments, the PNAG antigen is a pentas achharide (e.g., five glucosamine monomers). In other embodiments, the PNAG antigen is a nonasaccharide (e.g., nine glucosamine monomers).In certain aspects, provided herein are methods for preventing or treating a microbial infection in a subject. Such methods comprise administering to the subject a vaccine composition provided herein.In other aspects, provided herein are methods of preventing or treating opportunistic microbial infection in a subject. In some embodiments, the subject is immnocomprimised. Said methods comprise administering to the subject a vaccine composition provided herein.Brief Description of the DrawingsFig. 1 depicts Formula II, which represents an embodiment of the vaccine provided herein.Fig. 2 depicts a library of 32 pentasaccharides PNAG0-PNAG32 covering the free amine space of PNAG. The five-digit number in the bracket for each compound codes for free amine (0) or N- acetamide (1) at residues ABCDE from the non-reducing end to the reducing end of the pentasaccharide respectively. The five-digit number was then viewed as a binary number and converted to the decimal system as the compound number, e.g., 01010 in binary number is equivalent to 10 in the decimal system. Thus, the PNAG pentasaccharide bearing A-acetylation at units B and D only is named as PNAG10.Fig. 3 depicts the stuctural formula of compound 2.Fig. 4 depicts Scheme 1, which illustrates the synthesis of the reducing end of glucosamine building block 8.Fig. 5 depicts Scheme 2, which illustrates syntheses of a) compound 13 and b) compound 1.Fig. 6 depicts Scheme 3, which illustrates orthogonal deprotection of pentasaccharide 2.Fig. 7 depicts Scheme 4, which illustrates the divergent modifications made to the key pentasaccharide intermediates. Fig. 7A depicts the divergent syntheses of 16 PNAG pentasaccharides from the strategically protected pentasaccharide 1. Fig. 7B depicts the divergent syntheses of 16 PNAG pentasaccharides from the strategically protected pentasaccharide 2.Fig. 8 depicts Scheme 5, which illustrates the Syntheses of Fig. 8A) mQP-, Fig. 8B) BSA-, and Fig. 8C) TT-PNAG conjugates.Fig. 9 demonstrates the immunization of mice and rabbits with mQP-PNAG led to high levels and long lasting anti-PNAG IgG antibodies. Fig. 9A) C57B16 mouse antibody responses at day 35 after immunization. The EC50 value (the fold of serum dilution that gives half-maximal binding) of the IgG titers to the immunizing oligosaccharide were plotted with each symbol representing one animal and the horizonal line is the geometric mean value of the titers within the group. The ELISA titers were determined using the BSA-PNAG conjugate containing the same PNAG structure as the immunizing QP-PNAG construct. One-way ANOVA allowed for rejection of the null hypothesis that all groups have the same mean IgG titers (p < 0.0001). ****Statistical significance (p < 0.0001) by Dunnett's multiple comparisons post-hoc test. Fig. 9B) Anti-PNAGO IgG antibody responses of mice (n = 5) immunized with mQP-PNAGO monitored over time with mean titers plotted. The error bars represent the standard deviation of the titer numbers from the five mice. The arrows indicate days of vaccination (days 0, 14, 28, 360, and 655). The antibody responses could be boosted more than 650 days after prime vaccination. Fig. 9C) IgG antibody titers to the immunizing PNAG oligosaccharide in sera of rabbits on day 35 after prime vaccination. Fig. 9D) Rabbit antibody responses from mQB-conjugate immunized animals or to the 5G1CNH2-TT conjugate as well as titer of natural human IgG in pooled human serum to purified PNAG.Fig. 10 depicts the binding data for anti-PNAG mAb F598 to the 32 PNAG pentasaccharide-BSA conjugates immobilized onto a glycan microarray. Fig. 10A) Relative fluorescence unit (RFU) of F598 mAb binding with the library of 32 PNAG pentasaccharides. The glycans are grouped according to the number of NHAc units in the molecule. The error bars represent the standard deviations of five independent measurements. F598 generally prefers highly acetylated PNAG sequences. Both the location and the number of NHAc units are important determinants of F598 binding. Fig. 10B) Quantification of the preference of F598 for acetylation at each site of the PNAG pentasaccharide.Fig. 11 depicts the screening of post-immune sera on the PNAG microarray. Fig. 11A) Stacked bar graphs depicting the IgG signals at a serum dilution of 1:50,000 for each rabbit (n = 2) immunized with mQP-PNAGO, mQP-PNAGlO and mQP-PNAG26 as well as pre-immune sera respectively on the array. The complete microarray results are shown in Appendix 1. Fig. 11B) Normalized binding of the comprehensive library of PNAG pentasaccharides by IgG antibodies from post-immune sera of rabbits immunized with mQP-PNAGO, mQP-PNAGlO and mQP- PNAG26 respectively as well as pre-immune sera. PNAG sequences are grouped together according to the total number of acetamides in the molecules. The color scale bar is shown on the right with 100% indicating the strongest binding to a PNAG component and 0% indicating the weakest binder. For each antigen, the two rows represent sera from two rabbits per group immunized with the specific construct.Fig. 12 shows that post-immune rabbit sera bound well with 5. aureus cells as visualized by fluorescence microscopy. The scale bar is 10 pm.Fig. 13 illustrates the deposition of Clq onto purified PNAG polysaccharide by antibodies in sera from rabbits immunized with mQP-PNAG conjugates, the 5G1CNH2-TT conjugate or in pooled normal human sera. The mQP-PNAG10 and mQP-PNAG26 conjugates were more potent thanthe mQP-PNAGO and 5G1CNH2-TT conjugate in inducing Clq deposition onto purified PNAG. Note that normal human serum had no Clq depositing activity in spite of having a binding titer to PNAG (see Fig. 9) consistent with prior reports that naturally- acquired human antibody to PNAG is not functional due to the inability to activate the complement pathway.[6c’20a]Fig. 14 shows significantly higher levels of opsonic killing activities against .S'. aureus cells when using sera from rabbits immunized with mQP-PNAG conjugate led to.Fig. 15 shows that immunization with mQP-PNAGO effectively Fig. 15A) protected against .S'. aureus infection, and Fig. 15B) reduced bacterial count in mouse kidney. mQP-PNAGO was significantly better than TT-PNAGO in protecting mice and reducing disease burden (n = 20 for each group). Log rank test and a t test were performed for statistical analysis of Fig. 15A and B respectively. * p < 0.05; **** p < 0.0001.Fig. 16 shows the transfer of antisera from mQP-PNAG immunized rabbits to mice Fig. 16A) provided significant protection to mice against the lethal challenges by .S'. aureus ATCC 29213; and Fig. 16B) significantly reduced bacterial count in mouse kidneys. Each symbol represents one mouse with the horizontal bar representing the group mean. The combination of sera from mQP-PNAGO and mQP-PNAG26 immunized rabbits provided complete protection to mice. Statistical analysis for survival was performed using the logrank test. * p < 0.05; *** p < 0.001; **** p < 0.0001.Fig. 17 shows the transfer of anti-sera from mQP-PNAG immunized rabbits to mice Fig. 17A) provided significant protection to mice against the lethal challenges by MRS A strain 1058 (n = 10 per group); and Fig. 17B) significantly reduced bacterial count in mouse kidneys. Sera from mQP-PNAG26 immunized rabbits provided the highest protection to mice. Statistical analysis was performed with the logrank test and t test respectively. * p < 0.05.Fig. 18 illustrates the synthesis procedures for A) p-Tolyl 2-allyloxycarbonylamino-3,4-di-O- benzoyl-6-O-tert-butyldiphenylsilyl-2-deoxy-l-thio-P-D-glucopyranoside (3); B) 3-Azidopropyl 2-allyloxycarbonylamino-3,4-di-O-benzoyl-6-O-tert-butyldiphenylsilyl-2-deoxy-P-D- glucopyranoside (5); C) 3-Azidopropyl 2-acetamido-3,4-di-O-benzoyl-6-O-tert- butyldiphenylsilyl-2-deoxy-P-D-glucopyranoside (6); and D) 3-(3- (Acetylthio)propionylamino)propyl 2-acetamido-3,4-di-O-benzoyl-2-deoxy-P-D-glucopyranoside (8)Fig. 19 illustrates the synthesis procedures for A) p-Tolyl 3,4-di-O-benzoyl-6-O-tert- butyldiphenylsilyl-2-deoxy-l-thio-2-(2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranoside (10); B) p-Tolyl 2-allyloxycarbonylamino-3,4-di-O-benzoyl-2-deoxy-l-thio-P- D-glucopyranoside (11); C) p-Tolyl 3,4-di-O-benzoyl-6-O-tert-butyldiphenylsilyl-2-deoxy-2- (2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3.4-di-O-benzoyl-2-deoxy-l-thio-P-D-glucopyranoside (9); D) 3-(3- (Acetylthio)propionylamino)propyl 3,4-di-O-bcnzoyl-6-O- / c / 7-butyldiphcnylsilyl-2-dcoxy-2- (2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3.4-di-O-benzoyl-2-deoxy-P-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-O-benzoyl-2-deoxy- P-D-glucopyranoside (12); and E) 3-(3-(Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-2- deoxy-2-(2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl- 2-deoxy-2-fhiorenylmethyloxycarbonylamino-P-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di- O-benzoyl-2-deoxy-P-D-glucopyranoside (13).Fig. 20 illustrates the synthesis procedures for A) p-Tolyl 3,4-di-O-benzoyl-6-O-tert- butyldiphenylsilyl-2-tert-butyloxycarbonylamino-2-deoxy-P-D-glucopyranosyl-( 1— >6)-2- allyloxycarbonylamino-3,4-di-O-benzoyl-2-deoxy-l-thio-P-D-glucopyranoside (14); and B) 3-(3- (Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O-tert-butyldiphenylsilyl-2-tert- butyloxycarbonylamino-2-deoxy-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4-di-O- benzoyl-2-deoxy-P-D-glucopyranosyl-(l^-6)-3,4-di-O-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2-deoxy-2- fhiorenylmethyloxycarbonylamino-P-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-O-benzoyl-2- deoxy-P-D-glucopyranoside (1).Fig. 21 depicts the synthesis scheme of A) compound S5 and B) compound 2.Fig. 22 illustrates the synthesis procedures for A) 3-(3-(Acetylthio)propionylamino)propyl 3,4- di-O-benzoyl-2-deoxy-2-trifluoroacetamido-P-D-glucopyranoside (S3); B) 3-(3- (Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O-tert-butyldiphenylsilyl-2-deoxy-2- (2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3.4-di-O-benzoyl-2-deoxy-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2-deoxy-2- trifluoroacetamido-P-D-glucopyranoside (S4) and 3-(3-(Acetylthio)propionylamino)propyl 3,4- di-O-benzoyl-2-deoxy-2-(2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-( 1— >6)-3.4-di-O-benzoyl-2-deoxy-2-fluorenylmethyloxycarbonylamino-P-D-glucopyranosyl-( 1— >6)-3,4- di-O-benzoyl-2-deoxy-2-trifluoroacetamido-P-D-glucopyranoside (S5); and C) 3-(3- (Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O-tert-butyldiphenylsilyl-2-tert- butyloxycarbonylamino-2-deoxy-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4-di-O- benzoyl-2-deoxy-P-D-glucopyranosyl-( 1— >6)-2-deoxy-3,4-di-O-benzoyl-2-(2,2,2-trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2- fluorenylmethyloxycarbonylamino-2-deoxy-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2- deoxy-2-trifluoroacetamido-P-D-glucopyranoside (2).Fig. 23 illustrates the synthesis of 3-(3-(Acetylthio)propionylamino)propyl 6-O-acetyl-2- acetamido-3,4-di-O-benzoyl-2-deoxy-P-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4- di-O-benzoyl-2-deoxy-P-D-glucopyranosyl-( 1— >6)- 3,4-di-O-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2-deoxy-2- fluorenylmethyloxycarbonylamino-P-D-glucopyranosyl-(l— >6)-3,4-di-O-benzoyl-2-deoxy-2- trifluoroacetamido-P-D-glucopyranoside (15).Fig. 24 shows the formulas for A) PNAGO (00000), B) PNAG1 (00001), C) PNAG2 (00010), D) PNAG3 (00011), E) PNAG4 (00100), F) PNAG5 (00101), G) PNAG6 (00110), H) PNAG7 (00111), I) PNAG8 (01000), J) PNAG9 (01001), K) PNAG10 (01010), L) PNAG11 (01011), M) PNAG12 (01100), N) PNAG13 (01101), O) PNAG14 (OH IO), P) PNAG15 (01111), Q) PNAG16 (10000), R) PNAG17 (10001), S) PNAG18 (10010), T) PNAG19 (10011), U) PNAG20 (10100), V) PNAG21 (10101), W) PNAG22 (10110), X) PNAG23 (10111), Y) PNAG24 (11000), Z) PNAG25 (11001), Zi) PNAG26 (11010), Z2) PNAG27 (11011), Z3) PNAG28 (11100), Z4) PNAG29 (11101), Z5) PNAG30 (11110), and Z6) PNAG31 (11111).Fig. 25 shows the MALDI-TOF spectra of the mQP-PNAG conjugates. The average glycan loading per particle was calculated byL = 180where 180 is the number of subunits in each VLP, N is the number of glycan loading in a specific peak, and A is the relative abundance of the corresponding peak. The shoulder peaks showing at +113 position of the major peaks are due to +1 linker as a result of intrasubunit crosslinking. The average loading is around 250 glycans per mQP particle based on the MS results.Fig. 26 shows the MALDI-TOF mass spectra of representative BSA-PNAG conjugates. Average loadings of glycans per BSA are presented.Fig. 27 shows the MALDI-TOF MS characterization of TTHc conjugates. TTHc-SBAP: SBAP functionalized TTHc. The average glycan loading on the PNAGO conjugates was calculated to be 4.7 glycans per TTHc molecule.Fig. 28 depicts the immunization of CD1 mice with mQP-PNAG led to high levels of anti-PNAG IgG antibodies on day 35 after immunization. The ECso values (the fold of serum dilution that gives half-maximal binding) of the IgG titers were plotted with each symbol representing oneanimal and the horizontal line is the geometrical mean value of the titers within the group. The ELISA titers were determined using the BSA-PNAG conjugate containing the same PNAG structure as the immunizing mQP-PNAG construct. One-way Anova allowed for rejection of the null hypothesis that all groups have the same mean IgG titers (p < 0.0001). **** Statistical significance (p < 0.0001) by Dunnetf s multiple comparisons post-hoc test.Fig. 29 depicts post-immune rabbit sera binding well to .S'. aureus cells as visualized by fluorescence microscopy (Fig. 29A). The scale bar is 10 pm. Fig. 29B) Rabbit immune sera recognized clinically isolated MRSA strains, i.e., 41, 1055, 1056, 1057, 1058, 1059, and 1153. A control strain lacking PNAG expression with icaA gene knock out (954) showed negligible binding by the immune sera, suggesting the recognition is PNAG dependent. Scale bar: 10 pm.Fig. 30 depicts opsonic killing as dependent on all factors including bacterial specific serum antibody, active complement source, and phagocytic cells, wherein omitting each factor completely abrogates killing activity. Complement inactivation was performed under 56 °C for 30 mins. HL-60 omitted group and complement inactivated group were carried out with mQP- PNAG26 serum.Fig. 31 depicts Non-metric multidimensional scaling (NMDS) analysis of bacterial communities of controls and treatments showing no significant changes in the gut microbial community following mQP-PNAG immunization. Sampling times T1 = first sampling right before immunization, and T2 = control and treatment samples at 42 days after the prime immunization. Each of the four groups is delineated by a convex hull describing a minimum area. The stress test values for 2D and 3D NMDS were 0.141 and 0.109, respectively.Fig. 32 shows that Post-immune sera of Fig. 32A) mice and Fig. 32B) rabbits recognized the .S'. aureus cells well as determined by ELISA. The EC50 values (the fold of serum dilution that gives half-maximal binding) of the IgG titers were plotted with each symbol representing one animal and the horizontal line is the geometrical mean value of the titers within the group. The ELISA titers were determined against .S', aureus cell coated ELISA wells.Detailed Description of the InventionProvided herein is the synthesis of PNAG antigens, including methods for identifying PNAG antigens of particular utility and / or efficacy. A major challenge in vaccine design is the selection of a suitable antigen. The cell wall of .S', aureus as well as numerous other bacteria, fungal, and protozoal parasites contains the polysaccharide poly-P-(l-6)-A-acetylglucosamine (PNAG), which is composed of P~( 1— 6) linked glucosamine units of which 80-95% are N-acetylated.[6'9]Naturally occurring PNAG can vary in its chemical structures with the amino groups in some glucosamine existing as free amines rather than as / V-acctyl (NHAc).

[0010] The degree and position of free amines vs NHAc in PNAG can vary resulting in structural heterogeneity, and the precise PNAG structures from bacteria are not known.Present on the microbial cell surface and as an integral component of biofilms, PNAG has been found to be an important virulence factor that aids in evasion from the immune system.16, 11]The widespread expression of PNAG in multiple pathogenic microbes and its important roles in pathogenesis render it an attractive target for vaccine development.The immunological properties of PNAG have been investigated.!6, 121 PNAG pentasaccharides and nonasaccharides with all amine groups either free or fully acetylated have been synthesized and subsequently conjugated with an immunogenic protein carrier, tetanus toxoid (TT) J13-15] Mouse immunization with the conjugate showed that the TT-conjugates with the PNAG bearing all amines could induce protective immunity in mice, while the antibodies elicited by the TT conjugates with fully acetylated PNAG counterparts did not recognize the bacteria well and failed to mediate protective functions such as opsonic killing.

[0016] Despite this interesting finding, as PNAG-based vaccines investigated to date only contained the glycans with all amine groups either free or fully acetylated, it is not clear what PNAG structure would comprise the best epitope, and whether patterns of amine vs acetylation in PNAG can be designed to enhance vaccine efficacy.In view of the PNAG molecular variation, provided herein is the chemical synthesis of a comprehensive library of 32 PNAG pentasaccharides with all possible variations of the location and the number of free amines. The availability of such a library allowed the establishment of the amine / acetylation code (locations of the free amine / NHAc in PNAG) of an anti-PNAG monoclonal antibody (mAb). The fine patterns of free amine / NHAc were found to be critical, which guided the development of vaccines based on specific PNAG epitopes, providing highly effective protection in multiple mouse models against .S'. aureus infections, including those by MRSA. Accordingly, the PNAGs contemplated herein may have specific acetylation patterns due to substitution on the amino groups by acetate. PNAG produced in vitro by methods known in the art is almost fully substituted (e.g., acetylated) on amino groups (95-100%). Alternatively, a deacetylated PNAG can be used having less than 50%, 40%, 30%, 20%, 10% or 5% N- acetylation. In some embodiments, the PNAG is deacetylated so that less than 50%, 40%, 30%, 20%, 10% or 5% of amino groups are N-acetylated. In some preferred embodiments, specific glucosamine monomer units of the PNAG are N-acetylated, thus providing a particular free- amine / acetylation pattern or “code”.Provided herein are methods for generating the PNAG structures from just two strategically protected advanced pentasaccharide intermediates, thus greatly enhancing the overall synthetic efficiency. In addition, provided herein are methods for epitope screening a PNAG library using an anti-PNAG monoclonal, such as antibody F598. PNAG sequences with specific patterns of free amines capable of strongly binding F598 were identified. The location of the free amine in PNAG was found to be critical for F598 binding as is the total number of amines, establishing the amine / acetylation code.Also provided herein are new PNAG structures conjugated with mutant bacteriophage QP, a powerful immunogen carrier. Immunization of mice and rabbits with the QP conjugates generated high levels of IgG antibodies against PNAG, which strongly recognized Staphylococcus aureus (S. aureus), an important human pathogen. Furthermore, vaccination provided effective protection to mice against infections by .S'. aureus including methicillin- resistant .S', aureus (MRSA) in murine models of both active and passive immunization. Infection by a MRSA strain killed all mice within two days, while the antisera from rabbits immunized with a conjugate of QP and a specific PNAG glycan protected 90% of the mice from lethal MRSA challenge. Thus, the comprehensive PNAG pentasaccharide library is a useful tool to decipher the amine / acetylation code of PNAG, which can empower the design of potent new vaccines against infectious diseases.Aspects of the invention dislcosed herein include vaccine compositions comprising a poly-P-(l-6)-N-acetylglucosamine (PNAG) antigen. In some embodiments, said PNAG antigen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 glucosamine monomers. In some such embodiments, at least one glucosamine monomer is acetylated.In some aspects of the invention, the vaccine composition comprises a poly-P-(l-6)-A- acetylglucosamine (PNAG) antigen, wherein the PNAG antigen comprises at least 5, 6, 7, 8, 9, or 10 glucosamine monomers comprising the following Formula I:wherein Rl, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof.In some embodiments of the disclosed invention, the PNAG antigen is at least a disaccharide, e.g., comprising at least two glucosamine monomers. Thus, the PNAG antigen may comprise 2-10 glucosamine monomers. In certain embodiments, the PNAG antigen is apentas achharide (e.g., five glucosamine monomers). In other embodiments, the PNAG antigen is a nonasaccharide e.g., nine glucosamine monomers).In some embodiments, the PNAG antigens provided herein comprise free amines. For example, and without limitation, the PNAG antigen comprises at least 1 to 5 free amines. Thus, the PNAG antigen may comprise at least 1 free amine, at least 2 free amines, at least 3 free amines, at least 4 free amines, or at least 5 free amines. In other embodiments, the PNAG antigen does not comprise free amines.Without being bound by theory and purely for exemplary purposes, Ri, R2, R3, R4, and R5 of Formula I are each H. In other embodiments, at least R2 and / or R4 is / V- acetyl . In certain embodiments, Ri, R3, and R4 of Formula I are H; and R2 and R5 are A-acetyl. In other embodiments, R3 and R4 are H; and Ri, R2, and R5 are A-acetyl.In some embodiments, the antigen comprises a PNAG pentasaccharide set forth in Figure 2. Thus, the antigen may comprise a PNAG pentasaccharide selected from the library of 32 pentasaccharides (z.e., PNAG0-PNAG32) disclosed.In some embodiments, the vaccine composition comprises a PNAG antigen bearing a linker comprising a disulfide group. In some such embodiments, the PNAG antigen is conjugated to a protein carrier. Said PNAG antigen conjugated to a protein carrier may comprise the following Formula II:wherein RI, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof; wherein is 1 or 2; and wherein R’ is the protein carrier.In some embodiments, the protein carrier is a viral capsid. Preferably, the capsid comprises at least one non-natural mutation. In some embodiments the capsid comprises at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty mutations. Said at least one mutation may be a disulfide bond mutation.In some embodiments, the viral capsid is derived from a bacteriophage. The bacteriophage may be selected from the group consisting of (a): bacteriophage QP; (b) bacteriophage R17; (c) bacteriophage fr; (d) bacteriophage GA; I bacteriophage SP; (f)bacteriophage MS2; (g) bacteriophage Mi l; (h) bacteriophage MX1; (i) bacteriophage NL95; (j) bacteriophage f2; (k) bacteriophage PP7; (1) bacteriophage AP205; and (m) bacteriophage P22.In some preferred embodiments, the bacteriophage is bacteriophage Q beta (QP). In some such embodiments, the mutation comprises at least one mutation selected from N10K, A38K, A40C, A40S, T75K, D102C, D102S, or A117K, or combination thereof. Preferably, the mutation comprises A38K. More preferably, said capsid comprises at least two mutations selected from A40C / D102C, A40S / D102S, or A43C / Q98C.A. DefinitionsThe articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.The term “administering” means providing a pharmaceutical agent or composition to a subject, and includes, but is not limited to, administering by a medical professional and selfadministering.As used herein, “bacteriophage” refers to viruses that infect and replicate within bacterium. In certain embodiments, the bacteriophage is selected from, but not limited to, the group consisting of bacteriophage QP; (b) bacteriophage R17; (c) bacteriophage fr; (d) bacteriophage GA; (e) bacteriophage SP; (f) bacteriophage MS2; (g) bacteriophage Mi l; (h) bacteriophage MX1; (i) bacteriophage NL95; (j) bacteriophage f2; (k) bacteriophage PP7; (1) bacteriophage AP205; and (m) bacteriophage P22. As used herein, “bacteriophage QP” (also referred to as “bacteriophage Qb”, “QP” and “Qb”) is one of many small RNA bacteriophages infecting Escherichia coli.As used herein, the term “PNAG antigen” refers to classes of antigens comprising P- (l-6)-A-acetylglucosamine that elicit strong antibody responses. In some embodiments, the PNAG antigen comprises an oligosaccharide comprising a P-(l— >6)-glucosamine group and, without being bound by theory of convention, mimics a portion of the cell wall of PNAG-based microbes. The term “oligosaccharide comprising P-(l— >6)-glucosamine structures” or “oligosaccharide comprising P-(l-6)-A-acetylglucosamine structures” may refer to those structures found in the cell wall of microbes. The microbial wall contains a large number of these structures that are conserved across many microbial lines. These structures are found in the microbial cell wall and include those oligosaccharides wherein the majority of their units are P- (1— >6)-glucosamines (e.g., P-( l-6)-A-acetylglucosamine). The PNAG oligosaccharide antigens disclosed herein may be di-, tri-, tetra-, penta-, hexa-, hepta-, octa-, nona-, or dodecasaccharides, e.g., the PNAG oligosaccharide antigens comprise at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 glucosaminemonomers. In some embodiments, at least one monomer is acetylated. Preferably, in some such embodiments the PNAG oligosaccharide antigen comprises an amine / acetylation pattern contemplated herein. Thus, in some embodiments of the invention provided herein are included methods for providing immune protection against PNAG microbes by use of a vaccine comprising a 0-(l— >6)-glucosamine oligosaccharide-linked-capsid vaccine that provide effective immunity to a patient against, e.g., microbial infections wherein said microbe comprises -(l- 6)- A-acetylglucosamine structures in its cell walls. In some embodiments, the antibodies to the vaccine will bind to 0-(l— >6)-glucosamine structures, such as -(l-6)-A-acetylglucosamine. In some embodiments, said vaccine does not cross-react with a F-598 monoclonal antibody. In some embodiments, the vaccines generate antibodies that are complementary to F-598. That is, F-598 will selectively bind to acetylated 0-(l— >6)-glucosamine structures, i.e., N-acetyl glucosamine.In some embodiments, the invention disclosed herein provides for a vaccine against microbes comprising oligosaccharide 0-(l— >6)-glucosamine structures in their cell wall wherein said vaccine is represented by Formula II; wherein Rl, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof; wherein x is 1, 2, or 3; wherein R’ is the carrier capsid; and wherein A, B, C, D, and E each represent a “glucosamine unit”.The term “inhibit” or “inhibits” means to decrease, suppress, attenuate, diminish, arrest, or stabilize the development or progression of a disease, disorder, or condition, the activity of a biological pathway, or a biological activity, such as the growth of a solid malignancy, e.g., by at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 98%, 99%, or even 100% compared to an untreated control subject, cell, biological pathway, or biological activity or compared to the target, such as a growth of a solid malignancy, in a subject before the subject is treated. By the term “decrease” is meant to inhibit, suppress, attenuate, diminish, arrest, or stabilize a symptom of an infection, or disease, disorder, or condition thereof. It will be appreciated that, although not precluded, treating a disease, disorder or condition does not require that the disease, disorder, condition or symptoms associated therewith be completely eliminated.The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and / or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.The phrase "pharmaceutically-acceptable carrier" as used herein means a pharmaceutically-acceptable material, composition or vehicle, such as a liquid or solid filler,diluent, excipient, or solvent encapsulating material, involved in carrying or transporting the subject compound from one organ, or portion of the body, to another organ, or portion of the body. Each carrier must be "acceptable" in the sense of being compatible with the other ingredients of the formulation and not injurious to the patient. Some examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as com starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, ethyl cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, corn oil and soybean oil; (10) glycols, such as propylene glycol; (11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol; (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and / or poly anhydrides; and (22) other non-toxic compatible substances employed in pharmaceutical formulations.“Pharmaceutically-acceptable salts” refers to the relatively non-toxic, inorganic and organic acid addition salts of compounds.The terms “prevent,” “preventing,” “prevention,” “prophylactic treatment,” and the like refer to reducing the probability of developing a disease, disorder, or condition in a subject, who does not have, but is at risk of or susceptible to developing a disease, disorder, or condition.A “subject” can include a human subject for medical purposes, such as for the treatment of an existing disease, disorder, condition or the prophylactic treatment for preventing the onset of a disease, disorder, or condition or an animal subject for medical, veterinary purposes, or developmental purposes. Suitable animal subjects include mammals including, but not limited to, primates, e.g., humans, monkeys, apes, gibbons, chimpanzees, orangutans, macaques and the like; bovines, e.g., cattle, oxen, and the like; ovines, e.g., sheep and the like; caprines, e.g., goats and the like; porcines, e.g., pigs, hogs, and the like; equines, e.g., horses, donkeys, zebras, and the like; felines, including wild and domestic cats; canines, including dogs; lagomorphs, including rabbits, hares, and the like; and rodents, including mice, rats, guinea pigs, and the like. An animal may be a transgenic animal. In some embodiments, the subject is a human including, but not limited to, fetal, neonatal, infant, juvenile, and adult subjects. Further, a “subject” can include a patient afflicted with or suspected of being afflicted with a disease, disorder, or condition. Thus, the terms “subject” and “patient” are used interchangeably herein. Subjects also include animal disease models e.g., rats or mice used in experiments, and the like).The term “subject in need thereof’ means a subject identified as in need of a therapy or treatment.The term “immunocompromised” as used herein may include a subject who, prior to and / or at the time of pathogen exposure, has a pre-existing condition that reduces one or more mechanisms for normal defense against infection. The immunocompromised condition may be due to a defect or dysfunction of the immune system or to other factors that heighten susceptibility to infection (e.g., physical trauma and inflammation, immunosuppression or immunoablative / -depleting therapies, or immunodeficient conditions / diseases as are known in the art). Although such a categorization allows a conceptual basis for evaluation, immunocompromised individuals with infection often do not fit completely into one group or the other. More than one defect in the body's defense mechanisms may be affected. For example, individuals with a specific T-lymphocyte defect caused by HIV may also have neutropenia caused by drugs used for antiviral therapy or be immunocompromised because of a breach of the integrity of the skin and mucous membranes. An immunocompromised state can result from indwelling central lines or other types of impairment due to intravenous drug abuse; age-related immunologic impairment; or be caused by secondary malignancy, malnutrition, burn injury, or having been infected with other infectious agents such as tuberculosis or sexually transmitted diseases, e.g., syphilis or hepatitis.The terms “systemic administration,” “administered systemically,” “peripheral administration,” and “administered peripherally” mean the administration of a compound, drug or other material other than directly into the central nervous system, such that it enters the patient's system and, thus, is subject to metabolism and other like processes, for example, subcutaneous administration.The term “therapeutic agent” or “pharmaceutical agent” refers to an agent capable of having a desired biological effect on a host.The term “therapeutic effect” refers to a local or systemic effect in animals, particularly mammals, and more particularly humans, caused by a pharmacologically active substance.The terms “therapeutically-effective amount” and “effective amount” as used herein means that amount of a compound, material, or composition comprising a compound of the present invention which is effective for producing some desired therapeutic effect in at least a sub-population of cells in an animal at a reasonable benefit / risk ratio applicable to any medical treatment.The term “treating” a disease in a subject or “treating” a subject having a disease refers to subjecting the subject to a pharmaceutical treatment, e.g., the administration of a vaccinecomposition as described herein, such that at least one symptom of the disease is decreased, prevented from worsening, or delayed from worsening.B. CapsidsProvided herein are vaccine compositions comprising an antigen conjugated to a capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a wild type capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a wild type bacteriophage QP capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having a wild type or native sequence. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having a wild type or natural sequence set forth in SEQ ID NO: 1. In some embodiments, the vaccine composition comprises an antigen conjugated to a capsid having at least one mutation from the wild type capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having at least one mutation from the wild type bacteriophage QP capsid. In some embodiments, the at least one mutation comprises a non-natural mutation. In some embodiments, the non-natural mutation comprises a non-natural amino acid mutation. In some embodiments, the vaccine compositions provided herein comprises an antigen (e.g., carbohydrate antigen, polypeptides, peptides, proteins and small molecules) conjugated to a capsid (e.g., bacteriophage Qb), wherein said capsid comprises at least one mutation (e.g., at least one point mutation, or at least one non-natural disulfide bond). In some embodiments, the capsid are fragments or a portion of the capsid amino acid sequence of sufficient length, that when conjugated to the antigen, can elicit an enhanced and strong immune response. In certain embodiments, the capsid polypeptide also includes amino acids that do not correspond to the naturally occurring capsid amino acid sequence (e.g., comprising at least one point mutation, or at least one non-natural disulfide bond mutation, or a fusion protein comprising a capsid amino acid sequence and an amino acid sequence corresponding to a noncapsid protein or polypeptide).In some embodiments, the capsid is derived from bacteriophage. In certain embodiments, the bacteriophage is selected from the group consisting of bacteriophage QP; (b) bacteriophage R17; (c) bacteriophage fr; (d) bacteriophage GA; (e) bacteriophage SP; (f) bacteriophage MS2; (g) bacteriophage Mi l; (h) bacteriophage MX1; (i) bacteriophage NL95; (j) bacteriophage f2; (k) bacteriophage PP7; (1) bacteriophage AP205; and (m) bacteriophage P22.In some embodiments, the capsid has a sequence set forth in coat protein Table A. Table A Listing of capsid and associated GenBank numbers[Escherichia virus Qbeta] (GenBankAEQ25550.1)coat protein [Enterobacteria phage Mi l] (GenBank AAC06250.1) coat protein [Enterobacteria phage SP] (GenBank AEQ25562.1) coat protein [Enterobacteria phage FI sensu lato] (Genbank NP_695027.1) capsid protein [Escherichia virus FI] (Genbank ACT66758.1) coat protein [Escherichia virus Qbeta] (Genbank ACY07224.1)Chain A, Bacteriophage Q Beta Capsid (Genbank 1QBE_A) capsid protein [Escherichia virus Qbeta] (ACT66734.1) capsid protein [Escherichia virus Qbeta] (ACT66730.1) capsid protein [Escherichia virus Qbeta] (ACT66742.1) read-through protein [Escherichia virus Qbeta] (ACY07231.1)Al protein [Escherichia virus Qbeta] (AAA 16663.1) read-through protein [Escherichia virus Qbeta] (AEQ25545.1)Minor capsid protein Al (Q8LTE1.2) read-through protein [Escherichia virus Qbeta] (ACY07227.1) read-through protein [Escherichia virus Qbeta] (ACY07235.1) capsid protein [Escherichia virus Qbeta] (ACT66738.1) read-through protein [Escherichia virus Qbeta] (AEQ25549.1) read-through protein [Escherichia virus Qbeta] (AEQ25541.1) read-through protein [Escherichia virus Qbeta] (ACY07223.1) read-through protein [Escherichia virus Qbeta] (ACT66735.1)Chain A, Bacteriophage Qbeta Coat Protein In Complex With Rna Operator Hairpin (4L8H_A) read-through protein [Escherichia virus Qbeta] (ACT66731.1) read-through protein [Escherichia virus Qbeta] (ACT66743.1) read-through protein [Escherichia virus Qbeta] (ACT66739.1) capsid protein [Escherichia virus Qbeta] (ACT66746.1) major coat protein [Escherichia virus Qbeta] (NP_046751.1) read-through protein [Escherichia virus Qbeta] (ACT66747.1) minor coat protein [Escherichia virus Qbeta] (NP_046750.1) coat protein [Enterobacteria phage SP] (ACY07244.1) coat protein [Enterobacteria phage SP] (ACY07240.1)Al -protein [Enterobacteria phage Mi l] (AAC06251.1) coat protein [Enterobacteria phage SP] (AEQ25558.1) capsid protein [Escherichia virus FI] (ACT66762.1) major coat protein [Escherichia virus FI] (YP_009208147.1) capsid protein [Escherichia virus FI] (ACT66750.1)coat protein [Enterobacteria phage SP] (ACY07248.1) read- through protein [Enterobacteria phage SP] (ACY07251.1) read- through protein [Enterobacteria phage SP] (AEQ25561.1) read- through protein [Enterobacteria phage SP] ( AC Y07243.1)Minor capsid protein Al (P09677.1) readthrough protein [Enterobacteria phage FI sensu lato] (NP_695026.1) capsid protein [Escherichia virus FI] (ACT66754.1) read- through protein [Enterobacteria phage SP] (AEQ25553.1) read-through protein [Enterobacteria phage SP] (ACY07239.1) read-through protein [Enterobacteria phage SP] (AEQ25557.1) read-through protein [Escherichia virus FI] (ACT66759.1) major coat protein [Enterobacteria phage NL95] (AAC14703.1) read-through protein [Escherichia virus FI] (ACT66763.1) read- through protein [Enterobacteria phage SP] ( AC Y07247.1)Al-protein [Escherichia virus FI] (YP_009208146.1) read-through protein [Escherichia virus FI] (ACT66751.1) read-through protein [Escherichia virus FI] (ACT66755.1)Al-protein [Enterobacteria phage NL95] (AAC14704.1)In some embodiments, the bacteriophage QP capsid comprises a sequence set forth in SEQ ID NO: 1.(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVRT ELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 1)In some embodiments, the bacteriophage QP capsid comprises at least one mutation set forth in Table B.Table B. QP mutants reported that assemble to form the capsid.Note: Yield ++ : >80 mg / L, + : >20-80 mg / L, <20 mg / L, na : not applicable7a corresponds to Fiedler, J et al. Biomacromolecules 2012, 13 (8), 2339-2348; 42a corresponds to Prasuhn, D et al. JACS 2008, 130 (4), 1328-1334; 42b corresponds to Udit, A et al. ChemBioChem 2009, 10 (3), 503-510. 43 corresponds to Hovlid, M. L et al. The Scripps Research Institute, La Jolla, 2014.In certain embodiments, the capsid comprises at least two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty mutations. In some embodiments, the mutation is a non-natural disulfide bond mutation. In some embodiments, the capsid comprises, but not limited to, the following mutations (Table C).Table C: Listing of bacteriophage QP capsidN10K (Asparagine at position 10 of SEQ ID NO: 1 mutated to Lysine)K13R (Lysine at position 13 of SEQ ID NO: 1 mutated to Arginine)A38K (Alanine at position 38 of SEQ ID NO: 1 mutated to Lysine)A40C (Alanine at position 40 of SEQ ID NO: 1 mutated to Cysteine)A40S (Alanine at position 40 of SEQ ID NO: 1 mutated to Serine)T75K (Threonine at position 75 of SEQ ID NO: 1 mutated to Lysine)D102C (Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Cysteine)D102S (Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Serine)Al 17K (Alanine at position 117 of SEQ ID NO: 1 mutated to Lysine)A40C / D102C (double mutant wherein the Alanine at position 40 of SEQ ID NO: 1 mutated to Cysteine and the Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Cysteine)A40S / D102S (double mutant wherein the Alanine at position 40 of SEQ ID NO: 1 mutated to Serine and the Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Serine).A43C / N98C (double mutant wherein the Alanine at position 43 of SEQ ID NO: 1 mutated to Cysteine and the Glutamine at position 98 of SEQ ID NO: 1 mutated to Cysteine).A40C / D102C / K13R (triple mutant wherein the Alanine at position 40 of SEQ ID NO: 1 mutated to Cysteine, the Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Cysteine, and the Lysine at position 13 of SEQ ID NO: 1 mutated to Arginine)A38K / A40C / D102C (triple mutant wherein Alanine at position 38 of SEQ ID NO: 1 mutated to Lysine, the Alanine at position 40 of SEQ ID NO: 1 mutated to Cysteine, and the Aspartic acid at position 102 of SEQ ID NO: 1 mutated to Cysteine).Additional single point mutations may comprise, but not limited to, point mutations at K2, L4, V7, N10, K13, D14, K16, Q18, L20, A38, G40, E46, V67, T75, V85, Q99, E103, Al 17, Pl 19, L122, or D127 of SEQ ID NO: 1, or combinations thereof.N10K(M)AKLETVTLGKIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVSVSQPS RNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVRTELAA LLASPLLIDAIDQLNPAY (SEQ ID NO: 2)A38K(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQKGAVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 4)A40C(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGCVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 5)A40S(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGSVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 6)T75K(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACKANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 7)D102C(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTCEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 8)D102S(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTSEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 9)A117K(M)AKEETVTEGNIGKDGKQTEVENPRGVNPTNGVASESQAGAVPAEEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTDEERAFVR TEEAAEEKSPEEIDAIDQENPAY (SEQ ID NO: 10)A40C / D102C(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGCVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTCEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 11)A40S / D102S(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGCVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTSEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 12)A40C / D102C / K13R(M)AKLETVTLGNIGRDGKQTLVLNPRGVNPTNGVASLSQAGCVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTCEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 13)A43C / O98C(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQAGAVPCLEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTCYSTDEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 14)A38K / A40C / D102C(M)AKLETVTLGNIGKDGKQTLVLNPRGVNPTNGVASLSQKGCVPALEKRVTVS VSQPSRNRKNYKVQVKIQNPTACTANGSCDPSVTRQAYADVTFSFTQYSTCEERAFVR TELAALLASPLLIDAIDQLNPAY (SEQ ID NO: 15)In certain embodiments, bacteriophage Qb mutants may possess the following physical characteristics.Table D: Physical characteristics of QP mutants.A23K>A41"®132C 2S 12.5 22.55 3.020 4.55ASSS35132S 20 12.0 27.32 3.020 4,37AS3CmiO2C®3B 15 11.3 20M 3.113 4.S7In some embodiments, the bacteriophage Qb capsid comprises a polypeptide comprising amino acid sequences at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in SEQ ID NOs: 1-15. In some embodiments of the methods, compositions, and kits provided herein, the Qb mutant has at least one mutation set forth in Table C. In some embodiments, the bacteriophage QP capsid comprises a wild type or native sequence. In some embodiments, the bacteriophage QP capsid comprises a sequence consisting essentially of the sequence set forth in SEQ ID NO: 1. In some embodiments, the Qb mutant has an amino acid sequence that consists essentially of the mutations set forth in SEQ ID NOs: 2-15. In some embodiments, the capsid comprises a polypeptide comprising amino acid sequences at least 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to the amino acid sequence set forth in the GenBank numbers set forth in Table A.In some embodiments of the methods, compositions and kits provided herein, the vaccine composition comprises a capsid having an amino acid sequence that consists of 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95, 100, 105, 110, 115, 120, 125, 130 of SEQ ID Nos: 1-15, or biologically active variant thereof, or combinations thereof, or consecutive amino acids that are at least 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98% or 99% identical to an amino acid sequence for a capsid (e.g., SEQ ID NOs: 1-15, or any of the GenBank numbers set forth in Table A). In some embodiments, the bacteriophage QP capsid comprises a wild type or native sequence. In some embodiments, the bacteriophage QP capsid comprises a sequence consisting essentially of the sequence set forth in SEQ ID NO: 1.As is well-known to those skilled in the art, polypeptides having substantial sequence similarities can cause identical or very similar immune reactions in a host animal. Accordingly, in some embodiments, a derivative, equivalent, variant, fragment, or mutant of the Qb capsid, or fragment thereof, can also be suitable for the methods, compositions and kits provided herein.In some embodiments, the altered polypeptide may have an altered amino acid sequence, for example by conservative substitution, yet still elicits an enhanced immune response, and are considered functional equivalents. As used herein, the term “conservative substitution” denotes the replacement of an amino acid residue by another, biologically similar residue. It is well known in the art that the amino acids within the same conservative group can typically substitute for one another without substantially affecting the function of a protein. According to certain embodiments, the derivative, equivalents, variants, or mutants of the Qb are at least 85% homologous to a sequence set forth in SEQ ID NOs: 1-15, or biologically active variant thereof, or combinations thereof. In some embodiments, the homology is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, or at least 99%. In some embodiments, the bacteriophage QP capsid comprises a wild type or native sequence. In some embodiments, the bacteriophage QP capsid comprises a sequence consisting essentially of the sequence set forth in SEQ ID NO: 1.C. Vaccine compositions and pharmaceutical compositions / formulations of sameProvided herein are vaccine compositions comprising an antigen conjugated to a capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a wild-type capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a wild-type bacteriophage QP capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having a wild type or native sequence. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having a wild type or natural sequence set forth in SEQ ID NO: 1. In some embodiments, the vaccine composition comprises an antigen conjugated to a capsid having at least one mutation from the wild-type capsid. In some embodiments, the vaccine composition comprises an antigen conjugated to a bacteriophage QP capsid having at least one mutation from the wild-type bacteriophage QP capsid. In some embodiments, the at least one mutation comprises a non-natural mutation. In some embodiments, the non-natural mutation comprises a non-natural amino acid mutation. In some embodiments, the vaccine compositions provided herein comprises an antigen (e.g., carbohydrate antigen, polypeptides, peptides, proteins, and small molecules) conjugated to a capsid (e.g., bacteriophage Qb), wherein said capsid comprises at least one mutation (e.g., at least one point mutation, at least one non-natural amino acidmutation, or at least one non-natural disulfide bond mutation). The capsid is described in Section B, supra. In some embodiments, the antigens are fragments or a portion of the carbohydrate antigen of sufficient length, that when conjugated to the capsid, can elicit an enhanced and strong immune response. The capsid may be conjugated to a plurality of multiple antigens that are the same or different antigen. In certain embodiments, the antigen is a protein and peptide is selected from, but not limited to, TNFalpha, ILla, ILip, tau protein, PCSK9, or amyloid p. In some embodiments, the antigen is a small molecule selected from, but not limited to, nicotine, cocaine, or advanced glycation product.In some embodiments, the present invention provides pharmaceutically acceptable compositions which comprise a therapeutically effective amount of one or more vaccine compositions (e.g., one or more Qb wild type, or QP mutant, antigen conjugate as described above), formulated together with one or more pharmaceutically acceptable carriers (additives) and / or diluents. In another aspect, the compositions can be administered as such or in admixtures with pharmaceutically acceptable carriers and can also be administered in conjunction with other therapies. Conjunctive therapy thus includes sequential, simultaneous and separate, or coadministration of the composition, wherein the therapeutic effects of the first administered has not entirely disappeared when the subsequent compound is administered. In some embodiments, at least one vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) may be provided to the subject alone or in combination with at least one therapeutic drug, chemotherapeutic agents, scavenger compounds, antibiotics, antivirals, anti-fungals, anti-inflammatories, vasoconstrictors and anticoagulants, antigens useful for vaccine applications or corresponding pro-drugs.As described in detail below, the pharmaceutical compositions of the present invention may be specially formulated for administration in solid or liquid form, including those adapted for the following: (1) oral administration, for example, drenches (aqueous or non-aqueous solutions or suspensions), tablets, e.g., those targeted for buccal, sublingual, and systemic absorption, boluses, powders, granules, pastes for application to the tongue; (2) parenteral administration, for example, by subcutaneous, intramuscular, intravenous or epidural injection as, for example, a sterile solution or suspension, or sustained-release formulation; (3) topical application, for example, as a cream, ointment, or a controlled-release patch or spray applied to the skin; (4) intravaginally or intrarectally, for example, as a pessary, cream or foam; (5) sublingually; (6) ocularly; (7) transdermally; or (8) nasally.As set out above, certain embodiments of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) may contain a basicfunctional group, such as amino or alkylamino, and are, thus, capable of forming pharmaceutically acceptable salts with pharmaceutically-acceptable acids. These salts can be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting a purified compound of the invention in its free base form with a suitable organic or inorganic acid, and isolating the salt thus formed during subsequent purification. Representative salts include the hydrobromide, hydrochloride, sulfate, bisulfate, phosphate, nitrate, acetate, valerate, oleate, palmitate, stearate, laurate, benzoate, lactate, phosphate, tosylate, citrate, maleate, fumarate, succinate, tartrate, napthylate, mesylate, glucoheptonate, lactobionate, and laurylsulphonate salts and the like (see, for example, Berge et al. (1977) “Pharmaceutical Salts”, J. Pharm. Sci. 66:1-19).The pharmaceutically acceptable salts of the subject compounds include the conventional nontoxic salts or quaternary ammonium salts of the compounds, e.g., from non-toxic organic or inorganic acids. For example, such conventional nontoxic salts include those derived from inorganic acids such as hydrochloride, hydrobromic, sulfuric, sulfamic, phosphoric, nitric, and the like; and the salts prepared from organic acids such as acetic, propionic, succinic, glycolic, stearic, lactic, malic, tartaric, citric, ascorbic, palmitic, maleic, hydroxy maleic, phenylacetic, glutamic, benzoic, salicyclic, sulfanilic, 2-acetoxybenzoic, fumaric, toluenesulfonic, methanesulfonic, ethane disulfonic, oxalic, isothionic, and the like.In other cases, the one or more vaccine compositions e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) may contain one or more acidic functional groups and, thus, are capable of forming pharmaceutically acceptable salts with pharmaceutically acceptable bases. These salts can likewise be prepared in situ in the administration vehicle or the dosage form manufacturing process, or by separately reacting the purified compound in its free acid form with a suitable base, such as the hydroxide, carbonate or bicarbonate of a pharmaceutically acceptable metal cation, with ammonia, or with a pharmaceutically acceptable organic primary, secondary or tertiary amine. Representative alkali or alkaline earth salts include the lithium, sodium, potassium, calcium, magnesium, and aluminum salts and the like. Representative organic amines useful for the formation of base addition salts include ethylamine, diethylamine, ethylenediamine, ethanolamine, diethanolamine, piperazine and the like (see, for example, Berge et al., supra).Wetting agents, emulsifiers and lubricants, such as sodium lauryl sulfate and magnesium stearate, as well as coloring agents, release agents, coating agents, sweetening, flavoring and perfuming agents, preservatives and antioxidants can also be present in the compositions.Examples of pharmaceutically-acceptable antioxidants include: (1) water soluble antioxidants, such as ascorbic acid, cysteine hydrochloride, sodium bisulfate, sodium metabisulfite, sodium sulfite and the like; (2) oil-soluble antioxidants, such as ascorbyl palmitate, butylated hydroxyanisole (BHA), butylated hydroxytoluene (BHT), lecithin, propyl gallate, alpha-tocopherol, and the like; and (3) metal chelating agents, such as citric acid, ethylenediamine tetraacetic acid (EDTA), sorbitol, tartaric acid, phosphoric acid, and the like.The one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) may be formulations suitable for oral, nasal, topical (including buccal and sublingual), rectal, vaginal and / or parenteral administration. The formulations may conveniently be presented in unit dosage form and may be prepared by any methods well known in the art of pharmacy. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will vary depending upon the host being treated and the particular mode of administration. The amount of active ingredient which can be combined with a carrier material to produce a single dosage form will generally be that amount of the compound which produces a therapeutic effect.In certain embodiments, a formulation of one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) can comprise other carriers to allow more stability, to allow more stability, different releasing properties in vivo, targeting to a specific site, or any other desired characteristic that will allow more effective delivery of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) to a subject or a target in a subject, such as, without limitation, liposomes, microspheres, nanospheres, nanoparticles, bubbles, micelle forming agents, e.g., bile acids, and polymeric carriers, e.g., polyesters and poly anhydrides. In certain embodiments, an aforementioned formulation renders orally bioavailable a compound of the present invention.Liquid dosage formulations of one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) include pharmaceutically acceptable emulsions, microemulsions, solutions, suspensions, syrups and elixirs. In addition to the active ingredient, the liquid dosage forms may contain inert diluents commonly used in the art, such as, for example, water or other solvents, solubilizing agents and emulsifiers, such as ethyl alcohol, isopropyl alcohol, ethyl carbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propylene glycol, 1,3-butylene glycol, oils (in particular, cottonseed, groundnut, corn, germ, olive, castor and sesame oils), glycerol, tetrahydrofuryl alcohol, polyethylene glycols and fatty acid esters of sorbitan, and mixtures thereof. 1Besides inert diluents, the oral compositions can also include adjuvants such as wetting agents, emulsifying and suspending agents, sweetening, flavoring, coloring, perfuming and preservative agents.Suspensions, in addition to the active compounds, may contain suspending agents as, for example, ethoxylated isostearyl alcohols, polyoxyethylene sorbitol and sorbitan esters, microcrystalline cellulose, aluminum metahydroxide, bentonite, agar-agar and tragacanth, and mixtures thereof.Formulations suitable for oral administration may be in the form of capsules, cachets, pills, tablets, lozenges (using a flavored basis, usually sucrose and acacia or tragacanth), powders, granules, or as a solution or a suspension in an aqueous or non-aqueous liquid, or as an oil-in-water or water-in-oil liquid emulsion, or as an elixir or syrup, or as pastilles (using an inert base, such as gelatin and glycerin, or sucrose and acacia) and / or as mouth washes and the like, each containing a predetermined amount of an active ingredient. One or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) may also be administered as a bolus, electuary or paste.In solid dosage forms (e.g., capsules, tablets, pills, dragees, powders, granules and the like), the active ingredient is mixed with one or more pharmaceutically-acceptable carriers, such as sodium citrate or dicalcium phosphate, and / or any of the following: (1) fillers or extenders, such as starches, lactose, sucrose, glucose, mannitol, and / or silicic acid; (2) binders, such as, for example, carboxymethylcellulose, alginates, gelatin, polyvinyl pyrrolidone, sucrose and / or acacia; (3) humectants, such as glycerol; (4) disintegrating agents, such as agar-agar, calcium carbonate, potato or tapioca starch, alginic acid, certain silicates, and sodium carbonate; (5) solution retarding agents, such as paraffin; (6) absorption accelerators, such as quaternary ammonium compounds; (7) wetting agents, such as, for example, cetyl alcohol, glycerol monostearate, and non-ionic surfactants; (8) absorbents, such as kaolin and bentonite clay; (9) lubricants, such a talc, calcium stearate, magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate, and mixtures thereof; and (10) coloring agents. In the case of capsules, tablets and pills, the compositions may also comprise buffering agents. Solid compositions of a similar type may also be employed as fillers in soft and hard-shelled gelatin capsules using such excipients as lactose or milk sugars, as well as high molecular weight polyethylene glycols and the like.A tablet may be made by compression or molding, optionally with one or more accessory ingredients. Compressed tablets may be prepared using binder (for example, gelatin or hydroxypropylmethyl cellulose), lubricant, inert diluent, preservative, disintegrant (for example, sodium starch glycolate or cross-linked sodium carboxymethyl cellulose), surface-active ordispersing agent. Molded tablets may be made by molding in a suitable machine a mixture of the powdered compound moistened with an inert liquid diluent.The tablets, and other solid dosage forms, such as dragees, capsules, pills and granules, may optionally be scored or prepared with coatings and shells, such as enteric coatings and other coatings well known in the pharmaceutical-formulating art. They may also be formulated so as to provide slow or controlled release of the active ingredient therein using, for example, hydroxypropylmethyl cellulose in varying proportions to provide the desired release profile, other polymer matrices, liposomes and / or microspheres. Compositions may also be formulated for rapid release, e.g., freeze-dried. They may be sterilized by, for example, filtration through a bacteria-retaining filter, or by incorporating sterilizing agents in the form of sterile solid compositions which can be dissolved in sterile water, or some other sterile injectable medium immediately before use. These compositions may also optionally contain opacifying agents and may be of a composition that they release the active ingredient(s) only, or preferentially, in a certain portion of the gastrointestinal tract, optionally, in a delayed manner. Examples of embedding compositions which can be used include polymeric substances and waxes. The active ingredient can also be in micro-encapsulated form, if appropriate, with one or more of the abovedescribed excipients.Formulations for rectal or vaginal administration may be presented as a suppository, which may be prepared by mixing one or more compounds of the invention with one or more suitable nonirritating excipients or carriers comprising, for example, cocoa butter, polyethylene glycol, a suppository wax or a salicylate, and which is solid at room temperature, but liquid at body temperature and, therefore, will melt in the rectum or vaginal cavity and release the active compound.Formulations which are suitable for vaginal administration also include pessaries, tampons, creams, gels, pastes, foams or spray formulations containing such carriers as are known in the art to be appropriate.Dosage forms for the topical or transdermal administration of one or more vaccine compositions e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) include powders, sprays, ointments, pastes, creams, lotions, gels, solutions, patches and inhalants. The active compound may be mixed under sterile conditions with a pharmaceutically - acceptable carrier, and with any preservatives, buffers, or propellants which may be required.The ointments, pastes, creams and gels may contain, in addition to an active compound of this invention, excipients, such as animal and vegetable fats, oils, waxes, paraffins, starch,tragacanth, cellulose derivatives, polyethylene glycols, silicones, bentonites, silicic acid, talc and zinc oxide, or mixtures thereof.Powders and sprays can contain excipients such as lactose, talc, silicic acid, aluminum hydroxide, calcium silicates and polyamide powder, or mixtures of these substances. Sprays can additionally contain customary propellants, such as chlorofluorohydrocarbons and volatile unsubstituted hydrocarbons, such as butane and propane.Transdermal patches have the added advantage of providing controlled delivery to the body. Such dosage forms can be made by dissolving or dispersing the compound in the proper medium. Absorption enhancers can also be used to increase the flux of the compound across the skin. The rate of such flux can be controlled by either providing a rate controlling membrane or dispersing the compound in a polymer matrix or gel.Ophthalmic formulations, eye ointments, powders, solutions and the like, are also contemplated as being within the scope of this invention.Pharmaceutical compositions suitable for parenteral administration can comprise sterile isotonic aqueous or nonaqueous solutions, dispersions, suspensions or emulsions, or sterile powders which may be reconstituted into sterile injectable solutions or dispersions just prior to use, which may contain sugars, alcohols, antioxidants, buffers, bacteriostats, solutes which render the formulation isotonic with the blood of the intended recipient or suspending or thickening agents.Examples of suitable aqueous and nonaqueous carriers which may be employed in the pharmaceutical compositions of the invention include water, ethanol, polyols (such as glycerol, propylene glycol, polyethylene glycol, and the like), and suitable mixtures thereof, vegetable oils, such as olive oil, and injectable organic esters, such as ethyl oleate. Proper fluidity can be maintained, for example, by the use of coating materials, such as lecithin, by the maintenance of the required particle size in the case of dispersions, and by the use of surfactants.In certain embodiments, the above-described pharmaceutical compositions can be combined with other pharmacologically active compounds (“second active agents”) known in the art according to the methods and compositions provided herein. Second active agents can be large molecules (e.g., proteins) or small molecules (e.g., synthetic inorganic, organometallic, or organic molecules). In one embodiment, second active agents independently or synergistically help to treat infection.In another embodiment, the composition of the invention may comprise other biologically active substances, including therapeutic drugs or pro-drugs, for example, other chemotherapeutic agents, scavenger compounds, antibiotics, anti-virals, anti-fungals, anti-inflammatories,vasoconstrictors and anticoagulants, antigens useful for vaccine applications disclosed herein or corresponding pro-drugs.Exemplary scavenger compounds include, but are not limited to thiol-containing compounds such as glutathione, thiourea, and cysteine; alcohols such as mannitol, substituted phenols; quinones, substituted phenols, aryl amines and nitro compounds.Various forms of the chemotherapeutic agents and / or other biologically active agents may be used. These include, without limitation, such forms as uncharged molecules, molecular complexes, salts, ethers, esters, amides, and the like, which are biologically active.D. Therapeutic MethodsProvided herein are vaccine compositions for the treatment of and / or prevention of diseases and conditions for which an enhanced immune response may be beneficial. Such diseases include, but not limited to, pathogenic infections (e.g., bacterial, fungal, or protazoal infections). In some embodiments, the vaccine compositions described herein may be use as an immunotherapy .In certain aspects, provided herein are methods for preventing or treating a microbial infection in a subject. Such methods comprise administering to the subject a vaccine composition provided herein. In other aspects, provided herein are methods of preventing or treating opportunistic microbial infection in a subject. In some embodiments, the subject is immnocomprimised. Said methods comprise administering to the subject a vaccine composition provided herein.In some embodiments, the subject is elderly, a burn patient, a premature infant, undergoing chemotherapy, undergoing radiation therapy, receiving immunosuppressive or immunoablative / depleting therapy, or any combination thereof. In some embodiments, the subject is infected or at risk of infection by a PNAG-based microbe.In some embodiments, the vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) are useful in the treatment of diseases including, but not limited to, bacterial infections, fungal diseases, pathogenic diseases, persistent infectious disease, sexually transmitted diseases, gastro-intestinal diseases, pulmonary diseases, cardiovascular diseases, stress- and fatigue-related disorders.Bacterial, fungal and other pathogenic diseases include Aspergillus, Brugia, Candida, Chikungunya, Chlamydia, Coccidia, Cryptococcus, Dengue, Dirofilaria, Gonococcus, Histoplasma, Leishmania, Mycobacterium, Mycoplasma, Paramecium, Pertussis, Plasmodium, Pneumococcus, Pneumocystis, P. vivax in Anopheles mosquito vectors, Rickettsia, Salmonella, Shigella, Staphylococcus, Streptococcus, Toxoplasma and Vibriocholerae. Exemplary speciesinclude Neisseria gonorrhea, Mycobacterium tuberculosis, Candida albicans, Candida tropicalis, Trichomonas vaginalis, Haemophilus vaginalis, Group B Streptococcus sp., Microplasma hominis, Hemophilus ducreyi, Granuloma inguinale, Lymphopathia venereum, Treponema pallidum, Brucella abortus. Brucella melitensis, Brucella suis, Brucella canis, Campylobacter fetus, Campylobacter fetus intestinalis, Leptospira pomona, Listeria monocytogenes, Brucella ovis, Chlamydia psittaci, Trichomonas foetus, Toxoplasma gondii, Escherichia coli, Actinobacillus equuli, Salmonella abortus ovis, Salmonella abortus equi, Pseudomonas aeruginosa, Corynebacterium equi, Corynebacterium pyogenes, Actinobaccilus seminis, Mycoplasma bovigenitalium, Aspergillus fumigatus, Absidia ramosa, Trypanosoma equiperdum, Clostridium tetani, Clostridium botulinum', or, a fungus, such as, e.g., Paracoccidioides brasiliensis; or other pathogen, e.g., Plasmodium falciparum.Also included are National Institute of Allergy and Infectious Diseases (NIAID) priority pathogens. These include Category A agents, such as variola major (smallpox), Bacillus anthracis (anthrax), Yersinia pestis (plague), Clostridium botulinum toxin (botulism), Francisella tularensis (tularaemia), filo viruses (Ebola hemorrhagic fever, Marburg hemorrhagic fever), arenaviruses (Lassa (Lassa fever), Junin (Argentine hemorrhagic fever) and related viruses); Category B agents, such as Coxiella bumetti (Q fever), Brucella species (brucellosis), Burkholderia mallei (glanders), alphaviruses (Venezuelan encephalomyelitis, eastern & western equine encephalomyelitis), ricin toxin from Ricinus communis (castor beans), epsilon toxin of Clostridium perfringens; Staphylococcus enterotoxin B, Salmonella species, Shigella dysenteriae, Escherichia coli strain O157:H7, Vibrio cholerae, Cryptosporidium parvum', Category C agents, such as nipah virus, hantaviruses, yellow fever in Aedes mosquitoes, and multidrug-resistant tuberculosis; helminths, such as Schistosoma and Taenia', and protozoa, such as Leishmania e.g., L. mexicana) in sand flies, Plasmodium, Chagas disease in assassin bugs.Bacterial pathogens include, but are not limited to, bacterial pathogenic gram-positive cocci, which include but are not limited to: pneumococci; staphylococci; and streptococci. Pathogenic gram-negative cocci include: meningococci; and gonococci. Pathogenic enteric gramnegative bacilli include: enterobacteriaceae; pseudomonas, acinetobacteria and eikenella; melioidosis; salmonella; shigellosis; hemophilus; chancroid; brucellosis; tularemia; yersinia (pasteurella); streptobacillus moniliformis and spirilum; listeria monocytogenes; erysipelothrix rhusiopathiae; diphtheria; cholera; anthrax; and donovanosis (granuloma inguinale). Pathogenic anaerobic bacteria include; tetanus; botulism; other clostridia; tuberculosis; leprosy; and other mycobacteria. Pathogenic spirochetal diseases include: syphilis; treponematoses: yaws, pinta and endemic syphilis; and leptospirosis. Other infections caused by higher pathogen bacteria andpathogenic fungi include: actinomycosis; nocardiosis; cryptococcosis, blastomycosis, histoplasmosis and coccidioidomycosis; candidiasis, aspergillosis, and mucormycosis; sporotrichosis; paracoccidiodomycosis, petriellidiosis, torulopsosis, mycetoma and chromomycosis; and dermatophytosis. Rickettsial infections include rickettsial and rickettsioses. Examples of mycoplasma and chlamydial infections include: mycoplasma pneumoniae; lymphogranuloma venereum; psittacosis; and perinatal chlamydial infections. Pathogenic protozoans and helminths and infections eukaryotes thereby include: amebiasis; malaria; leishmaniasis; trypanosomiasis; toxoplasmosis; pneumocystis carinii; giardiasis; trichinosis; filariasis; schistosomiasis; nematodes; trematodes or flukes; and cestode (tapeworm) infections.Also included are biofilm-related infections and diseases. Biofilm-related infections are recognized in the art and may account for between 65-80% of all chronic, persistent (e.g., bacterial) infections, with the number of infections and conditions involving biofilms continuing to grow. For example, and without limitation, P. aeruginosa and the formation of highly -resistant biofilms are a dominant bacterial species in at least burn wound victims, chronic wound infections and cystic fibrosis (CF). Other bacteria such as, but not limited to; S. epidermis, S. aureus, and E. coli are also significant in biofilm formation, especially in relation to medicaldevice related infections.Aspergillus fumigatus, a non-limiting example of a fungal pathogen provided herein, is one of the most common Aspergillus species to cause disease in immuno-compromised individuals. Importantly, while A. fumigatus conidia account for less than 0.1% of all airborne fungal conidia, A. fumigatus conidia accounts for >80% of invasive infection in humans. Invasive aspergillosis is associated with higher morbidity and mortality, particularity in immunocompromised patients, e.g., in connection with bone marrow transplantation, liver transplant recipients, and patients with leukemia or lymphoma. A. fumigatus is the second most common cause of fungal infection found in hospitalized patients / healthcare settings after C. albicans.Thus, in some embodiments, the PNAG-based microbe is selected from Gram-positive bacteria, Gram-negative bacteria, fungal pathogens, protozoan pathogens.In some embodiments, the infection is a local infection. As non-limiting examples intended to be explanatory, the infection is limited to a region, organ, or system of the body of the subject, or otherwise encapsulted as is understood in the relevant art. In other emobdiments, the infection is a systemic infection.In some embodiments, provided herein are vaccines directed against multiple strains of a PNAG-based microbe. In some embodiments, the vaccine is directed against more than onePNAG-based microbe. In some embodiments, the more than one PNAG-based microbe is selected from Gram-positive bacteria, Gram-negative bacteria, fungal pathogens, protozoan pathogens, and any combination thereof.The uses for treating or inhibiting microbial / pathogenic infections (e.g., bacterial, fungal, and protozoal infections) may further comprise administering one or more antimicrobial agents to a subject having such an infection or at risk of developing such an infection. When an antimicrobial agent is included in the uses of the present invention, the antimicrobial agent may be administered prior to, concurrent with or after the vaccine formulation is administered to the subject. Where the antimicrobial agent is administered prior to or after the vaccine formulation, the period of time between when the antimicrobial agent and the vaccine formulation are administered may be a period of hours (such as 6, 12, 18 or 24 hours), days (such as 1, 2, 3, 4, 5, 6 or 7 days), weeks (such as 1, 2, 3 or 4 weeks) or months (such as 1, 2, 3, 4, 5, 6 or more months). The antimicrobial agent may be any known in the art to be generally effective in the treatment of the infection and may include, but is not limited to, an Aminoglycoside, such as Amikacin, Gentamicin, Kanamycin, Neomycin, Netilmicin, Streptomycin, Tobramycin or Paromomycin; a Carbacephem, such as Loracarbef; a Carbapenem, such as Ertapenem, Doripenem, Imipenem / Cilastatin or Meropenem; a Cephalosporin, such as Cefadroxil, Cefazolin, Cefalotin, Cefalexin, Cefaclor, Cefamandole, Cefoxitin, Cefprozil, Cefuroxime, Cefixime, Cefdinir, Cefditoren, Cefoperazone, Cefotaxime, Cefpodoxime, Ceftazidime, Ceftibuten, Ceftizoxime, Ceftriaxone, Cefepime or Ceftobiprole; a Glycopeptide, such as Teicoplanin or Vancomycin; a Macrolide, such as Azithromycin, Clarithromycin, Dirithromycin, Erythromycin, Erythroped, Roxithromycin, Troleandomycin, Telithromycin or Spectinomycin; a Monobactam, such as Aztreonam; a Penicillin, such as Amoxicillin, Ampicillin, Azlocillin, Carbenicillin, Cioxacillin, Dicloxacillin, Flucioxacillin, Mezlocillin, Meticillin, Nafcillin, Oxacillin, Penicillin, Piperacillin or Ticarcillin; a Polypeptide, such as Bacitracin, Colistin or Polymyxin B; a Quinolone, such as Ciprofloxacin, Enoxacin, Gatifloxacin, Levofloxacin, Lomefloxacin, Moxifloxacin, Norfloxacin, Ofloxacin or Trovafloxacin; a Sulfonamide, such as Mafenide, Prontosil (archaic), Sulfacetamide, Sulfamethizole, Sulfanilamide (archaic), Sulfasalazine, Sulfisoxazole, Trimethoprim or Trimethoprim-Sulfamethoxazole (Cotrimoxazole) (TMP-SMX); a Tetracycline, such as Demeclocycline, Doxycycline, Minocycline, Oxytetracycline or Tetracycline; as well as Chloramphenicol, Clindamycin, Lincomycin, Fusidic acid, Furazolidone, Linezolid, Metronidazole, Mupirocin, Nitrofurantoin, Macrobid, Platensimycin, Quinupristin / Dalfopristin, Rifampin or Rifampicin.The vaccine formulations provided herein may be administered in a pharmaceutically acceptable form and in substantially non-toxic quantities. The vaccine formulations may be administered to a subject using different dosing schedules, depending on the particular use to which the formulations are put (e.g., administration to the subject pre- or post-exposure to pathogen), the age and size of the subject, and the general health of the subject, to name only a few factors to be considered. In general, the vaccine formulations may be administered once, or twice, three times, four times, five times, six times or more, over a dosing schedule. The timing between each dose in a dosing schedule may range between a few hours, six, 12, or 18 hours, or 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 30 or more days. The same quantity of protein in the formulation may be administered in each dose of the dosing schedule, or the amounts in each dose may vary. The identity of the particular peptides and polypeptides in the formulation may also vary or remain the same in each dose in a dosing schedule.In some embodiments, the methods provided herein may comprise administering to a subject (e.g., a subject in need of treatment of or protection from infection), an effective amount of one or more vaccine compositions e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above). A subject in need thereof may include, for example, a subject who has been diagnosed with a microbial infection, at risk of microbial infection, or a subject who has been treated, including subjects that have been refractory to the previous treatment. For example, without being bound by theory or convention, subject contemplated herein include subjects with sepsis, septic shock, or at risk of either condition. Such subjects may also include, by way of example only, those who are elderly, burned subjects (particularly subjects having 20% or more burn coverage over their body), subjects with upcoming elected surgeries, those traveling to destinations where there is an outbreak of microbial infections, and the like. Thus, in some embodiments, the infection is resistant or refractory to drug treatment. In certain embodiments, the infection is resistant or refractory to one or more antibiotics.In some embodiments, the vaccines contemplated herein may be used therapeutically, particularly when the microbial infection is localized and / or non-life threatening. In such cases, a composition of the invention is administered to subjects suffering from a microbial infection arising from such microbes.The term “effective amount,” as in “a therapeutically effective amount,” of a therapeutic agent refers to the amount of the agent necessary to elicit the desired biological response. As will be appreciated by those of ordinary skill in this art, the effective amount of an agent may vary depending on such factors as the desired biological endpoint, the agent to be delivered, thecomposition of the pharmaceutical composition, the target tissue or cell, and the like. More particularly, the term “effective amount” refers to an amount sufficient to produce the desired effect, e.g., to reduce or ameliorate the severity, duration, progression, or onset of a disease, disorder, or condition, or one or more symptoms thereof; prevent the advancement of a disease, disorder, or condition, cause the regression of a disease, disorder, or condition; prevent the recurrence, development, onset or progression of a symptom associated with a disease, disorder, or condition, or enhance or improve the prophylactic or therapeutic effect(s) of another therapy.The compositions described herein may be delivered by any suitable route of administration, including orally, nasally, transmucosally, ocularly, rectally, intravaginally, parenterally, including intramuscular, subcutaneous, intramedullary injections, as well as intrathecal, direct intraventricular, intravenous, intra- articular, intra- sternal, intra- synovial, intra- hepatic, intralesional, intracranial, intraperitoneal, intranasal, or intraocular injections, intracisternally, topically, as by powders, ointments or drops (including eyedrops), including buccally and sublingually, transdermally, through an inhalation spray, or other modes of delivery known in the art.The terms “systemic administration,” “administered systemically,” “peripheral administration,” and “administered peripherally” as used herein mean the administration of the vaccine compositions described herein such that it enters the patient's system and, thus, is subject to metabolism and other like processes.The terms “parenteral administration” and “administered parenterally” as used herein mean modes of administration other than enteral and topical administration, usually by injection, and includes, without limitation, intravenous, intramuscular, intarterial, intrathecal, intracapsular, intraorbital, intraocular, intracardiac, intradermal, intraperitoneal, transtracheal, subcutaneous, subcuticular, intraarticular, subcapsular, subarachnoid, intraspinal and intrasternal injection and infusion.In some embodiments, the vaccine composition is administered systemically. Said systemic administration may be selected from the group consisting of oral, intravenous, intradermal, intraperitoneal, subcutaneous, and intramuscular administration.In certain embodiments the pharmaceutical compositions are delivered generally (e.g., via oral or parenteral administration). In certain other embodiments the pharmaceutical compositions are delivered locally through direct injection into a tumor or direct injection into the tumor’s blood supply (e.g., arterial or venous blood supply). In some embodiments, the pharmaceutical compositions are delivered by both a general and a local administration. For example, a subject with a tumor may be treated through direct injection of a compositioncontaining a composition described herein into the tumor or the tumor’s blood supply in combination with oral administration of a pharmaceutical composition of the present invention. If both local and general administration is used, local administration can occur before, concurrently with and / or after general administration.In some embodiments, the subject pharmaceutical compositions of the present invention will incorporate the substance or substances to be delivered in an amount sufficient to deliver to a patient a therapeutically effective amount of an incorporated therapeutic agent or other material as part of a prophylactic or therapeutic treatment. The desired concentration of the active compound in the particle will depend on absorption, inactivation, and excretion rates of the drug as well as the delivery rate of the compound. It is to be noted that dosage values may also vary with the severity of the condition to be alleviated. It is to be further understood that for any particular subject, specific dosage regimens should be adjusted over time according to the individual need and the professional judgment of the person administering or supervising the administration of the compositions. Typically, dosing will be determined using techniques known to one skilled in the art.Dosage may be based on the amount of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) per kg body weight of the patient. For example, a range of amounts of compositions or compound encapsulated therein are contemplated, including about 0.001, 0.01, 0.1, 0.5, 1, 10, 15, 20, 25, 50, 75, 100, 150, 200 or 250 mg or more of such compositions per kg body weight of the patient. Other amounts will be known to those of skill in the art and readily determined.In certain embodiments, the dosage of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) will generally be in the range of about 0.001 mg to about 250 mg per kg body weight, specifically in the range of about 50 mg to about 200 mg per kg, and more specifically in the range of about 100 mg to about 200 mg per kg. In one embodiment, the dosage is in the range of about 150 mg to about 250 mg per kg. In another embodiment, the dosage is about 200 mg per kg.In some embodiments the molar concentration of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) in a pharmaceutical composition will be less than or equal to about 2.5 M, 2.4 M, 2.3 M, 2.2 M, 2.1 M, 2 M, 1.9 M, 1.8 M, 1.7 M, 1.6 M, 1.5 M, 1.4 M, 1.3 M, 1.2 M, 1.1 M, 1 M, 0.9 M, 0.8 M, 0.7 M, 0.6 M, 0.5 M, 0.4 M, 0.3 M or 0.2 M. In some embodiments the concentration of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate asdescribed above) will be less than or equal to about 0.10 mg / ml, 0.09 mg / ml, 0.08 mg / ml, 0.07 mg / ml, 0.06 mg / ml, 0.05 mg / ml, 0.04 mg / ml, 0.03 mg / ml or 0.02 mg / ml.Actual dosage levels of the active ingredients in the compositions of the present invention may be varied so as to obtain an amount of the active ingredient which is effective to achieve the desired therapeutic response for a particular patient, composition, and mode of administration, without being toxic to the patient.The selected dosage level will depend upon a variety of factors including the activity of the particular therapeutic agent in the formulation employed, or the ester, salt or amide thereof, the route of administration, the time of administration, the rate of excretion or metabolism of the particular therapeutic agent being employed, the duration of the treatment, other drugs, compounds and / or materials used in combination with the particular compound employed, the age, sex, weight, condition, general health and prior medical history of the patient being treated, and like factors well known in the medical arts.A physician or veterinarian having ordinary skill in the art can readily determine and prescribe the effective amount of the pharmaceutical composition required. For example, the physician or veterinarian could prescribe and / or administer doses of the compounds of the invention employed in the pharmaceutical composition at levels lower than that required in order to achieve the desired therapeutic effect and gradually increase the dosage until the desired effect is achieved.In general, a suitable daily dose of a compound of the invention will be that amount of the compound which is the lowest dose effective to produce a therapeutic effect. Such an effective dose will generally depend upon the factors described above.If desired, the effective daily dose of the active compound may be administered as two, three, four, five, six or more sub-doses administered separately at appropriate intervals throughout the day, optionally, in unit dosage forms.The precise time of administration and amount of any particular compound that will yield the most effective treatment in a given patient will depend upon the activity, pharmacokinetics, and bioavailability of a particular compound, physiological condition of the patient (including age, sex, disease type and stage, general physical condition, responsiveness to a given dosage and type of medication), route of administration, and the like. The guidelines presented herein may be used to optimize the treatment, e.g., determining the optimum time and / or amount of administration, which will require no more than routine experimentation consisting of monitoring the subject and adjusting the dosage and / or timing.While the subject is being treated, the health of the patient may be monitored by measuring one or more of the relevant indices at predetermined times during a 24-hour period. All aspects of the treatment, including supplements, amounts, times of administration and formulation, may be optimized according to the results of such monitoring. The patient may be periodically reevaluated to determine the extent of improvement by measuring the same parameters, the first such reevaluation typically occurring at the end of four weeks from the onset of therapy, and subsequent reevaluations occurring every four to eight weeks during therapy and then every three months thereafter. Therapy may continue for several months or even years, with a minimum of one month being a typical length of therapy for humans. Adjustments, for example, to the amount(s) of agent administered and to the time of administration may be made based on these reevaluations.Treatment may be initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage may be increased by small increments until the optimum therapeutic effect is attained.Toxicity and therapeutic efficacy of subject compounds may be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 and the ED50. Compositions that exhibit large therapeutic indices are preferred. In some embodiments, the LD50 (lethal dosage) can be measured and can be, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% or more reduced for the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) described herein relative to the control. Similarly, the ED50 (i.e., the concentration which achieves a half-maximal inhibition of symptoms) can be measured and can be, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% or more increased for the one or more vaccine compositions (e.g., one or more QP wild type, QP mutant, antigen conjugate as described above) described herein relative to control. Also, similarly, the EC50 (i.e., the concentration which achieves half-maximal binding to or opsonic killing killing of targeted microbes) can be measured and can be, for example, at least 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 100%, 200%, 300%, 400%, 500%, 600%, 700%, 800%, 900%, 1000% or more increased for the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above)described herein relative to control. Although compounds that exhibit toxic side effects may be used, care should be taken to design a delivery system that targets the compounds to the desired site in order to reduce side effects.In some embodiments, the presently disclosed methods produce at least about a 10%, 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or even 100% protection from microbes by opsonic killing in an assay.In any of the above-described methods, the administering of the one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) can result in at least about a 10% , 15%, 20%, 25%, 30%, 35%, 40%, 45%, 50%, 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, or even 100% decrease in a solid malignancy in a subject, compared to the solid malignancy before administration of the vaccine compositions.In some embodiments, the therapeutically effective amount of one or more vaccine compositions (e.g., one or more QP wild type, or QP mutant, antigen conjugate as described above) is administered prophylactically to prevent a solid malignancy from forming in the subject.In some embodiments, the subject is human. In other embodiments, the subject is nonhuman, such as a mammal.The data obtained from the cell culture assays and animal studies may be used in formulating a range of dosage for use in humans. The dosage of any supplement, or alternatively of any components therein, lies preferably within a range of circulating concentrations that include the ED50 with little or no toxicity. The dosage may vary within this range depending upon the dosage form employed and the route of administration utilized. For agents of the present invention, the therapeutically effective dose may be estimated initially from cell culture assays. A dose may be formulated in animal models to achieve a circulating plasma concentration range that includes the ED50 as determined in cell culture. Such information may be used to more accurately determine useful doses in humans. Levels in plasma may be measured, for example, by high performance liquid chromatography.ExemplificationThe following Examples have been included to provide guidance to one of ordinary skill in the art for practicing representative embodiments of the presently disclosed subject matter. In light of the present disclosure and the general level of skill in the art, those of skill can appreciate that the following Examples are intended to be exemplary only and that numerous changes, modifications, and alterations can be employed without departing from the scope of the presently disclosed subject matter. The following Examples are offered by way of illustration and not by way of limitation.Example 1: Synthesis of the PNAG oligosaccharide library.To date, only fully acetylated PNAG and fully deacetylated PNAG have been investigated as immunogens for vaccine studies.3-15l The availability of a library of PNAG oligosaccharides with systematically varied numbers and locations of free amines can greatly aid in the understanding of the protective epitope structure. The proposed aim was to synthesize a comprehensive library of 32 pentasaccharides PNAG0-PNAG32 fully covering the free amine space of PNAG (Fig. 2). The reducing ends of the target pentasaccharides bear a linker terminated with a disulfide group, which could be reduced for chemo selective conjugation to a carrier protein.While several PNAG structures have been synthesized before,[17'20]a general method for expeditious construction of a comprehensive PNAG pentasaccharide library is lacking. To accelerate the library synthesis, rather than starting from monosaccharide building blocks for each targeted pentasaccharide, it was envisioned that the overall efficiency could be significantly enhanced with a divergent strategy. In the approach disclosed herein, the amine groups of strategically protected pentasaccharides were differentiated by orthogonal protective groups for selective deprotection and acetylation. After screening multiple synthetic intermediates, two key linchpin pentasaccharide intermediates (1 and 2) were designed, which bear four protective groups, i.e., tert-butyloxycarbonyl (Boc), allyloxycarbonyl (Alloc), 2,2,2- trichloroethoxycarbonyl (Troc), and fluorenylmethoxycarbonyl (Fmoc), on glucosamine units A, B, C and D. The reducing end glucosamine unit E is A- acetylated (for compound 1) or N- trifluoroacetylated (for compound 2).Based on the above design, synthesis commenced from thioglycoside 3, which glycosylated 3-azido-l -propanol 4 to provide compound 5 in 82% yield (Scheme 1 / Fig. 4). Upon removal of the Alloc group from 5 and A-acetylation, the resulting compound 6 was subjected to azide reduction, amidation of the free amine with carboxylic acid 7, and protective group adjustments leading to compound 8 in 45% yield for the four steps.Oligosaccharide assembly started from the CD disaccharide 9 containing A-Troc and A- Alloc groups (Scheme 2a / Fig. 5a). Thioglycoside donor 10 was preactivated with the p- TolSCl / AgOTf promoter system

[0021] at -78°C. Upon complete activation, the thioglycosyl acceptor 11 was added to the reaction mixture leading to disaccharide 9 in 83% yield (Scheme 2a). In order to extend the glycan chain, the glycosylation of disaccharide 9 with acceptor 8 was performed. When the reaction was first carried out under the pre-mix condition, i.e., 9 and 8 were mixed together followed by the addition of promoter (p-TolSCl / AgOTf or NIS / TfOH[22'23]), little desired trisaccharide 12 was obtained, which was likely due to the activation of the thioester(SAc) moiety by the thiophilic promoter. Next, the reaction was explored under the preactivation condition by activating 9 with the promoter p-TolSCl / AgOTf first, followed by the subsequent addition of acceptor 8. This change of the reaction protocol successfully produced trisaccharide 12 in 71% yield. Replacement of Alloc with Fmoc and removal of TBDPS group from 12 resulted in the trisaccharide 13. To extend 13 to a pentasaccharide, the Troc moiety of disaccharide 9 was replaced with Boc (disaccharide 14, Scheme 2b / Fig. 5b). Pre-activation based glycosylation of 14 and 13 produced pentasaccharide 1, which contains four different N- protective groups on units A, B, C and D. Analogously, pentasaccharide 2 was synthesized with four different A-protective groups on units A, B, C and D, and the A-TFA group on unit E (Fig. 21).With the two key pentasaccharides in hand, the orthogonal deprotection of pentasaccharides 1 and 2 was explored. As an example, the Boc and Alloc groups of compound 2 could be removed by 90% aqueous TFA and Pd(PPh3)4 / PhSiH3, while Troc and Fmoc deprotections were accomplished using Zn / AcOH and 20% piperidine in DMF respectively without affecting any other amine protective groups (Scheme 3 / Fig. 6). These results suggest that the four amine protective groups could be independently removed specifically.With the orthogonal deprotection conditions established, divergent modifications of the key pentasaccharide intermediates were carried out. Treatment of pentasaccharide 1 with 90% TFA cleaved both Boc and TBDPS groups (Scheme 4A / Fig. 7A). Upon acetylation of the newly liberated hydroxyl and amine moieties, the Alloc, Troc, and Fmoc groups were subsequently removed followed by deacylation with 20% hydrazine hydrate in MeOH, affording PNAG17 pentasaccharide in 48% overall yield bearing the A-acetylglucosamine (GlcNAc)-glucosamine (GlcN)-GlcN-GlcN-GlcNAc (10001) sequence. Alternatively, following TFA treatment of 1, the Fmoc group was cleaved, which was then acetylated with subsequent removal of Troc, Alloc and Bz moieties to produce pentasaccharide PNAG19 with the GlcNAc-GlcN-GlcN-GlcNAc- GlcNAc sequence (10011) in 51% overall yield. Similar divergent modification processes on the two key pentasaccharides 1 and 2 produced the full library of thirty-two PNAG pentasaccharides with all possible combinations of free amines in each glucosamine unit of the pentasaccharides (Scheme 4 / Fig. 7).Example 2: Superiority of the mQP-conjugate in inducing anti-PNAG antibodies compared to the corresponding tetanus toxoid conjugate.As carbohydrate antigens in general are T cell independent B cell antigens,

[0024] oligosaccharides <7 X 104Daltons are not immunogenic

[0015] and PNAG polysaccharides <100 KDa also are not immunogenic.[9]In order to boost the anti-carbohydrate antibody responses, theantigens were conjugated to an immunogenic carrier, such as the mutant bacteriophage QP (mQP).

[0025] [26'28]As PNAG oligosaccharides can potentially contain multiple free amine moieties, sulfhydryl chemistry for PNAG / mQP conjugation was employed. mQP was incubated with the bifunctional linker succinimidyl 3-(bromoacetamido)propionate (SBAP) 19 first to react with free amines on mQP surface (Scheme 5 A / Fig. 8A). Upon removal of the excess linker from SBAP functionalized mQP, PNAG pentasaccharide was added followed by quenching the unreacted bromoacetamide moieties on mQP with cysteine to avoid any potential side reactions of residual bromoacetamide in the vaccine upon storage or following vaccination. MALDI-TOF mass spectrometry (MS) analysis of the mQP-PNAG conjugate showed an average loading of around 250 copies of pentasaccharide per particle (Fig. 25). As each mQP particle consists of 180 copies of an identical coat protein,[25, 29]this corresponded to about 1.4 glycans per subunit on average.With the mQP-PNAG conjugates in hand, their abilities to induce anti-PNAG antibodies were evaluated. The conjugate of tetanus toxoid (TT) with the PNAG pentasaccharide bearing five free amines has undergone a phase 1 human clinical trial.

[0030] To benchmark this approach, PNAGO was covalently linked with the TT heavy chain (TTHc) using SBAP (Scheme 5C / Fig. 8C). (See Fig. 27.) Head-to-head comparison between the mQP and the TT conjugates was carried out. Groups of female C57B16 mice (n =5 per group) were immunized with freshly prepared mQP-PNAGO or the TT-PNAGO conjugate (8 nmol of PNAGO corresponding to 8 pg of PNAGO per injection) and monophosphoryl lipid A (MPLA, 20 pg) as the adjuvant on days 0, 14 and 28. A control group of mice received a mixture of mQP with PNAGO at equivalent amounts of mQP, PNAGO, and MPLA following the same immunization protocol. On day 35, sera were collected from all mice.To analyze the levels of antibodies generated, enzyme linked immunosorbent assay (ELISA) analyses were performed. In order to avoid the interference of anti-QP antibodies in the sera, the 32 PNAG pentasaccharides were conjugated with BSA (Fig. 26) individually and used as the ELISA coating antigen (Scheme 5B / Fig. 8B). As shown in Fig. 9A, mQP-PNAGO induced high anti-PNAG IgG titers (EC50 IgG titers GMT 75,613, measured against BSA- PNAGO) while the IgM titers were negligible (GMT < 1,000). The ability of mQP-PNAGO to produce anti-PNAG IgG antibodies suggests that the conjugate was able to activate helper T cells, leading to somatic hypermutation and isotype switching producing IgG antibodies. This is consistent with the idea that QP particles contain helper T cell epitopes that can activate these cells in both mice and humans.131 -32Furthermore, high levels of anti-PNAGO IgG responses were observed more than one year after the initial immunization (Fig. 9B). The IgG levels could beboosted back to near peak levels after nearly two years indicating that the mQP conjugate induced PNAGO specific memory B cells through immunization. Mice receiving mQP-PNAGO produced significantly higher anti-PNAGO IgG titers as compared to those immunized with the corresponding TT-PNAGO conjugate (GMT 4,765), indicating the superior immunogenicity of the mQP carrier. Mice immunized with the admixture of mQP and PNAGO did not produce any detectable levels of anti-PNAGO IgG (GMT < 1,000), highlighting the critical need to covalently conjugate mQP with PNAGO.As C57B16 mice were inbred, to enhance the rigor the immune analysis, outbred CD1 mice were immunized with mQP-PNAG conjugate following the same immunization protocol. mQP- PNAGO was comparably able to elicit high titers of anti-PNAGO IgG antibodies on day 35 in CD1 mice after the primary series of immunization (Fig. 28).Example 3: Probing antigen specificity of an Anti-PNAG mAb to guide vaccine design.The precise PNAG sequences synthesized by pathogens such as .S'. aureus are not known. Furthermore, the most abundant PNAG structure on cell surfaces that would encompass a fully acetylated pentasaccharide is not a protective epitope. O3] To guide vaccine design, it was envisioned that anti-PNAG mAb F598 could provide valuable information regarding optimal acetylation patterns in a PNAG pentasaccharide. Isolated from a patient who recovered from an .S'. aureus infection, F598 can protect mice against .S'. aureus inf ections.

[0033] The 32 PNAG pentasaccharide-BSA conjugates were immobilized onto a glycan micro array.

[0034] Following incubation of F598 with the microarray and washing, the amount of antibody remaining bound was quantified with a fluorescent secondary antibody. Interestingly, although F598 was initially identified due to binding to de-acetylated PNAG with - 15% N- acetylation,

[0033] glycan microarray showed that F598 actually preferred highly acetylated PNAG with the PNAG30 and PNAG31 having four or more consecutive GlcNAcs bound the strongest (Figure 10). No binding to PNAGO was observed on the microarray. Both the location and the number of NHAc were important for F598 binding, supporting the idea of amine / acetylation code. For example, despite having the same total number of NHAcs (4 in the molecules), PNAG23(10111) is a weak binder with apparent affinity less than 5% of that with PNAG30(l 1110). Out of the PNAGs with two or three GlcNAc residues, PNAG10 and PNAG26 are the strongest binders respectively.To better interpret the binding data, the GlcNAc binding preference of F598 was quantified by computing the preference index P for each unit of the pentasaccharide aswhere i (A-E) is the site of monosaccharide from the non-reducing end to the reducing end, j (0- 31) is the serial number of glycan, R is the intensity of the binding signal (RLU), and A is the code for amine vs acetylation (A = — 1 for free amine and A = 1 for NHAc). P value ranges from -1 to 1 with -1 and 1 indicating complete preference for free amine or NHAc respectively at the specific site. As shown in Fig. 10B, unit B position showed the highest P value of 0.91, indicating on average that there is a 91% chance to find an NHAc moiety rather than a free amine on saccharide B for ligand binding with F598. The P values for sites A, C and E were between 0.31-0.54 suggesting a moderate global preference for A-acetylation. There were almost no preferences for NHAc or free amine for site 5 as the P value at this site was close to 0.The importance of NHAc at unit B identified from microarray binding could be rationalized by the crystal structure of F598 complexed with fully acetylated PNAG oligosaccharides (PDB 6be4).

[0035] The binding pocket of F598 could accommodate PNAG with five GlcNAc residues. The NHAc groups on saccharides B and D in the binding pocket deeply inserted into the groove clamped by the heavy and the light chain of the mAb, forming multiple hydrogen bonds, while the NHAcs on units A, C and E only had weak to moderate interactions with the antibody. The carbonyl oxygen of the NHAc on saccharide B forms a hydrogen bonding with light chain A32 backbone amide while bridging with light chain R52 residue via a water molecule. The carbonyl oxygen of NHAc on saccharide D also formed hydrogen bonds with light chain A97 backbone amide and the hydroxyl of heavy chain Y50. Those interactions supported the relatively high dependence of NHAc on sites B and D for the binding of F598.Example 4: PNAG structure can be critical for the high immunogenicity of mQP-PNAG conjugates.Based on the microarray results and the report that fully acetylated PNAG antigen was poorly protective,[13'15]PNAG10 and PNAG26 were selected as new PNAG oligosaccharide antigens for evaluation. PNAG10 had the strongest binding to F598 in all PNAG structures with 2 or fewer NHAcs, and PNAG26 was the best binder among all structures with 3 or fewer NHAcs. Both PNAG10 and PNAG26 have NHAcs on glycan sites B and D. PNAG0 was utilized as a positive control since the corresponding TT-PNAG0 construct has entered clinical trials [ClinicalTrials.gov Identifier: NCT02853617].Mice were immunized with the mQP conjugates of PNAG10 or PNAG26 following the aforementioned immunization protocol (8 nmol PNAG, three injections on days 0, 14 and 28 with MPLA adjuvant). ELISA analysis of the immune sera showed significantly enhanced IgG antibody titers against the immunizing antigen (PNAG 10 or PNAG 26 respectively) with GMT of 191,141 and 227,064 ELISA units respectively as compared to pre-immune sera (Fig. 9A).To demonstrate the species generality of mQP conjugate, New Zealand white rabbits were immunized with mQP conjugates of PNAGO, PNAG10, and PNAG26 (8 nmol PNAG per injection) following a similar prime -boost protocol as that used in the mouse study. ELISA analysis of the post-immune sera showed that all 3 constructs induced strong anti-PNAG IgG responses with EC50 titers over 100,000 ELISA units (Fig. 9C), while those for the pre-immune sera were below 1,000 ELISA units. No side effects due to vaccinations were observed in either rabbits or mice.To evaluate the specificity of the immune sera, the post-immune sera was screened on the PNAG microarray, which included not only the 32 PNAG pentasaccharides, but also over 840 other components of common mammalian glycans and glycopeptides J34l As shown in Fig. 11A, serum IgG antibodies from rabbits immunized with mQP-PNAG were highly specific to PNAG structures with little binding to all non-PNAG microarray components including various GlcNAc containing glycans such as N-glycans and Lewis family glycans. Notably, none of the chitin oligosaccharides were bound by IgG antibodies in the sera suggesting that the antibodies well differentiated the pi,4-linked (chitin) from the pi,6-linked (PNAG) GlcNAcs.Analysis of the microarray binding by post-immune sera revealed selective PNAG epitope recognition by the post-immune sera (Fig. 11B). Rabbits immunized with mQP-PNAGO produced serum IgG antibodies exhibiting the strongest binding with the immunizing antigen structure PNAGO. Other good binders include PNAG1 and PNAG8, both having a single GlcNAc in the structure. PNAG4 with the sequence of GlcN-GlcN-GlcNAc-GlcN-GlcN, although it also only contains one GlcNAc, it had much lower binding with the sera (about 30% that to PNAG1). This suggests that greater than two consecutive GlcNs may be important for antibody binding.The mQP-PNAGlO immunized rabbits produced serum antibodies that preferentially bound to PNAG8(01000) and PNAGIO(OIOIO), which differ only by the GlcNAc in residue D indicating the non-reducing end GlcN-GlcNAc-GlcN may be the main epitope. Serum antibodies from mQP-PNAG26(l 1010) immunized rabbits preferentially bound to PNAG25(11001), PNAG26(11010), PNAG8(01000), and PNAG16(10000) suggesting GlcNAc-GlcNAc-GlcN and GlcNAc-GlcN-GlcN may be part of the epitopes being recognized.Example 5: Immune sera mediated bacterial recognition and protective functions.For an effective vaccine, it is important to establish that the post-immune sera not only bind the immunizing antigen, but also recognize the antigen on individual pathogen’s cells. .S'. aureus ATCC 29213 cells were reacted with rabbit immune sera. Bound antibodies were detected by a fluorescently labeled anti-rabbit IgG secondary antibody. As shown in Fig. 12, fluorescencemicroscopy images showed stronger binding to bacterial cells by IgG antibodies in mQP- PNAG10 and mQP-PNAG26 immune sera compared to the sera from mQP-PNAGO immunized rabbits or pre-immune sera.Whole cell ELISA was used to validate pathogen recognition and Fc mediated complement deposition. .S'. aureus cells were coated on ELISA plates, incubated with rabbit immune sera, and detected by secondary antibodies. For complement deposition, various immune sera were added to wells coated with purified PNAG isolated from Acinetobacter baumannii[6eialong with IgG / IgM depleted 2.5% human complement (Fig. 13 and Fig. 32). After incubation, the immobilized complement component Clq was detected by anti-Clq antibodies. As shown in Fig. 13, sera from mQP-PNAGlO and mQP-PNAG26 deposited significantly more Clq than those from mQP-PNAGO immunized rabbits, which in turn had more potent Clq binding than antibodies in sera from rabbits immunized with a conjugate of full length TT (150 kDa) and PNAGO (5GlcNH2-TT).[10a’10c]The abilities of the post-immune sera to kill bacteria in vitro were evaluated next via the opsonophagocytic killing (OPK) assay. .S', aureus cells were treated with pooled rabbit immune sera, followed by the addition of complement / phagocytic cells and quantification of the number of bacterial cells surviving the opsonic killing. As shown in Fig. 14, while the pre-immune sera were completely ineffective, all 3 constructs showed potent in vitro killing activity. mQP- PNAG26 (EC50: 2,534) and mQP-PNAGlO (EC50: 3,045) showed higher EC50 OPK titers as compared to mQP-PNAGO (EC50: 1,345). Omitting either immune sera, complement or phagocytic cells resulted complete loss of killing activity, suggesting the possible presence of classical complement activation pathway.Example 6: mQP-PNAG conjugate provided significant protection against .S', aureus induced death in both active and passive immunity studies.The efficacy of the vaccine in protecting against bacterial infection was tested in two mouse bacteremia challenge models. According to the CDC, bloodstream infections by .S', aureus are serious threats with nearly 20,000 death per year in the US.[4]For the in vitro study, first the mQP-PNAGO and TT-PNAG0 construct were compared. In the active protection model, mice were immunized three times with mQP-PNAGO or TT-PNAG0 at equivalent dose (8 nmol PNAGO) (n = 20 for each group) (Fig. 15). Another group of control mice received a mock injection of saline. Two weeks following the last vaccination, each mouse was challenged via tail vein with 10*LD50 of the .S'. aureus strain ATCC29213. Mice that received saline all died within 2 days of bacterial challenge. On the other hand, 95% of the mice receiving mQP-PNAGO were protected against death from this pathogen. The survival rate of the mQP vaccine group wassignificantly better than TT-PNAGO vaccinated group (p=0.0154) (Fig. 15A). A higher level of pathogen cells was detected in the kidneys of mice immunized with TT-PNAGO (35%), while mQP-PNAGO vaccination reduced the recovered levels of .S'. aureus from the kidneys to 5% of the infected mice (Fig. 15B). Contingency table analysis of the proportion of the 20 immunized mice in each group with or without detectable .S'. aureus by Fisher’s exact test showed significantly (p = 0.0436) fewer infected kidneys in the mQP-PNAGO immunized group, with a relative risk of 0.68 (95% CI= 0.45 to 0.93). Thus, disease burden evaluated by the levels of .S'. aureus in mouse kidneys was significantly better in mQP-PNAGO vaccinated group compared to those receiving the TT-PNAGO vaccine.As the mQP-PNAGO showed almost complete protection in the active protection model in mice, next a passive protection model was established to differentiate the various mQP-PNAG constructs, using rabbit sera transferred to mice. The passive model can be a more stringent test by using more dilute sera for protection. Rabbit sera were diluted 800 fold and administered intraperitoneally to mice, which were then challenged with 10*LD50 (200 million cells) of .S'. aureus ATCC29213 via the tail vein (Fig. 16). While all control mice receiving the pre-immune sera died within 3 days of this challenge, all post-immune sera from PNAG0, PNAG10, or PNAG26 immunized rabbits bestowed significant protection.Mice receiving sera from mQP-PNAG26 and mQP-PNAGlO immunized rabbits showed higher survival rates than those receiving PNAG0 sera (Fig. 16A) (60% and 50% respectively vs 30%) and lower pathogen load compared to mQP-PNAGO sera supporting the in vitro opsonic killing data (Fig. 16B). As PNAGs on cells can be heterogeneous in terms of the amine / acetylation patterns, next tested was the combination of two sera. Interestingly, administering the mixed PNAG26 and PNAG0 sera (1:1 ratio with each individual serum diluted 1,600 times, which is regarded equivalent to 1:800 dilution of a single serum) provided 100% protection to mice against the 10*LD50 challenges with .S', aureus (Fig. 16A). The kidneys of mice receiving the combination of PNAG26 and PNAG0 sera had no detectable bacteria (Fig. 16B).Example 7: mQP-PNAG vaccines are effective against MRS A challenges.The emergence of MRSA is a pressing public health concern)36! Effective vaccines can provide a complementary tool to combat .S', aureus infections and reduce the reliance on antibiotics. The post-immune rabbit sera were tested against clinically isolated MRSA strains first via immunofluorescent staining. All three mQP-PNAG sera recognized the six strains tested (Fig. 30) highlighting the breadth of immune recognition. Next, rabbit sera were diluted 800 times and administered to mice, which were then challenged with 10*LD50 (200 million cells) ofthe MRSA strain 1058 via the tail vein (Fig. 17A). Sera from mQP-PNAG26 immunized rabbits protected 90% of the mice from MRSA induced death, which was significantly higher than the 40% protection by mQP-PNAGO sera. Correspondingly, mice receiving mQP-PNAG26 rabbit sera had the lowest bacterial load in the kidneys of challenged mice (Fig. 17B).Example 8: Immunization with mQP-PNAG construct does not significantly alter the gut microbiome of mice.As PNAG is expressed on many types of bacteria, the effects of immunization on gut microbiome were explored. To analyze gut microbiome, mice were fully immunized with mQP- PNAG, and feces were collected on day 0 prior to immunization and day 35 following the initial prime immunization. The microbial species present in the droppings were analyzed via the 16S rRNA sequencing. Despite the significant amounts of anti-PNAG IgG produced in mouse sera, there were no significant changes in the microbial community present in the mouse gut. Amplicon sequence variant (ASV) datasets revealed similar patterns of ordination. When comparing treatment versus control within a timepoint, ANOSIM and PERMANOVA values of - 0.04 (Tl) and 1.518 (T2) indicated high similarity in the phylogenetic structure of the bacterial communities. (Fig. 31) Similar results were reported in a sponsored trial of the 5G1CNH2-TT vaccine and shown in the study of anti-PNAG therapy in the setting of graft-versus-host disease,

[0037] or in human subjects in phase 1 clinical trials of both the 5G1CNH2-TT vaccine or anti-PNAG mAb.[38'39]These observations corroborate that immunity to PNAG does not significantly alter the gut microbiome in immunized animals highlighting the potential safety of the vaccine.Example 9: DiscussionNumerous pathogens produce PNAG, rendering it a highly attractive target for vaccine development. However, the protective epitope of PNAG is not well understood, which severely hinders the development of effective vaccines. To elucidate epitope specificity, a comprehensive library of PNAG pentasaccharides covering all possible combinations of free amine and NHAc is provided herein. The synthesis is highlighted by a divergent design through the judicious choice of four amine protective groups, which can be orthogonally removed without affecting each other. The library of 32 PNAG pentasaccharides was synthesized from just two strategically protected pentasaccharide intermediates, thus significantly enhancing the overall synthetic efficiency.The library allowed probing of epitope specificity through a glycan microarray. Screening of an anti-PNAG mAb F598 on the microarray suggested that the NHAc at unit B plays a criticalrole in F598 binding. NHAc at unit D could further enhance the binding and led to the addition of two PNAG sequences (PNAG10 and PNAG26) beyond the fully deacetylated PNAG0 for vaccine studies.The three PNAGs were conjugated with carrier mQP, which elicited high levels of anti- PNAG IgG antibodies in both mice and rabbits. The mQP-PNAG conjugate was found to be superior to the corresponding PNAG conjugate with the benchmark carrier TT that has been evaluated in human clinical trials. Furthermore, the mQP-PNAG conjugate provided effective protection to mice against lethal challenges by .S'. aureus in both active and passive immunity models. Mice were also effectively protected from MRSA-induced death by the immune sera and significantly reduced the bacterial load in the kidneys. The vaccines are biocompatible with no adverse side effects and do not significantly disturb the gut microbiome of the immunized mice. PNAG based vaccine design guided by the well-defined synthetic library of PNAG is a powerful strategy to develop the next generation of vaccines to more effectively fight against pathogen infections including those by drug resistant strains.References[1] 2019 AR Threats Report, (cdc.gov / drugresistance / biggest-threats.html; accessed May 22nd, 2023).[2] T. M. Uddin, A. J. Chakraborty, A. Khusro, B. M. R. M. Zidan, S. Mitra, T. B. Emran, K. Dhama, M. K. H. Ripon, M. Gajdacs, M. U. K. Sahibzada, M. J. Hossain, N. Koirala, J. Infect. Public Health 2021, 14, 1750-1766.[3] F. Micoli, F. Bagnoli, R. Rappuoli, D. Serrato, Nat. Rev. Microbiol. 2021, 19, 287-302.[4] Deadly Staph Infections Still Threaten the U.S. (cdc.gov / media / releases / 2019 / p0305- deadly-staph-infections.html; accessed May 22nd, 2023).[5] R. C. Moellering, Jr, J. Antimicrob. Chemother. 2011, 67, 4-11.[6] T. Maira-Litran, A. Kropec, C. Abeygunawardana, J. Joyce, G. Mark, 3rd, D. A. Goldmann, G. B. Pier, Infect. Immun. 2002, 70, 4433-4440.[7] C. Cywes-Bentley, D. Skurnik, T. Zaidi, D. Roux, R. B. Deoliveira, W. S. Garrett, X. Lu, J. O'Malley, K. Kinzel, T. Zaidi, A. Rey, C. Perrin, R. N. Fichorova, A. K. Kayatani, T. Maira-Litran, M. L. Gening, Y. E. Tsvetkov, N. E. Nifantiev, L. O. Bakaletz, S. I. Pelton, D. T. Golenbock, G. B. Pier, Proc. Natl. Acad. Sci. U. S. A. 2013, 110, E2209-2218.[8] T. L. Nicholson, S. L. Brockmeier, N. Sukumar, A. E. Paharik, J. L. Lister, A. R. Horswill, M. E. Kehrli, Jr., C. L. Loving, S. M. Shore, R. Deora, Infect. Immun. 2017, 85.[9] K. E. Low, P. L. Howell, Curr. Opin. Struct. Biol. 2018, 53, 32-44.

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[0039] D. Vlock, J. C. Lee, A. Kropec-Huebner, G. B. Pier, in Abstracts of the 50th ICAAC 2010; Abstract Gl- 1654 / 329., 2010.Example 10: Synthesis Procedures -Tolyl 2-allyloxycarbonylamino-3.4-di-O-benzoyl-6-O- / (77-butyldiphenylsilyl-2-deoxy- 1- thio-p-D-glucopyranoside (3)To a solution of triol SI1(11.1 g, 30.0 mmol) in DCM (90 ml) was added imidazole (4.8 g, 2.5 equiv) and TBDPSC1 (8.6 ml, 1.1 equiv). (See Fig. 18A.) The reaction was stirred at room temperature for 6 h, after which it was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCOa and brine. The organic phase was dried (Na2SO4) followed by filtration, and thefiltrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (hexane s / ethyl acetate = 1 / 1) to afford the 3,4-diol compound (15.5g, 80% yield). The resulting diol was dissolved in pyridine (50 ml) followed by the addition of benzoyl chloride (6.1 ml, 2.2 equiv) under ice bath. The reaction was stirred at room temperature for 12 h, after which it was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCOa and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (hexanes / ethyl acetate = 5 / 1) to afford compound 3 (17.4 g, 91% yield). [a]o20= -21.2 (c 0.9, CH2CI2); ^-NMR (500 MHz, CDCh): 8. 7.93 (d, J =7.5 Hz, 2 H), 7.84 (d, J =8.0 Hz, 2 H), 7.72 (d, J =7.5 Hz, 2 H), 7.58 (d, J =7.5 Hz, 2 H), 7.52-7.47 (m, 4 H), 7.35-7.28 (m, 8 H), 7.18-7.15 (m, 2 H), 7.07 (d, J =7.5 Hz, 2 H), 5.80-5.73 (m, 1 H, -CH2CHCH2), 5.69-5.65 (m, 1 H, H-3), 5.59 (t, J =9.5 Hz, 1 H, H-4), 5.20 (d, J = 17.5 Hz, 1 H, -CH2CHCH2), 5.07-4.97 (m, 3 H, - CH2CHCH2, H-l and -NHAUoc), 4.49 (bs, 2 H, -CH2CHCH2), 3.92-3.79 (m, 4 H, H-2, H-5 and H-6), 2.33 (s, 3 H, SPI1-CH3), 1.03 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): A 165.0, 163.2, 155.5, 133.3 (2 C), 133.2, 132.9, 130.2, 130.0, 129.8, 129.7, 129.6 (2 C), 129.2, 129.0, 128.5, 128.3 (2 C), 127.7, 127.6, 87.1, 74.4, 68.8, 68.7, 62.8, 55.4, 26.6, 21.2, 19.1. HRMS: C47H49NNaO8SSi [M + Na]+calcd: 838.2840, obsd: 838.2830.3-Azidopropyl 2-allyloxycarbonylamino-3.4-di-O-benzoyl-6-O- / (77-butyldiphenylsilyl-2- deoxy-p-D-glucopyranoside (5)A solution of donor 3 (1.63 g, 2 mmol), acceptor 4 (0.18 ml, 2 mmol) and freshly activated molecular sieve MS 4 A in CH2Q2 (DCM) (15 mL) was stirred at room temperature for 30 minutes, and cooled to -78 °C, which was followed by the addition of AgOTf (6 mmol) dissolved in Et2<D / DCM without touching the wall of the flask. (See Fig. 18B.) After 5 minutes, orange colored p-TolSCI (290 pL, 2 mmol) was added to the solution through a microsyringe. The reaction mixture was warmed to -20 °C under stirring in 1 h. Then the mixture was diluted with DCM and filtered over Celite. The filtrate was extracted with ethyl acetate and washed with NaHCCh and brine. The organic phase was dried (NaoSCh) followed by filtration, and the filtrate was concentrated under reduced pressure and purified by silica gel flash chromatography (hexanes / ethyl acetate = 5 / 1) to provide compound 5 (1.3 g, 82% yield). [a]o20= -47.3 (c 1.0, CH2CI2); ^-NMR (500 MHz, CDCh): 8. 7.96 (d, J =8.0 Hz, 2 H), 7.86 (d, J =8.0 Hz, 2 H), 7.70 (d, J =8.0 Hz, 2 H), 7.59 (d, J =7.5 Hz, 2 H), 7.52-7.48 (m, 2 H), 7.39-7.31 (m, 8 H), 7.23 (t, J =7.5 Hz, 2 H), 5.82-5.76 (m, 1 H, -CH2CHCH2), 5.66-5.64 (m, 1 H, H-3), 5.61 (t, J =9.5 Hz, 1 H, H-4), 5.21 (d, 7 = 17.0 Hz, 1 H, -CH2CHCH2), 5.10-5.05 (m, 2 H), 4.71 (bs, 1 H), 4.53-4.50(m, 2 H, -CH2CHCH2), 4.02-3.98 (m, 1 H, -OCH2CH2CH2N3), 3.93-3.79 (m, 4 H), 3.65-3.61 (m, 1 H-OCH2CH2CH2N3), 3.45-3.36 (m, 2 H, -OCH2CH2CH2N3), 1.96-1.84 (m, 2 H, - OCH2CH2CH2N3), 1.04 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): A 166.6, 165.0, 155.7, 135.6, 135.5, 133.3, 133.2, 133.1, 133.0, 132.5, 130.0, 129.7 (2 C), 129.6, 129.2, 129.0, 128.3, 127.6 (2 C), 117.6, 101.4, 75.0, 73.0, 69.2, 66.1, 65.7, 62.8, 56.4, 48.2, 29.1, 26.6, 19.2. HRMS: C43H52N5O9Si [M + NH4]+calcd: 810.3529, obsd: 810.3518.3-Azidopropyl 2-acetamido-3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2-deoxy-|l-I)- glucopyranoside (6)To a solution of compound 5 (1.6 g, 2 mmol) in DCM (20 mL) was added Pd(PPh3)4(0.1 equiv) and phenylsilane (1 equiv). (See Fig. 18C.) The reaction was stirred at room temperature for 5 hours, after which it was extracted with ethyl acetate and washed with NaHCCh and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in pyridine (5 mL) followed by addition of acetic anhydride (2 ml). The reaction was stirred at room temperature for 1 hour, after which it was quenched by methanol and the mixture was concentrated under reduced pressure and purified by silica gel flash chromatography (hexanes / ethyl acetate = 2 / 1) to provide compound 6 (1.37 g, 91% yield). [α]D20= -101.9 (c 1.0, CH2CI2); ^-NMR (500 MHz, CDCh): A 7.96 (d, J =8.0 Hz, 2 H), 7.86 (d, J =8.5 Hz, 2 H), 7.71 (d, J =8.0 Hz, 2 H), 7.58 (d, J =8.0 Hz, 2 H), 7.51- 7.48 (m, 2 H), 7.40-7.29 (m, 8 H), 7.21 (t, J =7.5 Hz, 2 H), 5.86-5.82 (br, 1 H, -NHAc), 5.67- 5.61 (m, 2 H, H-3 and H-4), 4.75 (d, J =8.0 Hz, 1 H, H-l), 4.25-4.20 (m, 1 H, H-2), 4.02-3.98 (m, 1 H, -OCH2CH2CH2N3), 3.86-3.82 (m, 3 H), 3.65-3.60 (m, 1 H, -OCH2CH2CH2N3), 3.46- 3.36 (m, 2 H, -OCH2CH2CH2N3), 1.97-1.82 (m, 2 H, -OCH2CH2CH2N3), 1.03 (s, 9 H, - C(CH3)3).13C-NMR (125 MHz, CDCh): A 170.2, 167.0, 165.0, 135.7, 135.5, 133.5, 133.2,133.1, 133.0, 129.9, 129.7, 129.6, 129.2, 128.8, 128.5, 128.4, 127.6 (2 C), 101.3, 75.0, 73.4, 69.1,66.1, 65.7, 62.7, 54.8, 48.2, 29.1, 26.6, 23.4, 19.2. HRMS: C4iH47N4O8Si [M + H]+calcd: 751.3158, obsd: 751.3130.3-(3-(Acetylthio)propionylamino)propyl 2-acetamido-3.4-di-O-benzoyl-2-deoxy-|LI)- glucopyranoside (8)To a solution of compound 6 (1.5 g, 2 mmol) in MeOH / DCM (1:1, 16 mL) was added Pd(OH)2 (300 mg). (See Fig. 18D.) The mixture was hydrogenated with a hydrogen balloon for 5 hours. The reaction mixture was filtered through cotton and concentrated. The resultant amine and S-trityL3-mercaptopropionic acid 7 (765 mg, 2.2 mmol) were dissolved in DMF (8 ml).Upon addition of HATU (910 mg, 1.2 equiv) and DIPEA (1 ml, 3 equiv), the reaction was stirredat room temperature for 1 hour or until TLC indicated the reaction was complete. The reaction mixture was extracted by ethyl acetate and washed by 10% HC1, NaHCOa (sat) and brine. The organic phase was concentrated, and the mixture was purified by silica gel column chromatography (hexane s / ethyl acetate = 1 / 1) to afford the desired amide product. To a solution of the amide in THF (9 ml), acetic anhydride (3 ml) and acetic acid (6 mL) was added zinc dust (1.3 g). The reaction was stirred at room temperature for 5 hours, after which it was filtered over Celite and the filtrate was concentrated under reduced pressure. The residue was dissolved in pyridine (5 mL) and 70% HF*Py (1 mL) was added at 0 °C. The reaction was stirred at room temperature for 1 h, after which it was neutralized by sat. NaHCOa. The mixture was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCOa and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (toluene / acetone = 20 / 1) to afford compound 8 (554 mg, 45% yield). [α]D20= -78.9 (c 1.0, CH2CI2); ^-NMR (500 MHz, CDCh): . 7.90-7.88 (m, 4 H), 7.48-7.43 (m, 2 H), 7.37-7.26 (m, 4 H), 6.96 (d, J =8.5 Hz, 1 H, -NHAc), 6.74-6.71 (m, 1 H, -NHC=O), 5.74 (t, J = 10.0 Hz, 1 H, H-3), 5.44 (t, J =9.5 Hz, 1 H, H-4), 4.79 (d, J =8.0 Hz, 1 H, H-l), 4.30-4.24 (m, 1 H), 4.02-4.01 (m, 1 H), 3.83-3.69 (m, 3 H), 3.60-3.57 (m, 2 H, -OCH2CH2CH2N3), 3.32 (bs, 1 H), 3.15-3.11 (m, 3 H), 2.51-2.49 (m, 2 H, - NHC=OCH2-), 2.23 (s, 3 H, SAc), 1.88-1.70 (m, 5 H, NHAc and -OCH2CH2CH2NH-).13C-NMR (125 MHz, CDCh): A 196.2, 171.3, 171.0, 166.5, 166.0, 133.6, 133.4, 129.8, 129.0, 128.7, 128.5, 128.4, 101.5, 74.5, 73.2, 69.7, 67.6, 61.4, 54.4, 36.4, 36.0, 30.6, 29.4, 25.1, 23.2. HRMS: C30H37N2O10S [M + H]+calcd: 617.2169, obsd: 617.2190. -Tolyl 3.4-di-O-benzoyl-6-O- / (77-butyldiphenylsilyl-2-deoxy- l-thio-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranoside (10)To a solution of triol S21(9.2 g, 20.0 mmol) in DCM (60 ml) was added imidazole (3.2 g, 2.5 equiv) and TBDPSC1 (5.7 ml, 1.1 equiv). (See Fig. 19A.) The reaction was stirred at room temperature for 6 h, after which it was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCCh and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (hexane s / ethyl acetate = 1 / 1) to afford diol compound (11.9 g, 80% yield). The resulting compound was dissolved in pyridine (40 ml) followed by addition of benzoyl chloride (4.1 ml). The reaction was stirred at room temperature for 8 h, after which it was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCCh and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (hexanes / ethyl acetate = 5 / 1) to affordcompound 10 (11.6 g, 80% yield). [α]D20= -47.5 (c 1.5, CH2CI2); ^-NMR (500 MHz, CDCh): J 7.93 (d, J =8.0 Hz, 2 H), 7.84 (d, J =7.5 Hz, 2 H), 7.72 (d, J =7.0 Hz, 2 H), 7.58 (d, J =7.0 Hz, 2 H), 7.53-7.47 (m, 4 H), 7.35-7.28 (m, 8 H), 7.18-7.15 (m, 2 H), 7.08 (d, J =7.5 Hz, 2 H), 5.72 (t, J =10.0 Hz, 1 H, H-3), 5.62 (t, J =9.0 Hz, 1 H, H-4), 5.41 (d, J =9.0 Hz, 1 H, -NHTroc), 4.97 (d, J =10.5 Hz, 1 H, H-l), 4.74 (d, J =12.0 Hz, 1 H, -CH2CCI3), 4.62 (d, J =12.0 Hz, 1 H, - CH2CCI3), 3.99-3.93 (m, 1 H, H-2), 3.84-3.82 (m, 3 H, H-5 and H-6), 2.33 (s, 3 H, SPI1-CH3), 1.04 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): d. 166.5, 164.9, 154.0, 138.4, 135.7,135.5, 133.6, 133.4, 133.2, 132.8, 130.2, 130.0, 129.8, 129.7 (2 C), 129.6, 129.1, 128.8, 128.5, 128.4, 127.7, 127.6, 95.3, 87.1, 79.2, 74.4, 74.3, 68.7, 62.7, 55.5, 26.6, 21.2, 19.1. HRMS: C46H5oC13N208SSi [M + NH4]+calcd: 923.2123, obsd: 923.2120. -Tolyl 2-allyloxycarbonylamino-3,4-di-€>-benzoyl-2-deoxy-l-thio-p-D-glucopyranoside (11)To compound 3 (1.6 g, 2 mmol) dissolved in pyridine (5 mL) was added 70% HF*Py (1 mL) at 0 °C. (See Fig. 19B.) The reaction was stirred at room temperature for 1 h, after which it was neutralized by sat. NaHCCh. The mixture was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCCh and brine. The organic phase was dried (NajSCh) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (hexanes / ethyl acetate = 5 / 2) to afford compound 11 (831 mg, 72% yield). [α]D20= -52.9 (c 1.0, CH2CI2); ^-NMR (500 MHz, CDCh): 5. 7.90 (d, J =7.0 Hz, 4 H), 7.49-7.45 (m, 4 H), 7.34-7.21 (m, 4 H), 7.14-7.11 (m, 2 H), 5.81-5.70 (m, 2 H, NHAUoc and - CH2CHCH2), 5.43 (t, J =9.5 Hz, 1 H, H-3), 5.34 (d, J =9.5 Hz, 1 H, H-4), 5.16 (d, J = 17.0 Hz, 1 H, -CH2CHCH2), 5.05-5.01 (m, 2 H, H-l and -CH2CHCH2), 4.47 (bs, 2 H, -CH2CHCH2), 3.96- 3.88 (m, 1 H), 3.82-3.68 (m, 3 H), 2.59 (bs, 1 H, -OH), 2.33 (s, 3 H, SPI1CH3).13C-NMR (125 MHz, CDCh): A 166.5, 165.9, 155.6, 138.5, 133.6, 133.4, 132.5, 129.9 (2 C), 129.8, 128.9,128.6, 128.5, 128.4, 128.3, 117.4, 86.9, 78.5, 73.9, 69.4, 65.7, 61.7, 55.3, 21.2, 19.1. HRMS: C31H35N2O8S [M + NH4]+calcd: 595.2109, obsd: 595.2090. -Tolyl 3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2-deoxy-2-(2.2.2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4- di-O-benzoyl-2-deoxy-l-thio-p-D-glucopyranoside (9)A solution of donor 10 (905 mg, 1 mmol) and freshly activated molecular sieve MS 4 A (100 mg) in DCM (5 mL) was stirred at room temperature for 30 minutes, and cooled to -78 °C, which was followed by addition of AgOTf (3 mmol) dissolved in Et2<D / DCM without touching the wall of the flask. (See Fig. 19C.) After 5 minutes, orange colored p-TolSCl (144 pL, 1 mmol) was added to the solution through a microsyringe. After the donor was completely consumedaccording to TLC analysis (~ 10 minutes), a solution of acceptor 11 (519 mg, 0.9 mmol) in DCM (0.5 mL) was slowly added dropwise via a syringe. The reaction mixture was warmed to -20 °C under stirring in 1 h. Then the mixture was diluted with DCM and filtered over Celite. The filtrate was extracted with ethyl acetate and washed with NaHCOa and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure and purified by silica gel flash chromatography (hexanes / ethyl acetate = 4 / 1) to afford compound 9 (1 g, 83% yield). [α]D20= -59.1 (c 2.0, CH2CI2); ^-NMR (500 MHz, CDCh): A 7.96-7.87 (m, 8 H), 7.68 (d, J =6.5 Hz, 2 H), 7.56-7.47 (m, 8 H), 7.39-7.28 (m, 14 H), 7.20 (t, J =7.5 Hz, 2 H), 5.79-5.73 (m, 2 H, BH-3 and -CH2CHCH2), 5.57-5.47 (m, 2 H, AH-3 and AH-4), 5.39-5.36 (m, 1 H, BH-4), 5.20-5.03 (m, 5 H, BH-1, -NHAlloc, -NHTroc and -CH2CHCH2), 4.69 (d, J = 12.0 Hz, 1 H, -CH2CCI3), 4.62 (d, J =8.5 Hz, 1 H, AH-1), 4.56-4.50 (m, 3 H, - CH2CCI3 and -CH2CHCH2), 4.04-3.98 (m, 2 H), 3.94-3.91 (m, 1 H), 3.81-3.73 (m, 4 H), 3.70- 3.67 (m, 1 H), 2.36 (s, 3 H, SPI1CH3), 1.01 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): A166.3, 166.2, 165.5, 165.0, 155.3, 154.5, 138.9, 135.7, 135.5, 133.7 (2 C), 133.4, 133.3, 133.2, 133.1, 132.9, 132.4, 130.3, 130.0, 129.9, 129.8, 129.7, 129.6, 129.3, 129.0, 128.8, 128.6, 128.5,128.4, 128.3, 128.0, 127.6 (2 C), 117.5, 101.5, 95.5, 86.8, 78.5, 74.9, 74.3, 73.8, 73.2, 69.3, 69.2, 67.5, 65.8, 62.6, 56.5, 55.3, 26.6, 21.2, 19.1. HRMS: CvoHvaChlShOieSSi [M + NH4]+calcd: 1376.3541, obsd: 1376.3501.3-(3-(Acetylthio)propionylamino)propyl 3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2- deoxy-2-(2,2,2-trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-2- allyloxycarbonylamino-3,4-di-€)-benzoyl-2-deoxy-p-D-glucopyranosyl-(l— >6)-2-acetamido- 3,4-di-€>-benzoyl-2-deoxy-p-D-glucopyranoside (12)Following the general procedure for pre-activation based glycosylation, compound 12 was prepared from donor 9 and acceptor 8 (purified using toluene / acetone = 20 / 1, 71% yield). (See Fig. 19D.) [α]D20= -3.5 (c 0.2, CH2CI2); ^-NMR (500 MHz, CDCh): A 8.00-7.86 (m, 12 H), 7.54-7.11 (m, 28 H), 6.53 (d, J =6.5 Hz, 1 H, -NHC=O), 6.45 (bs, 1 H, -NHC=O), 6.19 (d, J =9.0 Hz, 1 H, -NHC=O), 5.69-5.59 (m, 5 H), 5.47-5.45 (m, 1 H), 5.29 (t, J =9.0 Hz, 1 H), 5.16 (d, J = 14.5 Hz, 1 H, -CH2CHCH2), 5.03 (d, J = 10.0 Hz, 1 H, -CH2CHCH2), 4.75-4.73 (m, 2 H, AH-1 and -CH2CCI3), 4.66 (d, J =7.0 Hz, 1 H, BH-1), 4.60 (d, J =7.0 Hz, 1 H, CH-1), 4.52-4.48 (m, 3 H, -CH2CCI3 and -CH2CHCH2), 4.24 (d, J =7.5 Hz, 1 H), 4.15-4.09 (m, 2 H), 4.04-4.00 (m, 3 H), 3.97-3.88 (m, 3 H), 3.82-3.70 (m, 4 H), 3.61 (bs, 3 H), 3.21-3.18 (m, 3 H, -CH2NH=O and -CH2SAC), 2.60-2.55 (m, 2 H, -NHC=OCH2-), 2.31 (s, 3 H, -SAc), 1.84-1.76 (m, 5 H, - NHAc and -OCH2CH2CH2NH-), 0.96 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): A 196.8,171.1, 170.7, 166.6, 166.3, 165.5, 164.9, 155.9, 154.7, 135.5, 135.4, 133.7, 133.6, 133.4, 133.3,133.2, 133.0 (2 C), 132.7, 132.6, 130.0, 129.9 (2 C), 129.7 (2 C), 129.6, 129.3, 129.0 (2 C), 128.9, 128.7, 128.6 (2 C), 128.5, 128.4 (2 C), 128.3 (2 C), 127.6, 127.5, 117.4, 102.4, 102.1,101.2, 95.6, 75.3, 74.1, 73.5, 73.2, 73.1, 70.4, 69.8, 69.3, 69.2, 67.9, 65.7, 64.0, 62.9, 56.6, 56.3, 54.6, 36.8, 35.9, 30.6, 29.7, 26.6, 25.3, 23.2, 19.0. HRMS: C93H98CI3N4O26SS1 [M + H]+calcd: 1851.5019, obsd: 1851.4938.3-(3-(Acetylthio)propionylamino)propyl 3,4-di-€>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- fluorenylmethyloxycarbonylamino-p-D-glucopyranosyl-( 1— >6)-2-acetamido-3,4-di-€L benzoyl-2-deoxy-p-D-glucopyranoside (13)To a solution of compound 12 (555 mg, 0.3 mmol) in DCM (5 mL) was added Pd(PPh3)4 (0.1 equiv) and phenylsilane (1 equiv). (See Fig. 19E.) The filtrate was extracted with ethyl acetate and washed with NaHCO3 and brine. The organic phase was dried (Na2SO4), filtered, and the filtrate was concentrated under reduced pressure. The residue was dissolved in pyridine (5 mL) followed by the addition of FmocCl (512 mg, 0.9 mmol) at 0°C. The reaction was stirred at room temperature for 5 hours, after which it was quenched by methanol. The mixture was extracted with ethyl acetate and washed with NaHCCL and brine. The organic phase was dried (Na2SO4) followed by filtration. The filtrate was concentrated under reduced pressure and purified by silica gel flash chromatography. The resultant compound was dissolved in pyridine (2 mL) followed by the addition of 70% HF*Py (0.5 mL) at 0 °C. The reaction was stirred at room temperature for 3 h, after which it was neutralized by sat. NaHCO3 ,. The mixture was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCCh and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (toluene / acetone = 12 / 1) to afford compound 13 (310 mg, 59% yield). [α]D20= -8.0 (c 0.3, CH2CI2); ^-NMR (500 MHz, CDCh): J. 7.99-7.86 (m, 12 H), 7.72 (d, J =7.5 Hz, 2 H), 7.68 (dd, J =7.5, 12.0 Hz, 1 H), 7.55-7.27 (m, 21 H), 7.20 (bs, 1 H), 7.12 (bs, 1 H), 6.66 (bs, 1 H, -NHC=O), 6.45 (bs, 1 H, -NHC=O), 6.13 (d, J =7.5 Hz, 1 H, -NHC=O), 5.82-5.68 (m, 4 H), 5.42-5.38 (m, 3 H), 4.92 (d, J =6.5 Hz, 1 H, AH- 1), 4.77-4.70 (m, 2 H, BH-1 and CH-1), 4.65 (d, J =12.0 Hz, 1 H, -CH2CCI3), 4.57 (d, J = 12.0 Hz, 1 H, -CH2CCI3), 4.51 (t, J =4.5 Hz, 1 H), 4.28-4.20 (m, 3 H), 4.12-3.88 (m, 8 H), 3.79-3.58 (m, 8 H), 3.20-3.14 (m, 3 H, -CH2NH=O and -CH2SAc), 2.52 (bs, 2 H, -NHC=OCH2-), 2.26 (s, 3 H, -SAc), 1.84 (s, 3 H, -NHAc), 1.77 (bs, 1 H, -OCH2CH2CH2NH-), 1.71 (bs, 1 H, - OCH2CH2CH2NH-).13C-NMR (125 MHZ, CDCh): 3. 196.7, 171.1, 170.9, 166.6, 166.1, 166.0, 165.7, 155.9, 154.6, 143.8, 141.1, 133.6, 133.4, 133.3, 133.2, 133.0, 132.1, 130.0 (3 C), 129.9 (2C), 129.8, 129.7, 128.9 (2 C), 128.8, 128.6, 128.5 (2 C), 128.4 (2 C), 128.3, 127.6, 127.0, 119.8, 101.7, 101.5, 101.3, 95.5, 74.7, 74.1, 73.4, 73.2, 72.6, 69.7, 66.8, 64.0, 61.2, 56.4, 54.4, 47.0,36.8, 35.9, 30.6, 29.3, 25.2, 23.2. HRMS: CssHseChN^S [M + H]+calcd: 1751.4311, obsd: 1751.4387. p-Tolyl 3,4-di-€>-benzoyl-6-€>- / er / -butyldiphenylsilyl-2-ter / -butyloxycarbonylamino-2-deoxy- P-D-glucopyranosyl-( 1— >6)-2-allyloxycarbonylamino-3,4-di-€)-benzoyl-2-deoxy- 1-thio-p-D- glucopyranoside (14)To a solution of disaccharide 9 (1.36 g, 1 mmol) in THF and acetic acid (3: 1, 12 mL) was added zinc dust (1.3 g, 20 mmol). (See Fig. 20A.)The reaction was stirred at room temperature for 6 hours, after which it was filtered by celite. The filtrate was extracted with ethyl acetate and washed with sat. NaHCOa (two times) and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The resultant residue and BOC2O (536 mg, 2 mmol) were dissolved in DCM (6 ml) followed by addition of DIPEA (0.2 ml) at 0°C. The reaction was stirred at room temperature for 5 hours. Then the solvent was removed and the resultant residue was purified by flash column chromatography (hexanes / ethyl acetate = 4 / 1) to afford compound 14 (1.2 g, 95% yield). [a]o20= -60.9 (c 1.5, CH2CI2); ^-NMR (500 MHz, CDCh): 8. 8.00 (d, J =7.0 Hz, 2 H), 7.94-7.89 (m, 6 H), 7.69-7.67 (m, 2 H), 7.58- 7.48 (m, 8 H), 7.40-7.26 (m, 14 H), 7.22 (t, J =7.5 Hz, 2 H), 5.80-5.74 (m, 2 H, BH-3 and - CH2CHCH2), 5.56-5.47 (m, 2 H, AH-3 and AH-4), 5.39 (t, J =9.5 Hz, 1 H, BH-4), 5.21 (d, J = 17.5 Hz, 1 H, -CH2CHCH2), 5.12-5.07 (m, 3 H, BH-1, -NHAUoc and -CH2CHCH2), 4.66 (d, J =7.5 Hz, 1 H, AH-1), 4.54-4.51 (m, 3 H, -NHBoc and -CH2CHCH2), 4.04 (d, J = 11.5 Hz, 1 H), 3.97-3.79 (m, 6 H), 3.71 (d, J =9.0 Hz, 1 H), 2.36 (s, 3 H, SPI1CH3), 1.30 (s, 9 H, -C(CH3)3), 1.02 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): 8 166.3, 165.4, 165.1, 155.5, 155.4,138.8, 135.7, 135.5, 133.7, 133.6, 133.4, 133.2, 133.1 (2 C), 132.9, 132.5, 130.2, 130.0, 129.9 (2 C), 129.8, 129.7, 129.6, 129.4 (2 C), 128.9, 128.7, 128.5, 128.3 (2 C), 128.1, 127.6 (2 C), 117.4, 101.7, 87.0, 79.6, 78.7, 74.9, 73.9, 73.7, 69.5, 69.4, 67.7, 65.8, 62.7, 55.8, 55.3, 28.2, 26.6, 21.2, 19.2. HRMS: C72H8oN30i6SSi [M + NH4]+calcd: 1302.5023, obsd: 1302.5016.3-(3-(Acetylthio)propionylamino)propyl 3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2- / (77- butyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-( 1 —>6)-2-allyloxycarbonylamino-3,4- di-€>-benzoyl-2-deoxy-p-D-glucopyranosyl-(1^6)-3,4-di-€>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- fluorenylmethyloxycarbonylamino-p-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-€)- benzoyl-2-deoxy-p-D-glucopyranoside (1)Following the general procedure for pre-activation based glycosylation, compound 1 was prepared from donor 14 and acceptor 13 (purified with toluene / acetone = 10 / 1, 69% yield). (See Fig. 20B.) [α]D20= -53.1 (c 0.8, CH2CI2); ^-NMR (500 MHz, CDCh): . 8.32 (d, J =7.5 Hz, 2 H), 8.27-8.24 (m, 4 H), 8.12-7.92 (m, 12 H), 7.87 (d, J =7.5 Hz, 1 H), 7.70-6.85 (m, 46 H), 6.80 (t, J =7.5 Hz, 1 H), 6.64 (bs, 1 H, -NHC=O), 6.01-5.89 (m, 3 H), 5.83 (t, J =9.5 Hz, 1 H), 5.71 (t, J =9.5 Hz, 1 H), 5.44-5.37 (m, 3 H, -CH2CHCH2), 5.25 (t, J = 10.0 Hz, 1 H), 5.18 (t, J =9.5 Hz, 1 H), 5.09 (d, J =8.5 Hz, 1 H, anomeric H), 4.96-4.73 (m, 7 H, two anomeric H and - CH2CHCH2), 4.59-4.50 (m, 2 H), 4.43-4.11 (m, 10 H, anomeric H), 3.96-3.87 (m, 3 H, anomeric H), 3.79-3.67 (m, 3 H), 3.62-3.55 (m, 3 H), 3.41-3.17 (m, 4 H), 2.68-2.64 (m, 2 H, -NHC=OCH2- ), 2.28 (s, 3 H, -SAc), 1.90-1.87 (m, 5 H, -NHAc and -OCH2CH2CH2NH-), 1.17 (s, 9 H, - C(CH3)3), 1.07 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): d. 195.9, 171.2 (2 C), 166.9,166.8, 166.7 (2 C), 166.4, 166.3, 166.0, 165.7, 165.0, 164.6, 156.6, 156.3, 156.0, 155.6, 144.2, 143.5, 141.0, 140.6, 135.6, 135.3, 133.4 (2 C), 133.3, 133.2, 133.1, 133.0, 132.9, 132.8, 132.1 (2 C), 132.0, 131.9, 131.8, 131.4, 130.5, 130.4, 130.3, 130.2, 130.0, 129.9, 129.8 (2 C), 129.7, 129.6 (2 C), 129.2 (2 C), 129.0, 128.9, 128.7 (2 C), 128.6, 128.5 (2 C), 128.4, 128.3 (2 C), 128.2,128.1, 28.0, 127.7, 127.3, 127.2, 127.1, 126.7, 125.3, 125.1, 119.4, 119.3, 117.7, 105.4, 105.3, 103.0, 102.3, 100.5, 95.4, 80.7, 74.6, 74.5, 74.2, 73.9, 73.4, 73.2, 72.7, 72.3, 72.1, 72.0, 71.7,70.8, 69.6, 69.2, 67.7, 66.5, 66.3, 64.6, 64.0, 57.4, 56.6, 56.3, 55.8, 54.2, 47.0, 37.0, 35.5, 30.5,29.1, 28.0, 26.8, 25.2, 22.7. HRMS: CissHissChNeC^SSi [M + 2H]2+calcd: 1456.4361, obsd: 1456.4336.3-(3-(Acetylthio)propionylamino)propyl 3.4-di-O-benzoyl-2-deoxy-2-trifluoroacetamido-|5- D-glucopyranoside (S3)Compound S3 was synthesized analogously as compound 8. (See Fig. 22A.)To a solution of compound 5 (1.6 g, 2 mmol) in DCM (20 mL) was added Pd(PPh3)4 (0.1 equiv) and phenylsilane (1 equiv). The reaction was stirred at room temperature for 5 hours, after which it was extracted with ethyl acetate and washed with NaHCO3and brine. The organic phase was dried (Na SCL) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was dissolved in DCM (10 mL) and DIPEA (1.7 ml) followed by addition of trifluoroacetic anhydride (0.3 ml, 2.2 mmol) at -78°C. The reaction was stirred at room temperature for 1 hour, after which it was quenched by methanol and the mixture was concentrated under reduced pressure and purified by silica gel flash chromatography (hexanes / ethyl acetate = 2 / 1) to provide the desired product. To a solution of above compound (1.6 g, 2 mmol) in MeOH / DCM (1:1, 16 mL) was added Pd(OH)2 (300 mg). The mixture was hydrogenated with a hydrogen balloon for 5 hours. The reaction mixture was filtered through cotton and concentrated. The resultant amine and S-trityl-3-mercaptopropionic acid 7 (765 mg, 2.2 mmol) were dissolved in DMF (8 ml). Upon addition of HATU (912 mg, 1.2 equiv) and DIPEA (1 ml, 3 equiv). The reaction was stirred at room temperature for 1 hour or until TLC indicated the reaction was complete. The reaction mixture was extracted by ethyl acetate and washed by 10% HC1, NaHCOa (sat) and brine. The organic phase was concentrated, and the mixture was purified by silica gel column chromatography (hexane s / ethyl acetate = 1 / 1) to afford the desired product. To a solution of the above compound in THF (9 ml), acetic anhydride (3 ml) and acetic acid (6 mL) was added zinc dust (1.3 g). The reaction was stirred at room temperature for 5 hours, after which it was filtered over Celite, and the filtrate was concentrated under reduced pressure. The residue was dissolved in pyridine (5 mL) followed by the addition of 70% HF*Py (1 mL) at 0 °C. The reaction was stirred at room temperature for 1 h, after which it was neutralized by sat. NaHCOa. The mixture was extracted with ethyl acetate and washed with 10% HC1, sat. NaHCOa and brine. The organic phase was dried (Na2SO4) followed by filtration, and the filtrate was concentrated under reduced pressure. The residue was purified by flash column chromatography (toluene / acetone = 20 / 1) to afford compound S3 (493 mg, 45% yield). [α]D20= -55.4 (c 1.0, CH2Q2); ^-NMR (500 MHz, CDCh): J. 8.21 (d, J =8.5 Hz, 1 H), 7.93-7.90 (m, 4 H), 7.53-7.47 (m, 2 H), 7.38-7.33 (m, 4 H), 6.18 (bs, 1 H, -NHTFA), 5.79 (t, J =10.0 Hz, 1 H, H-3), 5.49 (t, J =9.5 Hz, 1 H, H-4), 4.84 (d, J =7.5 Hz, 1 H, H-l), 4.34-4.29 (m, 1 H), 4.07-4.03 (m, 1 H), 3.86-3.71 (m, 3 H), 3.67-3.60 (m, 1 H), 3.55-3.51 (m, 1 H, -CH2NH=O), 3.19-3.10 (m, 3 H, -CH2NH=O and -CH2SAc), 2.56-2.44 (m, 2 H, -NHC=OCH2-), 2.31 (s, 3 H, -SAc), 1.85-1.82 (m, 1 H, -OCH2CH2CH2NH-), 1.67-1.63 (m, 1 H, -OCH2CH2CH2NH-).13C-NMR (125 MHZ, CDCh): A 196.7, 171.3, 166.5, 166.1, 158.4 (q, J =37.5 Hz, C=O of TFA), 133.8, 133.5, 129.9, 129.8, 128.6, 128.5 (2 C), 128.4, 119.1 (q, J =286 Hz, CF3of TFA), 101.0, 74.7, 72.7, 69.4, 67.9, 61.2, 55.0, 36.4, 36.1, 30.6, 29.6, 25.2. HRMS: C30H34F3N2O10S [M + H]+calcd: 671.1881, obsd: 671.1889.3-(3-(Acetylthio)propionylamino)propyl 3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2- deoxy-2-(2,2,2-trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-2- allyloxycarbonylamino-3,4-di-€)-benzoyl-2-deoxy-p-D-glucopyranosyl-(l— >6)-3,4-di-€)- benzoyl-2-deoxy-2-trifluoroacetamido-p-D-glucopyranoside (S4)Following the general procedure for pre-activation based glycosylation, compound S4 was prepared from donor 9 and acceptor S3 (See Fig. 21A and Fig. 22B.) (purified with toluene / acetone = 20 / 1, 65% yield). [α]D20= -2.9 (c 0.2, CH2CI2); ^-NMR (500 MHz, CDCh): A 8.00-7.86 (m, 12 H), 7.56-7.10 (m, 28 H), 6.17 (bs, 1 H, -NHC=O), 5.95 (bs, 1 H, -NHC=O), 5.80-5.43 (m, 5 H), 5.28-5.04 (m, 4 H), 4.84 (d, J =8.0 Hz, 1 H, CH-1), 4.71 (d, J =12.0 Hz, 1 H,-C / CCCh), 4.64 (d, J =8.0 Hz, 1 H, AH- 1 ), 4.55-4.48 (m, 4 H, BH-1, -CH2CCI3 and - CH2CHCH2), 4.37-4.31 (m, 1 H), 4.21-4.16 (m, 2 H), 4.05-3.86 (m, 5 H), 3.76-3.57 (m, 5 H), 3.21-3.18 (m, 3 H, -CH2NH=O and -CH2SAc), 2.58-2.52 (m, 2 H, -NHC=OCH2-), 2.32 (s, 3 H, - SAc), 1.86 (bs, 1 H, -OCH2CH2CH2NH-), 1.73 (bs, 1 H, -OCH2CH2CH2NH-), 0.99 (s, 9 H, - C(CH3)3).13C-NMR (125 MHz, CDCh): A 196.8, 171.1, 166.4, 166.2, 166.1, 165.5, 164.9, 158.3 (q, J =37.5 Hz, C=O of TFA), 155.9, 154.8, 135.5, 135.4, 133.8, 133.7, 133.4 (2 C), 133.3 (2 C), 133.0, 132.6, 130.0, 129.9 (2 C), 129.7 (2 C), 129.6, 129.2, 128.9 (2 C), 128.6 (2 C), 128.5, 128.4 (2 C), 128.3 (2 C), 127.6, 127.5, 119.1 (q, J =286.5 Hz, CF3of TFA), 117.4, 102.8, 102.4, 100.6,95.4, 75.7, 74.2, 73.5, 73.3, 73.0, 72.6, 70.4, 69.9, 69.3, 68.2, 65.7, 63.2, 56.7, 56.2, 54.9, 36.5, 36.1, 30.6, 29.4, 26.6, 25.2, 19.0. HRMS: CgsHgsChFsN^SSi [M + H]+calcd: 1905.4742, obsd: 1905.4798.3-(3-(Acetylthio)propionylamino)propyl 3,4-di-€>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- fluorenylmethyloxycarbonylamino-p-D-glucopyranosyl-( 1— >6)-3,4-di-€)-benzoyl-2-deoxy-2- trifluoroacetamido-p-D-glucopyranoside (S5)Following the procedure for synthesis of compound 13, compound S5 (toluene / acetone = 15 / 1, 49% yield) was prepared from compound S4. (See Fig. 21A and Fig. 22B.) [a]o20= -1.0 (c 0.1, CH2CI2); ^-NMR (500 MHz, CDCh): 8. 8.27 (bs, 1 H), 8.01-7.87 (m, 12 H), 7.72 (d, J =7.5 Hz, 2 H), 7.53-7.27 (m, 22 H), 7.21 (bs, 1 H), 7.12 (bs, 1 H), 6.25 (bs, 1 H, -NHC=O from linker), 5.97 (d, J =8.5 Hz, 1 H), 5.86-5.57 (m, 4 H), 5.44-5.40 (m, 3 H), 4.94 (d, J =5.5 Hz, 1 H, CH-1), 4.79 (bs, 2 H, AH-1 and BH-1), 4.60-4.55 (m, 2 H), 4.39-4.22 (m, 4 H), 4.07-3.98 (m, 6 H), 3.85-3.59 (m, 7 H), 3.17-3.14 (m, 3 H, -CH2NH=O and -CH2SAc), 2.50 (bs, 2 H, - NHC=OCH2-), 2.25 (s, 3 H, -SAc), 1.79 (bs, 1 H, -OCH2CH2CH2NH-), 1.68 (bs, 1 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, CDCh): A 196.8, 171.1, 166.4, 166.2, 158.3 (q, J =37.5 Hz, C=O of TFA), 155.9, 154.6, 143.9, 143.6, 141.1, 133.7 (2 C), 133.5, 133.4, 133.3, 129.9 (2 C), 129.8, 128.9, 128.8, 128.7, 128.6, 128.5, 128.4, 128.3, 127.6, 127.0, 125.1, 119.9, 119.2 (q, J =286.0 Hz, CF3of TFA), 101.9, 101.6, 100.7, 95.4, 74.6, 74.1, 73.6, 73.0, 72.8, 72.4,70.4, 69.8, 68.1, 66.8, 61.3, 56.8, 56.4, 54.7, 47.0, 36.4, 36.0, 30.6, 29.4, 25.1. HRMS: C88H83CI3F3N4O26S [M + H]+calcd: 1805.4034, obsd: 1805.4103.3-(3-(Acetylthio)propionylamino)propyl 3.4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2- / (77- butyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-( 1 —>6)-2-allyloxycarbonylamino-3,4- di-€>-benzoyl-2-deoxy-p-D-glucopyranosyl-(1^6)-2-deoxy-3,4-di-€>-benzoyl-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-fluorenylmethyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-( 1— >6)-3,4-di-€)-benzoyl-2- deoxy-2-trifluoroacetamido-p-D-glucopyranoside (2)Following the general procedure for pre-activation based glycosylation, compound 2 (toluene / acetone = 12 / 1, 61% yield) was prepared from donor 14 and acceptor S5. (See Fig. 21B and Fig. 22C.) [α]D20= -4.0 (c 0.2, CH2CI2); ^-NMR (500 MHz, CDCh): J. 8.63 (d, J =9.5 Hz,1 H, -NHC=O), 8.28-8.24 (m, 6 H), 8.11-8.05 (m, 6 H), 7.96-7.91 (m, 6 H), 7.87 (d, J =7.0 Hz, 2 H), 7.68-6.85 (m, 47 H), 6.78 (t, J =8.0 Hz, 1 H), 6.39 (bs, 1 H, -NHC=O), 6.05-5.99 (m, 2 H), 5.92-5.86 (m, 2 H), 5.73 (t, J =9.5 Hz, 1 H), 5.67 (t, J =9.5 Hz, 1 H), 5.44-5.38 (m, 3 H, - CH2CHCH2), 5.24-5.18 (m, 2 H, anomeric H), 5.14 (t, J = 10.0 Hz, 1 H), 4.96-4.64 (m, 9 H, two anomeric H and -CH2CHCH2), 4.53-4.19 (m, 11 H, anomeric H), 4.13-4.04 (m, 3 H), 3.95 (t, J =9.5 Hz, 1 H), 3.87-3.84 (m, 2 H, anomeric H), 3.79-3,50 (m, 7 H), 3.37-3.23 (m, 4 H, - CH2NH=O and -CH2SAc), 2.65-2.57 (m, 2 H, -NHC=OCH2-), 2.28 (s, 3 H, -SAc), 1.91-1.86 (m,2 H, -OCH2CH2CH2NH-), 1.15 (s, 9 H, -C(CH3)3), 1.06 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): A 195.9, 171.0, 166.9, 166.8, 166.7, 166.6, 166.4, 166.1, 166.0, 165.7, 164.7, 164.5, 158.5 (q, J =37.5 Hz, C=O of TFA), 156.6, 156.2, 156.0, 155.5, 144.2, 143.5, 141.0, 140.6,135.5, 135.3, 133.4 (2 C), 133.3, 133.2, 133.1 (2 C), 133.0, 131.9, 131.4, 130.5, 130.4, 130.3,130.2, 130.0, 129.9 (2 C), 129.7, 129.6, 129.4, 129.2, 129.0 (2 C), 128.8 (2 C), 128.7 (2 C),128.5, 128.4 (2 C), 128.2 (2 C), 128.1 (2 C), 127.7, 127.3, 127.2, 127.1, 126.7, 125.3, 125.1, 119.4, 119.3(2 C) (q, J =286.0 Hz, CF3of TFA), 117.6, 105.2, 103.0, 102.4, 99.7, 95.2, 80.7,74.5, 74.1, 73.8, 73.4, 73.2, 73.1, 72.7, 72.5, 72.4, 72.3 (2 C), 71.9, 71.8, 70.8, 69.6, 68.0, 66.5,66.2, 64.6, 57.5, 56.6, 56.3, 55.8, 54.9, 47.0, 36.8, 35.6, 30.5, 29.0, 27.9, 26.7, 25.1, 18.8. HRMS: Ci53Hi5iCl3F3N6O42SSi [M + H]+calcd: 2965.8367, obsd: 2965.8389.3-(3-(Acetylthio)propionylamino)propyl 6-O-acetyl-2-acetamido-3.4-di-O-benzoyl-2-deoxy- P-D-ghicopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4-di-€)-benzoyl-2-deoxy-p-D- glucopyranosyl-( 1 — >6)- 3,4-di-O>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- fhiorenylmethyloxycarbonylamino-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- trifluoroacetamido-p-D-glucopyranoside (15)Following the general procedure for Boc removal and acetylation, compound 15 was prepared in 85% yield for two steps. (See Fig. 23).1H-NMR (500 MHz, CDCh): A. 8.67 (m, 1 H), 8.27-7.89 (m, 20 H), 7.62-7.17 (m, 32 H), 7.11-7.03 (m, 3 H), 6.94-6.89 (m, 1 H), 6.82-6.78 (m, 1 H), 6.60-6.57 (m, 1 H, -NHC=O), 6.13-5.95 (m, 4 H), 5.84-5.30 (m, 12 H), 5.21-5.15 (m, 2 H), 4.97-4.75 (m, 8 H), 4.70-4.39 (m, 10 H), 4.32-4.08 (m, 11 H), 4.04-3.91 (m, 4 H), 3.83-3.55(m, 9 H), 3.52-3.29 (m, 2 H), 3.24-3.21 (m, 2 H, -CH2SAc), 2.67-2.56 (m, 2 H, -NHC=OCH2-), 2.27 (s, 3 H, -SAc), 1.96 (s, 3 H, -OAc), 1.94-1.83 (m, 2 H, -OCH2CH2CH2NH-), 1.72 (s, 3 H, - NHAc).13C-NMR (125 MHz, CDCh): A 171.1, 170.6 (2 C), 170.4, 167.2, 167.0, 166.7, 166.6, 166.4, 166.1, 166.0, 165.8, 165.5, 165.0 (2 C), 156.6, 156.2, 156.0, 143.9, 143.6, 143.5, 141.0, 140.7, 133.7, 133.6 (2 C), 133.5, 133.3, 133.2, 133.1 (2 C), 132.1, 131.9, 131.8, 130.3 (2 C),130.2, 130.1 (2 C), 130.0, 129.9 (2 C), 129.8 (2 C), 129.7, 129.6, 129.5, 129.4, 129.1, 128.9,128.6, 128.5, 128.3, 128.2 (2 C), 128.1, 127.3, 127.1, 126.7, 125.2, 125.1, 119.5, 119.4, 117.8,117.7, 117.6, 105.1, 103.7, 103.5, 102.2, 100.0, 95.5, 74.5, 74.0, 73.6, 73.4, 73.0, 72.7, 72.6,72.5, 72.4, 72.3 (2 C), 72.1, 71.8, 69.6, 68.5, 66.6, 66.1 (2 C), 57.1, 56.7, 56.1, 56.0, 54.8, 46.9, 36.8, 35.7, 30.5, 29.1, 25.8, 25.7, 25.1, 20.9. HRMS: CI36HI29C13F3N6O42S [M + H]+calcd: 2711.6876, obsd: 2711.6920.3-(3-(Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O- / (77-butyldiphenvlsilvl-2- / (77- butyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-€)-benzoyl- 2-deoxy-p-D-glucopyranosyl-(1^6)-3,4-di-€>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2- fluorenylmethyloxycarbonylamino-p-D-glucopyranosyl-( 1— >6)-3,4-di-€)-benzoyl-2-deoxy-2- trifluoroacetamido-p-D-glucopyranoside (16)Following the general procedure for Alloc removal and acetylation, compound 16 was prepared in 91% yield for two steps.1H-NMR (500 MHz, CDCh): 8. 8.64 (d, J =9.5 Hz, 1 H), 8.30-8.25 (m, 6 H), 8.12-8.05 (m, 6 H), 7.99-7.86 (m, 8 H), 7.73-7.70 (m, 1 H), 7.65-7.21 (m, 30 H), 7.16-7.01 (m, 8 H), 6.90-6.87 (m, 2 H), 6.83 (t, J =7.5 Hz, 1 H), 6.47-6.44 (m, 1 H, - NHC=O), 6.09-5.91 (m, 4 H), 5.80 (t, J =9.5 Hz, 1 H), 5.65 (t, J =9.5 Hz, 1 H), 5.46-5.37 (m, 2 H), 5.22 (d, J =9.0 Hz, 1 H), 5.16-5.12 (m, 1 H), 4.97-4.84 (m, 4 H), 4.76-4.64 (m, 3 H), 4.53- 4.50 (m, 2 H), 4.42-4.28 (m, 5 H), 4.18-4.03 (m, 5 H), 3.97 (d, J =8.5 Hz, 1 H), 3.92-3.90 (m, 1 H), 3.88-3.84 (m, 1 H), 3.81-3.74 (m, 2 H), 3.71-3.50 (m, 5 H), 3.37-3.31 (m, 2 H), 3.28-3.25 (m, 2 H, -CH2SAC), 2.69-2.58 (m, 2 H, -NHC=OCH2-), 2.29 (s, 3 H, -SAc), 1.91-1.86 (m, 2 H, - OCH2CH2CH2NH-),1.62 (S, 3 H, -NHAc), 1.16 (s, 9 H, -C(CH3)3), 1.07 (s, 9 H, -C(CH3)3).13C- NMR (125 MHz, CDCh): A 195.8, 171.1, 170.2, 167.0, 166.9, 166.5, 166.4, 166.1 (2 C), 165.7, 164.9, 164.5, 156.8, 156.1, 155.5, 144.3, 143.8, 141.0, 140.8, 135.5, 135.3, 133.7, 133.5, 133.4,133.2, 133.0, 132.9, 131.2, 130.6, 130.4 (2 C), 130.3, 130.2, 130.0 (2 C), 129.9, 129.7 (2 C), 129.4 (2 C), 129.3, 129.2, 129.0, 128.8, 128.7 (2 C), 128.5 (2 C), 128.4, 128.3 (2 C), 128.2,127.5, 127.4, 127.2, 127.1, 126.6 (2 C), 125.5, 125.2, 119.4, 119.3, 105.2, 105.1, 103.0, 102.3, 99.6, 95.2, 80.8, 77.2, 74.7, 74.5, 74.2, 74.0, 73.4, 73.1, 72.8, 72.6, 72.4, 72.2, 71.9, 71.8, 70.7,69.5, 69.2, 67.9, 66.5, 64.0, 57.5, 56.6, 55.9, 55.0, 54.3, 47.0, 36.7, 35.7, 30.5, 27.9, 26.7, 25.1, 22.9, 18.8. HRMS: C151H149CI3F3N6O41SS1 [M + H]+calcd: 2923.8266, obsd: 2953.8305.3-(3-(Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O- / (77-butyldiphenvlsilvl-2- / (77- butyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-(l— >6)-2-allyloxycarbonylamino-3,4- di-€)-benzoyl-2-deoxy-p-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-€)-benzoyl-2-deoxy-p- D-ghicopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2-fluorenylmethyloxycarbonylamino-p- D-ghicopyranosyl-(l— >6)-3,4-di-€)-benzoyl-2-deoxy-2-trifluoroacetamido-p-D- glucopyranoside (17)Following the general procedure for Troc removal and acetylation, compound 17 was prepared in 89% yield for two steps.1H-NMR (500 MHz, CDCI3): 8. 9.52 (d, J =9.5 Hz, 1 H, - NHC=O), 8.28-7.85 (m, 20 H), 7.66-6.98 (m, 44 H), 6.93-6.90 (m, 1 H), 6.87-6.84 (m, 2 H),6.76-6.73 (m, 1 H), 6.43-6.41 (m, 1 H, -NHC=O), 6.11 (t, 7 = 10.0 Hz, 1 H), 5.96-5.84 (m, 3 H),5.76-5.66 (m, 2 H), 5.45-5.37 (m, 3 H, -CH2CHCH2), 5.27-5.19 (m, 2 H, anomeric H), 5.13-5.09 (m, 1 H), 4.95-4.61 (m, 9 H, two anomeric H and -CH2CHCH2), 4.51-4.44 (m, 2 H), 4.36-4.03 (m, 10 H, anomeric H), 3.95-3.90 (m, 1 H), 3.87-3.83 (m, 1 H), 3.80-3.75 (m, 2 H), 3.60-3.50 (m, 5 H), 3.40-3.31 (m, 3 H), 3.25-3.22 (m, 2 H, -CH2SAc), 2.62-2.58 (m, 2 H, -NHC=OCH2-), 2.28 (s, 3 H, -SAc), 1.91-1.86 (m, 2 H, -OCH2CH2CH2NH-), 1.75 (s, 3 H, -NHAc), 1.17 (s, 9 H, - C(CH3)3), 1.06 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDC13): d. 195.9, 171.0, 170.7, 166.7 (2 C), 166.6, 166.4, 166.2, 166.0, 165.8, 164.9, 164.5, 158.7, 158.4, 156.6, 156.2, 155.6, 144.2, 143.5, 141.0, 140.6, 135.5, 135.3, 133.5, 133.4 (2 C), 133.3, 133.2 (2 C), 133.1, 132.9 (2 C), 131.9, 131.4, 130.5, 130.4, 130.3, 130.2, 130.1, 130.0, 129.9, 129.7, 129.6, 129.5, 129.2 (2 C), 129.0 (2 C), 128.9, 128.8 (2 C), 128.7, 128.5 (2 C), 128.3, 128.2, 127.7, 127.3, 127.2, 127.0, 126.7, 125.3, 125.1, 119.4, 119.3, 117.5, 105.3, 105.2, 103.3, 102.4, 99.7, 80.4, 77.5, 76.8, 74.5,73.6, 73.1, 73.0, 72.8, 72.7, 72.5, 72.4, 72.3, 72.2, 72.1, 71.7, 70.8, 69.4, 67.9, 66.5, 66.1, 64.5,56.6, 56.2, 56.0 (2 C), 54.9, 54.5, 53.8, 47.0, 36.9, 35.6, 30.5, 29.3, 28.8, 28.2, 27.8, 26.5, 25.1, 22.8, 18.8. HRMS: Ci52Hi52F3N6O4iSSi [M + H]+calcd: 2833.9430, obsd: 2833.9499.3-(3-(Acetylthio)propionylamino)propyl 3,4-di-O-benzoyl-6-O- / c / 7-butyldiphenylsilyl-2- / (77- butyloxycarbonylamino-2-deoxy-p-D-glucopyranosyl-( 1 —>6)-2-allyloxycarbonylamino-3,4- di-€>-benzoyl-2-deoxy-p-D-glucopyranosyl-(1^6)-3,4-di-€>-benzoyl-2-deoxy-2-(2,2,2- trichloroethyloxycarbonylamino)-p-D-glucopyranosyl-(l— >6)-2-acetamido-3,4-di-€)- benzoyl-2-deoxy-p-D-glucopyranosyl-( 1— >6)-3,4-di-€)-benzoyl-2-deoxy-2- trifluoroacetamido-p-D-glucopyranoside (18)Following the general procedure for Fmoc removal and procedure for acetylation, compound 18 was prepared in 63% yield for two steps.1H-NMR (500 MHz, CDCh): 8. 8.58 (d, J =9.5 Hz, 1 H, -NHC=O), 8.26-8.20 (m, 6 H), 8.16 (d, J =8.0 Hz, 2 H), 8.09 (d, J =8.0 Hz, 2 H), 8.03 (d, J =7.5 Hz, 2 H), 7.96-7.92 (m, 6 H), 7.87 (d, J =7.5 Hz, 2 H), 7.68-7.55 (m, 6 H), 7.51-7.11 (m, 28 H), 7.05-7.02 (m, 3 H), 6.97-6.94 (m, 1 H), 6.87-6.84 (m, 2 H), 6.42-6.39 (m, 1 H, -NHC=O), 6.03-5.96 (m, 2 H), 5.90 (t, J =9.5 Hz, 1 H), 5.78-5.67 (m, 2 H), 5.64-5.60 (m, 1 H), 5.54-5.46 (m, 1 H, -CH2CHCH2), 5.41-5.34 (m, 2 H), 5.18-5.09 (m, 3 H, anomeric H), 4.97- 4.74 (m, 6 H, two anomeric H and -CH2CHCH2), 4.68-4.56 (m, 3 H), 4.51-4.41 (m, 2 H), 4.35- 4.04 (m, 10 H, anomeric H), 3.85-3.83 (m, 1 H), 3.76-3.63 (m, 3 H, anomeric H), 3.58-3.49 (m, 3 H), 3.35-3.23 (m, 3 H, -CH2NH=O and -CH2SAc), 2.68-2.56 (m, 2 H, -NHC=OCH2-), 2.33 (s, 3 H, -CH2SAC), 1.90-1.83 (m, 2 H, -OCH2CH2CH2NH-), 1.71 (s, 3 H, -NHAc), 1.12 (s, 9 H, - C(CH3)3), 1.06 (s, 9 H, -C(CH3)3).13C-NMR (125 MHz, CDCh): 3. 195.9, 171.1, 170.5, 166.9, 166.7 (2 C), 166.5 (2 C), 166.1, 166.0, 165.7, 164.9, 164.5, 156.2, 156.0, 135.5, 135.3, 133.5 (2 C), 133.4, 133.2, 133.1 (2 C), 133.0, 131.8, 131.3, 130.4 (2 C), 130.3 (3 C), 130.0, 129.9, 129.8 (2 C), 129.7 (2 C), 129.6, 129.4, 129.2, 129.0 (4 C), 128.8, 128.7, 128.6, 128.5, 128.4 (2 C), 128.3 (2 C), 128.2, 128.1, 127.6, 127.3, 117.1 (2 C), 105.0, 104.7, 102.9, 102.2, 99.5, 95.2, 80.8, 74.5, 74.1, 73.9, 73.4, 73.2, 73.0, 72.7, 72.4 (2 C), 72.2, 72.1, 72.0, 71.8, 71.7, 70.7, 69.6, 67.8, 65.9, 64.6, 57.4, 56.4, 55.8, 54.9, 54.0, 36.7, 35.7, 30.6, 29.1, 27.9, 26.7, 25.1, 22.7, 18.8. HRMS: Ci4oHi43Cl3F3N604iSSi [M + H]+calcd: 2785.7791, obsd: 2785.7714.PNAG0 (00000): Fig. 24A: Compound 2 was subjected to Boc, Alloc, Troc, Fmoc, Bz and TFA removals according to the general procedures to afford PNAG0 (10 mg, 43% yield for 5 steps) [α]D20= -5.3 (c 0.4, MeOH). ^-NMR (500 MHz, D2O): 8. 4.67-4.62 (m, 4 H, anomeric H, obtained from HSQC NMR), 4.59 (d, J =8.5 Hz, 1 H, anomeric H), 4.14-4.08 (m, 4 H), 3.82- 3.72 (m, 6 H), 3.63-3.51 (m, 10 H), 3.44-3.23 (m, 7 H), 3.21-3.16 (m, 1 H, -OCH2CH2CH2NH-), 3.10-3.04 (m, 1 H, -OCH2CH2CH2NH-), 2.97-2.88 (m, 5 H, -CH-NH2), 2.80 (t, J =7.0 Hz, 2 H, - CH2S-), 2.52 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.70-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C- NMR (125 MHz, D2O): 3. 99.1 (4 C), 98.8, 76.1, 74.6, 71.6, 69.5, 68.3, 68.1, 67.5, 60.3, 60.1, 55.4, 35.9, 34.7, 33.0, 28.3 (obtained from HSQC NMR). HRMS: C72Hi36Ni2O44S2[M + 2 H]2+calcd: 968.4102, obsd: 968.4108.PNAG1 (00001): Fig. 24B: Compound 1 was subjected to Boc, Alloc, Troc, Fmoc, Bz removals according to the general procedures to afford PNAG1 (3.5 mg, 41% yield for 5 steps). [a]o20= - 4.4 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8 4.67-4.63 (m, 4 H, anomeric H, obtained from HSQC NMR), 4.37 (d, J =8.5 Hz, 1 H, anomeric H), 4.14-4.08 (m, 4 H), 3.83-3.71 (m, 6 H),3.63-3.30 (m, 18 H), 3.09-2.91 (m, 6 H, -OCH2CH2C / / 2NH- and -CH-NH2), 2.80 (t, J =6.5 Hz, 2 H, -CH2S-), 2.51 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.62-1.57 (m, 2H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.3, 99.2 (4 C), 76.1, 74.6, 74.4, 73.4, 71.6, 69.7, 69.5, 68.3, 67.5, 60.3, 55.4, 36.3, 34.9, 33.2, 28.3, 22.0 (obtained from HSQC NMR). HRMS: C76H140N12O46S2 [M + 2 H]2+calcd: 1010.4208, obsd: 1010.4211.PNAG2 (00010): Fig. 24C: Compound 2 was subjected to Fmoc removal followed by acetylation, removal of Boc, Alloc, Troc, Bz and TFA according to the general procedures to afford PNAG2 (2.8 mg, 53% yield for 6 steps). [a]o20= -3.6 (c 0.3, MeOH).1H-NMR (500 MHz, D2O): & 4.66-4.62 (m, 3 H, anomeric H obtained from HSQC NMR), 4.54 (d, J =8.0 Hz, 1 H, anomeric H), 4.39 (d, J =8.0 Hz, 1 H, anomeric H), 4.11-4.04 (m, 4 H), 3.81-3.76 (m, 5 H), 3.61-3.17 (m, 20 H), 3.09-3.03 (m, 1 H, -OCH2CH2CH2NH-), 2.96-2.90 (m, 3 H, -CH-NH2), 2.86 (t, J =9.5 Hz, 1 H, -CH-NH2), 2.79 (bs, 2H, -CH2S-), 2.52 (t, J =6.0 Hz, 2 H, -NHC=OCH2- ), 1.87 (s, 3 H, -NHAc), 1.69-1.64 (m, 2H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5, 99.1 (2 C), 99.0, 98.6, 76.1, 74.6, 74.4, 73.8, 71.8, 69.7, 69.5, 68.3, 68.1, 60.3, 55.4, 34.9, 33.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C76H140N12O46S2 [M + 2 H]2+calcd: 1010.4208, obsd: 1010.4200.PNAG3 (00011): Fig. 24D: Compound 1 was subjected to Fmoc removal followed by acetylation, removal of Boc, Alloc, Troc and Bz according to the general procedures to afford PNAG3 (1.1 mg, 37% yield for 6 steps). [α]D20= -2.6 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.67-4.62 (m, 3 H, anomeric H obtained from HSQC NMR), 4.42 (d, J =8.5 Hz, 1 H, anomeric H), 4.32 (d, J =8.5 Hz, 1 H, anomeric H), 4.13-4.00 (m, 4 H), 3.83-3.63 (m, 5 H), 3.63-3.21 (m, 16 H), 3.14-3.08 (m, 1 H), 3.04-2.91 (m, 4 H, -OCH2CH2CH2NH- and -CH-NH2), 2.81 (m, 1 H, - CH2S-), 2.64-2.59 (m, 1 H, -CH2S-), 2.53 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 6 H, - NHAc), 1.63-1.58 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5, 100.9, 99.1 (2 C), 99.0, 76.1, 74.6, 74.2, 73.8, 71.8, 71.6, 69.9, 69.7, 69.5, 68.7, 68.5, 68.3, 67.5, 60.3, 60.1, 55.4, 55.0, 36.1, 28.1, 22.0 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4316.PNAG4 (00100): Fig. 24E: Compound 2 was subjected to Troc removal followed by acetylation, removal of Boc, Alloc, Fmoc, Bz and TFA according to the general procedures to afford PNAG4 (2.0 mg, 48% yield for 6 steps). [a]o20= -1.2 (c 0.2, MeOH). ' H-NMR (500 MHz, D2O): 8. 4.64-4.62 (m, 2 H, anomeric H obtained from HSQC NMR), 4.60-4.57 (m, 2 H, anomeric H), 4.39 (d, J =8.5 Hz, 1 H, anomeric H), 4.11-4.04 (m, 4 H), 3.82-3.72 (m, 5 H), 3.62-3.30 (m, 18 H), 3.27 (t, J =9.0 Hz, 1 H), 3.20-3.16 (m, 1 H, -OCH2CH2CH2NH-), 3.10-3.04(m, 1 H, -OCH2CH2CH2NH-), 2.96-2.87 (m, 4 H, -CH-NH2), 2.80 (t, J =7.0 Hz, 2 H, -CH2S-), 2.52 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.69-1.64 (m, 2H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): <5. 101.7, 99.2 (2 C), 99.0 (2 C), 76.1, 74.8 (2 C), 74.4, 74.2, 71.8, 69.7, 69.5 (2 C), 68.3, 68.1, 67.9, 67.7, 60.3, 55.4, 34.9, 33.0, 28.5, 24.8, 22.2 (obtained from HSQC NMR). HRMS: C76H140N12O46S2 [M + 2 H]2+calcd: 1010.4208, obsd: 1010.4198.PNAG5 (00101): Fig. 24F: Compound 1 was subjected to Troc removal followed by acetylation, removal of Boc, Alloc, Fmoc and Bz according to the general procedures to afford PNAG5 (3.1 mg, 53% yield for 6 steps). [α]D20= -4.5 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.66-4.59 (m, 3 H, anomeric H obtained from HSQC NMR), 4.39 (d, J =8.0 Hz, 1 H, anomeric H), 4.36 (d, J =8.5 Hz, 1 H, anomeric H), 4.10-4.03 (m, 4 H), 3.80-3.68 (m, 5 H), 3.61-3.24 (m, 20 H), 3.11-3.07 (m, 1 H), 3.02-2.87 (m, 5 H, -OCH2CH2C / / 2NH- and -CH-NH2), 2.62-2.56 (m, 2 H, -CH2S-), 2.49 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.87 (s, 3 H, - NHAc), 1.61-1.56 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.4, 101.3, 99.0 (2 C), 98.9, 76.1, 74.6, 74.4, 73.4, 71.8, 69.5 (2 C), 68.5, 68.1, 67.7, 60.1, 55.4 (2 C), 55.2,46.6, 36.5, 36.3, 31.0, 29.1, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4304.PNAG6 (00110): Fig. 24G: Compound 2 was subjected to Troc and Fmoc removals followed by acetylation, removal of Boc, Alloc, Bz and TFA according to the general procedures to afford PNAG6 (2.3 mg, 51% yield for 6 steps). [α]D20= -2.6 (c 0.2, MeOH). ^-NMR (500 MHz, D2O): J. 4.67-4.62 (m, 2H, anomeric H obtained from HSQC NMR), 4.54 (d, J =9.0 Hz, 1 H, anomeric H), 4.41 (d, J =8.5 Hz, 1 H, anomeric H), 4.33 (d, J =9.0 Hz, 1 H, anomeric H), 4.11-3.99 (m, 4 H), 3.81-3.74 (m, 4 H), 3.62-3.17 (m, 22 H), 3.09-2.78 (m, 6 H, -OCH2CH2CH2NH-, -CH-NH2 and -CH2S-), 2.52-2.48 (m, 2 H, -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.5 (2 C), 99.2 (2 C),98.6, 76.1, 74.6, 74.3, 74.0, 71.8, 69.7 (2 C), 69.5 (2 C), 68.5, 68.3, 68.1, 67.3, 60.1, 55.4, 55.3, 36.1, 34.9, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4304.PNAG7 (00111): Fig. 24H: Compound 1 was subjected to Troc and Fmoc removals followed by acetylation, removal of Boc, Alloc and Bz according to the general procedures to afford PNAG7 (2.8 mg, 44% yield for 6 steps). [α]D20= -4.5 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): & 4.67- 4.62 (m, 2 H, anomeric H obtained from HSQC NMR), 4.42 (d, J =8.5 Hz, 1 H, anomeric H), 4.35 (d, J =8.0 Hz, 1 H, anomeric H), 4.31 (d, J =8.5 Hz, 1 H, anomeric H), 4.11-3.98 (m, 4 H),3.81-3.70 (m, 5 H), 3.62-3.20 (m, 21 H), 3.13-3.07 (m, 1 H, -OCH2CH2CH2NH-), 3.03-2.91 (m, 3 H, -CH-NH2 and -OCH2CH2CH2NH-), 2.80 (t, J =7.0 Hz, 2 H, -CH2S ), 2.51 (t, J =6.5 Hz, 2H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.87 (s, 6 H, -NHAc), 1.62-1.57 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.4, 101.2, 100.9, 99.0, 98.8, 76.1, 73.8, 71.8, 69.7, 69.5, 68.5, 68.3, 60.3, 55.4 (2 C), 55.2, 36.3, 36.2, 34.9, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C84HI48NI205OS2[M + 2 H]2+calcd: 1094.4419, obsd: 1094.4407.PNAG8 (01000): Fig. 241: Compound 2 was subjected to Alloc removal followed by acetylation, removal of Boc, Troc, Fmoc, Bz and TFA according to the general procedures to afford PNAG8 (3.1 mg, 40% yield for 6 steps). [a]o20= -2.4 (c 0.3, MeOH). ' H-NMR (500 MHz, D2O): & 4.65-4.57 (m, 4 H, anomeric H obtained from HSQC NMR), 4.38 (d, J =8.5 Hz, 1 H, anomeric H), 4.09-3.98 (m, 4 H), 3.78-3.71 (m, 5 H), 3.61-3.17 (m, 20 H), 3.08-3.04 (m, 1 H, -OCH2CH2CH2NH-), 2.95-2.87 (m, 4 H, -CH-NH2), 2.78 (bs, 2 H, -CH2S ), 2.50 (bs, 2 H, - NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.67 (bs, 2H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.8, 98.9, 98.8, 76.1, 74.6, 74.4 (2 C), 73.8, 71.8, 69.7, 69.5 (2 C), 68.3, 68.1 (2 C),67.7, 60.1, 55.4 (2 C), 34.7, 33.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C76Hi4oNi2046S2[M + 2 H]2+calcd: 1010.4208, obsd: 1010.4200.PNAG9 (01001): Fig. 24J: Compound 1 was subjected to Alloc removal followed by acetylation, removal of Boc, Troc, Fmoc and Bz according to the general procedures to afford PNAG9 (2.9 mg, 41% yield for 6 steps). [α]D20= -4.4 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.69-4.63 (m, 3 H, anomeric H obtained from HSQC NMR), 4.42-4.39 (m, 2 H, anomeric H), 4.13-4.06 (m, 4 H), 3.83-3.74 (m, 5 H), 3.65-3.28 (m, 21 H), 3.15-3.09 (m, 1 H, - OCH2CH2CH2NH-), 3.06-3.02 (m, 1 H, -OCH2CH2CH2NH-), 3.00-2.91 (m, 3 H, -CH-NH2), 2.83 (t, J =7.0 Hz, 2 H, -CH2S ), 2.54 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.92 (s, 3 H, -NHAc),I.91 (s, 3 H, -NHAc), 1.66-1.60 (m, 2H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3.101.7, 101.3, 99.0 (3 C), 75.7, 74.6, 74.4, 73.6, 71.8, 69.7 (2 C), 69.5, 68.3 (2 C), 68.1, 60.7, 60.3, 55.4, 55.2, 36.3 (2 C), 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C8oHi44Ni2048S2[M + 2 H]2+calcd: 1052.4313, obsd: 1052.4308.PNAG10 (01010): Fig. 24K: Compound 2 was subjected to Alloc and Fmoc removals followed by acetylation, removal of Boc, Troc, Bz and TFA according to the general procedures to afford PNAG10 (3.2 mg, 53% yield for 6 steps). [α]D20= -5.0 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.64-4.62 (m, 1 H, anomeric H obtained from HSQC NMR), 4.61 (d, J =8.5 Hz, 1 H, anomeric H), 4.54 (d, J =8.5 Hz, 1 H, anomeric H), 4.39 (d, J =8.5 Hz, 2 H, anomeric H), 4.09- 4.02 (m, 4 H), 3.79-3.70 (m, 4 H), 3.62-3.17 (m, 22 H), 3.09-3.04 (m, 1 H, -OCH2CH2CH2NH-),2.95-2.77 (m, 5 H, -CH-NH2and -CH2S ), 2.52 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.6 (2 C), 99.0 (2 C), 98.6, 76.1, 74.6, 74.5, 74.4, 73.6, 71.8 (2 C), 69.7 (2 C), 69.5,69.4, 68.3, 68.1, 67.9, 67.5, 60.1, 55.6, 55.4, 55.2, 34.9, 33.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C80Hi44N12O48S2[M + 2 H]2+calcd: 1052.4313, obsd: 1052.4318.PNAG11 (01011): Fig. 24L: Compound 1 was subjected to Alloc and Fmoc removals followed by acetylation, removal of Boc, Troc and Bz according to the general procedures to afford PNAG11 (2.8 mg, 50% yield for 6 steps). [α]D20= -3.7 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.65-4.61 (m, 2 H, anomeric H obtained from HSQC NMR), 4.42-4.39 (m, 2 H, anomeric H), 4.32 (d, J =8.5 Hz, 1 H, anomeric H), 4.11-4.02 (m, 4 H), 3.81-3.71 (m, 5 H), 3.62- 3.20 (m, 22 H), 3.13-3.08 (m, 1 H, -OCH2CH2CH2NH-), 3.04-2.99 (m, 1 H, -OCH2CH2CH2NH- ), 2.96-2.89 (m, 2 H, -CH-NH2), 2.81 (t, J =7.0 Hz, 2 H, -CH2S ), 2.52 (t, J =6.0 Hz, 2 H, - NHC=OCH2-), 1.90 (s, 3 H, -NHAc), 1.89 (s, 3 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.64-1.59 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5, 101.4, 100.9, 98.9 (2 C), 76.1,74.4, 74.1, 74.0, 73.5, 71.8, 69.7, 69.5, 69.4, 68.5, 68.3, 67.9, 60.1, 55.6, 55.5, 55.3, 36.3, 34.7, 33.2, 28.3, 22.0 (obtained from HSQC NMR). HRMS: C84H148N12O50S2[M + 2 H]2+calcd: 1094.4419, obsd: 1094.4407.PNAG12 (01100): Fig. 24M: Compound 2 was subjected to Alloc and Troc removals followed by acetylation, removal of Boc, Fmoc, Bz and TFA according to the general procedures to afford PNAG12 (2.7 mg, 49% yield for 6 steps). [α]D20= -1.4 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.64-4.56 (m, 3 H, anomeric H obtained from HSQC NMR), 4.39 (d, J =8.5 Hz, 1 H, anomeric H), 4.33 (d, J =8.5 Hz, 1 H, anomeric H), 4.09-3.98 (m, 4 H), 3.81-3.70 (m, 5 H), 3.60- 3.47 (m, 11 H), 3.41-3.16 (m, 10 H), 3.09-3.04 (m, 1 H, -OCH2CH2CH2NH-), 2.94-2.86 (m, 3 H, -CH-NH2), 2.80 (t, J =6.5 Hz, 2 H, -CH2S ), 2.52 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5 (2 C), 98.8, 98.7 (2 C), 75.9, 74.4, 74.2, 74.0, 73.6, 71.6, 69.7, 69.5, 69.3, 68.3, 68.1, 67.9, 67.7, 60.1, 36.1, 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C8oHi44Ni2048S2[M + 2 H]2+calcd: 1052.4313, obsd: 1052.4304.PNAG13 (01101): Fig. 24N: Compound 1 was subjected to Alloc and Troc removals followed by acetylation, removal of Boc, Fmoc and Bz according to the general procedures to afford PNAG13 (2.1 mg, 48% yield for 6 steps). [α]D20= -4.7 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.66-4.60 (m, 2 H, anomeric H obtained from HSQC NMR), 4.41 (d, J =8.5 Hz, 1 H, anomeric H), 4.37-4.33 (m, 2 H, anomeric H), 4.11-4.00 (m, 4 H), 3.80-3.69 (m, 4 H), 3.63-3.22(m, 22 H), 3.12-3.09 (m, 1 H, -OCH2CH2CH2NH-), 3.04-2.99 (m, 1 H, -OCH2CH2CH2NH-), 2.96-2.89 (m, 2 H, -CH-NH2), 2.81 (t, J =6.5 Hz, 2 H, -CH2S-), 2.52 (t, J =6.5 Hz, 2 H, - NHC=OCH2-), 1.90 (s, 3 H, -NHAc), 1.89 (s, 3 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.63-1.58 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.6, 101.5 (2 C), 99.0, 98.9, 76.1,74.5, 74.3, 74.0, 73.8, 73.7, 73.5, 71.8, 69.7, 69.6, 68.5, 68.1, 60.3, 55.4, 55.3, 36.3, 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C84H148N12O50S2 [M + 2 H]2+calcd: 1094.4419, obsd: 1094.4402.PNAG14 (OHIO): Fig. 240: Compound 2 was subjected to Alloc, Troc and Fmoc removals followed by acetylation, removal of Boc, Bz and TFA according to the general procedures to afford PNAG14 (3.2 mg, 44% yield for 6 steps). [a]o20= -5.0 (c 0.3, MeOH).1H-NMR (500 MHz, D2O): 8. 4.64-4.62 (m, 1 H, anomeric H obtained from HSQC NMR), 4.54 (d, J =8.0 Hz, 1 H, anomeric H), 4.41 (d, J =8.5 Hz, 1 H, anomeric H), 4.35-4.32 (m, 2 H, anomeric H), 4.10-3.98 (m, 4 H), 3.78-3.73 (m, 4 H), 3.61-3.20 (m, 24 H), 3.09-3.04 (m, 1 H, - OCH2CH2CH2NH-), 2.95 (t, J =9.0 Hz, 1 H, -CH-NH2), 2.87 (t, J =9.0 Hz, 1 H, -CH-NH2), 2.81 (t, J =6.0 Hz, 2 H, -CH2S-), 2.52 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.6, 101.5 (2 C), 99.0, 98.6, 76.1, 74.6, 74.4, 74.3, 74.0, 73.7, 71.9, 71.8, 69.9, 69.7, 69.6, 68.4, 68.1, 68.0, 67.5, 60.1, 55.4 (2 C), 55.2, 35.9, 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C84H148N12O50S2 [M + 2 H]2+calcd: 1094.4419, obsd: 1094.4413.PNAG15 (01111): Fig. 24P: Compound 1 was subjected to Alloc, Troc and Fmoc removals followed by acetylation, removal of Boc and Bz according to the general procedures to afford PNAG15 (2.2 mg, 39% yield for 6 steps). [α]D20= -3.1 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.65-4.63 (m, 1 H, anomeric H obtained from HSQC NMR), 4.43 (d, J =8.5 Hz, 1 H, anomeric H), 4.38-4.35 (m, 2 H, anomeric H), 4.30 (d, J =8.5 Hz, 1 H, anomeric H), 4.12 (d, J = 10.5 Hz, 1 H), 4.05-3.99 (m, 3 H), 3.80-3.70 (m, 4 H), 3.62-3.48 (m, 10 H), 3.44-3.34 (m, 10 H), 3.29-3.21 (m, 3 H), 3.16-3.10 (m, 1 H, -OCH2CH2CH2NH-), 3.04-2.98 (m, 1 H, - OCH2CH2CH2NH-), 2.97 (dd, J =8.5, 10.5 Hz, 1 H, -CH-NH2), 2.82 (t, J =6.5 Hz, 2 H, -CH2S-), 2.53 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.90 (s, 3 H, -NHAc), 1.89 (s, 3 H, -NHAc), 1.88 (s, 6 H, -NHAc), 1.64-1.59 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5, 101.3 (2 C), 100.9, 100.0, 76.1, 74.5, 74.2, 74.0, 73.8, 71.8, 69.9 (2 C), 69.5, 69.3, 68.5, 68.3, 68.2,67.5, 60.3, 55.6, 55.4, 55.2, 36.1, 34.7, 33.2, 28.1, 22.2 (obtained from HSQC NMR). HRMS: C88H152N12O52S2 [M + 2 H]2+calcd: 1136.4525, obsd: 1136.4507.PNAG16 (10000): Fig. 24Q: Compound 2 was subjected to Boe removal followed by acetylation, removal of Alloc, Troc, Fmoc, Bz and TFA according to the general procedures to afford PNAG16 (4.0 mg, 41% yield for 6 steps). [a]o20= -2.5 (c 0.3, MeOH).1H-NMR (500 MHz, D2O): 8. 4.65-4.63 (m, 2 H, anomeric H obtained from HSQC NMR), 4.61-4.57 (m, 2 H, anomeric H), 4.35 (d, J =8.5 Hz, 1 H, anomeric H), 4.11-4.01 (m, 4 H), 3.80-3.73 (m, 5 H), 3.65-3.46 (m, 12 H), 3.40-3.26 (m, 7 H), 3.21-3.17 (m, 1 H, -OCH2CH2CH2NH-), 3.09-3.05 (m, 1 H, -OCH2CH2CH2NH-), 2.95-2.87 (m, 4 H, -CH-NH2), 2.79 (t, J =6.5 Hz, 1 H, -CH2S-), 2.51 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.67-1.65 (m, 2H, -OCH2CH2CH2NH- ).13C-NMR (125 MHz, D2O): d. 101.7, 99.2 (2 C), 99.0, 98.8, 75.7, 74.6 (2 C), 74.4, 73.8, 71.8, 71.7, 69.7, 69.5, 68.3 (2 C), 68.1, 67.7, 60.5, 55.4, 55.3, 36.1, 34.7, 33.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C76H140N12O46S2 [M + 2 H]2+calcd: 1010.4208, obsd: 1010.4202.PNAG17 (10001): Fig. 24R: Compound 1 was subjected to Boc removal followed by acetylation, removal of Alloc, Troc, Fmoc and Bz according to the general procedures to afford PNAG17 (2.1 mg, 48% yield for 6 steps). [α]D20= -2.7 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.68-4.64 (m, 2 H, anomeric H obtained from HSQC NMR), 4.62 (d, J =9.0 Hz, 1 H, anomeric H), 4.37-35 (m, 2 H, anomeric H), 4.12-4.02 (m, 4 H), 3.78-3.73 (m, 5 H), 3.65-3.26 (m, 21 H), 3.11-2.99 (m, 2 H, -OCH2CH2CH2NH-), 2.99-2.88 (m, 3 H, -CH-NH2), 2.63-2.57 (m, 3 H, -CH2S- and -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.62-1.57 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.4, 101.2, 99.1, 99.0 (2 C), 75.7, 74.3 (2 C), 73.4, 71.6 (2 C), 69.7, 69.5 (2 C), 68.3 (2 C), 67.9, 60.5, 55.4, 55.2 (2 C), 36.1, 36.5, 29.1, 28.3, 22.0 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4302.PNAG18 (10010): Fig. 24S: Compound 2 was subjected to Boc and Fmoc removals followed by acetylation, removal of Alloc, Troc, Bz and TFA according to the general procedures to afford PNAG18 (2.5 mg, 47% yield for 6 steps). [α]D20= -2.7 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.67-4.60 (m, 2 H, anomeric H obtained from HSQC NMR), 4.55 (d, J =8.5 Hz, 1 H, anomeric H), 4.40-4.35 (m, 2 H, anomeric H), 4.11-4.02 (m, 4 H), 3.78-3.74 (m, 4 H), 3.65-3.17 (m, 22 H), 3.08-2.78 (m, 6 H, -OCH2CH2CH2NH-, -CH-NH2 and -CH2S-), 2.53-2.48 (m, 2 H, - NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.69-1.64 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.7, 101.5, 99.2, 99.0, 98.6, 75.7, 74.8, 74.6, 73.6 (2 C), 71.8, 71.7 (2 C), 69.7, 69.6, 68.2, 68.1, 67.3, 60.7, 55.7, 55.6, 55.2, 36.1, 32.8, 31.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4315.PNAG19 (10011): Fig. 24T: Compound 1 was subjected to Boe and Fmoc removals followed by acetylation, removal of Alloc, Troc and Bz according to the general procedures to afford PNAG19 (2.2 mg, 51% yield for 6 steps). [α]D20= -3.9 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.67-4.65 (m, 1 H, anomeric H obtained from HSQC NMR), 4.61 (d, J =8.5 Hz, 1 H, anomeric H), 4.41 (d, J =8.0 Hz, 1 H, anomeric H), 4.36 (d, J =8.5 Hz, 1 H, anomeric H), 4.31 (d, J =8.5 Hz, 1 H, anomeric H), 4.11-4.02 (m, 4 H), 3.88-3.69 (m, 4 H), 3.64-3.20 (m, 24 H), 3.13-3.07 (m, 1 H, -OCH2CH2CH2NH-), 3.02-2.88 (m, 3 H, -OCH2CH2CH2NH- and -CH-NH2),2.62-2.55 (m, 3 H, -C / / 2S- and -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.86 (ss, 6 H, -NHAc),1.62-1.57 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.5 (2 C), 101.1, 99.2, 99.0, 75.7, 74.6, 74.4, 73.7, 73.4, 71.9, 71.8, 69.7 (2 C), 69.6, 68.5, 68.3, 68.1, 67.7, 60.5, 55.4, 55.2 (2 C), 36.7, 36.3, 29.1, 28.1, 22.0 (obtained from HSQC NMR). HRMS: C84H148N12O50S2 [M + 2 H]2+calcd: 1094.4419, obsd: 1094.4404.PNAG20 (10100): Fig. 24U: Compound 2 was subjected to Boc and Troc removals followed by acetylation, removal of Alloc, Fmoc, Bz and TFA according to the general procedures to afford PNAG20 (3.3 mg, 53% yield for 6 steps). [α]D20= -2.5 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.62-4.57 (m, 3 H, anomeric H), 4.40- 4.35 (m, 2 H, anomeric H), 4.06-4.03 (m, 4 H), 3.80-3.70 (m, 5 H), 3.62-3.21 (m, 24 H), 3.06-2.99 (m, 2 H, -OCH2CH2CH2NH-), 2.92-2.86 (m, 3 H, -CH-NH2), 2.81 (t, J =7.0 Hz, 2 H, -CH2S-), 2.52-2.48 (m, 2 H, -NHC=OCH2-), 1.89 (s, 6 H, -NHAc), 1.70-1.65 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5, 101.2, 98.9, 98.8, 98.7, 75.5, 74.8, 74.2 (2 C), 73.6, 72.2, 71.6, 69.5, 69.3, 69.2, 68.3, 68.1, 67.5, 60.5, 55.4, 55.2, 46.8, 36.1, 31.2, 28.3, 22.4 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4298.PNAG21 (10101): Fig. 24V: Compound 1 was subjected to Boc and Troc removals followed by acetylation, removal of Alloc, Fmoc and Bz according to the general procedures to afford PNAG21 (2.8 mg, 51% yield for 6 steps). [α]D20= -4.0 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.67-4.65 (m, 2 H, anomeric H obtained from HSQC NMR), 4.40 (d, J =8.5 Hz, 1 H, anomeric H), 4.36 (d, J =8.5 Hz, 2 H, anomeric H), 4.06-3.99 (m, 4 H), 3.78-3.69 (m, 5 H), 3.64- 3.34 (m, 18 H), 3.31-3.25 (m, 4 H), 3.11-3.08 (m, 1 H, -OCH2CH2CH2NH-), 3.04-3.00 (m, 1 H, - OCH2CH2CH2NH-), 2.93-2.88 (m, 2 H, -CH-NH2), 2.80 (t, J =6.5 Hz, 2 H, -CH2S-), 2.51 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.81 (s, 6 H, -NHAc), 1.88 (s, 3 H, -NHAc), 1.62-1.57 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.7, 101.6 (2 C), 99.3, 99.2, 75.9, 74.8, 74.4, 73.8, 71.8, 69.8, 69.7, 68.3 (2 C), 68.1, 60.7, 55.6, 55.4, 36.3, 34.7, 33.2, 28.3, 22.2(obtained from HSQC NMR). HRMS: C84H148N12O50S2 [M + 2 H]2+calcd: 1094.4419, obsd: 1094.4408.PNAG22 (10110): Fig. 24W: Compound 2 was subjected to Boe, Troc and Fmoc removals followed by acetylation, removal of Alloc, Bz and TFA according to the general procedures to afford PNAG22 (2.8 mg, 51% yield for 6 steps). [a]o20= -2.8 (c 0.3, MeOH). ' H-NMR (500 MHz, D2O): 8. 4.61 (d, J =8.5 Hz, 1 H, anomeric H), 4.54 (d, J =8.5 Hz, 1 H, anomeric H), 4.41 (d, J =8.5 Hz, 1 H, anomeric H), 4.36 (d, J =8.5 Hz, 1 H, anomeric H), 4.33 (d, J =8.5 Hz, 1H, anomeric H), 4.07-3.99 (m, 4 H), 3.78-3.71 (m, 3 H), 3.63-3.40 (m, 18 H), 3.31-2.18 (m, 6 H), 3.09-3.05 (m, 1 H, -OCH2CH2CH2NH-), 2.93-2.78 (m, 4 H, -CH-NH2 and -CH2S-), 2.53 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 6 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.70-1.65 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.5 (2 C), 101.4, 99.2, 98.8, 75.7, 74.8, 74.2, 73.6, 69.8, 69.6 (2 C), 68.3, 68.1, 67.7, 67.3, 60.5, 55.4, 55.2, 35.9, 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C84H148N12O50S2 [M + 2 H]2+calcd: 1094.4419, obsd: 1092.4414.PNAG23 (10111): Fig. 24X: Compound 1 was subjected to Boc, Troc and Fmoc removals followed by acetylation, removal of Alloc and Bz according to the general procedures to afford PNAG23 (1.2 mg, 37% yield for 6 steps). [α]D20= -2.8 (c 0.3, MeOH).1H-NMR (500 MHz, D2O): 8. 4.62 (d, J =9.0 Hz, 1 H, anomeric H), 4.42 (d, J =8.5 Hz, 1 H, anomeric H), 4.37-4.30 (m, 3 H, anomeric H), 4.07-3.98 (m, 4 H), 3.78-3.72 (m, 4 H), 3.63-3.24 (m, 23 H), 3.13-3.07 (m, 1 H, -OCH2CH2CH2NH-), 3.04-2.98 (m, 1 H, -OCH2CH2CH2NH-), 2.81 (t, J =6.0 Hz, 1 H, - CH-NH2), 2.64-2.48 (m, 3 H, -C / / 2S- and -NHC=OCH2-), 1.89-1.87 (m, 12 H, -NHAc), 1.63-I.58 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.5, 101.4 (2 C), 101.1, 99.0, 75.7, 74.8, 74.2, 73.8, 71.9, 71.8, 69.7, 69.6, 68.5, 68.1 (2 C), 67.5, 60.5, 55.4, 55.2, 36.1, 32.8, 31.0, 28.1, 22.0 (obtained from HSQC NMR). HRMS: C88H152N12O52S2 [M + 2 H]2+calcd: 1136.4525, obsd: 1136.4517.PNAG24 (11000): Fig. 24Y: Compound 2 was subjected to Boc, Alloc removals followed by acetylation, removal of Troc, Fmoc, Bz and TFA according to the general procedures to afford PNAG24 (2.4 mg, 41% yield for 6 steps). [α]D20= -0.7 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.65-4.63 (m, 1 H, anomeric H obtained from HSQC NMR), 4.60-4.56 (m, 2 H, anomeric H), 4.36 (d, J =9.0 Hz, 1 H, anomeric H), 4.33 (d, J =8.5 Hz, 1 H, anomeric H), 4.08- 3.97 (m, 4 H), 3.82-3.71 (m, 4 H), 3.63-3.45 (m, 11 H), 3.42-3.34 (m, 5 H), 3.30-3.15 (m, 6 H), 3.09-3.03 (m, 1 H, -OCH2CH2CH2NH-), 2.95-2.86 (m, 3 H, -CH-NH2), 2.79 (t, J =7.0 Hz, 2 H, - CH2S-), 2.51 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 3 H, -NHAc), 1.87 (s, 3 H, -NHAc),1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D20): 3. 101.8, 101.6, 99.1, 99.0, 98.8, 75.7, 74.6, 74.4 (2 C), 73.5, 71.8, 69.7 (2 C), 69.5, 68.3 (2 C), 67.7 (2 C), 60.5, 55.4, 55.2, 36.1, 34.7, 33.0, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C80H144N12O48S2 [M + 2 H]2+calcd: 1052.4313, obsd: 1052.4309.PNAG25 (11001): Fig. 24Z: Compound 1 was subjected to Boe and Alloc removals followed by acetylation, removal of Troc, Fmoc and Bz according to the general procedures to afford PNAG25 (2.2 mg, 58% yield for 6 steps). [α]D20= -1.6 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.67-4.65 (m, 1 H, anomeric H obtained from HSQC NMR), 4.62 (d, J =8.5 Hz, 1 H, anomeric H), 4.39-4.34 (m, 3 H, anomeric H), 4.10-3.99 (m, 4 H), 3.79-3.73 (m, 4 H), 3.65-3.23 (m, 22 H), 3.13-3.08 (m, 1 H, -OCH2CH2CH2NH-), 3.05-3.00 (m, 1 H, -OCH2CH2CH2NH-), 2.97-2.89 (m, 2 H, -CH-NH2), 2.81 (t, J =6.5 Hz, 2 H, -CH2S ), 2.52 (t, J =6.5 Hz, 2 H, - NHC=OCH2-), 1.90 (s, 6 H, -NHAc), 1.89 (s, 3 H, -NHAc), 1.63-1.58 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.7, 101.5 (2 C), 99.0 (2 C), 75.7, 74.6, 74.4 (2 C), 74.1 (2 C), 73.6, 71.9, 71.8, 69.9 (2 C), 69.7 (2 C), 68.5, 68.4, 68.3, 60.7, 55.6, 55.4,36.3, 34.9, 33.4, 28.5, 22.2 (obtained from HSQC NMR). HRMS: C84HI48NI205OS2[M + 2 H]2+calcd: 1094.4419, obsd: 1092.4402.PNAG26 (11010): Fig. 24Zi: Compound 2 was subjected to Boc, Alloc and Fmoc removals followed by acetylation, removal of Troc, Bz and TFA according to the general procedures to afford PNAG26 (2.3 mg, 51% yield for 6 steps). [α]D20= -2.5 (c 0.3, MeOH). ' H-NMR (500 MHz, D2O): 8. 4.61 (d, J =8.5 Hz, 1 H, anomeric H), 4.54 (d, J =8.5 Hz, 1 H, anomeric H), 4.39-4.32 (m, 3 H, anomeric H), 4.06-3.99 (m, 4 H), 3.77-3.70 (m, 3 H), 3.63-3.18 (m, 24 H), 3.09-3.03 (m, 1 H, -OCH2CH2CH2NH-), 2.92-2.78 (m, 4 H, -CH-NH2and -CH2S ), 2.52 (t, J =7.0 Hz, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.88 (ss, 6 H, -NHAc), 1.69-1.64 (m, 2 H, - OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.4, 101.3 (2 C), 98.8, 98.4, 75.7, 74.5,74.4, 73.6, 71.6, 69.6 (2 C), 68.3, 68.1, 67.5, 60.5, 55.6, 55.4, 36.1, 34.9, 33.2, 28.5, 22.4 (obtained from HSQC NMR). HRMS: C84HI48NI2O50S2[M + 2 H]2+calcd: 1094.4419, obsd: 1094.4406.PNAG27 (11011): Fig. 24Z2: Compound 1 was subjected to Boc, Alloc and Fmoc removals followed by acetylation, removal of Troc and Bz according to the general procedures to afford PNAG27 (1.5 mg, 33% yield for 6 steps). [α]D20= -3.0 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): J. 4.62-4.60 (m, 1 H, anomeric H obtained from HSQC NMR), 4.41-4.30 (m, 4 H, anomeric H), 4.07-3.99 (m, 4 H), 3.78-3.70 (m, 4 H), 3.62-3.20 (m, 23 H), 3.13-3.07 (m, 1 H, - OCH2CH2CH2NH-), 3.03-2.98 (m, 1 H, -OCH2CH2CH2NH-), 2.93-2.89 (m, 1 H, -CH-NH2),2.80 (t, J =6.5 Hz, 1 H, -CH2S ), 2.63-2.49 (m, 3 H, -CH2S- and -NHC=OCH2-), 1.89 (s, 3 H, - NHAc), 1.88 (ss, 6 H, -NHAc), 1.87 (s, 3 H, -NHAc), 1.63-1.58 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): <5. 101.5, 101.4 (2 C), 101.3, 99.0, 75.7, 74.4, 74.2, 73.8, 71.8, 69.9 (2 C), 69.8, 68.5, 68.4, 68.1, 67.5, 60.5, 55.4, 55.2, 36.3, 34.9, 33.2, 28.1, 22.2 (obtained from HSQC NMR). HRMS: C88Hi52Ni2O52S2[M + 2 H]2+calcd: 1136.4525, obsd: 1136.4498.PNAG28 (11100): Fig. 24Zs: Compound 2 was subjected to Boe, Alloc and Troc removals followed by acetylation, removal of Fmoc, Bz and TFA according to the general procedures to afford PNAG28 (3.4 mg, 50% yield for 6 steps). [a]o20= -1.9 (c 0.3, MeOH). ' H-NMR (500 MHz, D2O): <5 4.60-4.56 (m, 2 H, anomeric H), 4.37-4.33 (m, 3 H, anomeric H), 4.03-3.97 (m, 4 H), 3.80-3.73 (m, 4 H), 3.62-3.35 (m, 17 H), 3.29-3.18 (m, 6 H), 3.10-3.05 (m, 1 H, - OCH2CH2CH2NH-), 2.91-2.86 (m, 2 H, -CH-NH2), 2.80 (t, J =6.5 Hz, 2 H, -CH2S ), 2.52 (t, J =8.5 Hz, 2 H, -NHC=OCH2-), 1.88 (s, 9 H, -NHAc), 1.69-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.9 (2 C), 101.7, 99.0, 98.9, 75.9, 74.8, 74.6, 74.5 (2 C), 73.8, 72.0, 71.8, 69.9, 69.7, 68.7 (2 C), 68.5, 67.9, 67.7, 60.5, 55.6, 55.4, 36.1, 34.9, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C84HI48NI205OS2[M + 2 H]2+calcd: 1094.4419, obsd: 1094.4414.PNAG29 (11101): Fig. 24Z4: Compound 2 was subjected to Boc, Alloc and Troc removals followed by acetylation, removal of Fmoc and Bz according to the general procedures to afford PNAG29 (1.5 mg, 36% yield for 6 steps). [α]D20= -2.2 (c 0.3, MeOH). ^-NMR (500 MHz, D2O): 8. 4.61 (d, J =9.0 Hz, 1 H, anomeric H), 4.38-4.34 (m, 4 H, anomeric H), 4.04-3.98 (m, 4 H), 3.77-3.68 (m, 4 H), 3.62-3.21 (m, 23 H), 3.13-3.07 (m, 1 H, -OCH2CH2CH2NH-), 3.02-2.99 (m, 1 H, -OCH2CH2CH2NH-), 2.92 (dd, J =9.0, 10.0 Hz, 1 H, -CH-NH2), 2.80 (t, J =7.0 Hz, 1H, -CH2S-), 2.62-2.47 (m, 3 H, -CH2S- and -NHC=OCH2-), 1.89-1.88 (m, 12 H, -NHAc), 1.62-I.57 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): d. 101.8, 101.5 (2 C), 101.3, 99.0, 75.7, 74.6, 74.3, 73.8, 72.0, 69.9 (2 C), 69.7, 68.3 (2 C), 67.9, 60.7, 55.6, 55.4, 36.7, 36.3, 29.1, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C88Hi52Ni2O52S2[M + 2 H]2+calcd: 1136.4525, obsd: 1136.4520.PNAG30 (11110): Fig. 24Zs: Compound 2 was subjected to Boc, Alloc, Troc and Fmoc removals followed by acetylation, removal of Bz and TFA according to the general procedures to afford PNAG30 (1.0 mg, 40% yield for 6 steps). [a]o20= -2.1 (c 0.2, MeOH). ' H-NMR (500 MHz, D2O): 8. 4.54 (d, J =8.5 Hz, 1 H, anomeric H), 4.38-4.32 (m, 4 H, anomeric H), 4.4.04-3.98 (m, 4 H), 3.77-3.73 (m, 3 H), 3.61-3.35 (m, 18 H), 3.29-3.18 (m, 7 H), 3.09-3.03 (m, 1 H, -OCH2CH2CH2NH-), 2.87 (t, J = 10.5 Hz, 1 H, -CH-NH2), 2.83-2.78 (m, 2 H, -CH2S ),2.53-2.49 (m, 2 H, -NHC=OCH2-), 1.89 (s, 3 H, -NHAc), 1.88 (s, 6 H, -NHAc), 1.87 (s, 3 H, - NHAc), 1.70-1.64 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.5 (2 C), 101.4 (2 C), 98.6, 75.7, 74.7, 74.0, 73.6, 71.8, 69.9, 69.7, 68.2, 67.8, 67.2, 60.5, 55.4, 55.2, 36.1, 34.7, 33.2, 28.3, 22.2 (obtained from HSQC NMR). HRMS: C88H152N12O52S2 [M + 2 H]2+calcd: 1136.4525, obsd: 1136.4508.PNAG31 (11111): Fig. 24Ze: Compound 1 was subjected to Boe, Alloc, Troc and Fmoc removals followed by acetylation, removal of Bz according to the general procedures to afford PNAG31 (3.5 mg, 38% yield for 6 steps). [α]D20= -2.4 (c 0.2, MeOH). ^-NMR (500 MHz, D2O): . 4.38-4.33 (m, 4 H, anomeric H), 4.30 (d, J =8.0 Hz, 1 H, anomeric H), 4.33 (d, J =9.0 Hz, 2 H), 4.04-3.98 (m, 4 H), 3.77-3.68 (m, 3 H), 3.61-3.47 (m, 10 H), 3.43-3.33 (m, 9 H), 3.29- 3.19 (m, 6 H), 3.14-3.09 (m, 1 H, -OCH2CH2CH2NH-), 3.03-2.97 (m, 1 H, -OCH2CH2CH2NH-), 2.80 (t, J =7.0 Hz, 2 H, -CH2S-), 2.51 (t, J =6.5 Hz, 2 H, -NHC=OCH2-), 1.88-1.86 (m, 15 H, - NHAc), 1.62-1.57 (m, 2 H, -OCH2CH2CH2NH-).13C-NMR (125 MHz, D2O): 3. 101.3 (4 C), 101.1, 75.7, 74.4, 73.5, 69.8, 69.7, 68.4, 68.3, 60.5, 55.4, 55.2, 36.1, 34.7, 33.0, 28.1, 22.0 (obtained from HSQC NMR). HRMS: C92H156N12O54S2 [M + 2 H]2+calcd: 1178.4630, obsd: 1178.4628.References:1. S. G. Hansen, T. Skrydstrup, Studies Directed to the Synthesis of Oligochitosans - Preparation of Building Blocks and Their Evaluation in Glycosylation Studies. Eur J. Org. Chem. 2007, 3392-3401 (2007).Appendicies include the following:Appendix 1: Glycan microarray screening results.Appendix 2: NMR spectra of new compounds.Incorporation by ReferenceAll publications, patent applications, patents, and other references mentioned in the specification are indicative of the level of those skilled in the art to which the presently disclosed subject matter pertains. All publications, patent applications, patents, and other references are herein incorporated by reference to the same extent as if each individual publication, patent application, patent, and other reference was specifically and individually indicated to be incorporated by reference. It will be understood that, although a number of patent applications, patents, and other references are referred to herein, such reference does not constitute an admission that any of these documents forms part of the common general knowledge in the art.EquivalentsAlthough the foregoing subject matter has been described in some detail by way of illustration and example for purposes of clarity of understanding, it will be understood by those skilled in the art that certain changes and modifications can be practiced within the scope of the appended claims.APPENDIX 1APPENDIX 24a6a8a9a10a11a13a14a15a16a17a18a19a20a21a22a23a24a25a26a27a28a29a30a32a33a34a35a36a38a39a40a41a42a43a44a45a46a47a48a49a50a51a52a53a54a55a56a57a58a59a60a61a62a63a64a65a66a67a68a69a70a71a72a73a74a75a76a77a

Claims

What is claimed is:

1. A vaccine composition comprising a poly-P-(l-6)-A-acetylglucosamine (PNAG) antigen, wherein the PNAG antigen comprises at least 2, 3, 4, 5, 6, 7, 8, 9, or 10 glucosamine monomers, wherein at least one glucosamine monomer is acetylated.

2. A vaccine composition comprising a poly-P-(l-6)-A-acetylglucosamine (PNAG) antigen, wherein the PNAG antigen comprises at least 5, 6, 7, 8, 9, or 10 glucosamine monomers comprising the following Formula I:wherein Rl, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof.

3. The vaccine composition of claim 1, wherein the PNAG antigen is a disaccharide.

4. The vaccine composition of any one of claims 1-3, wherein the PNAG antigen is a pentasaccharide or a nonasaccharide.

5. The vaccine composition of any one of claims 1-4, wherein the PNAG antigen comprises free amines.

6. The vaccine composition of any one of claims 1-5, wherein the PNAG antigen comprises 1 free amine, 2 free amines, 3 free amines, 4 free amines, or 5 free amines.

7. The vaccine composition of any one of claims 1-6, wherein the PNAG antigen does not comprise free amines.

8. The vaccine composition of any one of claims 2-7, wherein Ri, R2, R3, R4, and R5 are each H.

9. The vaccine composition of any one of claims 2-7, wherein at least R2 and / or R4 is N- acetyl.

10. The vaccine composition of any one of claims 2-6, wherein Ri, R3, and R4 are H; and wherein R2 and R5 are A- acetyl.

11. The vaccine composition of any one of claims 2-6, wherein R3 and R4 are H; and wherein Ri, R2, andRs are N- acetyl.

12. The vaccine composition of any one of claims 2-11, wherein the antigen comprises a PNAG pentasaccharide set forth in Figure 1.

13. The vaccine composition of any one of claims 1-13, wherein the PNAG antigen bears a linker comprising a disulfide group.

14. The vaccine composition of any one of claims 1-13, wherein the PNAG antigen is conjugated to a protein carrier.

15. The vaccine composition of claim 13, wherein the PNAG antigen conjugated to a protein carrier comprises the following Formula II:wherein Rl, R2, R3, R4, and R5 are each hydrogen (H), A-acetyl or any combination thereof; wherein is 1 or 2; and wherein R’ is the protein carrier.

16. The vaccine composition of claim 14 or 15, wherein the protein carrier is a viral capsid, and wherein said capsid comprises at least one non-natural mutation.

17. The vaccine composition of claim 16, wherein the capsid comprises at least one, two, three, four, five, six, seven, eight, nine, ten, eleven, twelve, thirteen, fourteen, fifteen, sixteen, seventeen, eighteen, nineteen, or twenty mutations.

18. The vaccine composition of claim 16 or 17, wherein the mutation is a disulfide bond mutation.

19. The vaccine composition of any one of claims 16-18, wherein the viral capsid is derived from a bacteriophage.

20. The vaccine composition of claim 19, wherein the bacteriophage is selected from the group consisting of (a): bacteriophage QP; (b) bacteriophage R17; (c) bacteriophage fr; (d) bacteriophage GA; I bacteriophage SP; (f) bacteriophage MS2; (g) bacteriophage Mi l; (h) bacteriophage MX1; (i) bacteriophage NL95; (j) bacteriophage f2; (k) bacteriophage PP7; (1) bacteriophage AP205; and (m) bacteriophage P22.

21. The vaccine composition of claim 19 or 20, wherein the bacteriophage is bacteriophage Q beta (QP).

22. The vaccine composition of any one of claims 16-21, wherein the mutation comprises at least one mutation selected from N10K, A38K, A40C, A40S, T75K, D102C, D102S, or A117K, or combination thereof.

23. The vaccine composition of any one of claims 16-22, wherein the mutation comprises A38K.

24. The vaccine composition of any one of claims 16-21, wherein said capsid comprises at least two mutations selected from A40C / D102C, A40S / D102S, or A43C / Q98C.

25. The vaccine composition of any one of claims 16-21, wherein said capsid comprises at least three mutations selected from A40C / D102C / K13R or A38K / A40C / D102C.

26. A method of preventing or treating a microbial infection in a subject, the method comprising administering to the subject the vaccine composition of any one of claims 1-25.

27. A method of preventing or treating opportunistic microbial infection in an immnocomprimised subject, the method comprising administering to the subject the vaccine composition of any one of claims 1-25.

28. The method of claim 26 or 27, wherein, the subject is elderly, a burn patient, a premature infant, undergoing chemotherapy, undergoing radiation therapy, receiving immunosuppressive or immunoablative / depleting therapy, or any combination thereof.

29. The method of any one of claims 26-27, wherein the subject is infected or at risk of infection by a PNAG-based microbe.

30. The method of claim 29, wherein the PNAG-based microbe is selected from Grampositive bacteria, Gram-negative bacteria, fungal pathogens, protozoan pathogens.

31. The method of any one of claims 26-30, wherein the infection is a local infection.

32. The method of any one of claims 26-30, wherein the infection is a systemic infection.

33. The method of any one of claims 26-32, wherein the infection is resistant or refractory to drug treatment.

34. The method of any one of claims 26-33, wherein the infection is resistant or refractory to one or more antibiotics.

35. The method of any one of claims 26-34, wherein the vaccine composition is administered systemically.

36. The method of claim 35, wherein the systemic administration is selected from the group consisting of oral, intravenous, intradermal, intraperitoneal, subcutaneous, and intramuscular administration.

37. The method of any one of claims 26-36, wherein the vaccine is directed against multiple strains of a PNAG-based microbe.

38. The method of any one of claims 26-37, wherein the vaccine is directed against more than one PNAG-based microbe.

39. The method of claim 38, wherein the more than one PNAG-based microbe is selected from Gram-positive bacteria, Gram-negative bacteria, fungal pathogens, protozoan pathogens, and any combination thereof.

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