Labeled oligosaccharides and methods of use

Abstract

Provided herein, inter alia, are methods for making [18F]-labeled oligosaccharides, methods of detecting a disease, and methods of treating the disease using the [18F] -labeled oligosaccharides.

Description

Attorney Docket No.: 048536-766001WO Client Ref. No. SF-2023-149-1-US LABELED OLIGOSACCHARIDES AND METHODS OF USE CROSS-REFERENCES TO RELATED APPLICATIONS

[0001] This application claims the benefit of U.S. Provisional Application No.63 / 503,141, filed May 18, 2023, which is incorporated herein by reference in its entirety and for all purposes. STATEMENT AS TO RIGHTS TO INVENTIONS MADE UNDER FEDERALLY SPONSORED RESEARCH AND DEVELOPMENT

[0002] This invention was made with government support under R01EB024014, R01EB025985, and R01EB030897, awarded by the National Institutes of Health, and under A132172, awarded by the Department of Defense. The government has certain rights in the invention. BACKGROUND

[0003] Clinicians often rely on positron emission tomography (PET) scans for detecting pathogens that cause infectious disease, where an injected radio-labeled tracer reveals evidence of disease. However, existing clinically used PET tracers largely detect the body’s response to infection, not the living pathogens in the body. Proper imaging and treatment of infected patients with equivocal magnetic resonance imaging (MRI) or computed tomography (CT) imaging has proved challenging. To date, there are no bacteria-specific imaging methods available to clinicians in the acute care setting, when patients present with signs and symptoms of bacterial infection. Many radiotracers in development are insensitive to bacterial pathogens, for example Staphylococcus aureus. Hence, there is an unmet need for developing PET tracers capable of detecting and visualizing active infectious pathogens. BRIEF SUMMARY

[0004] Provided herein, inter alia, are methods of producing [18F]-labeled oligosaccharides. The disclosed methods provide a rapid, one-step radiosynthesis using [18F]FDG, a phosphorylase, and phosphorylated monosaccharide to obtain living pathogen-sensitive [18F]- labeled oligosaccharides with high regioselectivity / stereoselectivity.

[0005] In embodiments, the disclosed methods comprise combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide in a reaction vessel to produce [18F]-labeled oligosaccharides. In embodiments, the [18F]-labeled oligosaccharides produced by the methods provided herein include [18F]-labeled disaccharides. In embodiments,the [18F]-labeled oligosaccharides produced by the methods provided herein include analpha-1,4 and beta-1,4 linked [18F] oligosaccharides.

[0006] In an aspect is provided a method of producing an [18F]-labeled oligosaccharide, the method including combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide selected from the group consisting of D-glucose-1-phosphate, D-galactose-1-phosphate, and D-mannose-1-phosphate in a reaction vessel, thereby producing said [18F]-labeled oligosaccharide.

[0007] Additionally provided herein are [18F]-labeled oligosaccharides suitable for use as PET tracers. Exemplary [18F]-labeled oligosaccharides include, but are not limited to, 2-deoxy-[18F]- fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro- cellobiose ([18F]FDC), 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), 2-deoxy-2-[18F]fluoro- laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 2-deoxy-2-[18F]fluoro-lactose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2- [18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro- glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2- deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro- cellotriose, 2-deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose,2-deoxy-[18F]- fluoro-cellohexaose.

[0008] In an aspect is provided a [18F]-labeled oligosaccharide selected from the group consisting of 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2- [18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro- glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2- deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro- cellotriose, 2-deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, and 2-deoxy- [18F]-fluoro-cellohexaose.

[0009] In an aspect is provided a pharmaceutical composition including an effective amount of the [18F]-labeled oligosaccharide provided herein including embodiments thereof and a pharmaceutically acceptable excipient.

[0010] Also provided herein are methods of detecting a disease in a subject in need thereof. The disclosed methods comprise injecting into the subject an effective amount of the [18F]- labeled oligosaccharides provided herein, imaging the subject and detecting the accumulation of the [18F]-labeled oligosaccharides in a tissue of the subject.

[0011] In an aspect is provided a method of detecting a disease in a subject in need thereof, the method including administering to the subject an effective amount of the [18F]-labeled oligosaccharide provided herein including embodiments thereof or the pharmaceutical composition provided herein including embodiments thereof, and detecting the [18F]-labeled oligosaccharide in the subject, thereby detecting the disease.

[0012] In embodiments, the disclosed methods may further comprise treating the disease by administering to the subject one or more anti-inflammatory agents, antibiotics, antifungal agents, or anti-cancer agents. BRIEF DESCRIPTION OF THE DRAWINGS

[0013] FIGS.1A-1B. Chemoenzymatic synthesis of disaccharides via reverse phosphorolysis using (FIG.1A) exemplary disaccharide phosphorylases. (FIG.1B) Reverse phosphorolysis of substrates using disaccharide phosphorylase with either α- or β-glucose-1-phosphate.

[0014] FIGS.2A-2C. Radiochemical syntheses of 2-deoxy-2-[18F]fluoro-maltose ([18F]FDM) and 2-deoxy-2-[18F]fluoro-sakebiose ([18F]FSK). (FIG.2A) Synthesis of ^-D-glucose-1- phosphate ( ^Glc1-P) precursor starting from S1. (FIG.2B) Enzymatic radiosynthesis of [18F]FDM and [18F]FSK from 2-deoxy-2-[18F]fluoro-D-glucose [18F]FDG and ^Glc1-P using maltose phosphorylase. (FIG.2C) Radio HPLC analysis of crude products using a YMC-Pack Polyamine II column.

[0015] FIGS.3A-3B. Chemoenzymatic radiosyntheses of (FIG.3A) [18F]FDT, [18F]FDL, [18F]FDC, [18F]FDM and [18F]FSK from [18F]FDG. All reactions were carried out at 37oC, stirring for 20 min, using 6 mg (0.020 mmol) of precursor, 0.3 mg of enzyme (3-6 units) and 10- 15mCi [18F]FDG in 0.5 mL of citrate buffer (0.1M, pH = 6.0). (FIG.3B) Radio HPLC analysis of each enzymatic reaction using a YMC-Pack Polyamine II column.

[0016] FIGS.4A-4D. In vitro bacteria uptake of (FIG.4A) [18F]FDG, [18F]FDS, [18F]FDM and [18F]FSK. (FIG.4B) In vitro bacteria uptake of [18F]FDM and [18F]FSK in gram-positive and gram-negative pathogens. (FIG.4C) Competition of [18F]FDM and [18F]FSK uptake withincreasing concentrations of unlabeled (cold) maltose in S. aureus. (FIG.4D) In vitro bacteria uptake of [18F]FDM and [18F]FSK in methicillin-resistant S. aureus (MRSA) clinical strains.

[0017] FIG.5. Living bacteria detection comparison by [18F]FDM, [18F]FSK and clinical [18F]FDG. μPET-CT imaging of MRSA myositis in mice with [18F]FDM, [18F]FSK, and [18F]FDG. The gray arrows indicate the site of inoculation with live bacteria, while the white arrows correspond to heat-killed bacteria. The corresponding bar graphs indicate region-of- interest (ROI) analysis. As reflected by the images, the mean [18F]FDM and [18F]FSK accumulation for tissues infected with live bacteria was respectively 6.1 and 6.5-fold higher than that seen for heat-killed inoculation (P <0.0001). In contrast, this difference was not seen for [18F]FDG.

[0018] FIGS.6A-6F. Imaging of S. aureus in vertebral discitis-osteomyelitis (VDO) rat models and A. baumannii in a myositis mouse model using [18F]FSK. (FIG.6A) Optical tomography image of rat tail showing bioluminescent signal from S. aureus Xen29 inoculation. (FIG.6B) Computed tomography study performed at 10 days highlights the similarity between rodent and human discitis osteomyelitis. (FIG.6C) PET / CT imaging of S. aureus Xen29 vertebral discitis-osteomyelitis (VDO) in rat (N = 5) with [18F]FSK. (FIG.6D) ROI analysis showing increased signal in segments inoculated with live bacteria versus background (P < 0.0001). (FIG.6E) PET / CT imaging of A. baumannii myositis in mice (N = 6) with [18F]FSK. The arrows on the left indicate the site of inoculation with live bacteria. The arrows on the right correspond to heat-killed bacteria. (FIG.6F) ROI analysis showing increased signal in infected muscle versus inflammation (P < 0.0001).

[0019] FIG.7. Schematic representation of the general synthesis of [18F] FDG-derived disaccharides via reverse phosphorolysis with an example of in vivo A. baumanii uptake in an infected mouse.

[0020] FIG.8. In vitro uptake of [18F]FDM and [18F]FSK in heat-killed bacteria and structures thereof.

[0021] FIG.9. In vitro bacteria uptake [18F]FDM and [18F]FSK in S. aureus and K. pneumoniae at 30 min, 60 min and 90 min.

[0022] FIG.10. Efflux experiment: plot showing the residual activity observed at 30 min post efflux of [18F]FDM and [18F]FSK in S. aureus and K. pneumoniae following an initial 30 minutes incubation.

[0023] FIG.11. Stability of [18F]FDM and [18F]FSK in human and mouse serum. Unlike its robustness in human serum, [18F]FDM and [18F]FSK degraded in mouse serum overtime due to presence of α-glucosidase, which can hydrolyze [18F]FDM and [18F]FSK.

[0024] FIG.12. Schematic of ^-glucosidase hydrolysis of [18F]FDM and [18F]FSK. In murine serum, ^-glucosidase catalyzes the decomposition of [18F]FDM and [18F]FSK into [18F]FDG and glucose.

[0025] FIG.13. Inhibitors of ^–glycosidase, including voglibose, acarbose, and miglitol, that have the potential for higher stability of [18F]FDM and [18F]FSK in mouse serum.

[0026] FIG.14. In vitro S. aureus uptake for [18F]FDM and [18F]FSK in the presence of the indicated ^-glucosidase inhibitors.

[0027] FIG.15. Dose response of [18F]FDM and [18F]FSK in mouse serum exposed to increasing concentrations of ^-glucosidase inhibitors (0.001 to 10 mM).

[0028] FIG.s 16A-16B. In vivo experiment: [18F]FDM (FIG.16A) and [18F]FSK (FIG.16B) in mouse, No bacteria, Inhibitor: Voglibose (1mg / inj), Injection: 200 uCi 18F-tracer, N = 5. Ex vivo data was obtained following tissue harvesting on a gamma counter.

[0029] FIG.s 17A-17C. In vivo experiment: [18F]FDG (FIG.17A), [18F]FDM (FIG.17B) and [18F]FSK (FIG.17C) in mouse, No bacteria, No inhibitor, Injection: ≈200uCi 18F-tracer, N = 5. Ex vivo data was obtained following tissue harvesting on a gamma counter.

[0030] FIG.18. In vivo experiment: [18F]FDM in mouse myositis model, S. aureus (MRSA 01), Inhibitor: Voglibose (1mg / inj), Injection: ≈200uCi 18F-tracer, N = 6. The arrows on the left indicate the site of inoculation with live bacteria. The arrows on the right indicate the site of inoculation with heat-killed bacteria. Ex vivo data was obtained following tissue harvesting on a gamma counter. ROI analysis, live vs HK: 6.1 fold excess, **** P value <0.0001, ex vivo analysis, live vs HK: 3.9 fold excess, ** P value = 0.001 (unpaired t-test).

[0031] FIG.19. In vivo experiment: [18F]FSK in mouse myositis model, S. aureus (MRSA 01), Inhibitor: Voglibose (1mg / inj), Injection: ≈200uCi 18F-tracer, N = 6. The arrows on the left indicate the site of inoculation with live bacteria. The arrows on the right indicate the site of inoculation with heat-killed bacteria. Ex vivo data was obtained following tissue harvesting on a gamma counter. ROI analysis, live vs HK: 6.5 fold excess, **** P value <0.0001, ex vivo analysis, live vs HK: 4.7 fold excess, **** P value < 0.0001 (unpaired t-test).

[0032] FIG.20. In vivo experiment: [18F]FDG in mouse myositis model, S. aureus (MRSA 01), Inhibitor: Voglibose (1mg / inj), Injection: ≈200uCi 18F-tracer, N = 5. The arrows on the left indicate the site of inoculation with live bacteria. The arrows on the right indicate the site of inoculation with heat-killed bacteria. Ex vivo data was obtained following tissue harvesting on a gamma counter. ROI analysis, live vs HK: 1.10 fold excess, ns, ex vivo analysis, live vs HK: 1.02 fold excess, ns (unpaired t-test).

[0033] FIGS.21A-21C. In vivo experiment: [18F]FSK in rat vertebral discitis-osteomyelitis (VDO), S. aureus (Xen29), Day 4, inhibitor: Voglibose (5mg / inj), Injection: ≈500uCi 18F- tracer, N = 5. The arrows on the top indicate the site of inoculation with live bacteria. The arrows on the bottom indicate the site of inoculation with heat-killed bacteria. (FIG.21A) PET / CT imaging of S. aureus Xen29 vertebral discitis-osteomyelitis (VDO) in rat (N = 5) with [18F]FSK. ROI analysis, live vs HK: 2.8 fold excess, **** P value <0.0001 (unpaired t-test). (FIG.21B) Computed tomography study performed at 10 days highlights the similarity between rodent and human discitis osteomyelitis. (FIG.21C) Optical tomography image of rat tail showing bioluminescent signal from S. aureus Xen29 inoculation.

[0034] FIG.22. In vivo experiment: [18F]FSK in rat vertebral discitis-osteomyelitis (VDO), S. aureus (Xen29), Day 10, inhibitor: Voglibose (5mg / inj), Injection: ≈500uCi 18F-tracer, N = 3. The arrows on the top indicate the site of inoculation with live bacteria. The arrows on the bottom indicate the site of inoculation with heat-killed bacteria. ROI analysis, live vs HK: 3.1 fold excess, **** P value <0.0001 (unpaired t-test).

[0035] FIG.23. In vivo experiment: [18F]FSK in mouse myositis model, A. baumanni (ATCC 19606), inhibitor: Voglibose (1mg / inj), Injection: ≈200uCi 18F-tracer, N = 6. The arrows on the left indicate the site of inoculation with live bacteria. The arrows on the right indicate the site of inoculation with heat-killed bacteria. Ex vivo data was obtained following tissue harvesting on a gamma counter. ROI analysis, live vs HK: 4.9 fold excess, **** P value <0.0001, ex vivo analysis, live vs HK: 3.9 fold excess, **** P value < 0.0001 (unpaired t-test).

[0036] FIG.24. Chromatograms for semi-prep HPLC purification of 2-deoxy-2[19F]-fluoro- maltose and 2-deoxy-2-[19F]-fluoro-sakebiose.

[0037] FIG.25. Chromatograms for semi-prep HPLC purification of 2-deoxy-2[19F]-fluoro- cellobiose.

[0038] FIG.26. Chromatograms for semi-prep HPLC purification of 2-deoxy-2[19F]-fluoro- trehalose.

[0039] FIG.27. Chromatograms for semi-prep HPLC purification of 2-deoxy-2[19F]-fluoro- laminaribiose.

[0040] FIG.28. Structures of ^- 2-deoxy-2[19F]-fluoro-maltose and ^ ^ 2-deoxy-2[19F]-fluoro- maltose.

[0041] FIG.29. Structure of ^-2-deoxy-2[19F]-fluoro-sakebiose and ^ ^ 2-deoxy-2[19F]-fluoro- sakebiose.

[0042] FIG.30. Structures of ^- 2-deoxy-2[19F]-fluoro-cellobiose and ^ ^ 2-deoxy-2[19F]- fluoro-cellobiose

[0043] FIG.31. Structure of 2-deoxy-2[19F]-fluoro-trehalose.

[0044] FIG.32. Structures of ^- 2-deoxy-2[19F]-fluoro-laminaribiose and ^ ^ ^ 2-deoxy-2[19F]- fluoro-laminaribiose.

[0045] FIG.33. Chemical structures of ^ / ^ glucose 1-phosphate, ^ / ^ galactose-1-phosphate, ^ mannose 1-phosphate.

[0046] FIG.34. Exemplary chemoenzymatic reactions described herein for producing oligosaccharides and [18F]-labeled derivatives thereof.

[0047] FIG.35. [18F]-Labeled Compounds produced by the reactions shown in FIG.34.

[0048] FIG.36. Reactions using maltodextrin phosphorylase.

[0049] FIG.37. [18F] Labelled Compounds produced by the reactions shown in FIG.36.

[0050] FIG.38. Reactions using cellodextrin phosphorylase.

[0051] FIG.39. [18F] Labelled Compounds produced by the reactions shown in FIG.38.

[0052] FIG.40. Reactions using α / β D-glucose-1-phosphate as precursor and [18F] Labelled Compounds produced by said reactions.

[0053] FIG.41. Reactions using α / β D-galactose-1-phosphate as precursor.

[0054] FIG.42. [18F] Labelled Compounds produced by the reactions shown in FIG.41.

[0055] FIG.43. Reactions using α D-mannose-1-phosphate as precursor and [18F] Labelled Compounds produced by said reactions.

[0056] FIG.s 44A-44B. Screening and optimization of [18F]FSK for imaging live bacteria. (FIG.44A) In vitro screening of S. aureus strains using multiple tracers. (FIG.44B) Condition optimization of radiosynthesis of [18F]FSK using the new sakebiose phosphorylase (nigerose phosphorylase, NP). DETAILED DESCRIPTION

[0057] Before the present invention is further described, it is to be understood that this invention is not strictly limited to particular embodiments described, as such may of course vary. It is also to be understood that the terminology used herein is for the purpose of describing particular embodiments only, and is not intended to be limiting, since the scope of the present invention will be limited only by the claims.

[0058] It must be noted that as used herein and in the appended claims, the singular forms “a,” “an,” and “the” include plural referents unless the context clearly dictates otherwise. It should further be understood that as used herein, the term “a” entity or “an” entity refers to one or more of that entity. For example, a nucleic acid molecule refers to one or more nucleic acid molecules. As such, the terms “a”, “an”, “one or more” and “at least one” can be used interchangeably. Similarly, the terms “comprising”, “including” and “having” can be used interchangeably.

[0059] Unless defined otherwise, technical and scientific terms used herein have the same meaning as commonly understood by a person of ordinary skill in the art. See, e.g., Singleton et al., Dictionary of Microbiology and Molecular Biology, 2nd ed., J. Wiley & Sons (New York, NY 1994); Sambrook et al., Molecular Cloning, A Laboratory Manual, Cold Springs Harbor Press (Cold Springs Harbor, NY 1989). Any methods, devices and materials similar or equivalent to those described herein can be used in the practice of this disclosure. The following definitions are provided to facilitate understanding of certain terms used frequently herein and are not meant to limit the scope of the present disclosure.

[0060] The abbreviations used herein have their conventional meaning within the chemical and biological arts. The chemical structures and formulae set forth herein are constructed according to the standard rules of chemical valency known in the chemical arts.

[0061] As used herein, the term “about” means a range of values including the specified value, which a person of ordinary skill in the art would consider reasonably similar to the specifiedvalue. In embodiments, about means within a standard deviation using measurements generally acceptable in the art. In embodiments, about means a range extending to + / - 10% of the specified value. In embodiments, about means the specified value.

[0062] It should be noted that throughout the application that alternatives are written in Markush groups. It is specifically contemplated that each member of the Markush group should be considered separately, thereby comprising another embodiment, and the Markush group is not to be read as a single unit.

[0063] A “cell” refers to a cell carrying out metabolic or other function sufficient to preserve or replicate its genomic DNA. A cell can be identified by well-known methods in the art including, for example, presence of an intact membrane, staining by a particular dye, ability to produce progeny or, in the case of a gamete, ability to combine with a second gamete to produce a viable offspring. Cells may include prokaryotic and eukaryotic cells. Prokaryotic cells include but are not limited to bacteria. Eukaryotic cells include but are not limited to yeast cells and cells derived from plants and animals, for example mammalian (e.g. human) cells. Cells may be useful when they are naturally nonadherent or have been treated not to adhere to surfaces, for example by trypsinization.

[0064] The term “microbe” is used in accordance with its plain ordinary meaning in the biological arts and refers generally to a microorganism. In embodiments, a microbe is a single cell or a group or colony of cells. A microbe may be a bacterium, fungus, archaea, protist, or virus. In embodiments, the microbe is a gram-positive bacteria. In embodiments, the microbe is a gram-negative bacteria. In embodiments, the microbe is a virus. In embodiments, the microbe is a fungus.

[0065] "Contacting" is used in accordance with its plain ordinary meaning and refers to the process of allowing at least two distinct species (e.g. an enzyme and a substrate (e.g. a phosphorylase and phosphorylated monosaccharide)) to become sufficiently proximal to react, interact, or physically touch. It should be appreciated; however, that the resulting reaction product can be produced directly from a reaction between the added reagents or from an intermediate from one or more of the added reagents which can be produced in the reaction mixture. In embodiments, the contacting occurs when combining the at least two distinct species in a solution. Thus, in the context of a reaction, in embodiments, “contacting” may be used interchangeably with “combining”.

[0066] The term "contacting" may include allowing two species to react, interact, or physically touch, wherein the two species may be, for example, a pharmaceutical composition as provided herein and a cell. In embodiments contacting includes, for example, allowing a composition including a [18F]-labeled oligosaccharide as described herein to interact with a cell (e.g. a bacterial cell).

[0067] The term “solution” is used in accordance to its ordinary meaning in the art and refers to a liquid mixture in which the minor component (e.g., a solute or compound) is uniformly distributed within the major component (e.g., a solvent). For example, in embodiments, the solution may include a solvent, [18F]FDG, a phosphorylase and a phosphorylated monosaccharide

[0068] As used herein, the term “salt” refers to acid or base salts of the compounds used in the methods of the present invention. Illustrative examples of acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, and the like) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid and the like) salts, quaternary ammonium (methyl iodide, ethyl iodide, and the like) salts.

[0069] The terms “bind” and “bound” as used herein is used in accordance with its plain and ordinary meaning and refers to the association between atoms or molecules. The association can be direct or indirect. For example, bound atoms or molecules may be bound, e.g., by covalent bond, linker (e.g. a first linker or second linker), or non-covalent bond (e.g. electrostatic interactions (e.g. ionic bond, hydrogen bond, halogen bond), van der Waals interactions (e.g. dipole-dipole, dipole-induced dipole, London dispersion), ring stacking (pi effects), hydrophobic interactions and the like). In embodiments, “join” or “joined” in the context of an association between atoms or molecules are used interchangeably with bind and bound.

[0070] As used herein, the term "conjugated” when referring to two moieties means the two moieties are bonded, wherein the bond or bonds connecting the two moieties may be covalent or non-covalent. In embodiments, the two moieties are covalently bonded to each other (e.g. directly or through a covalently bonded intermediary). In embodiments, the two moieties are non-covalently bonded (e.g. through ionic bond(s), van der waal’s bond(s) / interactions, hydrogen bond(s), polar bond(s), or combinations or mixtures thereof).

[0071] The term “non-nucleophilic base” as used herein refers to any sterically hindered base that is a poor nucleophile.

[0072] The term “nucleophile” as used herein refers to a chemical species that donates an electron pair to an electrophile to form a chemical bond in relation to a reaction. All molecules or ions with a free pair of electrons or at least one pi bond can act as nucleophiles.

[0073] The term “strong acid” as used herein refers to an acid that is completely dissociated or ionized in an aqueous solution. Examples of common strong acids include hydrochloric acid (HCl), nitric acid (HNO3), sulfuric acid (H2SO4), hydrobromic acid (HBr), hydroiodic acid (HI), perchloric acid (HClO4), or chloric acid (HClO3).

[0074] "Analog," "analogue," or "derivative" is used in accordance with its plain ordinary meaning within Chemistry and Biology and refers to a chemical agent that is structurally similar to another agent (i.e., a so-called "reference" agent) but differs in composition, e.g., in the replacement of one atom by an atom of a different element, or in the presence of a particular functional group, or the replacement of one functional group by another functional group, or the absolute stereochemistry of a chiral center of the reference agent. In some embodiments, a derivative may be a conjugate with a pharmaceutically acceptable agent, for example, phosphate or phosphonate.

[0075] As used herein, the term "salt" refers to acid or base salts of the agents used herein. Illustrative but non-limiting examples of acceptable salts are mineral acid (hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuric acid, and the like) salts, organic acid (acetic acid, propionic acid, glutamic acid, citric acid, and the like) salts, and quaternary ammonium (methyl iodide, ethyl iodide, and the like) salts.

[0076] The term “fluorine” as used herein refers to a chemical element with the symbol F and atomic number 9. In embodiment, the stable isotope19F occurs naturally in abundance. In embodiments, the synthesized isotope18F has a half-life of 109.77 minutes.

[0077] As used herein, the term “carbohydrate” refers to a saccharide, including a monosaccharide, oligosaccharide, or polysaccharide. Thus, in embodiments, carbohydrate may refer to a single monosaccharide unit. In embodiments, carbohydrate may refer to a plurality of monosaccharides joined by glycosidic linkages. For example, in embodiments, a carbohydrate may be an oligosaccharide including 2 to 6 monosaccharide units joined together by glycosidic bonds. in embodiments, carbohydrate may refer to an oligosaccharide. in embodiments, carbohydrate may refer to a polysaccharide.

[0078] As used herein, the term “polysaccharide” refers to a polymer composed of monosaccharide units (e.g. monomers) joined by glycosidic linkages. Polysaccharides include but are not limited to polymers of any length, from a disaccharide polymer to polymers including hundreds or thousands of monosaccharides. In embodiments, a polysaccharide is a disaccharide. In embodiments, a polysaccharide is an oligosaccharide. In embodiments, polysaccharides are linear. In embodiments, polysacchapolyrides are branched. In embodiments, exemplary polysaccharides include starch, glycogen, galactogen, cellulose, pectin, and chitin.

[0079] The term “oligosaccharide” is used in accordance to its plain ordinary meaning and refers to a saccharide polymer containing from 2 to 6 units of simple sugars or monosaccharide units (e.g. monomers) joined together by glycosidic bonds. In embodiments, oligosaccharides include disaccharides. In embodiments, oligosaccharides include trisaccharides. In embodiments, oligosaccharides include tetrasaccharides. In embodiments, oligosaccharides include pentasaccharides. In embodiments, oligosaccharides include hexasaccharides. In embodiments, the oligosaccharide includes 2 monosaccharide units. In embodiments, the oligosaccharide includes 3 monosaccharide units. In embodiments, the oligosaccharide includes 4 monosaccharide units. In embodiments, the oligosaccharide includes 5 monosaccharide units. In embodiments, the oligosaccharide includes 6 monosaccharide units.

[0080] As used herein, the term “monosaccharide” refers to a single sugar unit or monomer that serves as a building block of a polysaccharide. In embodiments, a monosaccharide is referred to as “saccharide monomer” or “saccharide monomer unit”. For example, in embodiments, a monosaccharide exists as a single unit without a glycosidic bond to other such monosaccharide units. In embodiments, monosaccharides are polyhydroxy aldehydes. In embodiments, monosaccharides are ketones. In embodiments, monosaccharides have the formula (CH2O)n. In embodiments, monosaccharides are molecules with more than one hydroxyl group (-OH), and a carbonyl group (C=O) either at the terminal carbon atom (aldose) or at the second carbon atom (ketose). In embodiments, monosaccharides are classified by the number of carbon atoms in the molecule. Dioses have two carbon atoms, trioses have three, tetroses four, pentoses five, hexoses six, and heptoses seven. In embodiments, monosaccharides include glucose, fructose, and glyceraldehyde. In embodiments, monosaccharides include glucose, galactose, and mannose.

[0081] In embodiments, monosaccharide (e.g. saccharide monomer, saccharide monomer unit) refers to the monovalent free radical structure of the monosaccharide. For example, inembodiments, monosaccharide (e.g. saccharide monomer, saccharide monomer unit) refers to the monovalent free radical structure obtained by removing a hydroxyl (-OH) group from the monosaccharide, or by removing a hydrogen atom from a hydroxyl group on the monosaccharide. In embodiments, monosaccharide refers to the monovalent free radical structure obtained by removing a hydroxyl (-OH) group from the hemiacetal (−CH(OH)O−) group in the monosaccharide. In embodiments, monosaccharide refers to the monovalent free radical structure obtained by removing a hydroxyl (-OH) group or the hydrogen atom from a hydroxyl group attached to the carbon-1 (C1), carbon-2 (C2), carbon-3 (C3), carbon-4 (C4), or carbon-6 (C6) position of the monosaccharide. For example, in embodiments, monosaccharide refers to the monovalent free radical structure obtained by removing a hydroxyl (-OH) attached to the carbon- 1 (C1), carbon-2 (C2), carbon-3 (C3), carbon-4 (C-4), or carbon-6 (C6) position of the monosaccharide. In embodiments, monosaccharide refers to the monovalent free radical structure obtained by removing a hydrogen atom from a hydroxyl group attached to the carbon-1 (C1), carbon-2 (C2), carbon-3 (C3), carbon-4 (C-4), or carbon-6 (C6) position of the monosaccharide. In embodiments, monosaccharide refers to the monovalent free radical structure obtained by removing an inorganic phosphate (e.g. (PO4)3-) from the monosaccharide. For example, in embodiments, monosaccharide refers to the monovalent free radical structure following removal of an inorganic phosphate from D-glucose-1-phosphate. In embodiments, monosaccharide refers to the monovalent free radical structure following removal of an inorganic phosphate from D- galactose-1-phosphate. In embodiments, monosaccharide refers to the monovalent free radical structure following removal of an inorganic phosphate from D-mannose-1-phosphate. In embodiments, the term monosaccharide (e.g. saccharide monomer, saccharide monomer unit) is interchangeable with “glycosyl”. In embodiments, the monomer is glucosyl, wherein glucosyl is a radical derived from glucose. In embodiments, the monomer is galactosyl, wherein galactosyl is a radical derived from galactose. In embodiments, the monomer is mannosyl, wherein mannosyl is a radical derived from mannose.

[0082] The term “glycosyl” as used herein refers to the monovalent free radical structure of a monosaccharide. In embodiments, a glycosyl may be formed by removal of a hemiacetal hydroxyl group or a hydrogen atom from the hydroxyl group of a monosaccharide. In embodiments, a glycosyl is formed by removal of an inorganic phosphate group from a monosaccharide. The resultant glycosyl structure is capable of forming a covalent bond with another glycosyl structure to form, for example, a polysaccharide. In embodiments, the glycosyl is glucosyl, galactosyl, or mannosyl.

[0083] As used herein, the term “disaccharide” refers to a sugar containing two monosaccharide units joined by a glycosidic linkage. In embodiments, a glycosidic linkage may be referred to as a glycosidic bond. In embodiments, the glycosidic linkage is in α position. In embodiments, the glycosidic linkage is in β position. In embodiments, disaccharides are soluble in water. In embodiments, disaccharides include sucrose, lactose, maltose, sakebiose / nigerose, trehalose, cellobiose, laminaribiose, isomaltose, gentiobiose, allolactose, melibiose, chitobiose, mannobiose, xyloobiose, sophorose, trehalulose, turanose, leucrose, isomaltuose, lactulose, rutinose, rutinulose, 1, 3- ^-galactosyl-glucose, 1,4- ^-mannosyl-glucose, and 1,3- ^-mannosyl- glucose. In embodiments, the disaccharide is sucrose. In embodiments, the disaccharide is lactose. In embodiments, the disaccharide is maltose. In embodiments, the disaccharide is sakebiose / nigerose. In embodiments, the disaccharide is trehalose. In embodiments, the disaccharide is cellobiose. In embodiments, the disaccharide is laminaribiose. In embodiments, the disaccharide is isomaltose. In embodiments, the disaccharide is gentiobiose. In embodiments, the disaccharide is allolactose. In embodiments, the disaccharide is melibiose. In embodiments, the disaccharide is chitobiose. In embodiments, the disaccharide is mannobiose. In embodiments, the disaccharide is xyloobiose. In embodiments, the disaccharide is sophorose. In embodiments, the disaccharide is trehalulose. In embodiments, the disaccharide is turanose. In embodiments, the disaccharide is leucrose. In embodiments, the disaccharide is isomaltuose. In embodiments, the disaccharide is lactulose. In embodiments, the disaccharide is rutinose. In embodiments, the disaccharide is rutinulose. In embodiments, the disaccharide is 1, 3- ^-galactosyl-glucose. In embodiments, the disaccharide is 1,4- ^-mannosyl-glucose. In embodiments, the disaccharide is 1,3- ^-mannosyl-glucose.

[0084] As used herein, the term “isomaltose” refers to a disaccharide including two molecules of glucose bounded via an ^-1,6-glycosidic linkage. In embodiments, the structure of isomaltose is as shown below:

[0085] As used herein, the term “gentiobiose” refers to a disaccharide including two molecules of glucose bounded via an α-1,6-glycosidic linkage. In embodiments, the structure of gentiobiose is as shown below:

[0086] As used herein, the term “1,3- ^-galactosyl-glucose” refers to a disaccharide including one molecule of galactose, and one molecule of glucose bounded via an ^-1,3-glycosidic linkage. In embodiments, the structure of 1,3- ^-galactosyl-glucose is as shown below:

[0087] As used herein, the term “allolactose” refers to a disaccharide including two molecules of glucose bounded via a ^-1,6-glycosidic linkage. In embodiments, the structure of allolactose is as shown below:

[0088] As used herein, the term “melibiose” refers to a disaccharide including one molecule of galactose, and one molecule of glucose bounded via an ^-1,6-glycosidic linkage. In embodiments, the structure of melibiose is as shown below:

[0089] As used herein, the term “1,4- ^-mannosyl-glucose” refers to a disaccharide including one molecule of mannose, and one molecule of glucose bounded via an ^-1,4-glycosidic linkage. In embodiments, the structure of 1,4- ^-mannosyl-glucose is as shown below:

[0090] As used herein, the term “1,3- ^-mannosyl-glucose” refers to a disaccharide including one molecule of mannose, and one molecule of glucose bounded via an ^-1,3-glycosidic linkage. In embodiments, the structure of 1,3- ^-mannosyl-glucose is as shown below:

[0091] As used herein,a disaccharide including two molecules of glucose bound via α-1,3-glycosidic linkage. In embodiments, the structure of sakebiose is as shown below:

[0092] Inof nigeran. In embodiments, nigeran is found in black mold. In embodiments, nigeran is found in rice mold. In embodiments, nigeran is found in L. mesenteroides.

[0093] As used herein, the term “lactose” refers to a disaccharide including one molecule of glucose and one molecule of galactose bounded via a β-1,4 glycosydic linkage. In embodiments, the structure of lactose is as shown below:

[0094] As used herein, the term “maltose” refers to a disaccharide including two molecules of glucose bounded via an α-1,4 linkage. In embodiments, the structure of maltose is as shown below:

[0095] As used herein, the term “trehalose” refers to a disaccharide including two molecules of glucose bounded via an α –1,1 linkage different from the one found in maltose. In embodiments, the structure of trehalose is as shown below:

[0096] As used herein, the term “cellobiose” refers to a disaccharide including two molecules of β-glucose bounded via a β –1,4 glycosidic linkage. In embodiments, the structure of cellobiose is as shown below:

[0097] As used her e including two molecules of D-glucose bounded via β-1,3 glycosidic linkage. In embodiments, the structure of laminaribiose is as shown below:

[0098] As used herein, the terms “maltotriose,” “maltotetraose”, “maltopentose”, and “maltohexaose” refer to oligosaccharides including 3, 4, 5, and 6 molecules (respectively) of D- glucose bounded via ^-1,4 glycosidic linkages. In embodiments, the structures are as shown below:

[0099] As used herein, the terms “cellotriose,” “cellotetraose”, “cellopentose”, and “cellohexaose” refer to oligosaccharides including 3, 4, 5, and 6 molecules (respectively) of D- glucose bounded via ^-1,4 glycosidic linkages. In embodiments, the structures of cellotriose, cellotetraose, cellopentose, and cellohexaose are as shown below:

[0100] As used herein, the term “α-glucan” refers to a polysaccharide including D-glucose monomers linked by α- glycosidic bonds. In embodiments, α-glucan is found in bacteria. In embodiments, α-glucan is found in yeast. In embodiments, α-glucan is found in plants.

[0101] As used herein, the term “β-glucan” refers to a polysaccharide including D-glucose monomers linked by β- glycosidic bonds. In embodiments, β-glucan is found in bacteria. In embodiments, β-glucan is found in yeast. In embodiments, β-glucan is found in mushrooms. In embodiments, β-glucan is found in plants.

[0102] As used herein, the term “glycosidic bond” or “glycosidic linkage” refers to a covalent bond joining a monosaccharide or polysaccharide to another molecule. In embodiments, a glycosidic linkage is a covalent bond joining two monosaccharide units. In embodiments, glycosidic bonds may be formed between a plurality of monosaccharide units to form a polysaccharide chain. In embodiments, monosaccharides, disaccharides, oligosaccharides and polysaccharides form O-glycosidic bonds with alcohols to produce sugar acetals. In embodiments, monosaccharides, disaccharides, oligosaccharides and polysaccharides form N- glycosidic bonds with amines to produce sugar nucleosides. In embodiments, an ^-type or ^-type glycosidic bond is formed between the carbon-1 (C1) of a first monosaccharide and a carbon of a second monosaccharide. For example, in embodiments, the glycosidic bond may be between the carbon-1 of a first monosaccharide and the carbon-1, carbon-3, carbon-4, or carbon-6 of a second monosaccharide. In embodiments, the 1,1-glycosidic bond is a 1,1 α-glycosidic bond. In embodiments, the 1,4-glycosidic bond is a 1,4 α-glycosidic bond. In embodiments, the 1,4- glycosidic bond is a 1,4 β-glycosidic bond. In embodiments, the 1,3-glycosidic bond is a 1,3 α- glycosidic bond. In embodiments, the 1,3-glycosidic bond is a 1,3 β-glycosidic bond. In embodiments, the 1,6-glycosidic bond is a 1,6 α-glycosidic bond. In embodiments, the 1,6- glycosidic bond is a 1,6 β-glycosidic bond.

[0103] As used herein, the terms “[18F]fluoro-D-glucose” or “[18F]FDG”, also referred to as “[18F]Fluorodeoxyglucose”, “2-deoxy-2-[18F]fluoroglucose” or “2-[18F]fluoro-2-deoxy-D- glucose” refers to a positron-emitting radioisotope. [18F]FDG has with a half-life (t1 / 2) of approximately 109.8 minutes and, in embodiments, may be used in imaging methods, including positron emission tomography (PET). In embodiments, [18F]FDG is accumulated in certain tissues allowing imaging of the tissue. For example, in embodiments, [18F]FDG uptake is indicative of glucose uptake. In embodiments, [18F]FDG uptake is indicative of metabolism. In embodiments, the structure of [18F]fluoro-D-glucose ([18F]FDG) is as shown below:F]FDG is used as radiotracer in positron emission tomography (PET).

[0105] As used herein, the term “phosphorylase” refers to an enzyme that is capable of catalyzing the addition of a phosphate group (e.g. (PO4)3-) to an acceptor or the reverse reaction where a phosphate group is released from a phosphorylated molecule. In embodiments, the addition of the phosphate group to the acceptor is referred to as phosphorolysis. For example, a phosphorylase may catalyze the addition of a phosphate group to a glucose molecule. For example, a phosphorylase may catalyze attack by an inorganic phosphate group on the terminal glycosyl residue at the nonreducing end of a glycogen molecule, thereby breaking a glycosidic bond and producing a molecule of glucose 1-phosphate. In embodiments, the removal of a phosphate group from a phosphorylated molecule is referred to as reverse phosphorolysis. In embodiments, an example of reverse phosphorylysis is shown in FIG.2B. In embodiments, a phosphorylase is capable of catalyzing reverse phosphorolysis, wherein a phosphate group is released from a phosphorylated molecule. In embodiments, a phosphorylase enzyme catalyzes reverse phosphorolysis, wherein a phosphate group is released from a phosphorylated monosaccharide. Thus, in embodiments, the phosphorylase catalyzes the release of a phosphate group from a monosaccharide. For example, in embodiments, a phosphorylase enzyme catalyzes reverse phosphorolysis, wherein a phosphate group is released from a phosphorylated monosaccharide, thereby allowing formation of an O-glycosidic bond between the dephosphorylated monosaccharide and a labeled monosaccharide (e.g. ([18F]FDG), thereby forming a labeled oligosaccharide.

[0106] In embodiments, phosphorylases include allosteric enzymes that are capable of catalyzing the reversible reaction of an inorganic phosphate and a glucan, including starch, glycogen, or maltodextrin to produce glucose-1-phosphate, galactose-1-phosphate or maltose-1- phosphate. In embodiments, the phosphorylase is a glycosyltransferase. In embodiments, theglycosyltransferase catalyzes the breakdown of glycogen into glucose-1-phosphate. In embodiments, the glycosyltransferase is purine nucleoside phosphorylase. In embodiments, the phosphorylase is a nucleotidyltransferase. In embodiments, a nucleotidyltransferase breaks a phosphodiester bond. In embodiments, the nucleotidyltransferase is a tRNA nucleotidyltransferase (RNase PH. In embodiments, the nucleotidyltransferase polynucleotide phosphorylase (PNPase).

[0107] The term "maltose phosphorylase " as used herein includes any of the recombinant or naturally-occurring forms of maltose phosphorylase, also known as maltose:phosphate 1-beta-D- glucosyltransferase, or variants or homologs thereof that maintain maltose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to maltose phosphorylase). In embodiments, the term “maltose phosphorylase” refers to an enzyme that is capable of catalyzing the reversible conversion of maltose and inorganic phosphate into beta-D-glucose-1-phosphate and glucose. In embodiments, the conversion does not require any cofactors, such as pyridoxal phosphate. In embodiments, the maltose phosphorylase catalyzes the removal of the inorganic phosphate group from β-D-glucose-1-phosphate to produce maltose and an inorganic phosphate. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring maltose phosphorylase protein. In embodiments, the maltose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number Q7SIE1 or a variant or homolog having substantial identity thereto.

[0108] The term "sakebiose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of maltose phosphorylase, also known as nigerose phosphorylase, 3-O- alpha-D-glucopyranosyl-D-glucopyranose:phosphate beta-D-glucosyltransferase, or variants or homologs thereof that maintain sakebiose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to sakebiose phosphorylase). In embodiments, the term “sakebiose phosphorylase” or “nigerose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of sakebiose and inorganic phosphate into beta- D-glucose-1-phosphate and glucose. In embodiments, sakebiose phosphorylase catalyzes the removal of the inorganic phosphate from β-D-glucose-1-phosphate to produce sakebiose and an inorganic phosphate group. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to anaturally occurring sakebiose phosphorylase protein. In embodiments, the sakebiose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number A9KT32 or a variant or homolog having substantial identity thereto.

[0109] The term "trehalose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of trehalose phosphorylase, or variants or homologs thereof that maintain trehalose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to trehalose phosphorylase). In embodiments, the term “trehalose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of trehalose and inorganic phosphate into β-D-glucose-1-phosphate and glucose. In embodiments, trehalose phosphorylase catalyzes the catalyzes the removal of the inorganic phosphate from β- D-glucose-1-phosphate to produce trehalose and an inorganic phosphate group. In embodiments, the term “trehalose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of trehalose and inorganic phosphate into α-D-glucose-1-phosphate and glucose. In embodiments, trehalose phosphorylase catalyzes the catalyzes the removal of the inorganic phosphate from α-D-glucose-1-phosphate to produce trehalose and an inorganic phosphate group. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring trehalose phosphorylase protein. In embodiments, the trehalose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number O75003 or a variant or homolog having substantial identity thereto.

[0110] The term "cellobiose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of cellobiose phosphorylase, also known as cellobiose:phosphate alpha-D-glucosyltransferase, or variants or homologs thereof that maintain cellobiose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to cellobiose phosphorylase). In embodiments, the term “cellobiose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of cellobiose and inorganic phosphate into α-D-glucose-1-phosphate and glucose. In embodiments, cellobiose phosphorylase catalyzes the catalyzes the removal of the inorganic phosphate from α-D-glucose- 1-phosphate to produce cellobiose and an inorganic phosphate group. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring cellobiose phosphorylaseprotein. In embodiments, the cellobiose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number B9K7M or a variant or homolog having substantial identity thereto.

[0111] The term "laminaribiose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of laminaribiose phosphorylase, also known as 3-beta-D-glucosyl- D-glucose:phosphate alpha-D-glucosyltransferase, or variants or homologs thereof that maintain laminaribiose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to laminaribiose phosphorylase). In embodiments, the term “laminaribiose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 3- β-D-glucosyl-D-glucose and inorganic phosphate into α-D-glucose-1-phosphate and glucose. In embodiments, laminaribiose phosphorylase catalyzes the removal of inorganic phosphate from α- D-glucose-1-phosphate to produce 3-β-D-glucosyl-D-glucose. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring laminaribiose phosphorylase protein. In embodiments, the laminaribiose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number D7UT17 or a variant or homolog having substantial identity thereto.

[0112] The term "isomaltose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms isomaltose phosphorylase, or variants or homologs thereof that maintain isomaltose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to isomaltose phosphorylase). In embodiments, the term “isomaltose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 6- ^-D-glucosyl-D-glucose and inorganic phosphate into β-D-glucose-1-phosphate and glucose. In embodiments, isomaltose phosphorylase catalyzes the removal of inorganic phosphate from β- D-glucose-1-phosphate to produce 6- ^-D-glucosyl-D-glucose.

[0113] The term "gentiobiose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms gentiobiose phosphorylase, or variants or homologs thereof that maintain gentiobiose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to gentiobiose phosphorylase). In embodiments, the term “gentiobiose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 6- ^-D-glucosyl-D-glucose and inorganic phosphate into α-D-glucose-1-phosphate and glucose.In embodiments, gentiobiose phosphorylase catalyzes the removal of inorganic phosphate group from α-D-glucose-1-phosphate to produce 6- ^-D-glucosyl-D-glucose.

[0114] The term "lactose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms lactose phosphorylase, or variants or homologs thereof that maintain lactose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to lactose phosphorylase). In embodiments, the term “lactose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 4- ^-D-galactosyl- D-glucose and inorganic phosphate into α-D-galactose-1-phosphate and glucose. In embodiments, lactose phosphorylase catalyzes the removal of an inorganic phosphate group from α-D-galactose-1-phosphate to produce 4- ^-D-galactosyl-D-glucose.

[0115] The term "1,3- ^-galactosyl-glucose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms lactose phosphorylase, or variants or homologs thereof that maintain 1,3- ^-galactosyl-glucose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to 1,3- ^-galactosyl-glucose phosphorylase). In embodiments, the term “1,3- ^-galactosyl-glucose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 3- ^-D-galactosyl-D-glucose and inorganic phosphate into α-D-galactose-1-phosphate and glucose. In embodiments, 1,3- ^-galactosyl- glucose phosphorylase catalyzes the removal of an inorganic phosphate from α-D-galactose-1- phosphate to produce 3- ^-D-galactosyl-D-glucose.

[0116] The term "1,6- ^-galactosyl-glucose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms lactose phosphorylase, or variants or homologs thereof that maintain 1,6- ^-galactosyl-glucose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to 1,6- ^-galactosyl-glucose phosphorylase). In embodiments, the term “1,6- ^-galactosyl-glucose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of 6- ^-D-galactosyl-D-glucose (allolactose) and inorganic phosphate into α-D-galactose-1-phosphate and glucose. In embodiments, 1,6- ^- galactosyl-glucose phosphorylase catalyzes the removal of inorganic phosphate from α-D- galactose-1-phosphate to produce 6- ^-D-galactosyl-D-glucose.

[0117] The term "1,6- ^-galactosyl-glucose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms 1,6- ^-galactosyl-glucose phosphorylase, or variants or homologs thereof that maintain 1,6- ^-galactosyl-glucose phosphorylase activity (e.g. within atleast 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to 1,6- ^- galactosyl-glucose phosphorylase). In embodiments, the term “1,6- ^-galactosyl-glucose phosphorylase” refers to an enzyme that catalyzes the conversion of 6- ^-D-glucosyl-D-glucose (melibiose) and inorganic phosphate into β- D-galactose-1-phosphate and glucose. In embodiments, 1,6- ^-galactosyl-glucose phosphorylase catalyzes the removal of inorganic phosphate from β- D-galactose-1-phosphate to produce 6- ^-D-glucosyl-D-glucose.

[0118] The term "1,4- ^-mannosyl-glucose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of 1,4- ^-mannosyl-glucose phosphorylase, also known as mannosylglucose phosphorylase, 4-O-beta-D-mannopyranosyl-D-glucopyranose:phosphate alpha-D-mannosyltransferase, or variants or homologs thereof that maintain 1,4- ^-mannosyl- glucose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to 1,4- ^-mannosyl-glucose phosphorylase). In embodiments, the term “1,4- ^-mannosyl-glucose phosphorylase” refers to the enzyme that catalyzes the reversible reaction of 4-O-beta-D-mannopyranosyl-D-glucopyranose and an inorganic phosphate to produce D-glucose and α-D-mannose 1-phosphate. In embodiments, 1,4- ^-mannosyl-glucose phosphorylase catalyzes the removal of a phosphate group from α-D-mannose-1-phosphate in the presence of D-glucose to produce 4-O-beta-D-mannopyranosyl-D-glucopyranose. In some aspects, the variants or homologs have at least 90%, 95%, 96%, 97%, 98%, 99% or 100% amino acid sequence identity across the whole sequence or a portion of the sequence (e.g. a 50, 100, 150 or 200 continuous amino acid portion) compared to a naturally occurring 1,4- ^-mannosyl- glucose phosphorylase protein. In embodiments, the 1,4- ^-mannosyl-glucose phosphorylase protein is substantially identical to the protein identified by the UniProt reference number E6UIS7 or a variant or homolog having substantial identity thereto.

[0119] The term "1,3- ^-mannosyl-glucose phosphorylase" as used herein includes any of the recombinant or naturally-occurring forms of 1,3- ^-mannosyl-glucose phosphorylase, also known as β-1,3-oligoglucan:orthophosphate glucosyltransferase II, β-1,3-oligoglucan phosphorylase, or variants or homologs thereof that maintain 1,3- ^-mannosyl-glucose phosphorylase activity (e.g. within at least 50%, 80%, 90%, 95%, 96%, 97%, 98%, 99% or 100% activity compared to 1,3- ^- mannosyl-glucose phosphorylase). In embodiments, the term “1,3- ^-mannosyl-glucose phosphorylase” refers to an enzyme that catalyzes the reversible conversion of a (1,3-β-D- glucosyl)n and inorganic phosphate group to α-D-glucose 1-phosphate and (1,3-β-D-glucosyl)n-1. In embodiments, the 1,3- ^-mannosyl-glucose phosphorylase catalyzes the removal of a phosphate from α-D-glucose 1-phosphate generate an polysaccharide having the formula (1,3-β- D-glucosyl)n.

[0120] As used herein, the term “α-glucan phosphorylase” refers to an enzyme that catalyzes the reversible cleavage of α-1-4-linked glucose polymers, such as glycogen, starch, and maltodextrins, into α-D-glucose-1-phosphate. In embodiments, a α-glucan phosphorylase catalyzes the removal of an inorganic phosphate from α-D-glucose-1-phosphate to produce a polysaccharide including α-1-4- linked glucose monomer units. In embodiments, the polysaccharide is a branched polysaccharide. In embodiments, the polysaccharide is a linear polysaccharide. In embodiments, α-glucan phosphorylases are found in bacteria. In embodiments, α-glucan phosphorylases are found in yeast. In embodiments, α-glucan phosphorylases are found in plants. In embodiments, α-glucan phosphorylases are found in vertebrates.

[0121] As used herein, the term “β-glucan phosphorylase” refers to an enzyme that catalyzes the reversible cleavage of β-1-4-linked glucose polymers, such as cellulose, into β -D-glucose-1- phosphate. In embodiments, a β -glucan phosphorylase catalyzes the removal of an inorganic phosphate from β -D-glucose-1-phosphate to produce a polysaccharide including β -1-4- linked glucose monomer units. In embodiments, β -glucan phosphorylases are found in bacteria. In embodiments, β -glucan phosphorylases are found in yeast. In embodiments, β -glucan phosphorylases are found in plants. In embodiments, β -glucan phosphorylases are found in vertebrates.

[0122] As used herein, the term “D-glucose-1-phosphate” refers to a glucose molecule containing a phosphate group on the first carbon (e.g. carbon-1, C1). In embodiments, D- glucose-1-phosphate is produced by the phosphorylase-catalyzed breakdown of glycogen. In embodiments, the D-glucose-1-phosphate is α- D-glucose-1-phosphate. In embodiments, the structure of α- D-glucose-1-phosphate is as shown below:In embodiments, the D-glucose-1-phosphate is β- D-glucose-1-phosphate. In embodiments, the structure of β- D-glucose-1-phosphate is as shown below:galactose-1-phosphate” refers to a galactose molecule containing a phosphate group on the first carbon. In embodiments, the D-galactose-1-phosphate is α- D-galactose-1-phosphate. In embodiments, the structure of α- D-galactose-1-phosphate is as shown below:1-phosphate is β- D-galactose-1-phosphate. In embodiments, the structure of β- D-galactose-1-phosphate is as shown below:

[0124] As used herein, the term “D-mannose-1-phosphate” refers to a mannose molecule containing a phosphate group on the first carbon. In embodiments, the D-mannose-1-phosphate is α- D-mannose-1-phosphate. In embodiments, the structure of α- D-mannose-1-phosphate is as shown below:In embodiments, the D-mannose-1-phosphate is β - D-mannose-1-phosphate. In embodiments, the structure of β- D-mannose-1-phosphate is as shown below:used in accordance with its plain and ordinary meaning and refers to an identification or likelihood of the presence of a disease (e.g., cancer, a microbe infection) or outcome in a subject. In embodiments, diagnosis refers to identification or likelihood of the presence of a disease caused by infection by a microbe. In embodiments, diagnosis includes detection of the [18F]-labeled oligosaccharide provided herein including embodiments thereof in a subject. In embodiments, diagnosis includes detection of the [18F]- labeled oligosaccharide provided herein including embodiments thereof in a biological sample obtained from the subject.

[0126] The terms “treating” or “treatment” are used in accordance with their plain and ordinary meaning and broadly includes any approach for obtaining beneficial or desired results in a subject’s condition, including clinical results. Beneficial or desired clinical results can include, but are not limited to, alleviation or amelioration of one or more symptoms or conditions, diminishment of the extent of a disease, stabilizing (i.e., not worsening) the state of disease, prevention of a disease’s transmission or spread, delay or slowing of disease progression, amelioration or palliation of the disease state, diminishment of the reoccurrence of disease, and remission, whether partial or total and whether detectable or undetectable. In other words, “treatment” as used herein includes any cure, amelioration, or prevention of a disease. Treatment may prevent the disease from occurring; inhibit the disease’s spread; relieve the disease’s symptoms, fully or partially remove the disease’s underlying cause, shorten a disease’s duration, or do a combination of these things.

[0127] The terms “treating” and “treatment” may include prophylactic treatment. Treatment methods include administering to a subject a therapeutically effective amount of an active agent. The administering step may consist of a single administration or may include a series of administrations. The length of the treatment period depends on a variety of factors, such as the severity of the risk or condition, the age of the patient, the concentration of active agent, the activity of the compositions used in the treatment, or a combination thereof. It will also beappreciated that the effective dosage of an agent used for the treatment or prophylaxis may increase or decrease over the course of a particular treatment or prophylaxis regime. Changes in dosage may result and become apparent by standard diagnostic assays known in the art. In embodiments, chronic administration is required. For example, the therapeutic agents are administered to the subject in an amount and for a duration sufficient to treat the patient.

[0128] The term “prevent” is used in accordance with its plain and ordinary meaning and refers to a decrease in the occurrence of disease symptoms in a patient. The prevention may be complete (no detectable symptoms) or partial, such that fewer symptoms are observed than would likely occur absent treatment.

[0129] An “effective amount” is an amount sufficient to accomplish a stated purpose (e.g. achieve the effect for which it is administered, treat a disease). An example of an “effective amount” is an amount sufficient to contribute to the treatment, prevention, or reduction of a symptom or symptoms of a disease, which could also be referred to as a “therapeutically effective amount.” A “reduction” of a symptom or symptoms (and grammatical equivalents of this phrase) means decreasing of the severity or frequency of the symptom(s), or elimination of the symptom(s). A “prophylactically effective amount” of a drug is an amount of a drug that, when administered to a subject, will have the intended prophylactic effect, e.g., preventing or delaying the onset (or reoccurrence) of a disease, pathology or condition, or reducing the likelihood of the onset (or reoccurrence) of a disease, pathology, or condition, or their symptoms. The full prophylactic effect does not necessarily occur by administration of one dose, and may occur only after administration of a series of doses. Thus, a prophylactically effective amount may be administered in one or more administrations. The exact amounts will depend on the purpose of the treatment, and will be ascertainable by one skilled in the art using known techniques.

[0130] For any compound described herein, the therapeutically effective amount can be initially determined from cell culture assays. Target concentrations will be those concentrations of active compound(s) that are capable of achieving the methods described herein, as measured using the methods described herein or known in the art.

[0131] As is well known in the art, therapeutically effective amounts for use in humans can also be determined from animal models. For example, a dose for humans can be formulated to achieve a concentration that has been found to be effective in animals. The dosage in humans can be adjusted by monitoring compounds effectiveness and adjusting the dosage upwards ordownwards, as described above. Adjusting the dose to achieve maximal efficacy in humans based on the methods described above and other methods is well within the capabilities of the ordinarily skilled artisan.

[0132] Dosages may be varied depending upon the requirements of the patient and the compound being employed. The dose administered to a patient, in the context of the present disclosure, should be sufficient to effect a beneficial therapeutic response in the patient over time. The size of the dose also will be determined by the existence, nature, and extent of any adverse side-effects. Determination of the proper dosage for a particular situation is within the skill of the practitioner. Generally, treatment is initiated with smaller dosages which are less than the optimum dose of the compound. Thereafter, the dosage is increased by small increments until the optimum effect under circumstances is reached. Dosage amounts and intervals can be adjusted individually to provide levels of the administered compound effective for the particular clinical indication being treated. This will provide a therapeutic regimen that is commensurate with the severity of the individual’s disease state.

[0133] As used herein, the term “administering” means oral administration, administration as a suppository, topical contact, intravenous, parenteral, intraperitoneal, intramuscular, intralesional, intrathecal, intranasal or subcutaneous administration, or the implantation of a slow-release device, e.g., a mini-osmotic pump, to a subject. Administration is by any route, including parenteral and transmucosal (e.g., buccal, sublingual, palatal, gingival, nasal, vaginal, rectal, or transdermal). Parenteral administration includes, e.g., intravenous, intramuscular, intra-arteriole, intradermal, subcutaneous, intraperitoneal, intraventricular, and intracranial. Other modes of delivery include, but are not limited to, the use of liposomal formulations, intravenous infusion, transdermal patches, etc. In embodiments, administering includes injection of a solution including the [18F]-labeled oligosaccharide provided herein including embodiments thereof to a subject. In embodiments, the administering does not include administration of any active agent other than the recited active agent.

[0134] “Biological sample” or “sample” refer to materials obtained from or derived from a subject or patient. A biological sample includes sections of tissues such as biopsy and autopsy samples, and frozen sections taken for histological purposes. Such samples include bodily fluids such as blood and blood fractions or products (e.g., serum, plasma, platelets, red blood cells, and the like), sputum, tissue, cultured cells (e.g., primary cultures, explants, and transformed cells), stool, urine, synovial fluid, joint tissue, synovial tissue, immune cells, hematopoietic cells,fibroblasts, macrophages, T cells, etc. A biological sample is typically obtained from a eukaryotic organism, such as a mammal such as a primate e.g., chimpanzee or human; cow; dog; cat; a rodent, e.g., guinea pig, rat, mouse; rabbit; or a bird; reptile; or fish.

[0135] The terms “fine-needle aspiration” refer to diagnostic procedure used to investigate lumps or masses. In this procedure a thin, hollow needle and a syringe are used to extract cells, fluid or tissue from a suspicious lump or other abnormal area of the body. The material is then examined under a microscope or tested in the laboratory to determine the cause of the abnormality. The sampling and biopsy considered together are called fine-needle aspiration biopsy or fine-needle aspiration cytology (the latter to emphasize that any aspiration biopsy involves cytopathology, not histopathology).

[0136] The term “biopsy” refers to a medical test which involves extraction of sample cells or tissues for examination to determine the presence or extent of a disease in a subject. The extracted tissue is generally examined under a microscope by a pathologist, and it may also be analyzed chemically. When an entire lump or suspicious area is removed, the procedure is called an excisional biopsy. An incisional biopsy or core biopsy samples a portion of the abnormal tissue without attempting to remove the entire lesion or tumor. When a sample of tissue or fluid is removed with a needle in such a way that cells are removed without preserving the histological architecture of the tissue cells, the procedure is called a needle aspiration biopsy. The terms “biopsy material” refer to the sample extracted from the subject. The terms “tissue biopsy” refer to the extraction of tissue from a subject.

[0137] As used herein, the term “tissue sample” is used in accordance with its plain ordinary meaning and refers to a piece of tissue removed from an organism for examination, analysis, or propagation.

[0138] The terms “DNA test” or “genetic test” refer to test of DNA material obtained from a subject or sample, which is used to identify changes in DNA sequence or chromosome structure. Genetic testing can also include measuring the results of genetic changes, such as RNA analysis as an output of gene expression, or through biochemical analysis to measure specific protein output. In a medical setting, genetic testing can be used to diagnose or rule out suspected genetic disorders, predict risks for specific conditions, or gain information that can be used to customize medical treatments based on an individual’s genetic makeup. Genetic testing can also be used to determine biological relatives, such as a child’s parentage (genetic mother and father) through DNA paternity testing, or be used to broadly predict an individual’s ancestry.

[0139] The terms “blood test” refer to a laboratory analysis performed on a blood sample. Blood tests are often used in health care to determine physiological and biochemical states, such as disease, mineral content, pharmaceutical drug effectiveness, and organ function. Blood tests can involve different tests on the blood sample, such as biochemal analyses, molecular profiling, and cellular evaluation.

[0140] A “control” or “standard control” refers to a sample, measurement, or value that serves as a reference, usually a known reference, for comparison to a test sample, measurement, or value. For example, a test sample can be taken from a patient suspected of having a given disease (e.g. cancer, microbe infection) and compared to a known normal (non-diseased) individual (e.g. a standard control subject). A standard control can also represent an average measurement or value gathered from a population of similar individuals (e.g. standard control subjects) that do not have a given disease (i.e. standard control population), e.g., healthy individuals with a similar medical background, same age, weight, etc. A standard control value can also be obtained from the same individual, e.g. from an earlier-obtained sample from the patient prior to disease onset. For example, a control can be devised to compare therapeutic benefit based on pharmacological data (e.g., half-life) or therapeutic measures (e.g., comparison of side effects). Controls are also valuable for determining the significance of data. For example, if values for a given parameter are widely variant in controls, variation in test samples will not be considered as significant. One of skill will recognize that standard controls can be designed for assessment of any number of parameters (e.g. uptake of the [18F]-labeled oligosaccharide provided herein, etc).

[0141] One of skill in the art will understand which standard controls are most appropriate in a given situation and be able to analyze data based on comparisons to standard control values. Standard controls are also valuable for determining the significance (e.g. statistical significance) of data. For example, if values for a given parameter are widely variant in standard controls, variation in test samples will not be considered as significant.

[0142] The terms “patient” or “subject” are used in accordance with its plain and ordinary meaning and refer to a living organism suffering from or prone to a disease that can be treated by administration of a pharmaceutical composition, such as anti-cancer agents and chemotherapeutic agents. Non-limiting examples include humans, other mammals, bovines, rats, mice, dogs, cats, monkeys, and other non-mammalian animals. In embodiments, a patient ishuman. In embodiments, the patient is a human with an infectious disease (e.g. a disease caused by infection by a bacteria). In embodiments, the subject is a human with cancer.

[0143] A "label" or a "detectable moiety" is a composition detectable by spectroscopic, photochemical, biochemical, immunochemical, chemical, or other physical means. In embodiments, the label is a positron-emitting radioisotope (e.g. radionuclide). For example, the label (e.g. radioisotope) may be detected using positron emission tomography (PET). In embodiments, the label is fluorine-18 (18F). The label may be covalently attached to a monosaccharide as provided herein including embodiments thereof. For example, the label may be part of [18F]FDG molecule. In embodiments, the label (e.g. fluorine-18 (18F)) is substituted for the hydroxy group at the C-2 position of a glucose molecule (e.g.2-deoxy-2-[18F]fluoro-D- glucose). In embodiments, labels include 32P, fluorescent dyes, electron-dense reagents, enzymes (e.g., as commonly used in an ELISA), biotin, digoxigenin, or haptens and proteins or other entities which can be made detectable, e.g., by incorporating a radiolabel into a peptide or antibody specifically reactive with a target peptide.

[0144] The term "isolated", when applied to an oligosaccharide (e.g. [18F]-labeled oligosaccharide), denotes that the oligosaccharide is essentially free of other components with which it is associated in the chemoenzymatic reaction. The oligosaccharide may be, for example, isolated from starting materials (e.g. reactants), intermediate compounds, or other products resulting from a chemoenzymatic reaction. In embodiments, the oligosaccharide may be isolated from the solvent in which the reaction took place. Alternatively, the oligosaccharide may be the predominant component that is distributed in a solution (e.g. solvent). Thus, in embodiments, an isolated oligosaccharide may be the predominant solute species in an aqueous solution (e.g. solvent). In embodiments, the oligosaccharide is stored in a dry form. In embodiments, an isolated oligosaccharide is the predominant species present in a preparation is substantially purified. In embodiments, purity and homogeneity may be using analytical chemistry techniques such mass spectrometry, nuclear magnetic resonance, and / or high performance liquid chromatography.

[0145] The terms “computed tomography scan” or “CT scan” refer to a medical imaging technique that uses computer-processed combinations of multiple X-ray measurements taken from different angles to produce tomographic (cross-sectional) images (virtual “slices”) of a body, allowing the user to see inside the body without cutting. CT imaging is an extremely valuable tool for the evaluation of pancreatic lesions. The CT can provide the precise location ofthe lesion, size, and, most importantly, any involvement of any surrounding structures. CT imaging can really help determine the resectability of the tumor. A pancreatic CT protocol involves triphasic cross-sectional imaging with arterial, late arterial, and venous phases. This type of imaging allows identification of the majority of pancreatic cancers in the delayed arterial phase / venous phase, as many are hypovascular in nature. The arterial phase can identify many other pancreatic lesions, as well. CT imaging has a reported sensitivity ranging from 76-92% for diagnosing pancreatic cancer and surgical resectability up to 90%.

[0146] The term “X-ray” or “X-radiation” refers to a penetrating form of high-energy electromagnetic radiation. Most X-rays have a wavelength ranging from 10 picometers to 10 nanometers, corresponding to frequencies in the range 30 petahertz to 30 exahertz (30×1015Hz to 30×1018 Hz) and energies in the range 124 eV to 124 keV. X-ray wavelengths are shorter than those of UV rays and typically longer than those of gamma rays.

[0147] The terms “PET”, “PET scan”, “positron emission tomography”, or “positron emission tomography scan” refer to a functional imaging technique that uses radioactive substances known as radiotracers to visualize and measure changes in metabolic processes, and in other physiological activities including blood flow, regional chemical composition, and absorption. Different tracers are used for various imaging purposes, depending on the target process within the body. PET scan is a common imaging technique, a medical scintillography technique used in nuclear medicine. A radiopharmaceutical – a radioisotope attached to a drug is injected into the body as a tracer. Gamma rays are emitted and detected by gamma cameras to form a three- dimensional image, in a similar way that an X-ray image is captured. PET scans have conflicting evidence for the regular use in diagnosing and staging of pancreatic disease. The sensitivity depends on the lesion’s size, so it is susceptible to miss small areas of disease. In embodiments, PET scans include micro PET-CT / MRI scans for imaging animals. In embodiments, PET scans include PET-CT / MRI scans for imaging humans. Elevated serum glucose levels can cause false negatives and cause missed metastatic disease. When CT or MRI and PET scans are combined, this can increase the overall accuracy while increasing the sensitivity and specificity, especially for lesions that were missed on CT or MRI imaging. Overall, combined CT and PET scans and combined MRI and PET scans are more useful in finding and confirming diseases.

[0148] The term “MRI” or “magnetic resonance imaging” refer to a medical imaging technique used in radiology to form pictures of the anatomy and the physiological processes of the body. MRI scanners use strong magnetic fields, magnetic field gradients, and radio waves togenerate images of the organs in the body. MRI does not involve X-rays or the use of ionizing radiation, which distinguishes it from CT and PET scans. MRI is a medical application of nuclear magnetic resonance (NMR) which can also be used for imaging in other NMR applications, such as NMR spectroscopy. MRI also has a role in diagnosing pancreatic lesions. MRI is an excellent modality for determining pancreaticobiliary anatomy and pathology. MRCP can identify pancreatic duct abnormalities, such as strictures. It is better overall at identifying the biliary and pancreatic ductal systems. MRI is also used for diagnosing and following lesions post-operatively as well.

[0149] The term “disease” or “condition” refers to a state of being or health status of a patient or subject that is being treated with the compounds or methods provided herein. The disease may be a cancer. The disease may be an autoimmune disease. The disease may be an inflammatory disease. The disease may be an infectious disease. In some further instances, “cancer” refers to human cancers and carcinomas, sarcomas, adenocarcinomas, lymphomas, leukemias, etc., including solid and lymphoid cancers, kidney, breast, lung, bladder, colon, ovarian, prostate, pancreas, stomach, brain, head and neck, skin, uterine, testicular, glioma, esophagus, and liver cancer, including hepatocarcinoma, lymphoma, including B-acute lymphoblastic lymphoma, non-Hodgkin’s lymphomas (e.g., Burkitt’s, Small Cell, and Large Cell lymphomas), Hodgkin’s lymphoma, leukemia (including AML, ALL, and CML), or multiple myeloma.

[0150] The term “infection” or “infectious disease” refers to a disease or condition that can be caused by infection by an organism such as a bacterium, virus, fungi or any other pathogenic microbial agents. In embodiments, the infectious disease is caused by a pathogenic bacteria. Pathogenic bacteria are bacteria which cause diseases (e.g., in humans). In embodiments, the infectious disease is a bacteria associated disease (e.g., tuberculosis, which is caused by Mycobacterium tuberculosis). Non-limiting bacteria associated diseases include pneumonia, which may be caused by bacteria such as Streptococcus and Pseudomonas; or foodborne illnesses, which can be caused by bacteria such as Shigella, Campylobacter, and Salmonella. Bacteria associated diseases also includes tetanus, typhoid fever, diphtheria, syphilis, and leprosy. In embodiments, the disease is Bacterial vaginosis (i.e. bacteria that change the vaginal microbiota caused by an overgrowth of bacteria that crowd out the Lactobacilli species that maintain healthy vaginal microbial populations) (e.g., yeast infection, or Trichomonas vaginalis); Bacterial meningitis (i.e. a bacterial inflammation of the meninges); Bacterial pneumonia (i.e. a bacterial infection of the lungs); Urinary tract infection; Bacterial gastroenteritis; or Bacterial skin infections (e.g. impetigo, or cellulitis). In embodiments, the infectious disease is aCampylobacter jejuni, Enterococcus faecalis, Haemophilus influenzae, Helicobacter pylori, Klebsiella pneumoniae, Legionella pneumophila, Neisseria gonorrhoeae, Neisseria meningitides, Staphylococcus aureus, Streptococcus pneumonia, or Vibrio cholera infection.

[0151] As used herein, the term “inflammatory disease” refers to a disease or condition characterized by aberrant inflammation (e.g. an increased level of inflammation compared to a control such as a healthy person not suffering from a disease). Examples of inflammatory diseases include traumatic brain injury, arthritis, rheumatoid arthritis, psoriatic arthritis, juvenile idiopathic arthritis, multiple sclerosis, systemic lupus erythematosus (SLE), myasthenia gravis, juvenile onset diabetes, diabetes mellitus type 1, Guillain-Barre syndrome, Hashimoto’s encephalitis, Hashimoto’s thyroiditis, ankylosing spondylitis, psoriasis, Sjogren’s syndrome,vasculitis, glomerulonephritis, auto-immune thyroiditis, Behcet’s disease, Crohn’s disease, ulcerative colitis, bullous pemphigoid, sarcoidosis, ichthyosis, Graves ophthalmopathy, inflammatory bowel disease, Addison’s disease, Vitiligo,asthma, asthma, allergic asthma, acne vulgaris, celiac disease, chronic prostatitis, inflammatory bowel disease, pelvic inflammatory disease, reperfusion injury, sarcoidosis, transplant rejection, interstitial cystitis, atherosclerosis, and atopic dermatitis.

[0152] As used herein, the term “myositis” refers to a rare disease causing inflammation in the muscles. Myositis may occur in subjects with lupus, scleroderma, and rheumatoid arthritis. Myositis is typically characterized by weak, painful, or aching muscles. In embodiments, inflammatory conditions causing myositis include dermatomyositis, polymyositis, lupus, scleroderma, rheumatoid arthritis, viral infections, bacterial infections, fungal infections.

[0153] As used herein, the term “discitis” refers to an infection of a disc between the vertebrae and the spine. In embodiments, the destruction of the disc along with the inflammation from the infection causes severe back pain. In embodiments, discitis is caused by a bacterial infection.

[0154] As used herein, the term “vertebral osteomyelitis” refers to a rare spine infection. In embodiments, vertebral osteomyelitis is caused by Staphylococcus aureus. n embodiments, vertebral osteomyelitis is caused by Mycobacterium tuberculosis. In embodiments, vertebral osteomyelitis is caused by the spreading of discitis to an adjacent bone.

[0155] As used herein, the term “pneumonia” refers to an infection of one or both the lungs. In embodiments, pneumonia is caused by bacteria. In embodiments, pneumonia is caused by Streptococcus pneumoniae. In embodiments, pneumonia is caused by bacteria. In embodiments, pneumonia is caused by viruses. In embodiments, pneumonia is caused by fungi. Inembodiments, pneumonia is caused by Micoplasma pneumoniae. In embodiments, pneumonia is caused by chemical irritants.

[0156] As used herein, the term “microorganism” or “microbe” refers to a single-cell or a multiple cell organism of microscopic size. In embodiments, microorganisms include archaea, bacteria, protists, fungi, and embryophytes.

[0157] As used herein, the term “gram-positive bacteria” refers to bacteria that give a positive result in the Gram stain test. In embodiments, the Gram stain test is used to classify bacteria according to the type of cell wall. In embodiments, gram positive bacteria retain gram stain in the peptidoglycan layer in the cell wall. In embodiments, some gram-positive bacteria are pathogenic. In embodiments, pathogenic gram-positive bacteria include, but not limited to, Streptococcus, Staphylococcus, Corynebacterium, Listeria, Bacillus, Clostridium, Catabacter, Rathybacter, Leifsonia, and Clavibacter.

[0158] As used herein, the term “gram-negative bacteria” refers to bacteria that give a negative result in the Gram stain test. In embodiments, gram-negative bacteria cause infections including pneumonia, bloodstream infections, wound or surgical site infections, and meningitis. In embodiments, gram-negative bacteria are resistant to multiple drugs. In embodiments, gram- negative bacteria are resistant to antibiotics. In embodiments, gram-negative infections include those caused by Klebsiella, Acinetobacter, Pseudomonas aeruginosa, and E. coli.

[0159] As used herein, the term “fungal infection” refers to infections caused by fungi, yeast, or molds. In embodiments, fungal infections are localized. In embodiments, fungal infections are systemic. In embodiments, systemic fungal infections affect the lungs. In embodiments, fungal infections affect the eye. In embodiments, systemic fungal infections affect the liver. In embodiments, systemic fungal infections affect the brain. In embodiments, systemic fungal infections are opportunistic infections that take advantage of a weakened immune system. In embodiments, opportunistic systemic fungal infections include aspergillosis, candidiasis and demucormycosis. In embodiments, systemic fungal infections are primary fungal infections. In embodiments, primary fungal infections include histoplasmosis, blastomycosis, coccidioidomycosis, and paracoccidioidomycosis.

[0160] As used herein, the terms “live infection” or “living infection” (e.g. live microbe infection, or living microbe infection) refer to in vivo detection, visualization, and monitoring of pathogen infection stages during the imaging process. In embodiments, the pathogen is a microbe. In embodiments, the microbe is a bacterium, fungus, archaea, protist, or virus. Forexample, in embodiments, live infection refers to visualizing and monitoring the spread of a pathogen throughout a biological sample obtained in a subject. In embodiments, live infection refers to visualizing or monitoring the spread of a pathogen throughout the tissue or to different organs in a subject. In embodiments, the live infection includes identifying the tissue or organ where a pathogen infection has occurred. In embodiments, live infection does not include detection of a dead pathogen. In embodiments, live infection refers to detecting an absence of pathogen infection. For example, the absence of pathogen may be indicative that a treatment has the desired clinical results.

[0161] As used herein, the term “cancer” is used in accordance with its plain ordinary meaning and refers to all types of cancer, neoplasm or malignant tumors found in mammals (e.g. humans), including leukemias, lymphomas, carcinomas and sarcomas. Exemplary cancers that may be treated with a compound or method provided herein include brain cancer, glioma, glioblastoma, neuroblastoma, prostate cancer, colorectal cancer, pancreatic cancer, Medulloblastoma, melanoma, cervical cancer, gastric cancer, ovarian cancer, lung cancer, cancer of the head, Hodgkin’s Disease, and Non-Hodgkin’s Lymphomas. Exemplary cancers that may be treated with a compound or method provided herein include cancer of the thyroid, endocrine system, brain, breast, cervix, colon, head & neck, liver, kidney, lung, ovary, pancreas, rectum, stomach, and uterus. Additional examples include, thyroid carcinoma, cholangiocarcinoma, pancreatic adenocarcinoma, skin cutaneous melanoma, colon adenocarcinoma, rectum adenocarcinoma, stomach adenocarcinoma, esophageal carcinoma, head and neck squamous cell carcinoma, breast invasive carcinoma, lung adenocarcinoma, lung squamous cell carcinoma, non-small cell lung carcinoma, mesothelioma, multiple myeloma, neuroblastoma, glioma, glioblastoma multiforme, ovarian cancer, rhabdomyosarcoma, primary thrombocytosis, primary macroglobulinemia, primary brain tumors, malignant pancreatic insulanoma, malignant carcinoid, urinary bladder cancer, premalignant skin lesions, testicular cancer, thyroid cancer, neuroblastoma, esophageal cancer, genitourinary tract cancer, malignant hypercalcemia, endometrial cancer, adrenal cortical cancer, neoplasms of the endocrine or exocrine pancreas, medullary thyroid cancer, medullary thyroid carcinoma, melanoma, colorectal cancer, papillary thyroid cancer, hepatocellular carcinoma, or prostate cancer.

[0162] The term “carcinoma” refers to a malignant new growth made up of epithelial cells tending to infiltrate the surrounding tissues and give rise to metastases. Exemplary carcinomas that may be treated with a compound or method provided herein include, for example, medullary thyroid carcinoma, familial medullary thyroid carcinoma, acinar carcinoma, acinous carcinoma,adenocystic carcinoma, adenoid cystic carcinoma, carcinoma adenomatosum, carcinoma of adrenal cortex, alveolar carcinoma, alveolar cell carcinoma, basal cell carcinoma, carcinoma basocellulare, basaloid carcinoma, basosquamous cell carcinoma, bronchioalveolar carcinoma, bronchiolar carcinoma, bronchogenic carcinoma, cerebriform carcinoma, cholangiocellular carcinoma, chorionic carcinoma, colloid carcinoma, comedo carcinoma, corpus carcinoma, cribriform carcinoma, carcinoma en cuirasse, carcinoma cutaneum, cylindrical carcinoma, cylindrical cell carcinoma, duct carcinoma, carcinoma durum, embryonal carcinoma, encephaloid carcinoma, epiermoid carcinoma, carcinoma epitheliale adenoides, exophytic carcinoma, carcinoma ex ulcere, carcinoma fibrosum, gelatiniforni carcinoma, gelatinous carcinoma, giant cell carcinoma, carcinoma gigantocellulare, glandular carcinoma, granulosa cell carcinoma, hair-matrix carcinoma, hematoid carcinoma, hepatocellular carcinoma, Hurthle cell carcinoma, hyaline carcinoma, hypernephroid carcinoma, infantile embryonal carcinoma, carcinoma in situ, intraepidermal carcinoma, intraepithelial carcinoma, Krompecher’s carcinoma, Kulchitzky-cell carcinoma, large-cell carcinoma, lenticular carcinoma, carcinoma lenticulare, lipomatous carcinoma, lymphoepithelial carcinoma, carcinoma medullare, medullary carcinoma, melanotic carcinoma, carcinoma molle, mucinous carcinoma, carcinoma muciparum, carcinoma mucocellulare, mucoepidermoid carcinoma, carcinoma mucosum, mucous carcinoma, carcinoma myxomatodes, nasopharyngeal carcinoma, oat cell carcinoma, carcinoma ossificans, osteoid carcinoma, papillary carcinoma, periportal carcinoma, preinvasive carcinoma, prickle cell carcinoma, pultaceous carcinoma, renal cell carcinoma of kidney, reserve cell carcinoma, carcinoma sarcomatodes, schneiderian carcinoma, scirrhous carcinoma, carcinoma scroti, signet- ring cell carcinoma, carcinoma simplex, small-cell carcinoma, solanoid carcinoma, spheroidal cell carcinoma, spindle cell carcinoma, carcinoma spongiosum, squamous carcinoma, squamous cell carcinoma, string carcinoma, carcinoma telangiectaticum, carcinoma telangiectodes, transitional cell carcinoma, carcinoma tuberosum, tuberous carcinoma, verrucous carcinoma, or carcinoma villosum.

[0163] The term “metastasis” or the plural form “metastases” refers to the development of secondary malignant growths at a distance from a primary site of cancer. The condition refers to when cancer cells break away from the main tumor and enter the bloodstream or lymphatic system. The terms “metastasis,” “metastatic,” and “metastatic cancer” can be used interchangeably and refer to the spread of a proliferative disease or disorder, e.g., cancer, from one organ or another non-adjacent organ or body part.

[0164] The term “diagnosis” refers to the identification of the nature and cause of a certain phenomenon. Diagnosis is used in many different disciplines, with variations in the use of logic, analytics, and experience, to determine “cause and effect”. In medicine, a “diagnosis”, a “diagnostic procedure” or “medical diagnosis” is the process of determining which disease or condition explains a person’s symptoms and signs. It is most often referred to as diagnosis with the medical context being implicit. The information required for diagnosis is typically collected from a history and physical examination of the person seeking medical care. Often, one or more diagnostic procedures, such as medical tests, are also done during the process. Sometimes posthumous diagnosis is considered a kind of medical diagnosis. The term “to diagnose” refers to the act of realizing a diagnosis. The terms “confirmatory diagnostic procedure” or “confirmatory diagnosis procedure” refer to a process of confirming a diagnosis.

[0165] The term “distant metastasis” or “distant metastasis tumor” refers to a cancer that has spread from the original (primary) tumor to distant organs or distant lymph nodes.

[0166] As used herein, the term “pharmaceutically acceptable” is used synonymously with “physiologically acceptable” and “pharmacologically acceptable”. A pharmaceutical composition will generally comprise agents for buffering and preservation in storage, and can include buffers and carriers for appropriate delivery, depending on the route of administration.

[0167] "Pharmaceutically acceptable excipient" and "pharmaceutically acceptable carrier" refer to a substance that aids the administration of an active agent to and absorption by a subject and can be included in the compositions of the present invention without causing a significant adverse toxicological effect on the patient. For example, in embodiments, a pharmacuetically acceptable excipeint allows intravenous delivery of the [18F]-labeled oligosaccharide provided herein including embodiments thereof. Non-limiting examples of pharmaceutically acceptable excipients include water, NaCl, normal saline solutions, lactated Ringer’s, normal sucrose, normal glucose, binders, fillers, disintegrants, lubricants, coatings, sweeteners, flavors, salt solutions (such as Ringer's solution), alcohols, oils, gelatins, carbohydrates such as lactose, amylose or starch, fatty acid esters, hydroxymethycellulose, polyvinyl pyrrolidine, and colors, and the like. Such preparations can be sterilized and, if desired, mixed with auxiliary agents such as lubricants, preservatives, stabilizers, wetting agents, emulsifiers, salts for influencing osmotic pressure, buffers, coloring, and / or aromatic substances and the like that do not deleteriously react with the compounds of the invention. One of skill in the art will recognize that other pharmaceutical excipients are useful in the present invention.

[0168] As used herein, the term “neoadjuvant therapy” is used in accordance with its plain ordinary meaning and refers to a treatment given as a first step to shrink a tumor before the main treatment, which is usually surgery, is given.

[0169] The term “radiation therapy” or “radiotherapy” refers to a therapy that uses ionizing radiation, and is generally provided as part of cancer treatment to control or kill malignant cells. Radiation therapy is normally delivered by a linear accelerator. Radiation therapy may be curative in a number of types of cancer if they are localized to one area of the body. It may also be used as part of adjuvant therapy, to prevent tumor recurrence after surgery that removes a primary malignant tumor. Radiation therapy is synergistic with chemotherapy, and has been used before, during, and after chemotherapy in susceptible cancers.

[0170] The term “chemotherapy” refers to a type of cancer treatment that uses one or more anti-cancer drugs (chemotherapeutic agents) as part of a standardized chemotherapy regimen. Chemotherapy may be given with a curative intent (which almost always involves combinations of drugs), or it may aim to prolong life or to reduce symptoms (palliative chemotherapy). Chemotherapy drugs include, but are not limited to, alkylating agents, nitrosoureas, antimetabolites, alkaloids, antitumor antibiotics, hormonal agents and biological response modifiers. Examples of chemotherapy drugs include, but are not limited to, cyclophosphamide, melphalan, temozolomide, carboplatin, cisplatin, oxaliplatin, 5-fluorouracil, 6-mercaptopurine, cytarabine, gemcitabine, methotrexate, actimycin-D, blemycin, daunorubicin, doxorubicin, docetaxel, estramustine, paclitaxel, vinblastine, etoposide, irinotecan, teniposide, topotecan, prednisone, methylprednisolone and dexamethasone. Traditional chemotherapeutic agents are cytotoxic by means of interfering with cell division (mitosis) but cancer cells vary widely in their susceptibility to these agents. Many of the side effects of chemotherapy can be traced to damage to normal cells that divide rapidly and are thus sensitive to anti-mitotic drugs such as, but not limited to, cells in the bone marrow, digestive tract and hair follicles. In embodiments, the chemotherapy includes administration of an effective amount of an anticancer agent as set forth herein.

[0171] The term “targeted therapy” refers to a method for treating cancer that blocks the growth of cancer cells by interfering with specific targeted molecules needed for carcinogenesis and tumor growth, rather than by simply interfering with all rapidly dividing cells. Exemplary forms of targeted therapy include, but are not limited to, antibody-drug conjugates, nano- engineered enzymes that bind to a tumor cell, and chemical entities that target or preferentiallytarget a protein or enzyme that carries a mutation or other genetic alteration that is specific to cancer cells and is not found in normal host tissue. Exemplary drugs that are used in targeted therapy include, but are not limited to, trastuzumab and bevacizumab. In embodiments, the targeted therapy includes administration of an effective amount of an anticancer agent as set forth herein.

[0172] The term “immunotherapy” refers to methods of treating cancer that are based on the stimulation of the patient’s immune system. Cancer immunotherapy exploits the fact that cancer cells often have tumor antigens that can be detected and bound by the antibodies of the immune system. Clinical success of cancer immunotherapy is highly variable between different forms of cancer. Examples of immunotherapy include, but are not limited to, therapeutic cancer vaccines, CAR-T cell, and targeted antibody therapies. Examples of drugs used in immunotherapy include, but are not limited to, ipilimumab, pembrolizumab, nivolumab and atezolizumab. In embodiments, the immunotherapy includes administration of an effective amount of an anticancer agent as set forth herein.

[0173] The term “hormonal therapy” refers to a type of a cancer treatment that slows or stops the growth of cancer that uses hormones to grow. Hormonal therapy may be used alone as the main treatment or with other treatments. It may be used before surgery or radiation therapy to shrink the tumor. Hormonal therapy may be given in addition to main treatments such as surgery, radiation therapy or chemotherapy to lower the risk of cancer recurrence. Hormonal therapy includes, but is not limited to, removing the gland or organ that makes the hormone, irradiating the gland or organ to destroy hormone-producing cells, and administration of drugs that suppress hormonal production. In embodiments, the hormonal therapy includes administration of an effective amount of an anticancer agent as set forth herein.

[0174] As used herein, the term “synthetic lethality” or “synthetic lethality therapy” is used in accordance with its plain ordinary meaning and refers to a combination of deficiencies in the expression of two or more genes that lead to cell death, whereas a deficiency in only one of these genes does not. The deficiencies can arise through mutations, epigenetic alterations or inhibitors of one of the genes. “Synthetic lethality therapy” has utility for purposes of molecular targeted cancer therapy, with the first example of a molecular targeted therapeutic exploiting a synthetic lethal exposed by an inactivated tumor suppressor gene (BRCA1 and 2) receiving FDA approval in 2016 (PARP inhibitor).[1] A sub-case of synthetic lethality, where vulnerabilities are exposedby the deletion of passenger genes rather than tumor suppressor is the so-called collateral lethality.

[0175] The term “angiogenesis inhibitor administration therapy” refers to methods of treating cancer that block the growth of blood vessels that support tumor growth rather than blocking the growth of tumor cells themselves. Exemplary angiogenesis inhibitors include, but are not limited to, monoclonal antibodies that specifically recognize and bind to vascular endothelial growth factor (VEGF) and thus block activation of the VEGF receptor, and immunomodulatory drugs that stimulate or suppress the immune system.

[0176] “Anti-cancer agent” and “anticancer agent” are used in accordance with their plain ordinary meaning and refers to a composition (e.g. compound, drug, antagonist, inhibitor, modulator) having antineoplastic properties or the ability to inhibit the growth or proliferation of cells. In some embodiments, an anti-cancer agent is a chemotherapeutic. In some embodiments, an anti-cancer agent is an agent identified herein having utility in methods of treating cancer. In some embodiments, an anti-cancer agent is an agent approved by the FDA or similar regulatory agency of a country other than the USA, for treating cancer. Examples of anti-cancer agents include, but are not limited to, MEK (e.g. MEK1, MEK2, or MEK1 and MEK2) inhibitors (e.g. XL518, CI-1040, PD035901, selumetinib / AZD6244, GSK1120212 / trametinib, GDC-0973, ARRY-162, ARRY-300, AZD8330, PD0325901, U0126, PD98059, TAK-733, PD318088, AS703026, BAY 869766), alkylating agents (e.g., cyclophosphamide, ifosfamide, chlorambucil, busulfan, melphalan, mechlorethamine, uramustine, thiotepa, nitrosoureas, nitrogen mustards (e.g., mechloroethamine, cyclophosphamide, chlorambucil, meiphalan), ethylenimine and methylmelamines (e.g., hexamethlymelamine, thiotepa), alkyl sulfonates (e.g., busulfan), nitrosoureas (e.g., carmustine, lomusitne, semustine, streptozocin), triazenes (decarbazine)), anti- metabolites (e.g., 5- azathioprine, leucovorin, capecitabine, fludarabine, gemcitabine, pemetrexed, raltitrexed, folic acid analog (e.g., methotrexate), or pyrimidine analogs (e.g., fluorouracil, floxouridine, Cytarabine), purine analogs (e.g., mercaptopurine, thioguanine, pentostatin), etc.), plant alkaloids (e.g., vincristine, vinblastine, vinorelbine, vindesine, podophyllotoxin, paclitaxel, docetaxel, etc.), topoisomerase inhibitors (e.g., irinotecan, topotecan, amsacrine, etoposide (VP16), etoposide phosphate, teniposide, etc.), antitumor antibiotics (e.g., doxorubicin, adriamycin, daunorubicin, epirubicin, actinomycin, bleomycin, mitomycin, mitoxantrone, plicamycin, etc.), platinum-based compounds (e.g. cisplatin, oxaloplatin, carboplatin), anthracenedione (e.g., mitoxantrone), substituted urea (e.g., hydroxyurea), methyl hydrazine derivative (e.g., procarbazine), adrenocortical suppressant (e.g.,mitotane, aminoglutethimide), epipodophyllotoxins (e.g., etoposide), antibiotics (e.g., daunorubicin, doxorubicin, bleomycin), enzymes (e.g., L-asparaginase), inhibitors of mitogen- activated protein kinase signaling (e.g. U0126, PD98059, PD184352, PD0325901, ARRY- 142886, SB239063, SP600125, BAY 43-9006, wortmannin, or LY294002, Syk inhibitors, mTOR inhibitors, antibodies (e.g., rituxan), gossyphol, genasense, polyphenol E, Chlorofusin, all trans-retinoic acid (ATRA), bryostatin, tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), 5-aza-2’-deoxycytidine, all trans retinoic acid, doxorubicin, vincristine, etoposide, gemcitabine, imatinib (Gleevec.RTM.), geldanamycin, 17-N-Allylamino-17- Demethoxygeldanamycin (17-AAG), flavopiridol, LY294002, bortezomib, trastuzumab, BAY 11-7082, PKC412, PD184352, 20-epi-1, 25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone; aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TK antagonists; altretamine; ambamustine; amidox; amifostine; aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole; andrographolide; angiogenesis inhibitors; antagonist D; antagonist G; antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen, prostatic carcinoma; antiestrogen; antineoplaston; antisense oligonucleotides; aphidicolin glycinate; apoptosis gene modulators; apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; arginine deaminase; asulacrine; atamestane; atrimustine; axinastatin 1; axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatin III derivatives; balanol; batimastat; BCR / ABL antagonists; benzochlorins; benzoylstaurosporine; beta lactam derivatives; beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor; bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistratene A; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine; calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2; capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRest M3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinase inhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins; chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine; clomifene analogues; clotrimazole; collismycin A; collismycin B; combretastatin A4; combretastatin analogue; conagenin; crambescidin 816; crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A; cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate; cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B; deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil; diaziquone; didemnin B; didox; diethylnorspermine; dihydro-5-azacytidine; 9-dioxamycin; diphenyl spiromustine; docosanol; dolasetron; doxifluridine; droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine; edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin; epristeride; estramustine analogue;estrogen agonists; estrogen antagonists; etanidazole; etoposide phosphate; exemestane; fadrozole; fazarabine; fenretinide; filgrastim; finasteride; flavopiridol; flezelastine; fluasterone; fludarabine; fluorodaunorunicin hydrochloride; forfenimex; formestane; fostriecin; fotemustine; gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix; gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam; heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid; idarubicin; idoxifene; idramantone; ilmofosine; ilomastat; imidazoacridones; imiquimod; immunostimulant peptides; insulin-like growth factor-1 receptor inhibitor; interferon agonists; interferons; interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact; irsogladine; isobengazole; isohomohalicondrin B; itasetron; jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide; leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole; leukemia inhibiting factor; leukocyte alpha interferon; leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole; linear polyamine analogue; lipophilic disaccharide peptide; lipophilic platinum compounds; lissoclinamide 7; lobaplatin; lombricine; lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine; lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides; maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysin inhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone; meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone; miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone; mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growth factor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonal antibody, human chorionic gonadotrophin; monophosphoryl lipid A+myobacterium cell wall sk; mopidamol; multiple drug resistance gene inhibitor; multiple tumor suppressor 1- based therapy; mustard anticancer agent; mycaperoxide B; mycobacterial cell wall extract; myriaporone; N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip; naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin; nemorubicin; neridronic acid; neutral endopeptidase; nilutamide; nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn; O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone; ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin; osaterone; oxaliplatin; oxaunomycin; palauamine; palmitoylrhizoxin; pamidronic acid; panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase; peldesine; pentosan polysulfate sodium; pentostatin; pentrozole; perflubron; perfosfamide; perillyl alcohol; phenazinomycin; phenylacetate; phosphatase inhibitors; picibanil; pilocarpine hydrochloride; pirarubicin; piritrexim; placetin A; placetin B; plasminogen activator inhibitor; platinum complex; platinum compounds; platinum- triamine complex; porfimer sodium; porfiromycin; prednisone; propyl bis-acridone;prostaglandin J2; proteasome inhibitors; protein A-based immune modulator; protein kinase C inhibitor; protein kinase C inhibitors, microalgal; protein tyrosine phosphatase inhibitors; purine nucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine; pyridoxylated hemoglobin polyoxyethylerie conjugate; raf antagonists; raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors; ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re 186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide; rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol; saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics; semustine; senescence derived inhibitor 1; sense oligonucleotides; signal transduction inhibitors; signal transduction modulators; single chain antigen-binding protein; sizofuran; sobuzoxane; sodium borocaptate; sodium phenylacetate; solverol; somatomedin binding protein; sonermin; sparfosic acid; spicamycin D; spiromustine; splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-cell division inhibitors; stipiamide; stromelysin inhibitors; sulfinosine; superactive vasoactive intestinal peptide antagonist; suradista; suramin; swainsonine; synthetic glycosaminoglycans; tallimustine; tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium; tegafur; tellurapyrylium; telomerase inhibitors; temoporfin; temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine; thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic; thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroid stimulating hormone; tin ethyl etiopurpurin; tirapazamine; titanocene bichloride; topsentin; toremifene; totipotent stem cell factor; translation inhibitors; tretinoin; triacetyluridine; triciribine; trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinase inhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenital sinus-derived growth inhibitory factor; urokinase receptor antagonists; vapreotide; variolin B; vector system, erythrocyte gene therapy; velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine; vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; zinostatin stimalamer, Adriamycin, Dactinomycin, Bleomycin, Vinblastine, Cisplatin, acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin; aldesleukin; altretamine; ambomycin; ametantrone acetate; aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase; asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa; bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin; bleomycin sulfate; brequinar sodium; bropirimine; busulfan; cactinomycin; calusterone; caracemide; carbetimer; carboplatin; carmustine; carubicin hydrochloride; carzelesin; cedefingol; chlorambucil; cirolemycin; cladribine; crisnatol mesylate; cyclophosphamide; cytarabine; dacarbazine; daunorubicin hydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguanine mesylate; diaziquone; doxorubicin; doxorubicinhydrochloride; droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin; edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin; enpromate; epipropidine; epirubicin hydrochloride; erbulozole; esorubicin hydrochloride; estramustine; estramustine phosphate sodium; etanidazole; etoposide; etoposide phosphate; etoprine; fadrozole hydrochloride; fazarabine; fenretinide; floxuridine; fludarabine phosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium; gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicin hydrochloride; ifosfamide; iimofosine; interleukin I1 (including recombinant interleukin II, or rlL.sub.2), interferon alfa-2a; interferon alfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-1a; interferon gamma-1b; iproplatin; irinotecan hydrochloride; lanreotide acetate; letrozole; leuprolide acetate; liarozole hydrochloride; lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol; maytansine; mechlorethamine hydrochloride; megestrol acetate; melengestrol acetate; melphalan; menogaril; mercaptopurine; methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide; mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper; mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazoie; nogalamycin; ormaplatin; oxisuran; pegaspargase; peliomycin; pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan; piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium; porfiromycin; prednimustine; procarbazine hydrochloride; puromycin; puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol; safingol hydrochloride; semustine; simtrazene; sparfosate sodium; sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin; streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium; tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone; testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin; tirapazamine; toremifene citrate; trestolone acetate; triciribine phosphate; trimetrexate; trimetrexate glucuronate; triptorelin; tubulozole hydrochloride; uracil mustard; uredepa; vapreotide; verteporfin; vinblastine sulfate; vincristine sulfate; vindesine; vindesine sulfate; vinepidine sulfate; vinglycinate sulfate; vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate; vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicin hydrochloride, agents that arrest cells in the G2-M phases and / or modulate the formation or stability of microtubules, (e.g. Taxol.TM (i.e. paclitaxel), Taxotere.TM, compounds comprising the taxane skeleton, Erbulozole (i.e. R-55104), Dolastatin 10 (i.e. DLS-10 and NSC-376128), Mivobulin isethionate (i.e. as CI-980), Vincristine, NSC-639829, Discodermolide (i.e. as NVP-XX-A-296), ABT-751 (Abbott, i.e. E-7010), Altorhyrtins (e.g. Altorhyrtin A and Altorhyrtin C), Spongistatins (e.g. Spongistatin 1, Spongistatin 2, Spongistatin 3, Spongistatin 4, Spongistatin 5,Spongistatin 6, Spongistatin 7, Spongistatin 8, and Spongistatin 9), Cemadotin hydrochloride (i.e. LU-103793 and NSC-D-669356), Epothilones (e.g. Epothilone A, Epothilone B, Epothilone C (i.e. desoxyepothilone A or dEpoA), Epothilone D (i.e. KOS-862, dEpoB, and desoxyepothilone B), Epothilone E, Epothilone F, Epothilone B N-oxide, Epothilone A N-oxide, 16-aza-epothilone B, 21-aminoepothilone B (i.e. BMS-310705), 21-hydroxyepothilone D (i.e. Desoxyepothilone F and dEpoF), 26-fluoroepothilone, Auristatin PE (i.e. NSC-654663), Soblidotin (i.e. TZT-1027), LS-4559-P (Pharmacia, i.e. LS-4577), LS-4578 (Pharmacia, i.e. LS- 477-P), LS-4477 (Pharmacia), LS-4559 (Pharmacia), RPR-112378 (Aventis), Vincristine sulfate, DZ-3358 (Daiichi), FR-182877 (Fujisawa, i.e. WS-9885B), GS-164 (Takeda), GS-198 (Takeda), KAR-2 (Hungarian Academy of Sciences), BSF-223651 (BASF, i.e. ILX-651 and LU-223651), SAH-49960 (Lilly / Novartis), SDZ-268970 (Lilly / Novartis), AM-97 (Armad / Kyowa Hakko), AM-132 (Armad), AM-138 (Armad / Kyowa Hakko), IDN-5005 (Indena), Cryptophycin 52 (i.e. LY-355703), AC-7739 (Ajinomoto, i.e. AVE-8063A and CS-39.HCl), AC-7700 (Ajinomoto, i.e. AVE-8062, AVE-8062A, CS-39-L-Ser.HCl, and RPR-258062A), Vitilevuamide, Tubulysin A, Canadensol, Centaureidin (i.e. NSC-106969), T-138067 (Tularik, i.e. T-67, TL-138067 and TI- 138067), COBRA-1 (Parker Hughes Institute, i.e. DDE-261 and WHI-261), H10 (Kansas State University), H16 (Kansas State University), Oncocidin A1 (i.e. BTO-956 and DIME), DDE-313 (Parker Hughes Institute), Fijianolide B, Laulimalide, SPA-2 (Parker Hughes Institute), SPA-1 (Parker Hughes Institute, i.e. SPIKET-P), 3-IAABU (Cytoskeleton / Mt. Sinai School of Medicine, i.e. MF-569), Narcosine (also known as NSC-5366), Nascapine, D-24851 (Asta Medica), A-105972 (Abbott), Hemiasterlin, 3-BAABU (Cytoskeleton / Mt. Sinai School of Medicine, i.e. MF-191), TMPN (Arizona State University), Vanadocene acetylacetonate, T- 138026 (Tularik), Monsatrol, lnanocine (i.e. NSC-698666), 3-IAABE (Cytoskeleton / Mt. Sinai School of Medicine), A-204197 (Abbott), T-607 (Tuiarik, i.e. T-900607), RPR-115781 (Aventis), Eleutherobins (such as Desmethyleleutherobin, Desaetyleleutherobin, lsoeleutherobin A, and Z-Eleutherobin), Caribaeoside, Caribaeolin, Halichondrin B, D-64131 (Asta Medica), D- 68144 (Asta Medica), Diazonamide A, A-293620 (Abbott), NPI-2350 (Nereus), Taccalonolide A, TUB-245 (Aventis), A-259754 (Abbott), Diozostatin, (-)-Phenylahistin (i.e. NSCL-96F037), D-68838 (Asta Medica), D-68836 (Asta Medica), Myoseverin B, D-43411 (Zentaris, i.e. D- 81862), A-289099 (Abbott), A-318315 (Abbott), HTI-286 (i.e. SPA-110, trifluoroacetate salt) (Wyeth), D-82317 (Zentaris), D-82318 (Zentaris), SC-12983 (NCI), Resverastatin phosphate sodium, BPR-OY-007 (National Health Research Institutes), and SSR-250411 (Sanofi)), steroids (e.g., dexamethasone), finasteride, aromatase inhibitors, gonadotropin-releasing hormoneagonists (GnRH) such as goserelin or leuprolide, adrenocorticosteroids (e.g., prednisone), progestins (e.g., hydroxyprogesterone caproate, megestrol acetate, medroxyprogesterone acetate), estrogens (e.g., diethlystilbestrol, ethinyl estradiol), antiestrogen (e.g., tamoxifen), androgens (e.g., testosterone propionate, fluoxymesterone), antiandrogen (e.g., flutamide), immunostimulants (e.g., Bacillus Calmette-Guérin (BCG), levamisole, interleukin-2, alpha- interferon, etc.), monoclonal antibodies (e.g., anti-CD20, anti-HER2, anti-CD52, anti-HLA-DR, and anti-VEGF monoclonal antibodies), immunotoxins (e.g., anti-CD33 monoclonal antibody- calicheamicin conjugate, anti-CD22 monoclonal antibody-pseudomonas exotoxin conjugate, etc.), immunotherapy (e.g., cellular immunotherapy, antibody therapy, cytokine therapy, combination immunotherapy, etc.), radioimmunotherapy (e.g., anti-CD20 monoclonal antibody conjugated to 111In, 90Y, or 131I, etc.), immune checkpoint inhibitors (e.g., CTLA4 blockade, PD-1 inhibitors, PD-L1 inhibitors, etc.), triptolide, homoharringtonine, dactinomycin, doxorubicin, epirubicin, topotecan, itraconazole, vindesine, cerivastatin, vincristine, deoxyadenosine, sertraline, pitavastatin, irinotecan, clofazimine, 5-nonyloxytryptamine, vemurafenib, dabrafenib, erlotinib, gefitinib, EGFR inhibitors, epidermal growth factor receptor (EGFR)-targeted therapy or therapeutic (e.g. gefitinib (Iressa ™), erlotinib (Tarceva ™), cetuximab (Erbitux™), lapatinib (Tykerb™), panitumumab (Vectibix™), vandetanib (Caprelsa™), afatinib / BIBW2992, CI-1033 / canertinib, neratinib / HKI-272, CP-724714, TAK- 285, AST-1306, ARRY334543, ARRY-380, AG-1478, dacomitinib / PF299804, OSI- 420 / desmethyl erlotinib, AZD8931, AEE788, pelitinib / EKB-569, CUDC-101, WZ8040, WZ4002, WZ3146, AG-490, XL647, PD153035, BMS-599626), sorafenib, imatinib, sunitinib, dasatinib, or the like.

[0177] As used herein, the term “anti-inflammatory agent” refers to a drug or substance that reduces inflammation. In embodiments, anti-inflammatory agents include non-steroidal anti- inflammatory drugs. In embodiments, anti-inflammatory agents include steroidal anti- inflammatory drugs.

[0178] As used herein, the term “antifungal” refers to a drug or substance that treats or reduces fungal infection. In embodiments, antifungals include clotrimazole, econazole, miconazole, terbinafine, fluconazole, ketoconazole, nystatin.

[0179] “Remission” means that the clinical signs and symptoms of cancer have been significantly diminished or have disappeared entirely based on clinical diagnostics, although cancerous cells can still exist in the body. Thus, it is contemplated that remission encompassespartial and complete remission. Remission can occur for any period of time, such as from one month to several years or more.

[0180] “Relapse” or “Recurrence” refers to the clinical diagnosis of a return of cancer after a period of remission.

[0181] “Relapse-free survival” or “Recurrence-free survival” or “RFS” refers to the length of time from the date of primary treatment for a cancer ends that the patient survives without any signs or symptoms of that cancer until the date of relapse. In clinical trials, measuring the relapse-free survival is a way to determine the efficacy of a new treatment.

[0182] “Good prognosis” refers to a normal risk of relapse, a reduced risk of relapse, an increased chance for remission, an increased relapse-free survival time, or a high survival rate. In embodiments, a “good prognosis” refers to a reduced risk of relapse, an increased chance for remission, an increased relapse-free survival time, or a high survival rate. In embodiments, a “good prognosis” refers to a reduced risk of relapse. In embodiments, a “good prognosis” refers to an increased chance for remission. In embodiments, a “good prognosis” refers to an increased relapse-free survival time. In embodiments, a “good prognosis” refers to a high survival rate. In embodiments, a high survival rate refers to a 5-year survival rate greater than 50%. In embodiments, a high survival rate refers to a 5-year survival rate greater than 60%, greater than 70%, greater than 80%, or greater than 90%. In embodiments, “good prognosis” is an increased likelihood of a good prognosis.

[0183] As used herein, the term “cancer recurrence” is used in accordance with its plain ordinary meaning and refers to cancer that has recurred (come back), usually after a period of time during which the cancer could not be detected. The cancer may come back to the same place as the original (primary) tumor or to another place in the body.

[0184] As used herein, the term “periodically monitored” is used in accordance with its plain ordinary meaning and refers to direct observation and assessment of the circumstances at least once a day or the time period assigned.

[0185] As used herein, the term “surgery” is used in accordance with its plain ordinary meaning and refers to medical intervention using operative manual and instrumental techniques on a person to investigate or treat a pathological condition such as a disease or injury, to help improve bodily function, appearance, or to repair unwanted ruptured areas.

[0186] As used herein, the term “sensitivity” is used in accordance with its plain ordinary meaning and refers to the measurement of the proportion of positives that are correctly identified (i.e. the proportion of those who have some condition (affected) who are correctly identified as having the condition).

[0187] As used herein, the term “specificity” is used in accordance with its plain ordinary meaning and refers to the measurement of the proportion of negatives that are correctly identified (i.e. the proportion of those who do not have the condition (unaffected) who are correctly identified as not having the condition).

[0188] “Assaying” or “detecting” means using an analytical procedure to qualitatively assess or quantitatively measure a disease or a live infection in a subject in need thereof. In embodiments, the detecting includes or is assaying, which includes wet lab analysis, physical steps and / or physical manipuatlion of the sample, for example in a laboratory setting involving physical assaying techniques. METHODS OF PRODUCING [18F]-LABELED OLIGOSACCHARIDES

[0189] Provided herein, inter alia, are methods of producing [18F]-labeled oligosaccharides from 2-[18F] fluoro-2-deoxy-d-glucose ([18F] FDG) in one step via chemoenzymatic syntheses. In embodiments, the disclosed methods produce α- and β-linked [18F]-labeled disaccharides of high biomedical interest for pathogen-specific imaging. In embodiments, the disclosed methods can be used to produce positron-labeled (e.g.18F-labeled) derivatives of oligosaccharides, including maltose, sakebiose, trehalose, laminaribiose and cellobiose for use as PET radiotracers to detect infections and other diseases.

[0190] In an aspect is provided a method of producing an [18F]-labeled oligosaccharide, the method including combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide selected from the group consisting of D-glucose-1-phosphate, D-galactose-1-phosphate, and D-mannose-1-phosphate in a reaction vessel, thereby producing said [18F]-labeled oligosaccharide. In embodiments, the terms “combine”, “combining” and the like may be used interchangeably with “contact” or “contacting”. For example, in embodiments, combining [18F]FDG, a phosphorylase and a phosphorylated monosaccharide refers to contacting a [18F]FDG, a phosphorylase and a phosphorylated monosaccharide. In embodiments, the [18F]FDG, a phosphorylase and a phosphorylated monosaccharide are combined (e.g. contacted) in a solvent suitable for reverse phosphorolysis chemoenzymatic reaction.

[0191] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined (e.g. contacted) simultaneously in the reaction vessel. For example, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide may be simultaneously combined (e.g. contacted) in a reaction vessel. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined (e.g. contacted) sequentially in the reaction vessel. In embodiments, the phosphorylase and phosphorylated monosaccharide are combined (e.g. contacted) in a reaction vessel prior to adding the [18F]FDG to the reaction vessel. In embodiments, the phosphorylase and [18F]FDG are combined (e.g. contacted) in a reaction vessel prior to adding the phosphorylated monosaccharide to the reaction vessel.

[0192] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined in an acidic solution. For example, the solution may be at a pH of about 5.0 to about 6.0. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined in a solution at a pH of about 6.0. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined in solution at a pH of 6.0.

[0193] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 10 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 20 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 30 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 40 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 50 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 60 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 70 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 80 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 90 minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 100minute to about 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 110 minute to about 120 minutes.

[0194] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 110 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 100 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 90 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 80 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 70 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 60 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 50 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 40 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 30 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 20 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1 minute to about 10 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 1, 10, 20, 30, 40, 50, 60, 70, 80, 90, 100, 110, or 120 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for about 20 minutes. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined for 20 minutes.

[0195] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 22 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 24 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 26 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 28 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combinedat a temperature of about 30 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 32 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 34 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 36 °C to about 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 38 °C to about 40 °C.

[0196] In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 38 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 36 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 34 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 32 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 30 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 28 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 26 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 24 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C to about 22 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 20 °C, 22 °C, 24 °C, 26 °C, 28 °C, 30 °C, 32 °C, 34 °C, 36 °C, 38 °C, or 40 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of about 37 °C. In embodiments, the [18F]FDG, phosphorylase, and phosphorylated monosaccharide are combined at a temperature of 37 °C.

[0197] In embodiments, the [18F]-labeled oligosaccharide provided herein including embodiments thereof is isolated. For example, the [18F]-labeled oligosaccharide may be isolated from non-reacted starting materials (e.g. reactants ([18F]FDG, phosphorylated monosaccharide)), intermediate compounds, secondary products, and / or the phosphorylase. The [18F]-labeled oligosaccharide may be isolated (e.g. purified) from the reaction components using any method well known to a person having ordinary skill in the chemical arts. In embodiments, the [18F]-labeled oligosaccharide is isolated using high-performance liquid chromatography (HPLC) and / or silica gel chromatography. In embodiments, the isolated [18F]-labeled oligosaccharide may be stored in its aqueous form in a solution. In embodiments, the isolated [18F]-labeled oligosaccharide is stored in its dry form..

[0198] In embodiments, the phosphorylated monosaccharide is D-glucose-1-phosphate. In embodiments, the phosphorylated monosaccharide is D-galactose-1-phosphate. In embodiments, the phosphorylated monosaccharide is D-mannose-1-phosphate. In embodiments, the D-glucose- 1-phosphate is α- D-glucose-1-phosphate. In embodiments, the D-glucose-1-phosphate is a β- D- glucose-1-phosphate. In embodiments, the phosphorylated monosaccharide is an α- D-mannose- 1-phosphate. In embodiments, the phosphorylated monosaccharide is a β- D-mannose-1- phosphate. In embodiments, the phosphorylated monosaccharide is an α- D-galactose-1- phosphate. In embodiments, the phosphorylated monosaccharide is a β- D-galactose-1- phosphate.

[0199] For the method provided herein, in embodiments, phosphorylated monosaccharide is an α- or a β- phosphorylated monosaccharide. In embodiments, the D-glucose-1-phosphate is α- D- glucose-1-phosphate or β- D-glucose-1-phosphate. In embodiments, the D-glucose-1-phosphate is α- D-glucose -1-phosphate. In embodiments, the D-glucose-1-phosphate is β-D-glucose-1- phosphate. In embodiments, the D-galactose-1-phosphate is α- D-galactose-1-phosphate or β- D-galactose-1-phosphate. In embodiments, the D-galactose-1-phosphate is α- D-galactose-1- phosphate. In embodiments, the D-galactose-1-phosphate is β- D-galactose-1-phosphate. In embodiments, the D-mannose-1-phosphate is α - D-mannose-1-phosphate or β- D-mannose-1- phosphate. In embodiments, the D-mannose-1-phosphate is α - D-mannose-1-phosphate. In embodiments, the D-mannose-1-phosphate is β- D-mannose-1-phosphate.

[0200] For the method provided herein, in embodiments, the [18F]-labeled oligosaccharide is the product of a chemoenzymatic reaction. In embodiments, the chemoenzymatic reaction is a reverse phosphorylysis reaction. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 1 to 5 additional saccharide monomer units. In embodiments, the 1 to 5 additional saccharide monomer units have been dephosphorylated in a reverse phosphorolysis reaction catalyzed by the phosphorylase provided herein including embodiments thereof. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 2 to 5 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a[18F]FDG and 3 to 5 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 4 to 5 additional saccharide monomer units.

[0201] In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 1 to 4 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 1 to 3 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 1 to 2 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 1 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 2 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 3 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 4 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and 5 additional saccharide monomer units. In embodiments, the 1 to 5 additional saccharide monomer units include D- glucose, D-galactose, or D-mannose. In embodiments, the D-glucose is α- D- glucose or β- D- glucose. In embodiments, the D-glucose is α- D- glucose. In embodiments, the D-glucose is β- D-glucose. In embodiments, the D-galactose is α- D-galactose or is β- D-galactose. In embodiments, the D-galactose is α- D-galactose. In embodiments, the D-galactose is β- D- galactose. In embodiments, the D-mannose is α - D-mannose or β- D-mannose. In embodiments, the D-mannose is α - D-mannose. In embodiments, the D-galactose is β- D-mannose.

[0202] In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes α- D- glucose, β- D-glucose, α- D-galactose, β- D-galactose, α - D-mannose, β- D-mannose, or a combination thereof. In embodiments, the additional saccharide monomer unit(s) includes α- D- glucose. In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes β- D- glucose. In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes α- D- galactose. In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes β- D- galactose. In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes α - D- mannose. In embodiments, the 1 to 5 additional saccharide monomer unit(s) includes β- D- mannose.

[0203] In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and α- D- glucose. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and β- D- glucose. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and α- D- galactose. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and β- D-galactose. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and α - D- mannose. In embodiments, the [18F]-labeled oligosaccharide includes a [18F]FDG and β- D- mannose.

[0204] In embodiments, the 1 to 5 additional monomer units include the same monomer unit.

[0205] In embodiments, the [18F]-labeled oligosaccharide includes 2 to 6 saccharide monomers, and wherein the [18F]-labeled oligosaccharide includes a product of reverse phosphorolysis of D-glucose-1-phosphate and 1 to 5 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a product of reverse phosphorolysis of D-glucose-1-phosphate and 2 to 5 additional saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes a product of reverse phosphorolysis of D-glucose-1- phosphate and additional saccharide monomer units. In embodiments, the [18F]- oligosaccharide includes a product of reverse phosphorolysis of D-glucose-1-phosphate and 4 to 5 saccharide monomer reactants. In embodiments, one of the additional saccharide monomer units is [18F]- fluoro-D-glucose ([18F]FDG).

[0206] In embodiments, the [18F]-labeled oligosaccharide includes 2 to 6 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 3 to 6 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 4 to 6 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 5 to 6 saccharide monomer units.

[0207] In embodiments, the [18F]-labeled oligosaccharide includes 2 to 5 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 2 to 4 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 2 to 3 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 2, 3, 4, 5, or 6 saccharide monomers. In embodiments, the [18F]-labeled oligosaccharide includes 2 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 3 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 4 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 5 saccharide monomer units. In embodiments, the [18F]-labeled oligosaccharide includes 6 saccharide monomer units. In embodiments, the saccharide monomer units include [18F]FDG and α- D- glucose, β- D-glucose, α- D-galactose, β- D-galactose, α - D-mannose, β- D-mannose, or a combination thereof.

[0208] In embodiments, the [18F]-labeled oligosaccharide is an [18F]-labeled disaccharide. In embodiments, the [18F]-labeled disaccharide is [18F]FDM. In embodiments, the [18F]-labeleddisaccharide is [18F]FSK. In embodiments, the [18F]-labeled disaccharide is [18F]FDT. In embodiments, the [18F]-labeled disaccharide is [18F]FDL. In embodiments, the [18F]-labeled disaccharide is [18F]FDC.

[0209] In embodiments, the [18F]-labeled oligosaccharide is a trisaccharide, tetrasaccharide, pentasaccharide, or hexasaccharide. In embodiments, the [18F]-labeled oligosaccharide is a trisaccharide. In embodiments, the [18F]-labeled oligosaccharide is a tetrasaccharide. In embodiments, the [18F]-labeled oligosaccharide is a pentasaccharide. In embodiments, the [18F]- labeled oligosaccharide is a hexasaccharide. “Trisaccharide” is used in accordance with its plain ordinary meaning in the art and refers to an oligosaccharide including three monosaccharides with two glycosidic bonds connecting them. In embodiments, the glycosidic bond is formed between any hydroxyl group on the component monosaccharides. In embodiments, the monosaccharides may be the same monosaccharide unit or different monosaccharide units. “Tetrasaccharide” is used in accordance to its plain ordinary meaning and refers to an oligosaccharide including four monosaccharide units wherein the monosaccharide units may be of the same or different monosaccharides. The monosaccharide units are connected via glycosidic bonds. Similarly, “pentasaccharide” and “hexasaccharide” refer to oligosaccharides including five or six monosaccharide units, respectively, wherein the monosaccharide units may be the same monosaccharide unit or different monosaccharide units.

[0210] In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltohexaose.

[0211] In embodiments, the [18F]-labeled oligosaccharide is [18F]FDC. In embodiments, the [18F]-labeled oligosaccharide is [18F]FDL. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-cellotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy- [18F]-fluoro-cellotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]- fluoro-cellopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- cellohexaose.

[0212] In embodiments, the disclosed methods make use of a phosphorylase enzyme. In embodiments, the phosphorylase enzyme catalyzes the addition of an inorganic phosphate group to a carbohydrate acceptor with the breaking of an O-glycosidic bond.

[0213] For the methods provided herein, in embodiments, the phosphorylase enzyme catalyzes a reverse phosphorolysis reaction. In embodiments, the phosphorylase enzyme catalyzes a reverse phosphorolysis reaction to release an inorganic phosphate group from a phosphorylated monosaccharide provided herein including embodiments thereof. In embodiments, the phosphorylated monosaccharide is D-glucose-1-phosphate, D-galactose-1-phosphate, or D- mannose-1-phosphate. In embodiments, the phosphorylase enzyme catalyzes a reverse phosphyolysis reaction to release an inorganic phosphate group from a phosphorylated monosaccharide and form an O-glycosidic bond between the resulting dephosphorylated monosaccharide and a labeled monosaccharide (e.g. [18F]FDG). In embodiments, the labeled monosaccharide is [18F]-fluoro-D-glucose ([18F]FDG). In embodiments, the phosphorylase enzyme catalyzes a reverse phosphyolysis reaction to release an inorganic phosphate group from a phosphorylated monosaccharide and form an O-glycosidic bond between the resulting dephosphorylated monosaccharide and a labeled monosaccharide, thereby producing a labeled oligosaccharide.

[0214] In embodiments, the labeled oligosaccharide is an [18F]-labeled oligosaccharide. In embodiments, the [18F]-labeled oligosaccharide is an [18F]-labeled disaccharide. In embodiments, phosphorylated monosaccharide is D-glucose-1-phosphate. In embodiments, the phosphorylated monosaccharide is an α- D-glucose-1-phosphate. In embodiments, the phosphorylated monosaccharide is a β- D-glucose-1-phosphate. In embodiments, the phosphorylated monosaccharide is D-galactose-1-phosphate. In embodiments, the phosphorylated monosaccharide is an α- D-galactose-1-phosphate. In embodiments, the phosphorylated monosaccharide is a β- D-galactose-1-phosphate. In embodiments, the phosphorylated monosaccharide is D-mannose-1-phosphate. In embodiments, the phosphorylated monosaccharide is an α- D-mannose-1-phosphate. In embodiments, the phosphorylated monosaccharide is a β- D-mannose-1-phosphate.

[0215] For the method provided herein, in embodiments, the [18F]-labeled oligosaccharide incudes [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-4 glycosidic bond or a β- ^1-4 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide incudes [18F]-fluoro-D- glucose linked to D-glucose through an ^- ^1-4 glycosidic bond. In embodiments, the [18F]- labeled oligosaccharide incudes [18F]-fluoro-D-glucose linked to D-glucose through a β- ^1-4 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D- glucose linked to D-glucose through an ^- ^1-3 glycosidic bond or a β- ^1-3 glycosidic bond. Inembodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D- glucose through an ^- ^1-3 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-glucose through a β- ^1-3 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D- glucose through an ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-6 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D- glucose linked to D-glucose through an β- ^1-6 glycosidic bond. In embodiments, the [18F]- labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-1 glycosidic bond. In embodiments, [18F]-fluoro-D-glucose is linked to 1, 2, 3, 4 or 5 D-glucose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 1 D-glucose monomer unit. In embodiments, [18F]-fluoro-D-glucose is linked to 2 D-glucose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 3 D-glucose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 4 D-glucose monomer units. In embodiments, [18F]-fluoro-D- glucose is linked to 5 D-glucose monomer units.

[0216] In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-galactose through a β- ^1-3 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-galactose through a β- ^1-4 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D- glucose linked to D-galactose through a ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D- galactose through a ^- ^1-6 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-galactose through a β- ^1-6 glycosidic bond. In embodiments, [18F]-fluoro-D-glucose is linked to 1, 2, 3, 4, or 5 D-galactose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 1 D-galactose monomer unit. In embodiments, [18F]-fluoro-D-glucose is linked to 2 D-galactose monomer units. In embodiments, [18F]-fluoro- D-glucose is linked to 3 D-galactose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 4 D-galactose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 5 D- galactose monomer units.

[0217] In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-mannose through an β- ^1-3 glycosidic bond. In embodiments, the [18F]-labeled oligosaccharide includes [18F]-fluoro-D-glucose linked to D-mannose through a β- ^1-4 glycosidicbond. In embodiments, [18F]-fluoro-D-glucose is linked to 1, 2, 3, 4, or 5 D-mannose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 1 D-mannose monomer unit. In embodiments, [18F]-fluoro-D-glucose is linked to 2 D-mannose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 3 D-mannose monomer units. In embodiments, [18F]-fluoro- D-glucose is linked to 4 D-mannose monomer units. In embodiments, [18F]-fluoro-D-glucose is linked to 5 D-mannose monomer units.

[0218] In embodiments, the phosphorylase enzyme is maltose phosphorylase, sakebiose phosphorylase, trehalose phosphorylase, cellobiose phosphorylase, laminaribiose phosphorylase, α-glucan phosphorylase, β-glucan phosphorylase, isomaltose phosphorylase, gentiobiose phosphorylase, lactose phosphorylase, 1,3-β-galactosyl glucose phosphorylase, 1,6-β-galactosyl glucose phosphorylase, 1,6-α-galactosyl glucose phosphorylase, 1,4-β-mannosyl glucose phosphorylase, or 1,3-β-mannosyl glucose phosphorylase. In embodiments, the phosphorylase enzyme is maltose phosphorylase. In embodiments, the phosphorylase enzyme is sakebiose phosphorylase. In embodiments, the phosphorylase enzyme is trehalose phosphorylase. In embodiments, the phosphorylase enzyme is cellobiose phosphorylase. In embodiments, the phosphorylase enzyme is laminaribiose phosphorylase. In embodiments, the phosphorylase enzyme is α-glucan phosphorylase. In embodiments, the phosphorylase enzyme is β-glucan phosphorylase. In embodiments, the phosphorylase enzyme is isomaltose phosphorylase. In embodiments, the phosphorylase enzyme is gentiobiose phosphorylase. In embodiments, the phosphorylase enzyme is lactose phosphorylase. In embodiments, the phosphorylase enzyme is 1,3-β-galactosyl glucose phosphorylase. In embodiments, the phosphorylase enzyme is 1,6-β- galactosyl glucose phosphorylase. In embodiments, the phosphorylase enzyme is 1,6-α- galactosyl glucose phosphorylase. In embodiments, the phosphorylase enzyme is 1,4-β-mannosyl glucose phosphorylase. In embodiments, the phosphorylase enzyme is or 1,3-β-mannosyl glucose phosphorylase.

[0219] In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC), 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 2-deoxy-2- [18F]fluoro-lactose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro- allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2- deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, or 2-deoxy-[18F]-fluoro- cellohexaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltose ([18F]FDM). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]-fluoro- sakebiose ([18F]FSK). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2- [18F]fluoro-cellobiose ([18F]FDC). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy- 2-[18F]fluoro-trehalose ([18F]FDT). In embodiments, the [18F]-labeled oligosaccharide is 2- deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL) . In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-isomaltose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-gentobiose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-lactose. In embodiments, the [18F]-labeled oligosaccharide is 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose. In embodiments, the [18F]- labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-allolactose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-melibiose. In embodiments, the [18F]-labeled oligosaccharide is 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose. In embodiments, the [18F]- labeled oligosaccharide is 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotriose. In embodiments, the [18F]- labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-cellotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-cellotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-cellopentaose. In embodiments, the [18F]-labeled oligosaccharide is or 2-deoxy-[18F]-fluoro-cellohexaose.

[0220] In embodiments, the methods disclosed herein produce living bacteria-sensitive tracers. In embodiments, living bacteria-sensitive tracers include [18F]-labeled α-1,4 and α-1,3 oligosaccharides. In embodiments, the [18F]-labeled α-1,4 and α-1,3 oligosaccharides are [18F]- labeled α-1,4 and α-1,3 disaccharides In embodiments, [18F]-labeled α-1,4 and α-1,3 disaccharides include, but are not limited to, 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy- 2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), 2-deoxy-[18F]- fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose.

[0221] In embodiments, the methods disclosed herein produce living fungi-sensitive tracers. In embodiments, living fungi-sensitive tracers include [18F]-labeled β-1,4 and β-1,3oligosaccharides. In embodiments, [18F]-labeled β-1,4 and β-1,3 oligosaccharides are [18F]- labeled β-1,4 and β-1,3 disaccharides. In embodiments, [18F]-labeled β-1,4 and β-1,3 disaccharides include, but are not limited to, 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC), 2- deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]- fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, and 2-deoxy-[18F]-fluoro-cellohexaose. [18F]-LABELED OLIGOSACCHARIDES

[0222] Provided herein, inter alia, are compositions including an [18F]-labeled oligosaccharide provided herein including embodiments thereof. In an aspect is provided an [18F]-labeled oligosaccharide selected from the group consisting of 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy- 2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^- mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2- deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro- maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]-fluoro-cellotetraose, 2-deoxy- [18F]-fluoro-cellopentaose, and 2-deoxy-[18F]-fluoro-cellohexaose. In embodiments, the [18F]- labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltose ([18F]FDM). In embodiments, the [18F]- labeled oligosaccharide is 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT). In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL) . In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-isomaltose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-gentobiose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-lactose. In embodiments, the [18F]-labeled oligosaccharide is 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro- glucose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-allolactose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-2-[18F]fluoro-melibiose. In embodiments, the [18F]-labeled oligosaccharide is 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro- glucose. In embodiments, the [18F]-labeled oligosaccharide is 1,3- ^-mannosyl-2-deoxy-2- [18F]fluoro-glucose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- maltotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- maltotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- maltopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-cellotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- cellotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- cellopentaose. In embodiments, the [18F]-labeled oligosaccharide is or 2-deoxy-[18F]-fluoro- cellohexaose.

[0223] The compositions provided herein including embodiments thereof may be suitable for administration to a subject. For example, in embodiments, the compositions including [18F]- labeled oligosaccharides provided herein may be provided in a pharmaceutically acceptable excipient for injection into a subject. As described throughout the specification, in embodiments, the compositions may be administered to a subject for detecting living or diagnosing a disease caused by infection by the bacteria. In embodiments, the compositions may be administered to a subject for detecting a fungus or diagnosing a disease caused by the fungus. Thus, in an aspect is provided a pharmaceutical composition including an effective amount of an [18F]-labeled oligosaccharide provided herein including embodiments thereof in a pharmaceutically acceptable excipient.

[0224] In embodiments, the [18F]-labeled oligosaccharides provided herein including embodiments thereof are living bacteria-sensitive tracers. In embodiments, living bacteria- sensitive tracers include [18F]-labeled α-1,4 and α-1,3 oligosaccharides. In embodiments, the [18F]-labeled α-1,4 and α-1,3 oligosaccharides are [18F]-labeled α-1,4 and α-1,3 disaccharides. In embodiments, [18F]-labeled α-1,4 and α-1,3 disaccharides include, but are not limited to, 2- deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), or 2- deoxy-2-[18F]fluoro-trehalose ([18F]FDT). In embodiments, [18F]-labeled α-1,4 and α-1,3 oligosaccharides include 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, or 2-deoxy-[18F]-fluoro-maltopentaose.

[0225] In embodiments, the [18F]-labeled oligosaccharides is a [18F]-labeled disaccharide. In embodiments, the [18F]-labeled disaccharide is [18F]FDM. In embodiments, the [18F]-labeled disaccharide is [18F]FSK. In embodiments, the [18F]-labeled disaccharide is [18F]FDM. In embodiments, the [18F]-labeled disaccharide is [18F]FDT. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro-maltohexaose.

[0226] In embodiments, the [18F]-labeled oligosaccharides provided herein including embodiments thereof are living fungi-sensitive tracers. In embodiments, living fungi-sensitive tracers include [18F]-labeled β-1,4 and β-1,3 oligosaccharides. In embodiments, [18F]-labeled β- 1,4 and β-1,3 oligosaccharides are [18F]-labeled β-1,4 and β-1,3 disaccharides. In embodiments, [18F]-labeled β-1,4 and β-1,3 disaccharides include 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC) or 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL). In embodiments, [18F]-labeled β-1,4 and β- 1,3 oligosaccharides include 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, or 2-deoxy-[18F]-fluoro-cellohexaose.

[0227] In embodiments, the [18F]-labeled disaccharide is [18F]FDC. In embodiments, the [18F]- labeled disaccharide is [18F]FDL. In embodiments, the [18F]-labeled disaccharide is 2-deoxy- [18F]-fluoro-cellotriose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]- fluoro-cellotetraose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- cellopentaose. In embodiments, the [18F]-labeled oligosaccharide is 2-deoxy-[18F]-fluoro- cellohexaose.

[0228] In embodiments, the composition provided herein includes an effective amount of an ^–glycosidase inhibitor. In embodiments, the ^–glycosidase inhibitor includes voglibose, acarbose, miglitol, or a combination thereof. In embodiments, the ^–glycosidase inhibitor is voglibose. In embodiments, the ^–glycosidase inhibitor is acarbose. In embodiments, the ^– glycosidase inhibitor is miglitol. In embodiments, an effective amount of a ^–glycosidase inhibitor decreases or inhibits degradation of a [18F]-labeled oligosaccharide by ^–glycosidase into monomer saccharide units. In embodiments, the ^–glycosidase inhibitor decreases or inhibits degradation of a [18F]-labeled oligosaccharide by at least 40%, 50%, 60%, 70%, 80% 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, or 99% relative to degradation of the [18F]- labeled oligosaccharide in the absence of the ^–glycosidase inhibitor. DETECTION METHODS

[0229] Provided herein, inter alia, are methods for detecting or diagnosing a disease in a subject in need thereof. For example, the composition provided herein including embodiments thereof may be used to detect an infection (e.g. bacterial or fungal infection) or a disease caused by the infection in a subject in need thereof. In embodiments, the composition provided herein including embodiments thereof may be used to diagnose a subject with an infection or a disease caused by the infection in a subject in need thereof. In embodiment, the subject is suspected of having an infection or a disease. In embodiment, the subject has the infection or disease. Inembodiments, the disclosed methods include injecting into the subject an effective amount of a [18F]-labeled oligosaccharide as provided herein, imaging the subject by µPET-CT or PET-MR, and detecting the accumulation of the [18F]-labeled oligosaccharide in a tissue of the subject, thereby detecting the infection or disease. Thus, in an aspect is provided a method of detecting a disease in a subject in need thereof, the method including injecting into the subject an effective amount of an [18F]-labeled oligosaccharide provided herein including embodiments thereof, imaging the subject by positron emission tomography–computed tomography (PET-CT) or positron emission tomography–magnetic resonance imaging (PET-MRI), and detecting an accumulation of the [18F]-labeled oligosaccharide in a tissue of the subject, thereby detecting the disease.

[0230] In another aspect a method of detecting a disease in a subject in need thereof is provided, the method including administering to the subject an effective amount of the [18F]- labeled oligosaccharide provided herein including embodiments thereof or the pharmaceutical composition provided herein including embodiments thereof and detecting the [18F]-labeled oligosaccharide in said subject, thereby detecting the disease. In embodiments, the [18F]-labeled oligosaccharide is detected in a tissue or an organ of the subject. In embodiments, the method includes administering to the subject an effective amount of the [18F]-labeled oligosaccharide provided herein including embodiments thereof. In embodiments, the method includes administering to the subject an effective amount of the pharmaceutical composition provided herein including embodiments thereof.

[0231] In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro- laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro-allolactose, 2-deoxy-2- [18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy- 2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2- deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]-fluoro- cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, 2-deoxy-[18F]-fluoro-cellohexaose or a combination thereof. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]- fluoro-maltose ([18F]FDM). In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy- 2-[18F]-fluoro-sakebiose ([18F]FSK) . In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL) . In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-2-[18F]fluoro-isomaltose. In embodiments, the [18F]-labeledoligosaccharide includes 2-deoxy-2-[18F]fluoro-gentobiose. In embodiments, the [18F]-labeled oligosaccharide includes 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-2-[18F]fluoro-allolactose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-2-[18F]fluoro-melibiose. In embodiments, the [18F]-labeled oligosaccharide includes 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose. In embodiments, the [18F]-labeled oligosaccharide includes 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro- glucose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- maltotriose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- maltotetraose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- maltopentaose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- cellotriose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- cellotetraose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- cellopentaose. In embodiments, the [18F]-labeled oligosaccharide includes 2-deoxy-[18F]-fluoro- cellohexaose. In embodiments, the [18F]-labeled oligosaccharide includes a combination of two or more [18F]-labeled oligosaccharides. In embodiments, the [18F]-labeled oligosaccharide includes [18F]FDM and [18F]FSK.

[0232] In embodiments, the imaging includes positron emission tomography–computed tomography (PET-CT). In embodiments, the imaging includes positron emission tomography– magnetic resonance imaging (PET-MRI). In embodiments, the administration includes intravenous infusion of a composition including the [18F]-labeled oligosaccharide provided herein including embodiments thereof. In embodiments, the administration includes injecting the [18F]-labeled oligosaccharide provided herein including embodiments thereof into the subject.

[0233] The compositions provided herein including embodiments thereof are contemplated to be useful for detecting and / or diagnosing a disease in a subject in need thereof. In embodiments, the disease is a microbe infection. In embodiments, the microbe infection is a bacterial infection or a fungal infection. For example, in embodiments, the [18F]-labeled oligosaccharide provided herein including embodiments thereof is metabolized by fungi or incorporated into the cell wall of a fungi, thereby allowing detection of the fungal infection in a subject. In embodiments, the [18F]-labeled oligosaccharide provided herein including embodiments thereof is metabolized by a bacteria or incorporated into the cell wall of a bacteria, thereby allowing detection of a bacterial infection in a subject.

[0234] In embodiments, the composition provided herein including embodiments thereof is useful for detecting or diagnosing cancer in a subject in need thereof. For example, in embodiments, a cancer cell may have increased glucose uptake and / or glycolysis, thereby allowing detection or diagnosis of a cancer in a subject. In embodiments, the detection or diagnosis occurs prior to onset of symptoms.

[0235] In embodiments, the [18F]-labeled oligosaccharides provided herein including embodiments thereof may incorporate into the cell wall of bacteria, thereby allowing sensitive and specific detection of the bacteria. Thus, in embodiments, the [18F]-labeled oligosaccharides provided herein including embodiments thereof are contemplated to be useful for detecting bacterial infections and diseases caused by the bacterial infections. In embodiments, the [18F]- labeled oligosaccharides provided herein including embodiments thereof is localized to the bacterial in a subject (e.g. the tissue or an organ of the subject). For the methods provided herein, in embodiments, the disease is a microbe infection. In embodiments, the microbe infection is a bacterial infection.

[0236] In embodiments, the [18F]-labeled oligosaccharides are capable of detecting living microbes and do not detect dead microbes. For example, the [18F]-labeled oligosaccharide is not metabolized or incorporated into the cell wall of a dead microbe. For example, in embodiments, the [18F]-labeled oligosaccharide when contacted with dead microbes produce a signal that is less than 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold of a background signal. In embodiments, the [18F]-labeled oligosaccharide when contacted with dead microbes produce a signal that is less than about 50%, 40%, 35% 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% greater than a control. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a sample (e.g. cell culture, tissue sample) that does not include the microbe to be detected. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a healthy subject (e.g. a subject who does not have an infection or a disease caused by the infection).

[0237] For the method provided herein, in embodiments, the microbe infection is a live microbe infection. In embodiments, the bacterial infection is a living bacterial infection. In embodiments, the bacterial infection is a living fungal infection.

[0238] In embodiments, the bacterial infection is a gram-positive bacterial infection. In embodiments, the living bacterial infection is a gram-positive living bacterial infection. In embodiments, the gram-positive bacterial infection is infection by one or more bacteria includingincluding Streptococcus, Staphylococcus, Corynebacterium, Listeria, Bacillus, Clostridium, Catabacter, Rathybacter, Leifsonia, or Clavibacter. In embodiments, the infection is by one or more bacteria, including Streptococcus, Staphylococcus, Corynebacterium, Listeria, Bacillus, Clostridium, Catabacter, Rathybacter, Leifsonia, or Clavibacter. In embodiments, the bacterial infection is a Streptococcus infection. In embodiments, the bacterial infection is a Streptococcus infection. In embodiments, the bacterial infection is a Staphylococcus infection. In embodiments, the bacterial infection is a Corynebacterium infection. In embodiments, the bacterial infection is a Listeria infection. In embodiments, the bacterial infection is a Bacillus infection. In embodiments, the bacterial infection is a Clostridium infection. In embodiments, the bacterial infection is a Catabacter infection. In embodiments, the bacterial infection is a Rathybacter infection. In embodiments, the bacterial infection is a Leifsonia infection. In embodiments, the bacterial infection is a Clavibacter infection.

[0239] In embodiments, the living bacterial infection is a gram-negative bacterial infection. In embodiments, the gram-negative bacterial infection is infection by one or more bacterial including Klebsiella, Acinetobacter, Pseudomonas aeruginosa, or E. coli. In embodiments, the infection is by one or more bacteria, including Klebsiella, Acinetobacter, Pseudomonas aeruginosa, or E. coli. In embodiments, the bacterial infection is a Klebsiella infection. In embodiments, the bacterial infection is an Acinetobacter infection. In embodiments, the bacterial infection is a Pseudomonas aeruginosa infection. In embodiments, the bacterial infection is an E. coli infection. In embodiments, the disease is pneumonia. In embodiments, the disease is a bloodstream infection. In embodiments, the disease is a wound infection. In embodiments, the disease is a surgical site infection. In embodiments, the disease is meningitis.

[0240] In embodiments, the bacterial infection is caused by one or more of Staphylococcus epidermis, Acinetobacter baumannii, Streptococcus pneumonia, Enterobacter cloacae, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Proteus mirabilis Escherichia coli, Klebsiella pneumoniae Staphylococcus aureus, Staphylococcus epidermidis, or Enterococcus faecalis. In embodiments, the bacterial infection is caused by Staphylococcus epidermis. In embodiments, the bacterial infection is caused by Acinetobacter baumannii. In embodiments, the bacterial infection is caused by Streptococcus pneumonia. In embodiments, the bacterial infection is caused by Enterobacter cloacae. In embodiments, the bacterial infection is caused by Listeria monocytogenes. In embodiments, the bacterial infection is caused by Pseudomonas aeruginosa. In embodiments, the bacterial infection is caused by Salmonella typhimurium. In embodiments, the bacterial infection is caused by Proteus mirabilis. Inembodiments, the bacterial infection is caused by Escherichia coli. In embodiments, the bacterial infection is caused by Klebsiella pneumoniae. In embodiments, the bacterial infection is caused by Staphylococcus aureus. In embodiments, the bacterial infection is caused by Staphylococcus epidermidis. In embodiments, the bacterial infection is caused by Enterococcus faecalis.

[0241] In embodiments, the living microbe infection is a living fungal infection. In embodiments, the living fungal infection is an infection by one or more of Mycobacterium tuberculosis, Candida auris, Candida albicans, Aspergillus sp, Pneumocystis jirovecii, Criptococcus neoformans, and Taloromyces sp. In embodiments, the fungal infection is a Mycobacterium tuberculosis infection. In embodiments, the fungal infection is a Candida auris infection. In embodiments, the fungal infection is a Candida albicans infection. In embodiments, the fungal infection is an Aspergillus sp. Infection. In embodiments, the fungal infection is a Pneumocystis jirovecii infection. In embodiments, the fungal infection is a Criptococcus neoformans infection. In embodiments, the fungal infection is a Taloromyces sp. infection.

[0242] In embodiments, the subject has a disease. In embodiments, the disease is an infection. In embodiments, the disease is myositis, vertebral discitis-osteomyelitis, pneumonia, or cancer. In embodiments, the disease is myositis. In embodiments, the disease is vertebral discitis- osteomyelitis. In embodiments, the disease is pneumonia. In embodiments, the disease is a cancer.

[0243] In embodiments, the subject is suspected of having a disease. In embodiments, the subject is suspected of having an infection. In embodiments, the subject is suspected of having myositis, vertebral discitis-osteomyelitis, pneumonia, or cancer. In embodiments, the subject is suspected of having myositis. In embodiments, the subject is suspected of having vertebral discitis-osteomyelitis. In embodiments, the subject is suspected of having pneumonia. In embodiments, the subject is suspected of having a cancer.

[0244] In embodiments, the method provided herein including embodiments thereof include preventing a disease. For example, in embodiments, the method includes detecting the [18F]- labeled oligosaccharide provided herein including embodiments thereof prior to onset of disease symptoms. In embodiments, the method includes detecting the [18F]-labeled oligosaccharide provided herein including embodiments at the onset of a microbe infection. Thus, in embodiments, the method includes administering a therapeutic, vaccine or prophylactic to the subject thereby preventing the disease.

[0245] In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize α-1,3 linkages and α-1,4 linkages in α-D-glucans (ADG) found in bacteria. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize α-1,3 linkages. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize α-1,4 linkages. “Glucan” is used in accordance to its plain ordinary meaning in the art and refers to a polysaccharide including glucose monomer units. In embodiments, a glucan includes glucose monomer units and no other monosaccharide units.

[0246] In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively detect α-1,3 linkages and α-1,4 linkages in α-D-glucans (ADG) found in bacteria. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively detect α-1,3 linkages found in bacteria. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively found in bacteria α-1,4 linkages. For example, and as described throughout the specification, in embodiments, the18F-labeled oligosaccharides provided herein including embodiments thereof are metabolized by bacteria or incorporated into the cell wall of bacteria including α-1,3 linkages and α-1,4 linkages in α-D-glucans (ADG), thereby allowing detection of α-1,3 linkages and α-1,4 linkages in α-D-glucans.

[0247] In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize ^ ^-1,3 and ^-1,4 linkages in ^-D-glucans (BDG), found in the cell walls of fungi. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize ^ ^-1,3 linkages. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively recognize ^ ^-1,4 linkages.

[0248] In embodiments, the the18F-labeled oligosaccharides provided herein specifically and selectively detect ^ ^-1,3 and ^-1,4 linkages in ^-D-glucans (BDG), found in the cell walls of fungi. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively detect ^ ^-1,3 linkages. In embodiments, the18F-labeled oligosaccharides provided herein specifically and selectively detect ^ ^-1,4 linkages. For example, and as described throughout the specification, in embodiments, the18F-labeled oligosaccharides provided herein including embodiments thereof are metabolized or incorporated into the cell wall of fungi including ^ ^-1,3 and ^-1,4 linkages in ^-D-glucans (BDG), thereby allowing detection of ^ ^-1,3 and ^-1,4 linkages in ^-D-glucans.

[0249] In embodiments, the [18F]-labeled oligosaccharide specifically and selectively recognize ^ ^-1,4 linkages when contacted with α-1,3 oligosaccharides produce a signal that is less than 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold of that of the background signal. In embodiments, the [18F]-labeled oligosaccharide specifically and selectively recognizes ^ ^-1,4 linkages when contacted with α-1,3 oligosaccharide produce a signal that is less than about 40%, 35% 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% greater of that of a control. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a sample (e.g. cell culture, tissue sample) that does not include a detectable amount of a ^-1,4 oligosaccharide. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a sample (e.g. cell culture, tissue sample) that does not include a bacteria or fungus including a detectable amount of an ^-1,4 oligosaccharide in its cell wall.

[0250] In embodiments, the [18F]-labeled oligosaccharide specifically and selectively recognize α-1,3 linkages when contacted with ^ ^-1,4 oligosaccharides produce a signal that is less than 10-fold, 9-fold, 8-fold, 7-fold, 6-fold, 5-fold, 4-fold, 3-fold, 2-fold, or 1-fold of that of the background signal. In embodiments, the [18F]-labeled oligosaccharide specifically and selectively recognizes α-1,3 linkages when contacted with ^ ^-1,4 oligosaccharide produce a signal that is less than about 40%, 35% 30%, 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, or 1% greater of that of a control. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a sample (e.g. cell culture, tissue sample) that does not include a detectable amount of a α-1,3 oligosaccharide. In embodiments, the control is the signal of the [18F]-labeled oligosaccharide in a sample (e.g. cell culture, tissue sample) that does not include a bacteria or fungus including a detectable amount of an α-1,3 oligosaccharide in its cell wall.

[0251] In embodiments, the method further includes administering to the subject an ^– glycosidase inhibitor. In embodiments, the ^–glycosidase inhibitor includes voglibose, acarbose, miglitol, or a combination thereof. In embodiments, the ^–glycosidase inhibitor is voglibose. In embodiments, the ^–glycosidase inhibitor is acarbose. In embodiments, the ^–glycosidase inhibitor is miglitol. In embodiments, the structures of voglibose, acarbose, miglitol are as shown in FIG.13.

[0252] In embodiments, the methods provided herein further include treating the disease detected by the described imaging methods in a subject in need thereof. In embodiments, themethods comprise administering to the subject neoadjuvant therapy, if positron emission tomography (PET) imaging detects the presence of a disease in the subject. In embodiments, the disease is an infection. In embodiments, the disease is myositis. In embodiments, the disease is vertebral discitis-osteomyelitis. In embodiments, the disease is pneumonia. In embodiments, the disease is a cancer.

[0253] In embodiments, the method further includes treating the disease by administering to the subject one or more of an anti-inflammatory agent, an antibiotic, an antifungal agent, and an anti-cancer agent. In embodiments, the methods comprise administering to the subject an effective amount of an anti-inflammatory agent. In embodiments, the methods comprise administering to the subject an effective amount of an antibiotic. In embodiments, the methods comprise administering to the subject an effective amount of an anti-fungal agent. In embodiments, the methods comprise administering to the subject an effective amount of an anticancer agent.

[0254] In embodiments, the methods comprise administering to the subject an effective amount of radiation therapy. In embodiments, the methods comprise administering to the subject an effective amount of chemotherapy. In embodiments, the methods comprise administering to the subject an effective amount of targeted therapy. In embodiments, the methods comprise administering to the subject an effective amount of immunotherapy. In embodiments, the methods comprise administering to the subject an effective amount of hormonal therapy. In embodiments, the methods comprise administering to the subject an effective amount of angiogenesis inhibitor. In embodiments, the methods comprise administering to the subject an effective amount of synthetic lethality therapy. In embodiments, the methods comprise administering to the subject an effective amount of any combination of anti-inflammatory agent, antibiotic, antifungal agent, an anticancer agent, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, angiogenesis inhibitor administration therapy, and synthetic lethality therapy.

[0255] In embodiments, the neoadjuvant therapy comprises effective amounts of one or more active agents selected from the group consisting of anti-inflammatory agent, antibiotic, anti- fungal agent, antiviral agent, anticancer agent, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, angiogenesis inhibitor administration therapy, synthetic lethality therapy, and any combination thereof. In embodiments, the methods comprise administering to the subject an effective amount of an anti-inflammatory agent. In embodiments,the methods comprise administering to the subject an effective amount of an antibiotic. In embodiments, the methods comprise administering to the subject an effective amount of an antifungal agent. In embodiments, the methods comprise administering to the subject an effective amount of an anticancer agent. In embodiments, the methods comprise administering to the subject an effective amount of radiation therapy. In embodiments, the methods comprise administering to the subject an effective amount of chemotherapy. In embodiments, the methods comprise administering to the subject an effective amount of targeted therapy. In embodiments, the methods comprise administering to the subject an effective amount of immunotherapy. In embodiments, the methods comprise administering to the subject an effective amount of hormonal therapy. In embodiments, the methods comprise administering to the subject an effective amount of angiogenesis inhibitor. In embodiments, the methods comprise administering to the subject an effective amount of synthetic lethality therapy. In embodiments, the methods comprise administering to the subject an effective amount of any combination of an anticancer agent, radiation therapy, chemotherapy, targeted therapy, immunotherapy, hormonal therapy, angiogenesis inhibitor administration therapy, and synthetic lethality therapy.

[0256] In embodiments, the methods further comprise surgically removing all or portions of the cancer and infected tissues in the subject after completion of one or more rounds of neoadjuvant therapy.

[0257] It is understood that the examples and embodiments described herein are for illustrative purposes only and that various modifications or changes in light thereof will be suggested to persons skilled in the art and are to be included within the spirit and purview of this application and scope of the appended claims. All publications, patents, and patent applications cited herein are hereby incorporated by reference in their entirety for all purposes. P EMBODIMENTS

[0258] P Embodiment 1. A method of producing an [18F]-labeled oligosaccharide, the method comprising combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide selected from the group consisting of D-glucose-1-phosphate, D-galactose-1- phosphate, and D-mannose-1-phosphate in a reaction vessel, thereby producing said [18F]-labeled oligosaccharide.

[0259] P Embodiment 2. The method of P Embodiment 1, wherein said [18F]-labeled oligosaccharide is an [18F]-labeled disaccharide.

[0260] P Embodiment 3. The method of P Embodiment 1 or 2, wherein said [18F]-labeled oligosaccharide comprises 2 to 6 saccharide monomers and wherein the oligosaccharide is comprised of D-glucose-1-phosphate and 1 to 5 saccharide unit reactants.

[0261] P Embodiment 4. The method of any one of P Embodiments 1-3, wherein said [18F]- labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an α-1-4 glycosidic bond.

[0262] P Embodiment 5. The method of any one of P Embodiments 1-4, wherein the D- glucose-1-phosphate is an α- D-glucose-1-phosphate.

[0263] P Embodiment 6. The method of any one of P Embodiments 1-5, wherein the D- glucose-1-phosphate is a β- D-glucose-1-phosphate.

[0264] P Embodiment 7. The method of any one of P Embodiments 1-6, wherein the D- galactose-1-phosphate is a ^- D-galactose-1-phosphate.

[0265] P Embodiment 8. The method of any one of P Embodiments 1-6, wherein the D- galactose-1-phosphate is a β- D-galactose-1-phosphate.

[0266] P Embodiment 9. The method of any one of P Embodiments 1-6, wherein the D- mannose-1-phosphate is a β- D-mannose-1-phosphate.

[0267] P Embodiment 10. The method of any one of P Embodiments 1-9, wherein the phosphorylase enzyme is selected from the group consisting of maltose phosphorylase, sakebiose phosphorylase, trehalose phosphorylase, cellobiose phosphorylase, laminaribiose phosphorylase, α-glucan phosphorylase, β-glucan phosphorylase, isomaltose phosphorylase, gentiobiose phosphorylase, lactose phosphorylase, 1,3- ^-galactosyl glucose phosphorylase, 1,6- ^-galactosyl glucose phosphorylase, 1,6- ^-galactosyl glucose phosphorylase, 1,4- ^-mannosyl glucose phosphorylase, and 1,3- ^-mannosyl glucose phosphorylase.

[0268] P Embodiment 11. The method of any one of P Embodiments 1-10, wherein the [18F]- labeled oligoisaccharide is 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro- sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC), 2-deoxy-2-[18F]fluoro- trehalose ([18F]FDT), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro- isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 2-deoxy-2-[18F]fluoro-lactose, 1,3- ^-galactosyl-2- deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose,2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2-deoxy-[18F]-fluoro- maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]-fluoro-cellotetraose, 2-deoxy- [18F]-fluoro-cellopentaose, or 2-deoxy-[18F]-fluoro-cellohexaose.

[0269] P Embodiment 12. An [18F]-labeled oligosaccharide selected from the group consisting of 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2- deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2- [18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro- allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]- fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2- deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, and 2-deoxy-[18F]-fluoro- cellohexaose.

[0270] P Embodiment 13. A method of detecting a disease in a subject in need thereof, the method comprising injecting into the subject an effective amount of the [18F]-labeled oligosaccharide of P Embodiment 12, imaging the subject, and detecting the accumulation of the [18F]-labeled oligosaccharide in a tissue of the subject, thereby detecting the disease.

[0271] P Embodiment 14. The method of P Embodiment 13, wherein the disease is a microbe infection.

[0272] P Embodiment 15. The method of P Embodiment 14, wherein the microbe infection is a bacterial infection.

[0273] P Embodiment 16. The method of P Embodiment 15, wherein the bacterial infection is a gram-positive live bacterial infection.

[0274] P Embodiment 17. The method of P Embodiment 15, wherein the bacterial infection is a gram-negative live bacterial infection.

[0275] P Embodiment 18. The method of any one of P Embodiments 13-17, wherein the infection is caused by one or more of Staphylococcus epidermis, Acinetobacter baumannii, Streptococcus pneumonia, Enterobacter cloacae, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, or Proteus mirabilis.

[0276] P Embodiment 19. The method of P Embodiment 14, wherein the microbe infection is a live fungal infection.

[0277] P Embodiment 20. The method of P Embodiment 19, wherein the live infection is caused by one or more of Mycobacterium tuberculosis, Candida auris, Candida albicans, Aspergillus sp, Pneumocystis jirovecii, Criptococcus neoformans, or Taloromyces sp.

[0278] P Embodiment 21. The method of any one of P Embodiments 13-20, wherein the disease is myositis, vertebral discitis-osteomyelitis, pneumonia, or cancer.

[0279] P Embodiment 22. The method of any one of P Embodiments 13-21, wherein the method further comprises treating the disease by administering to the subject one or more of an anti-inflammatory agent, an antibiotic, an antifungal agent, and an anti-cancer agent. EMBODIMENTS

[0280] Embodiment 1. A method of producing an [18F]-labeled oligosaccharide, the method comprising combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide selected from the group consisting of D-glucose-1-phosphate, D-galactose-1- phosphate, and D-mannose-1-phosphate in a reaction vessel, thereby producing said [18F]-labeled oligosaccharide.

[0281] Embodiment 2. The method of Embodiment 1, wherein the phosphorylated monosaccharide is D-glucose-1-phosphate.

[0282] Embodiment 3. The method of Embodiment 2, wherein the D-glucose-1-phosphate is α- D-glucose-1-phosphate.

[0283] Embodiment 4. The method of Embodiment 2, wherein the D-glucose-1-phosphate is β- D-glucose-1-phosphate.

[0284] Embodiment 5. The method of Embodiment 1, wherein the phosphorylated monosaccharide is D-galactose-1-phosphate.

[0285] Embodiment 6. The method of Embodiment 5, wherein the D-galactose-1-phosphate is α- D-galactose-1-phosphate.

[0286] Embodiment 7. The method of Embodiment 5, wherein the D-galactose-1-phosphate is β- D-galactose-1-phosphate.

[0287] Embodiment 8. The method of Embodiment 1, wherein the phosphorylated monosaccharide is D-mannose-1-phosphate.

[0288] Embodiment 9. The method of Embodiment 8, wherein the D-mannose-1-phosphate is α- D-mannose-1-phosphate.

[0289] Embodiment 10. The method of Embodiment 8, wherein the D-mannose-1-phosphate is β- D-mannose-1-phosphate.

[0290] Embodiment 11. The method of any one of Embodiments 1 to 10, wherein the [18F]- labeled oligosaccharide comprises [18F]-fluoro-D-glucose and 1 to 5 additional saccharide monomer units.

[0291] Embodiment 12. The method of Embodiment 11, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose and 1 additional saccharide monomer unit.

[0292] Embodiment 13. The method of Embodiment 11 or 12, wherein said 1 to 5 additional saccharide monomer units comprise D-glucose, D-galactose, or D-mannose.

[0293] Embodiment 14. The method of Embodiment 13, wherein the D-glucose is α- D- glucose or β- D-glucose.

[0294] Embodiment 15. The method of Embodiment 13, wherein the D-galactose is α- D- galactose or β- D- galactose.

[0295] Embodiment 16. The method of Embodiment 13, wherein the D-mannose is α- D- mannose or β- D- mannose.

[0296] Embodiment 17. The method of any one of Embodiments 1 to 5 or 11 to 14, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-4 glycosidic bond or a β- ^1-4 glycosidic bond.

[0297] Embodiment 18. The method of any one of Embodiments 1 to 5 or 11 to 14, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-3 glycosidic bond or a β- ^1-3 glycosidic bond.

[0298] Embodiment 19. The method of any one of Embodiments 1 to 5 or 11 to 14, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond.

[0299] Embodiment 20. The method of any one of Embodiments 1 to 5 or 11 to 14, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-1 glycosidic bond.

[0300] Embodiment 21. The method of any one of Embodiments 1, 5 to 7, 11 to 13, or 15, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D- galactose through a β- ^1-3 glycosidic bond.

[0301] Embodiment 22. The method of any one of Embodiments 1, 5 to 7, 11 to 13, or 15, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D- galactose through a β- ^1-4 glycosidic bond.

[0302] Embodiment 23. The method of any one of Embodiments 1, 5 to 7, 11 to 13, or 15, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D- galactose through a ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond.

[0303] Embodiment 24. The method of any one of Embodiments 1, 8 to 13 or 16, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-mannose through an β- ^1-3 glycosidic bond.

[0304] Embodiment 25. The method of any one of Embodiments 1, 8 to 13 or 16, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-mannose through a β- ^1-4 glycosidic bond.

[0305] Embodiment 26. The method of any one of Embodiments 1 to 25, wherein said [18F]- labeled oligosaccharide is an [18F]-labeled disaccharide.

[0306] Embodiment 27. The method of Embodiment 26, wherein the [18F]-labeled disaccharide is 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), or 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC), 2-deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2- [18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, or 1,3- ^-mannosyl-2-deoxy-2- [18F]fluoro-glucose.

[0307] Embodiment 28. The method of Embodiment 26 or 27, wherein the [18F]-labeled disaccharide is [18F]FDM, [18F]FSK, or [18F]FDT.

[0308] Embodiment 29. The method of Embodiment 26 or 27, wherein the [18F]-labeled disaccharide is [18F]FDM and [18F]FSK.

[0309] Embodiment 30. The method of any one of Embodiments 1 to 25, wherein the [18F]- labeled oligosaccharide is [18F]FDM, [18F]FSK, [18F]FDC, [18F]FDT, 2-deoxy-2-[18F]fluoro-[18F]FDL, 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 2-deoxy-2- [18F]fluoro-lactose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro- allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]- fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2- deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, or 2-deoxy-[18F]-fluoro- cellohexaose.

[0310] Embodiment 31. The method of any one of Embodiments 1 to 30, wherein the phosphorylase enzyme is selected from the group consisting of maltose phosphorylase, sakebiose phosphorylase, trehalose phosphorylase, cellobiose phosphorylase, laminaribiose phosphorylase, α-glucan phosphorylase, β-glucan phosphorylase, isomaltose phosphorylase, gentiobiose phosphorylase, lactose phosphorylase, 1,3-β-galactosyl glucose phosphorylase, 1,6-β-galactosyl glucose phosphorylase, 1,6-α-galactosyl glucose phosphorylase, 1,4-β-mannosyl glucose phosphorylase, and 1,3-β-mannosyl glucose phosphorylase.

[0311] Embodiment 32. An [18F]-labeled oligosaccharide selected from the group consisting of 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2- deoxy-2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2- [18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro- allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]- fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2- deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, and 2-deoxy-[18F]-fluoro- cellohexaose.

[0312] Embodiment 33. A pharmaceutical composition comprising an effective amount of the [18F]-labeled oligosaccharide of Embodiment 32 and a pharmaceutically acceptable excipient.

[0313] Embodiment 34. The pharmaceutical composition of Embodiment 33, further comprising an effective amount of an ^–glycosidase inhibitor.

[0314] Embodiment 35. The pharmaceutical composition of Embodiment 34, wherein the ^– glycosidase inhibitor is voglibose, acarbose, miglitol or a combination thereof.

[0315] Embodiment 36. A method of detecting a disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the [18F]-labeledoligosaccharide of Embodiment 32 or the pharmaceutical composition of any one of Embodiments 33 to 35, and detecting the [18F]-labeled oligosaccharide in said subject, thereby detecting the disease.

[0316] Embodiment 37. The method of Embodiment 36, wherein the detecting comprises positron emission tomography–computed tomography (PET-CT) or positron emission tomography–magnetic resonance imaging (PET-MRI).

[0317] Embodiment 38. The method of Embodiment 36 or 37, wherein the disease is a microbe infection.

[0318] Embodiment 39. The method of Embodiment 38, wherein the microbe infection is a bacterial infection.

[0319] Embodiment 40. The method of Embodiment 39, wherein the bacterial infection is a gram-positive bacterial infection.

[0320] Embodiment 41. The method of Embodiment 39, wherein the bacterial infection is a gram-negative bacterial infection.

[0321] Embodiment 42. The method of any one of Embodiments 39 to 41, wherein the bacterial infection is caused by one or more of Staphylococcus epidermis, Acinetobacter baumannii, Streptococcus pneumonia, Enterobacter cloacae, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Proteus mirabilis, Escherichia coli, Klebsiella pneumoniae Staphylococcus aureus, Staphylococcus epidermidis, or Enterococcus faecalis.

[0322] Embodiment 43. The method of Embodiment 38, wherein the microbe infection is a fungal infection.

[0323] Embodiment 44. The method of Embodiment 43, wherein the fungal is caused by one or more of Mycobacterium tuberculosis, Candida auris, Candida albicans, Aspergillus sp, Pneumocystis jirovecii, Criptococcus neoformans, or Taloromyces sp.

[0324] Embodiment 45. The method of any one of Embodiments 36 to 44, wherein the microbe infection is a live microbe infection.

[0325] Embodiment 46. The method of any one of Embodiments 36 to 45, wherein the disease is myositis, vertebral discitis-osteomyelitis, pneumonia, or cancer.

[0326] Embodiment 47. The method of any one of Embodiments 36 to 46, wherein the method further comprises administering to the subject an effective amount of an ^–glycosidase inhibitor.

[0327] Embodiment 48. The method of Embodiment 47, wherein the ^–glycosidase inhibitor is voglibose, acarbose, miglitol or a combination thereof.

[0328] Embodiment 49. The method of any one of Embodiments 36 to 48, wherein the method further comprises treating the disease by administering to the subject one or more of an anti- inflammatory agent, an antibiotic, an antifungal agent, and an anti-cancer agent. EXAMPLES Introduction to Exemplary Studies

[0329] Biocatalysis is now frequently used in chemical synthesis, both for the development of new building blocks and late-stage modification of complex molecules1. Chemoenzymatic syntheses of polysaccharides are particularly appealing, given the challenges of regioselectivity and stereoselectivity using standard organic methods2,3. Glucose-based polysaccharides are amenable to enzymatic incorporation of unnatural monosaccharide units, suggesting the possibility of using the common clinical imaging tracer 2-deoxy-[18F]-fluoro-D-glucose ([18F]FDG) as a synthon for building complex glycans compatible with in vivo positron emission tomography (PET) imaging. Both chemical and enzymatic transformations of [18F]FDG represent promising ways to develop new imaging tools, given the wide availability of this tracer. For example, [18F]FDG can (1) be reduced with NaBH4 to produce the Enterobacteriaceae-targeted PET radiotracer 2-deoxy-[18F]-fluoro-D-sorbitol ([18F]FDS)4,5, (2) be derivatized with amines and other substituents to yield analyte-sensitive “caged” prodrugs6,7and (3) be employed as a prosthetic group to label drugs and peptide structures8,9. These approaches can be used to easily transform [18F]FDG into tracers targeting bacterial metabolism, the tumoral microenvironment, or specific oncologic targets, for example the prostate-specific membrane antigen (PSMA) found in prostate cancer.

[0330] Our previous studies describing molecular tools for bacterial infection have focused on PET tracers labeling the bacterial cell wall, including D-[3-11C]alanine10and D-[methyl-11C]methionine11–13, as well as the folic acid pathway using ^ ^-[11C]PABA14. In addition to maltose-derived PET radiotracers, there have been several elegant methods in the last decade applied to bacteria using siderophore-derived probes15,16, radiolabeled trimethoprim derivatives17, and [18F] sugar alcohols including 2-deoxy-2-[18F]fluorosorbitol ([18F]FDS4) and 2-deoxy-[18F]-fluoro-D-mannitol18. The most advanced radiotracer in bacteria-specific PETimaging is [18F]FDS, which has been studied in numerous advanced preclinical models of infection as well as in infected patients19. A recent innovation in [18F]FDS is its rapid, kit-based, and on-demand radiosynthesis from [18F]FDG20. One limitation of [18F]FDS is lack of sensitivity for gram-positive pathogens, which are major causes of musculoskeletal infections including vertebral discitis-osteomyelitis (VDO)21,22. Therefore, we and others have a sustained interest in developing [18F]-labeled PET tracers targeting S. aureus and other gram-positive organisms.

[0331] In the current study, we used readily available [18F]FDG to construct [18F]-labeled dimers in one step via chemoenzymatic syntheses. This approach contrasts with most chemical methods reported to generate [18F]glycans, which typically use a structurally complex / protected precursor, and [18F]fluoride incorporation via SN2 displacement at sterically amenable sites23. Based on prior work, a phosphorylase-catalyzed approach appeared feasible24–28, with several glucose-derived dimers highlighted in FIG.1A. Phosphorylases (glycosyltransferases, E.C.2.4) are enzymes that catalyze the addition of an inorganic phosphate group to a carbohydrate acceptor with breaking an O-glycosidic bond. This reaction can be run “in reverse” to construct disaccharides from D-glucose-1-phosphate and a glucose derivative (FIG.1B). This strategy might therefore be used to rapidly fabricate [18F]-labeled disaccharides from [18F]FDG. An analogous method is used for α- or β-galactose-1-phosphate, or α-mannose-1-phosphate.

[0332] The first radiotracer we aimed to synthesize using this enzymatic method was the maltose derived tracer 2-deoxy-[18F]-fluoro-maltose ([18F]FDM). Our interest in this probe was based on the potential specificity of microbial maltose metabolism29–31and the recent development of maltodextrin transporter-targeted imaging methods32–35. Maltodextrin (D-glucose units with α-1,4 glycosidic linkages) and its structural relatives are important energy sources for bacteria. These oligosaccharides are taken up by the maltodextrin transporter, which are present in both gram-positive and gram-negative bacterial species but not found in mammalian cells. In order to enzymatically obtain [18F]FDM from [18F]FDG, we focused on commercially available maltose phosphorylase (E.C.2.4.1.8), which has been used to synthesize a variety of disaccharides36–40.

[0333] In this work, we report the one step radiosyntheses of [18F]FDM and α-1,3-product, 2- deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), from easily accessible [18F]FDG, using maltose phosphorylase as catalyst. Both newly reported [18F]-labelled radiotracers were accumulated by important human pathogens including S. aureus and were specific to bacterial infection in vivo. This phosphorylase-catalyzed method was extended to additional positron-labeled disaccharidesof high biomedical interest including 2-deoxy-2-[18F]fluoro-trehalose ([18F]FDT), 2-deoxy-2- [18F]fluoro-laminaribiose ([18F]FDL), and 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC). Example 1: Maltose phosphorylase-catalyzed radiosynthesis of [18F]FDM from clinical [18F]FDG showed the α-1,3-product [18F]FSK as a minor product

[0334] Our initial goal was to demonstrate that the phosphorylase-catalyzed chemoenzymatic synthesis of an [18F] disaccharide from [18F]FDG was a viable alternative to conventional radiochemical approaches. Although several bacteria-specific glycans could be constructed chemoenzymatically, we focused on maltose-derived PET radiotracers given the strong literature precedent33–35. We therefore synthesized the [18F]FDG-derived, 2-position labeled [18F]FDM to target the bacterial maltose receptor. First, the precursor, ^-D-glucose-1-phosphate ( ^Glc1-P) was synthesized in three steps starting from acetobromo- ^-D-glucose with 75% overall yield (FIG.2A). Cyclotron-produced [18F]FDG was then added to a mixture of ^Glc1-P and maltose phosphorylase in citrate buffer (0.1M, pH=6.0), and stirred at 37oC for 20 minutes. The reaction led to the formation of [18F]FDM with 72% ± 4% decay-corrected radiochemical yield (RCY) and the ^-1,3-linked product [18F]FSK with 15%± 3% decay-corrected RCY (N = 25) (FIG.2B). Each [18F]-labelled product was isolated using semi-preparatory HPLC (FIG.2C) followed by formulation for subsequent in vitro and in vivo studies. Example 2: Phosphorylase-catalyzed radiosyntheses of [18F]FDT, [18F]FDL and [18F}FDC from clinical [18F]FDG

[0335] Several additional positron-labeled disaccharides might be used to image important human pathogens in vivo. The unique metabolism of the ^ ^ ^-1,1 linked disaccharide trehalose by Mycobacterium tuberculosis has been previously targeted for therapy and imaging41–44. The ^- linked disaccharides laminaribiose and cellobiose could potentially be leveraged for the metabolic imaging of fungal infections given the presence of ^-1,3 and ^-1,4 linkages in ^-D- glucans (BDG), found in the cell walls of fungi45–47. An existing clinical assay (serum and cerebrospinal fluid) detects fungal ^-1,3 linkages via the coagulation cascade of the horseshoe crab to detect invasive fungal infections caused by Aspergillus and Candida species48. With these potential imaging applications in mind, we attempted [18F]-radiosyntheses of the additional disaccharides indicated in FIG.1A. The use of trehalose-, cellobiose- and laminaribiose- phosphorylase in combination with [18F]FDG and adequate ^ ^ ^or ^-D-glucose-1-phosphate precursor led to the formation of [18F]FDT (decay-corrected RCY 62% ± 4%; N = 5), [18F]FDL (decay-corrected RCY 96% ± 3%; N = 5), and [18F]FDC (decay-corrected RCY 81% ± 7%; N =5) (FIG.s 3A-3B). Of note, direct radiosynthesis of [18F]FSK via sakebiose phosphorylase produced only a modest yield of the desired product (decay-corrected RCY 5% ± 3%; N = 5). The [18F]FSK tracer was therefore obtained via maltose phosphorylase as described above, for subsequent in vitro and in vivo studies. Example 3: In vitro studies using [18F]FDM and [18F]FSK showed high tracer accumulation in key human pathogens including S. aureus and A. baumannii

[0336] We hypothesized that the disaccharides [18F]FDM and [18F]FSK would have different microbial sensitivity from previously reported tracers, with accumulation in the key human pathogen S. aureus. We therefore studied these tracers in vitro to assess incorporation in a variety of clinically relevant bacteria and establish microbial specificity. The gram-positive bacteria S. aureus and the gram-negative Enterobacteriaceae Escherichia coli and Klebsiella pneumoniae were used to assess assimilation of [18F]FDM and [18F]FSK and compare to their uptake to that of reported PET tracers [18F]FDG and [18F]FDS (FIG.4A). The glucose derivative [18F]FDG, used frequently in the clinic for oncologic and neuroimaging applications, was incorporated into all three bacteria. In contrast, as expected [18F]FDS accumulated in E. coli and K. pneumoniae but not S. aureus4. Both [18F]FDM and [18F]FSK showed high incorporation in S. aureus and K. pneumoniae, similar to that of [18F]FDG but low uptake in E. coli. Further bacteria pathogens were tested with [18F]FDM and [18F]FSK (FIG.4B). Overall, the two radiotracers demonstrated similar sensitivities towards gram-positive and gram-negative bacteria. They showed high uptake in Staphylococcus epidermis, A. baumannii and Enterobacter cloacae, but low uptake in Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium and Proteus mirabilis. There was only background level of incorporation of [18F]FDM and [18F]FSK into heat-killed bacteria for all species studied. (FIG.8). [18F]FDM and [18F]FSK demonstrated increased bacterial accumulation over time in bacterial cultures (FIG.9), and showed a high degree of retention in “efflux” experiments (FIG.10). To determine if the differential uptake of [18F]FDM and [18F]FSK by bacteria reflected differential use of sugar transporters, S. aureus cultures were incubated with [18F]FDM and [18F]FSK in the presence of increasing concentrations of unlabeled maltose (FIG.4C), suggesting that they both use a maltodextrin transporter for uptake. To determine if there was strain-to-strain variability in tracer accumulation, we compared uptake in 4 different clinical isolates of methicillin-resistant S. aureus (MRSA) (FIG.4D). Three of the isolates exhibited similar levels of uptake for both tracers, whereas one isolate showed diminished uptake of [18F]FDM.Example 4: [18F]FDM and [18F]FSK is stable in human serum and its degradation in mouse serum can be abrogated by use of an α-glucosidase inhibitor.

[0337] In preparation for in vivo studies in mice and humans, we assessed the stability of [18F]FDM and [18F]FSK incubated with mouse and human serum using radio HPLC. In mouse serum, both radiotracers exhibited increasing time-dependent hydrolysis to [18F]FDG, while they remained stable in human serum. (FIG.11). This observation may be explained by the increased abundance of ^-glucosidase, which has been reported to hydrolyze maltodextrin-based tracers49, in murine versus human serum. The ^-glucosidase enzyme is predicted to degrade maltose from the non-reducing end, hydrolyzing [18F]FDM and [18F]FSK to glucose and [18F]FDG (FIG.12). In contrast, ^-amylase (EC 3.2.1.1), which is present in human serum, degrades maltodextrin from the reducing end. Unlike longer chain maltodextrin-based tracers (>3 units), [18F]FDM and [18F]FSK would be anticipated to be resistant to ^-amylase and thus stable in human serum49.

[0338] We therefore tested the ability of the ^-glucosidase inhibitors voglibose, acarbose and miglitol (FIG.13), which are commonly used as diabetes treatment, to prevent the ^- glucosidase-mediated degradation of [18F]FDM and [18F]FSK in mouse serum. First, we verified that voglibose and miglitol did not affect accumulation of [18F]FDM and [18F]FSK in culture- grown S. aureus, whereas acarbose did (FIG.14). We then compared inhibitor potency in mouse serum and found that voglibose was the most potent inhibitor to prevent [18F]FDM and [18F]FSK degradation (FIG.15). Together, these studies suggested that the concurrent administration of voglibose with [18F]FDM and [18F]FSK in murine studies allowed a better approximation of future human performance. Example 5: In vivo analysis of [18F]FDM and [18F]FSK in a murine model of bacterial infection demonstrated higher signal to background for the sakebiose-derived tracer

[0339] We first tested [18F]FDM and [18F]FSK in non-infected mice, to both assess tracer stability and to detect potential contributions of the normal microbiome to tracer signals. When voglibose was added to tracer injection, in vivo analysis of [18F]FDM and [18F]FSK in conventionally-raised mice10(N = 5), which have bacteria colonization of their gut, showed background signal only in kidney and bladder (FIG.s 16A-16B). In contrast, when voglibose was omitted from the injection of [18F]FDM and [18F]FSK, the resulted signal was similar to [18F]FDG, with high heart and brain uptake, demonstrating the utility of the inhibitor (FIG.s 17A-17C).

[0340] We next evaluated a murine model of acute bacterial infection to test whether [18F]FDM and [18F]FSK uptake could detect live bacteria in in vivo mouse models of infection. We chose the MRSA myositis model as it has been studied extensively in tracer development and used to compare tracer accumulation in infected tissues (harboring live bacteria) versus sterile inflammation (reflecting the host immune response)4,10,11,14. Mice were inoculated with live MRSA in the left shoulder and with a 10-fold higher dose of heat-killed MRSA in the right shoulder. Following tracer injection in the presence of voglibose, both [18F]FDM and [18F]FSK accumulated at the site of injection of live MRSA but not of heat-killed MRSA (N = 6 for each tracer) (FIG.5). The region-of-interest (ROI) analysis revealed [18F]FDM and [18F]FSK uptake at the side of live MRSA injection was 6.1 and 6.5-fold higher (P <0.0001) respectively than at the side of heat-killed MRSA injection. These results were further corroborated by ex vivo analysis (tissue harvesting and gamma counting) which showed that the mean [18F]FDM and [18F]FSK accumulation in tissues inoculated with live MRSA was respectively 3.8 and 4.7-fold higher than that seen for tissues inoculated with heat-killed bacteria (P = 0.001 for [18F]FDM and P value < 0.0001 for [18F]FSK) (FIG.s 18 and 19). In contrast, the glucose-transporter targeted parent tracer [18F]FDG accumulated equally at injection sites inoculated with either live or heat- killed MRSA, as quantified by in vivo and ex vivo analysis (FIG.20). We thus conclude that both the [18F]FDM and [18F]FSK tracers are specific for live versus heat-killed MRSA and have the potential to distinguish bacterial inflammation from sterile inflammation. However, as [18F]FSK proved to have better in vivo performance than [18F]FDM, it was chosen for testing in additional preclinical models. Example 6: Preclinical models of vertebral discitis-osteomyelitis and A. baumannii myositis suggest that [18F]FSK could be used in challenging clinical settings.

[0341] 2-deoxy-2-[18F]fluoroglucose, abbreviated [18F]FDG or FDG, is a radiopharmaceutical compound commonly used in positron emission tomography (PET).[1-3][18F]FDG can also serve as a central synthon for building glycans or disaccharide complexes, which are analogs for bacterial cell wall components[4]or energy sources for bacteria, respectively.[5-6]Given the wide availability of [18F]FDG and well-established enzymatic transformations of glucose analogues,[7-8]we recently studied the chemoenzymatic transformation of FDG to18F-radiolabeled disaccharide derivatives, including but not limited to ^-1,4 linked 2-deoxy-[18F]-fluoro-maltose ([18F]FDM), ^-1,3 linked 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), ^-1,3 linked 2-deoxy-2- [18F]-fluoro-laminaribiose ([18F]FDL). An efficient one-pot chemoenzymatic method wasutilized to obtain these derivatives with moderate to excellent yields (15-96%) via reverse phosphorolysis.

[0342] Stability assessment of [18F]FSK showed it stays intact in human serum after incubation for 2 hours. In vitro studies showed high accumulation in common human pathogens including S. aureus (ca.60 Bq per 106cells) and A. baumannii (ca.90 Bq per 106cells). Moreover, in contrast to [18F]FDG, which accumulated equally at injection sites inoculated with either live or heat-killed MRSA, [18F]FSK uptake at the site of live MRSA injection was 6.5-fold higher (P<0.0001) than on the side of the heat-killed MRSA injection.

[0343] Based on our encouraging results with MRSA in vitro and in the murine myositis model, we next studied a rat model of vertebral discitis-osteomyelitis (VDO) using [18F]FSK. Rats were inoculated with live S. aureus (Xen-29, bioluminescent strain) in the third intervertebral space from the base of the tail and with heat-killed S. aureus (Xen-29) in the fifth intervertebral space. The tail was imaged by luminescence emission using a Xenogen IVIS ^ 50 to verify the location and presence of the bacterial infection (FIG.6A). After 4 days, CT was used to image damage to the disc and bone, revealing the development of disc-space narrowing and end-plate degeneration, which mimics human bacterial spinal infections (FIG.6B). Imaging of the tail using [18F]FSK was performed at day 4 (N = 5) and day 10 (N = 3) following inoculation of bacteria (FIG.6C, FIG.s 21A-21C, and FIG.22). In both cases [18F]FSK accumulated at the infection site, with ROI analyses demonstrating 2.8-fold higher signals at day 4 (P <0.0001) and 3.1-fold higher signals at day 10 (P <0.0001), in the third intervertebral space versus the fifth intervertebral space (FIG.6D and FIG.22).

[0344] After showing the efficacy of [18F]FSK in vivo to detect S. aureus infection, we further studied A. baumannii in the murine myositis model, as it is a common cause of soft tissue infections in the battlefield. We injected A. baumannii as described above, and performed ^PET / CT following [18F]FSK injection. The tracer accumulated at the site of live A. baumannii injection into the left shoulder but not at the site of heat-killed A. baumannii injection into the right shoulder (FIG.6E). ROI analysis revealed [18F]FSK accumulation in the live A. baumannii injected muscle was 4.9-fold higher than at the heat-killed A. baumannii injected muscle (P <0.0001) (FIG.6F). Ex vivo analysis revealed that tracer uptake at the site of live A. baumannii injection was 3.9-fold higher compared to the site of heat-killed A. baumannii injection (P <0.0001) (FIG.23). Together, these studies employing two clinically important human bacterial pathogens, S. aureus and A. baumannii, in two different murine models that mimic challengingto treat human infections, demonstrate the potential of using [18F]FSK to image human bacterial infections in the clinic.

[0345] The bacterial strains included in this study are listed in Table 1. Table 1. Bacterial strains Strain Phenotype or Genotype Source or Reference S. aureus Wild-type ATCC 12600 of of of of

[0346] Discussion

[0347] Bacteria-specific metabolic pathways have been exploited by antimicrobial agents for decades, with infection imaging a more recent application of this approach. In recent years, numerous compelling methodologies have been further validated in patients most notably[18F]FDS, which is highly sensitive for Enterobacteriaceae and which can be efficiently radiosynthesized from the common tracer [18F]FDG20, making it a more practical tool for potential clinical use. To further impact infectious disease management in the clinic, we will require imaging tools with both straightforward radiosyntheses, and applicability to a broader range of bacteria, including the common human gram-positive bacteria S. aureus.

[0348] Tracers labeled with radionuclides with a shorter half-life [11C; t1 / 2 = 20 min]10–12,14or studied primarily in academic centers [89Zr; t1 / 2 = 78 hrs]50face significant challenges in the acute care / emergency setting, highlighting the need for infection imaging methods that might be more broadly useful. We therefore developed a chemoenzymatic method for pathogen-targeted PET radiotracers, benefitting from (1) the efficiency of chemoenzymatic reactions, versus standard PET radiochemical methods and (2) the general availability of [18F]FDG as a synthon. A major challenge in the chemical synthesis of positron-labeled carbohydrates is the short [18F] radionuclide half-life (109.7 min), requiring complex precursors, late-stage SN2 radiofluorination via [18F]-fluoride, and less sterically hindered labeling sites23. In contrast, the current report describes a rapid (20 min), one-step radiosynthesis using [18F]FDG and commercially available precursors, with high regioselectivity / stereoselectivity and potential metabolic advantages conferred via 2-position [18F] labeling. With the goal of developing an on- demand S. aureus-sensitive tracer, we used maltose phosphorylase to dimerize [18F]FDG into the α-1,4 and α-1,3 disaccharides [18F]FDM and [18F]FSK. The sakebiose-derived tracer showed outstanding performance characteristics in its ability to detect living bacteria in vivo including S. aureus.

[0349] Sakebiose (also known as nigerose) has been described as an “uncommon sugar,” and investigated for its potential as an alternative sweetener, oral probiotic and immunopotentiating therapy51–53. Most published work on sakebiose has focused on production of the disaccharide via enzymatic degradation of dextrans or synthesis using phosphorylases, whose activity may be modified via mutagenesis54–57. Little is known about its transport and metabolism, although the current study suggests that sakebiose (at least the described 2-[18F] derivative) shares transport mechanism(s) with maltose for the bacteria studied, since [18F]FDM and [18F]FSK incorporation were inhibited by cold maltose to a similar degree in vitro. Further studies are needed to better understand its mechanism of bacterial incorporation, which may drive the discovery of additional metabolic imaging tools. The described radiosynthesis of [18F]FSK does indicate that the chemoenzymatic construction of complex PET radiotracers from common synthons including [18F]FDG is feasible, to the extent that the required tracers, glycan-derived “acceptors,” andenzymes are available. Furthermore, the reported engineering of disaccharide phosphorylases suggests that changing the distribution of [18F] products obtained may be possible. In the current case, maltose phosphorylase could be altered to increase the radiochemical yield of the desired [18F]FSK tracer.

[0350] As stated previously, we believe that [18F]FDM and [18F]FSK have outstanding potential as clinical PET tracers, especially for the detection and monitoring of S. aureus infections. Both can be produced quickly and efficiently from widely available [18F]FDG, without the need for regioselective precursor modification and chemical protection / deprotection. Most interestingly, the 2-position18F substituted maltose derivative appears to confer numerous advantages over the 6-position 18F derivative originally reported by Namavari et al.34,58, in terms of stability and microorganism export. Although the 6-position is more chemically accessible, the corresponding 6-[18F] derivative is both more vulnerable to defluorination, and prohibits 6- position phosphorylation, which is a major mechanism of [18F]FDG retention (via hexokinase, E.C.2.7.1.1). Indeed, in terms of in vitro stability, bacterial export, and in vivo performance [18F]FDM and [18F]FSK more closely mimic the “second generation” PET tracer 6”-[18F]- fluoromaltotriose35,59. Explicit comparison between positron-labelled alpha-1,4 linked oligomers (maltose, maltotriose, and maltodextrin) would be helpful to guide clinical implementation and is the basis of ongoing laboratory efforts.

[0351] In conclusion, we have developed chemoenzymatic method for the radiosyntheses of [18F]-labelled disaccharides from the readily available precursor [18F]FDG. The strategy was used to generate both α- and β-linked [18F] disaccharides of high biomedical interest for pathogen-specific imaging, specifically positron-labeled derivatives of maltose, sakebiose, trehalose, laminaribiose and cellobiose. We anticipate that this approach may be used to construct other complex [18F] glycans and facilitate the on-demand radiosynthesis of PET radiotracers for infection and other diseases. Example 7: Materials and Methods

[0352] General Chemistry and Radiochemistry

[0353] Unless otherwise noted, all reagents were obtained commercially and used without further purification. Radioisotopes were generated in the UCSF radiopharmaceutical facility.

[0354] Synthesis of ^-D-glucose-1-phosphate ( ^Glc1-P)

[0355] ^-D-glucose-1-phosphate ( ^Glc1-P) was synthesized in three steps from 1-bromo-α-D- glucose tetraacetate (75 % overall yield, gram scale). For detailed methods and characterization of each compound, please see the Supporting Information.

[0356] General enzymatic radiosynthesis of [18F]FDC, [18F]FDL, [18F]FDT, [18F]FDM and [18F]FSK

[0357] In a 4 mL borosilicate vial containing a PTFE stir bar, phosphorylase (0.3-0.5 mg, 3-6 units) and ^ ^ ^or ^ ^ ^Glc1-P (6 mg, 20 mmol) were added. A solution of [18F]FDG (15-30 mCi) in citrate buffer (pH=6.0) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through a C18 light cartridge, and subsequently purified via semi-preparative HPLC using a YMC column Polyamine Pack II, 10 mm (73% MeCN / 27 % H2O). The [18F]-labelled disaccharide product was isolated in 5-7 mL fractions. The fractions were then diluted with MeCN (40 mL) before being passed through a Sep-pak Plus NH2 Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2gas, the tracer was eluted using a saline solution for direct formulation for in vitro or in vivo use. Radiochemical yields and purity of [18F]-product were confirmed by analytical HPLC. The synthesis and characterization of cold standards are described in the Supporting Information.

[0358] Uptake of [18F]FDM and [18F]FSK in gram-positive and gram-negative bacteria in vitro

[0359] S.S. epidermidis, E. faecalis, K. pneumoniae, E. coli, P. aeruginosa, A. baumannii, S. typhimurium, P. mirabilis and E. cloacae were grown overnight in lysogeny broth (LB) in a shaking incubator at 37 °C. Overnight cultures were diluted to an optical density at 600 nm (OD600) of 0.05 and grown to exponential phase (~ 0.4- 0.6). Bacterial cultures were incubated with 24 µCi of [18F]FDM and [18F]FSK at 37 °C for 90 minutes for uptake studies and also at 30 min and 60 min for temporal evaluation. After tracer incubation, 500 µL of the bacterial cultures were transferred to Spin-X LC 1.5 mL tubes (0.22 µm) and were centrifuged (6 min, 13200 rpm) to separate bacterial cells and supernatant. Bacterial cells were then washed 1x with phosphate buffered saline (PBS) to remove any tracer not taken up by bacteria. Heat-killed bacterial samples used as control were prepared by incubating the bacterial cultures at 90 °C for 30 min. Retained radiotracer within samples was then counted using an automated gamma-counter (Hidex). Blocking experiments were performed by adding cold maltose (0.01 mM to 10 mM) together with 24 µCi of [18F]FDM and [18F]FSK and following the same protocol. Efflux experiments were performed by incubating the bacteria with 24 µCi of[18F]FDM and [18F]FSK for 30 minutes, then pelleting the bacteria and replacing the media with fresh LB. The cultures were then incubated for an additional 30 min, then similar method was used to separate bacteria cells and supernatant. Radioactivity for both was counted using a gamma-counter (HIDEX) to obtain residual activity.

[0360] Animal experiments

[0361] All animal procedures were approved by the UCSF Institutional Animal Care and Use Committee and were performed in accordance with UCSF guidelines. CBA / J mice (female, 8-10 weeks old) and Sprague / Dawley rats (female, 10–12 weeks old) were used for the experiments. Mice and rats were housed in individually ventilated cages under normal diet in groups of 3 rats or 5 mice, with ad libitum access to food and water throughout the experiment. Prior to infection and during imaging the animals were anesthetized with 5% isoflurane. Mice and rats were inoculated with S. aureus Xen29, MRSA, K. pneumoniae, or heat-killed bacteria as described previously10. At different time points further specified below, the mice and rats were imaged using a Xenogen IVIS ^ 50 instrument or Inveon µPET-CT following the injection of [18F]FDM, [18F]FSK or [18F]FDG.

[0362] In vivo [18F]FDM and [18F]FSK dynamic imaging in a myositis mouse model

[0363] Mice inoculated with MRSA or A. baumannii (~ 2x107CFU) in the left deltoidmuscle and 10-fold higher bacterial load of heat-killed bacterial in the right deltoid muscle. After 12h, [18F]FDM or [18F]FSK was injected via teil vain (~ 100 μL, 200 μCi, containing 1 mg voglibose). The mice were then imaged by µPET-CT using the same protocol previously described10(90 min dynamic PET scan, 5 min CT). The resulting µPET-CT images were analyzed with Amide’s Medical Image Data Examiner as described below.

[0364] In vivo [18F]FSK dynamic imaging in a VDO rat model

[0365] S. aureus Xen29 was used to induce discitis in 5 Sprague / Dawley Rats (Charles River). Xen29 is a bioluminescent S. aureus strain that carries a stable copy of the Photorhabdus luminescens lux operon (luxABCDE) and was used in the study to verify successful bacterial inoculation in the third intervertebral space. Xen29 was grown overnight in LB containing 100 μg / L of kanamycin as aforementioned described and diluted in PBS to obtain the desired bacterial load for infection (~ 2x107colony forming units, CFU). Rats (N = 5) were inoculated with Xen29 live and heat-killed (10-fold higher bacterial load) in the third and fifth intervertebral space respectively. At days 0 and 2, the rats were imaged using a Xenogen IVIS ^ 50 imagingsystem to detect bioluminescence signal and confirm the infection. At days 4 and 10, [18F]FSK injection (~ 200 μL, 500 μCi, 5 mg Voglibose) was performed using a tail vein catheter. One hour post tracer injection, rats were transferred to the µPET-CT system (Siemens) and imaged using a 90 min dynamic PET acquisition scan followed by a 5 min µCT scan for attenuation correction and anatomical co-registration. Anesthesia was maintained during bioluminescence and µPET-CT imaging using 5% isofluorane. Resulting µPET-CT images were analyzed using AMIDE and %ID / cc was used for quantitative comparison. %ID / cc values were established via 8 mm3region of interest ROI’s using the spherical tool. Region of interest (ROI) analysis from resulting µPET-CT images was used to compare tracer performance. Resulting bioluminescence images were analyzed with Living Image Software 3.2.

[0366] Data analysis and statistical considerations

[0367] For synthesis, radiochemical yield incorporates decay-correction for18F (t1 / 2=109.7 min). In vitro data was normalized to CFU’s for sensitivity analysis to account for differential growth rates between organisms. All in vivo PET data were viewed using open-source AMIDE software. Quantification of uptake was performed by drawing spherical regions of interest (5-8 mm3) over indicated organs on the CT portion of the exam and expressed as percent injected dose per gram. All statistical analysis was performed using GraphPad Prism v 9. Data were analyzed using an unpaired two-tailed Student’s t-test. All graphs are depicted with error bars corresponding to the standard error of the mean.

[0368] Synthetic Procedures

[0369] All chemical reagents were purchased from commercial sources (Acros Organics, Alfa Aesar, AK Scientific & Sigma-Aldrich) and used without further purification unless otherwise stated. All separatory cartridges were purchased from Waters. Reactions were monitored by thin layer chromatography (TLC) on precoated (250 μm) silica gel 60 F254 aluminum sheets and visualized under a UV-254 lamp followed by staining with potassium permanganate. Flash chromatography was performed on silica gel (60A pore size).1H,13C,31P and19F NMR spectra were obtained on a Bruker Avance III HD 400 mHz instrument at the UCSF Nuclear Magnetic Resonance Laboratory and data were processed using MestReNova. Abbreviations are as follows: s (singlet), d (doublet), t (triplet), q (quartet), m (multiplet). High resolution mass spectra (HRMS) services were provided by University of California, Berkeley Spectrometry Facility. The18F labeled compounds were characterized by developing the compounds in different solvent systems on silica gel TLC plates on glass followed by imaging on a radio TLCscanner (Bioscan AR2000). Analytical HPLC was performed using a Waters pump equipped with a manual Rheodyne injector (1 mL loop), a refracted index (RI) detector and a RAD detector. The stationary phase was YMC-Pack Polyamine II column and the mobile phase of 73:27 acetonitrile / H2O at a flowrate of 1 mL / min. For semi prep HPLC, a YMC-Pack Polyamine II stationary phase was used with a mobile phase of 73:27 acetonitrile / H2O at a flowrate of 4 mL / min, The radioactivity of the bacterial pallets and filtrate were counted on a γ counter (Hidex Automatic Gamma Counter).

[0370] Synthesis of precursor: b-D-glucose-1-phosphate (bGlc1-P)

[0372] In a 250 mL Erlenmeyer flask, dibenzyl phosphate (6.8 g, 24.4 mmol) and tetrabutylammonium hydrogen sulfate (4.1 g, 12.2 mmol) were added and mixed with 120 mL of saturated NaHCO3aqueous solution. After stirring at room temperature for 10 min, the mixture was added to a 500 mL round bottom flask containing bromide S1 (5.0 g, 12.2 mmol) in DCM (120 mL). The biphasic reaction mixture was capped then stirred vigorously at room temperature for 72 hrs. Over the reaction time, the pH of the aqueous phase was maintained at pH 8-9 via addition of saturated NaHCO3 solution. The organic phase was extracted using ethyl acetate, before being washed with saturated NaHCO3, water, and brine. The organic phase was then dried with sodium sulfate, filtered, and evaporated under reduced pressure. The residue was purified via silica flash chromatography using 1% t-butanol in DCM (with a few drops of Et3N) as eluant to give pure phosphate S2 (5.2 g, 71% yield).

[0373] 1H NMR (400 MHz, CDCl3) δ 7.46 – 7.30 (m, 10H), 5.38 (t, J = 7.6 Hz, 1H), 5.23 (d, J = 9.4 Hz, 1H), 5.16 (d, J = 7.8 Hz, 1H), 5.14 (d, J = 6.3 Hz, 1H), 5.11 (d, J = 7.4 Hz, 2H), 5.04 (d, J = 7.1 Hz, 2H), 4.26 (dd, J = 12.5, 4.8 Hz, 1H), 4.14 (dd, J = 12.4, 2.1 Hz, 1H), 3.83 (ddd, J = 10.0, 4.8, 2.2 Hz, 1H), 2.06 (s, 3H), 2.03 (s, 6H), 1.92 (s, 3H).

[0374] 31P NMR (162 MHz, CDCl3) δ -3.20 (dd, J = 14.5, 7.3 Hz). The NMR values matched the reported literature.60

[0375] b-D-Glucose-1-phosphate (bGlc1-P)

[0376] In a 100 mL round bottom flask, phosphate S2 (0.5 mg, 0.82 mmol) was hydrogenated (14.7 psi) over 5% Pd / C (94.3 mg), 14 mL of EtOH and 9 mL of 10% NaHCO3for 16 hrs at room temperature. The mixture was then filtered then concentrated. The residue was transferred to a 25 mL round bottom flask, to which was added Et3N (0.4 mL) and a 1:1 mixture of MeOH / H2O (5.5 mL). The mixture was stirred for 16 hrs before being concentrated then diluted with H2O. The pH was lowered using Dowex 50W-X8 [H+] resin until pH 7, before residue was filtered then lyophilized to obtain b-D-glucose-1-phosphate (bGlc1-P), (201 mg, 80 % yield).

[0377] 1H NMR (400 MHz, D2O) δ 4.90 (t, J = 7.7 Hz, 1H), 3.91 (dd, J = 12.3, 2.1 Hz, 1H), 3.68 (dd, J = 12.3, 6.9 Hz, 1H), 3.56 – 3.47 (m, 2H), 3.34 (t, 1H), 3.31 (t, 1H).

[0378] 31P NMR (162 MHz, D2O) δ 2.29 (d, J = 7.7 Hz). The NMR values matched the reported literature.2

[0379] Synthesis of19F Standards:

[0380] 2-deoxy-2-[19F]fluoro-maltose and 2-deoxy-2-[19F]fluoro-sakebiose

[0381] , - glucose (30 mg, 0.16 mmol) were added to a 10 mL round bottom flask containing maltose phosphorylase (EC 2.4.1.8, Sigma Aldrich), (0.6 mg, 6 units) in 3 mL of aqueous citrate buffer solution (pH = 6.0). The mixture was stirred for 24 hrs at 37oC. The residue was diluted with MeCN then purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using 70% MeCN / 30 % H2O to yield 2-deoxy-2[19F]-fluoro-maltose (22 mg, 41% yield) and 2-deoxy-2- [19F]-fluoro-sakebiose (8 mg, 14% yield).

[0383] 2-deoxy-2-[19F]fluoro-cellobiose^ ^ ^ ^ ^ ^ ^ ^-D-glucose-1-phosphate ( ^Glc1-P) (30 mg, 0.1 mmol), 2-deoxy-2[19F]-fluoro-glucose (10 mg, 0.05 mmol) were added to a 5 mL round bottom flask containing cellobiose phosphorylase (EC 2.4.1.20), (0.4 mg, 6 units) in 1 mL of aqueous citrate buffer solution (pH = 6.0). The mixture was stirred for 24 hrs at 37oC. The residue was diluted with MeCN then purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using 70% MeCN / 30 % H2O to yield 2-deoxy-2[19F]-fluoro-cellobiose (10 mg, 59% yield).

[0385] 2-deoxy-2-[19F]fluoro-trehalose

[0386] ^ D- mg, , - (10 mg, 0.05 mmol) were added to a 5 mL round bottom flask containing trehalose phosphorylase (EC 2.4.1.64), (0.4 mg, 6 units) in 1 mL of aqueous citrate buffer solution (pH = 6.0). The mixture was stirred for 24 hrs at 37oC. The residue was diluted with MeCN then purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using 70% MeCN / 30 % H2O to yield 2-deoxy-2[19F]-fluoro-trehalose (8 mg, 47% yield).

[0388] 2-deoxy-2-[19F]fluoro-laminaribiose^ ^ ^ ^ ^ ^ ^ ^-D-glucose-1-phosphate ( ^Glc1-P) (30 mg, 0.1 mmol), 2-deoxy-2[19F]-fluoro-glucose (10 mg, 0.05 mmol) were added to a 5 mL round bottom flask containing laminaribiose phosphorylase (EC 2.4.1.31), (0.4 mg, 6 units) in 1 mL of aqueous citrate buffer solution (pH = 6.0). The mixture was stirred for 24 hrs at 37oC. The residue was diluted with MeCN then purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using 70% MeCN / 30 % H2O to yield 2-deoxy-2[19F]-fluoro-laminaribiose (5 mg, 29% yield).

[0390] NMR and HRMS analysis:maltose

[0392] 1H NMR (400 MHz, D2O) δ 5.39 – 5.30 (m, 3H), 4.83 (dd, J = 7.7, 2.3 Hz, 1H), 4.36 (ddd, J = 49.2, 9.6, 3.9 Hz, 1H), 4.20 – 3.96 (m, 3H), 3.89 (ddd, J = 10.0, 4.4, 2.4 Hz, 1H), 3.86 – 3.78 (m, 2H), 3.77 – 3.73 (m, 2H), 3.72 – 3.53 (m, 11H), 3.49 (ddd, J = 9.9, 3.9, 0.8 Hz, 2H), 3.34 (td, J = 9.5, 1.4 Hz, 2H).

[0393] 13C NMR (100 MHz, D2O) δ 99.57 (C1’ ^), 99.41 (C1’ ^), 93.39 (d, J = 23.1 Hz, C1 ^), 92.67 (d, J = 183.6 Hz, C2 ^), 89.95 (d, J = 186.0 Hz, C2 ^), 89.43 (d, J = 21.3 Hz, C1 ^), 75.96 (d, J = 14.6 Hz, C4 ^), 75.88 (d, J = 14.9 Hz, C4 ^), 74.58 (d, J = 6.5 Hz, C5 ^), 74.49 (d, J = 24.2 Hz, C3 ^), 72.77 (C5’ ^), 72.74 (C5’ ^), 72.67 (C3’ ^), 72.66 (C3’ ^), 71.66 (C2’ ^) ^ ^71.57 (d, J = 18.2 Hz, C3 ^), 71.56 (C2’ ^) ^ 69.71(C5 ^), 69.26 (C4’ ^, C4’ ^) ^ 60.49 - 60.31 (C6 ^, C6’ ^ ^ ^C6 ^ ^ ^C6’ ^).

[0394] 19F NMR (376 MHz, D2O) δ -199.90 – -200.12 (m), -200.24 (dd, J = 49.2, 13.7 Hz). The NMR values matched the reported literature.62HRMS (ESI) m / z calculated for C12H21FO10(M+Na) 367.1, found 367.1.[ o-sakebiose

[0396] 1H NMR (400 MHz, D2O) δ 5.37 (d, J = 3.9 Hz, 1H), 5.23 (t, J = 4.3 Hz, 2H), 4.85 (dd, J = 7.9, 2.7 Hz, 1H), 4.44 (ddd, J = 48.9, 9.4, 3.9 Hz, 1H), 4.13 (dt, J = 53.2, 9.6 Hz, 1H), 4.00 (dt, J = 13.1, 10.5 Hz, 1H), 3.87 (m, 1H), 3.83 – 3.57 (m, 15H), 3.46 (m, 3H), 3.35 (t, J = 9.4 Hz, 2H).

[0397] 13C NMR (100 MHz, D2O) δ 99.15 (C1’ ^), 99.09 (C1’ ^), 93.57 (d, J = 23.7 Hz, C1 ^), 91.43 (d, J = 185.7 Hz, C2 ^), 89.64 (d, J = 21.7 Hz, C1 ^), 88.71 (d, J = 188.2 Hz, C2 ^), 79.65 (d, J = 17.0 Hz, C3 ^), 77.06 (d, J = 17.0 Hz, C3 ^), 75.75 (C5 ^), 72.84 (C3’ ^), 72.83 (C3’ ^), 71.75 (C5’ ^), 71.71 (C5’ ^), 71.51 (C2’ ^), 71.46 (C2’ ^), 71.05 (C5 ^), 69.71 (d, J = 8.0 Hz, C4 ^), 69.53 (d, J = 7.7 Hz, C4 ^), 69.31 (C4’ ^ +C4’ ^), 60.34 - 60.10 (C6 ^, C6’ ^ ^ ^C6 ^ ^ ^C6’ ^).

[0398] 19F NMR (376 MHz, D2O) δ -196.49 (dd, J = 50.8, 15.1 Hz), -196.72 (dd, J = 48.9, 13.1 Hz). HRMS (ESI) m / z calculated for C12H21FO10(M+Na) 367.1, found 367.1.-

[0400] 1H NMR (400 MHz, D2O) δ 5.45 (d, J = 3.9 Hz, 1H), 4.93 (dd, J = 7.8, 2.4 Hz, 1H), 4.54 (d, J = 7.8 Hz, 2H), 4.48 (ddd, J = 49.5, 9.5, 3.9 Hz, 1H), 4.24 – 4.08 (m, 2H), 4.02 – 3.86 (m, 7H), 3.86 – 3.78 (dd, J = 12.4, 5.0 Hz, 1H), 3.78 – 3.69 (m, 4 H), 3.65 (ddd, J = 9.8, 4.9, 2.1 Hz, 1H), 3.58 – 3.38 (m, 6H), 3.33 (m, 2H).

[0401] 13C NMR (100 MHz, D2O) δ 102.45 (C1’ ^+ C1’ ^), 93.45 (C1 ^), 93.22 (C2 ^), 89.90 (C2 ^), 89.32 (C1 ^), 77.93 (C4 ^+C4 ^), 75.94 (C5’ ^+C5’ ^), 75.42 (C3’ ^+C3’ ^), 74.86 (C5 ^), 73.07 (C2’ ^+C2’ ^), 72.66 (C3 ^), 69.92 (C5 ^), 69.54 (C3 ^), 69.44 (C4’ ^+C4’ ^), 60.56 (C6’ ^+C6’ ^), 59.80 (C6 ^), 59.63 (C6 ^).

[0402] 19F NMR (376 MHz, D2O) δ -199.14 (ddd, J = 51.2, 15.2, 2.3 Hz), -199.33 (dd, J = 49.3, 13.5 Hz). The NMR values matched the reported literature.63HRMS (ESI) m / z calculated for C12H21FO10 (M+Na) 367.1, found 367.1.trehalose

[0404] 1H NMR (400 MHz, D2O) δ 5.44 (d, J = 3.9 Hz, 1H), 5.22 (d, J = 3.7 Hz, 1H), 4.51 (ddd, J = 49.0, 9.6, 3.9 Hz, 1H), 4.13 (dt, J = 13.2, 9.4 Hz, 1H), 3.95 – 3.72 (m, 7H), 3.67 (dd, J = 9.9, 3.7 Hz, 1H), 3.52 (t, J = 9.6 Hz, 1H), 3.46 (t, J = 9.4 Hz, 1H).

[0405] 13C NMR (100 MHz, D2O) δ 94.00 (C1’), 91.19 (d, J = 21.6 Hz, C1), 89.47 (d, J = 187.7 Hz, C2), 72.58 (C3’), 72.19 (C5+C5’), 71.08 (d, J = 17.4 Hz, C3), 70.93 (C2’), 69.55 (C4’), 69.02 (d, J = 8.0 Hz, C4), 60.46 (C6), 60.26 (C6’).

[0406] 19F NMR (376 MHz, D2O) δ -201.15 (dd, J = 49.0, 13.3 Hz). HRMS (ESI) m / z calculated for C12H21FO10 (M+Na) 367.1, found 367.1.-

[0408] 1H NMR (400 MHz, D2O) δ 5.46 (d, J = 3.8 Hz, 1H), 4.94 (dd, J = 7.9, 2.5 Hz, 1H), 4.68 (dd, J = 12.1, 8.0 Hz, 2H), 4.61 (ddd, J = 49.5, 9.4, 3.9 Hz, 1H), 4.30 (dt, J = 53.2, 9.1 Hz, 1H), 4.19 (dt, J = 12.6, 9.3 Hz, 1H), 4.09 (dt, J = 14.6, 8.7 Hz, 1H), 3.98 – 3.86 (m, 4H), 3.86 – 3.70 (m, 5H), 3.62 – 3.46 (m, 7H), 3.46 – 3.39 (m, 2H), 3.39 – 3.32 (m, 2H).

[0409] 13C NMR (100 MHz, D2O) δ 102.45 (C1’ ^+ C1’ ^), 93.45 (C1 ^), 92.87 (C2 ^), 90.17 (C2 ^), 89.61 (C1 ^), 81.65 (C3 ^), 79.23 (C3 ^), 75.89 (C5’ ^+C5’ ^), 75.70 (C5 ^), 75.54(C3’ ^+C3’ ^), 73.19 (C2’ ^+C2’ ^), 71.08 (C5 ^), 69.56 (C4’ ^+C4’ ^), 67.67 (C4 ^+C4 ^), 60.66 (C6’ ^+C6’ ^), 60.50 (C6 ^), 60.28 (C6 ^).

[0410] 19F NMR (376 MHz, D2O) δ -198.87 (dd, J = 51.0, 14.6 Hz), -199.20 (dd, J = 49.1, 12.7 Hz). HRMS (ESI) m / z calculated for C12H21FO10 (M+Na) 367.1, found 367.1.

[0411] Radiochemistry

[0412] Radiochemical procedure for 2-deoxy-[18F]-fluoro-D-glucose ([18F]FDG) production:

[0413] 2-deoxy-[18F]-fluoro-D-glucose ([18F]FDG) was produced in aqueous solution using standard methods.64

[0414] Radiochemical procedure for 2-deoxy-[18F]-fluoro-D-sorbitol ([18F]FDS) production:

[0415] 2-deoxy-[18F]-fluoro-D-sorbitol ([18F]FDS) was synthesized using procedure reported by Weinstein et al.65

[0416] Radiosynthesis of 2-deoxy-[18F]-fluoro-maltose ([18F]FDM) and 2-deoxy-2-[18F]- fluoro-sakebiose ([18F]FSK):

[0417] In a 4 mL borosilicate vial containing PTFE stir bar, maltose phosphorylase (EC 2.4.1.8, Sigma Aldrich), (0.3 mg, 3 units) and bGlc1-P (6 mg, 0.020 mmol) were added. A dose of [18F]FDG (10-15 mCi) in citrate buffer (0.1M, pH=6.0, 0.4-0.5 mL) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through C18 light cartridge, before being purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using mobile phase 73% MeCN / 27 % H2O. Both [18F]FDM and [18F]FSK were isolated in 5-7mL fractions. The fractions were then diluted with MeCN (40 mL) before being passed through Sep-pak Plus NH2 Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2 gas, they were eluted using saline solution for direct formulation before use in vitro or in vivo. [18F]FDM (RCY= 72% ± 4%, RCP=99%),[18F]FSK (15%± 3%, RCP=99%) (N=25). Chemical purity of both [18F]FDM and [18F]FSK were confirmed by analytical HPLC.

[0418] Radiosynthesis of 2-deoxy-2[18F]-fluoro-cellobiose ([18F]FDC):2.4.1.20, Prof. Bernd Nidetzky lab, Graz University of Technology), (0.3 mg, 3 units) and aGlc1- P (6 mg, 0.020 mmol) were added. A dose of [18F]FDG (10-15 mCi) in citrate buffer (0.1M, pH=6.0, 0.4-0.5 mL) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through C18 light cartridge, before being purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using mobile phase 73% MeCN / 27 % H2O. [18F]FDC was isolated in 5-7mL fractions. The fractions were then diluted with MeCN (40 mL) before being passed through Sep-pak Plus NH2 Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2gas, the tracer was eluted using saline solution for further analysis. [18F]FDC (RCY= 81% ± 7%, RCP=99%), (N=5). Chemical purity of [18F]FDC was confirmed by analytical HPLC.

[0420] Radiosynthesis of 2-deoxy-2[18F]-fluoro-trehalose ([18F]FDT):

[0421] In a 4 mL borosilicate vial containing PTFE stir bar, trehalose phosphorylase (EC 2.4.1.64, Prof. Tom Desmet lab, Ghent University), (0.3 mg, 3 units) and bGlc1-P (6 mg, 0.020mmol) were added. A dose of [18F]FDG (10-15 mCi) in citrate buffer (0.1M, pH=6.0, 0.4-0.5 mL) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through C18 light cartridge, before being purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using mobile phase 73% MeCN / 27 % H2O. [18F]FDT was isolated in 5-7mL fractions. The fractions were then diluted with MeCN (40 mL) before being passed through Sep-pak Plus NH2 Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2gas, the tracer was eluted using saline solution for further analysis. [18F]FDT (RCY= 62% ± 4%, RCP=99%), (N=5). Chemical purity of [18F]FDT was confirmed by analytical HPLC.

[0422] Radiosynthesis of 2-deoxy-2[18F]-fluoro-laminaribiose ([18F]FDL):(EC 2.4.1.31, Prof. Tom Desmet lab, Ghent University), (0.3 mg, 3 units) and aGlc1-P (6 mg, 0.020 mmol) were added. A dose of [18F]FDG (10-15 mCi) in citrate buffer (0.1M, pH=6.0, 0.4-0.5 mL) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through C18 light cartridge, before being purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using mobile phase 73% MeCN / 27 % H2O. [18F]FDL was isolated in 5-7mL fractions. The fractions were then diluted with MeCN (40 mL) before being passed through Sep-pak Plus NH2Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2 gas, the tracer was eluted using saline solution for further analysis. [18F]FDL (RCY= 96% ± 3%, RCP=99%), (N=5). Chemical purity of [18F]FDL was confirmed by analytical HPLC.

[0424] Radiosynthesis of 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK) with sakebiose phosphorylase:C 2.4.1.279, Creative Enzymes), (0.3 mg, 3 units) and bGlc1-P (6 mg, 0.020 mmol) were added. A dose of [18F]FDG (10-15 mCi) in citrate buffer (0.1M, pH=6.0, 0.4-0.5 mL) was directly transferred to the vial and the mixture was stirred at 37 °C for 20 min. The mixture was diluted with MeCN then filtered through C18 light cartridge, before being purified via semi prep HPLC (YMC-Pack Polyamine II, 250 X 10 mm) using mobile phase 73% MeCN / 27 % H2O. [18F]FSK was isolated in 5 mL fraction. The fraction was then diluted with MeCN (40 mL) before being passed through Sep-pak Plus NH2 Cartridge at 5 mL / min to trap each dimer product. After flushing the cartridge with air and N2gas, the tracer was eluted using saline solution for further analysis. [18F]FSK (RCY= 5% ± 2%, RCP=99%), (N=3). Chemical purity of [18F]FSK was confirmed by analytical HPLC.

[0426] Control Experiment: Reaction without enzyme:was same except presence of phosphorylase.Example 8: Sensing living bacterial in vivo using 2-deoxy-2[18F]-fluorosakebiose ([18F]FSK)

[0428] To show [18F]FSK uptake in numerous S. aureus strains, an ATCC (12600) organism and eight clinical isolates (4 MSSA; 4 MRSA) were studied using the same protocol as described in Example 6. The positive control [18F]FDG showed high uptake and negative control [18F]FDS showed no uptake. [18F]FSK showed the best performance among all chemoenzymatic tracers and higher accumulation than the positive control FDG (FIG.44A).

[0429] [18F]FSK was previously isolated by using either maltose phosphorylase with a radiochemical yield (RCY) of 15% as a side product, or commercial sakebiose phosphorylase (also designated as nigerose phosphorylase, NP) with a lower RCY of 5%. However recently we are able to prepare [18F]FSK exclusively with a significantly improved RCY of 43% (FIG.44B), using an NP provided by the Desmet lab. Due to the synthetic ease of [18F]FSK radiosynthesis, and the encouraging preclinical data obtained, we focus on rapid cartridge—based methods for [18F]FSK purification and automation. REFERENCES (1) Winkler, C. K.; Schrittwieser, J. H.; Kroutil, W. Power of Biocatalysis for Organic Synthesis. ACS Cent. Sci.2021, 7 (1), 55–71. (2) Kadokawa, J. Precision Polysaccharide Synthesis Catalyzed by Enzymes. Chem. Rev. 2011, 111 (7), 4308–4345. (3) Li, J.; Amatuni, A.; Renata, H. Recent Advances in the Chemoenzymatic Synthesis of Bioactive Natural Products. Curr. Opin. Chem. Biol.2020, 55, 111–118. 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Claims

WHAT IS CLAIMED IS:

1. A method of producing an [18F]-labeled oligosaccharide, the method comprising combining [18F]-fluoro-D-glucose ([18F]FDG), a phosphorylase and a phosphorylated monosaccharide selected from the group consisting of D-glucose-1-phosphate, D-galactose-1- phosphate, and D-mannose-1-phosphate in a reaction vessel, thereby producing said [18F]-labeled oligosaccharide.

2. The method of claim 1, wherein the phosphorylated monosaccharide is D- glucose-1-phosphate.

3. The method of claim 2, wherein the D-glucose-1-phosphate is α- D- glucose-1-phosphate.

4. The method of claim 2, wherein the D-glucose-1-phosphate is β- D- glucose-1-phosphate.

5. The method of claim 1, wherein the phosphorylated monosaccharide is D- galactose-1-phosphate.

6. The method of claim 5, wherein the D-galactose-1-phosphate is α- D- galactose-1-phosphate.

7. The method of claim 5, wherein the D-galactose-1-phosphate is β- D- galactose-1-phosphate.

8. The method of claim 1, wherein the phosphorylated monosaccharide is D- mannose-1-phosphate.

9. The method of claim 8, wherein the D-mannose-1-phosphate is α- D- mannose-1-phosphate.

10. The method of claim 8, wherein the D-mannose-1-phosphate is β- D- mannose-1-phosphate.

11. The method of claim 1, wherein the [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose and 1 to 5 additional saccharide monomer units.

12. The method of claim 11, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose and 1 additional saccharide monomer unit.

13. The method of claim 11, wherein said 1 to 5 additional saccharide monomer units comprise D-glucose, D-galactose, or D-mannose.

14. The method of claim 13, wherein the D-glucose is α- D-glucose or β- D- glucose.

15. The method of claim 13, wherein the D-galactose is α- D- galactose or β- D- galactose.

16. The method of claim 13, wherein the D-mannose is α- D- mannose or β- D- mannose.

17. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-4 glycosidic bond or a β- ^1-4 glycosidic bond.

18. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-3 glycosidic bond or a β- ^1-3 glycosidic bond.

19. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond.

20. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-glucose through an ^- ^1-1 glycosidic bond.

21. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-galactose through a β- ^1-3 glycosidic bond.

22. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-galactose through a β- ^1-4 glycosidic bond.

23. The method of any one of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-galactose through a ^- ^1-6 glycosidic bond or a β- ^1-6 glycosidic bond.

24. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-mannose through an β- ^1-3 glycosidic bond.

25. The method of claim 1, wherein said [18F]-labeled oligosaccharide comprises [18F]-fluoro-D-glucose linked to D-mannose through a β- ^1-4 glycosidic bond.

26. The method of claim 1, wherein said [18F]-labeled oligosaccharide is an [18F]-labeled disaccharide.

27. The method of claim 26, wherein the [18F]-labeled disaccharide is 2- deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy- 2-[18F]fluoro-trehalose ([18F]FDT), or 2-deoxy-2-[18F]fluoro-cellobiose ([18F]FDC), 2-deoxy-2- [18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro- gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro-allolactose, 2-deoxy-2-[18F]fluoro-melibiose, or 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose.

28. The method of claim 26, wherein the [18F]-labeled disaccharide is [18F]FDM, [18F]FSK, or [18F]FDT.

29. The method of claim 26, wherein the [18F]-labeled disaccharide is [18F]FDM and [18F]FSK.

30. The method of claim 1, wherein the [18F]-labeled oligosaccharide is [18F]FDM, [18F]FSK, [18F]FDC, [18F]FDT, 2-deoxy-2-[18F]fluoro- [18F]FDL, 2-deoxy-2- [18F]fluoro-isomaltose, 2-deoxy-2-[18F]fluoro-gentobiose, 2-deoxy-2-[18F]fluoro-lactose, 1,3- ^- galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro-allolactose, 2-deoxy-2- [18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy- 2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]-fluoro-maltotetraose, 2- deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2-deoxy-[18F]-fluoro- cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, or 2-deoxy-[18F]-fluoro-cellohexaose.

31. The method claim 1, wherein the phosphorylase enzyme is selected from the group consisting of maltose phosphorylase, sakebiose phosphorylase, trehalose phosphorylase, cellobiose phosphorylase, laminaribiose phosphorylase, α-glucan phosphorylase, β-glucan phosphorylase, isomaltose phosphorylase, gentiobiose phosphorylase, lactose phosphorylase, 1,3-β-galactosyl glucose phosphorylase, 1,6-β-galactosyl glucose phosphorylase, 1,6-α-galactosyl glucose phosphorylase, 1,4-β-mannosyl glucose phosphorylase, and 1,3-β- mannosyl glucose phosphorylase.

32. An [18F]-labeled oligosaccharide selected from the group consisting of 2- deoxy-[18F]-fluoro-maltose ([18F]FDM), 2-deoxy-2-[18F]-fluoro-sakebiose ([18F]FSK), 2-deoxy- 2-[18F]fluoro-laminaribiose ([18F]FDL), 2-deoxy-2-[18F]fluoro-isomaltose, 2-deoxy-2- [18F]fluoro-gentobiose, 1,3- ^-galactosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-2-[18F]fluoro- allolactose, 2-deoxy-2-[18F]fluoro-melibiose, 1,4- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 1,3- ^-mannosyl-2-deoxy-2-[18F]fluoro-glucose, 2-deoxy-[18F]-fluoro-maltotriose, 2-deoxy-[18F]- fluoro-maltotetraose, 2-deoxy-[18F]-fluoro-maltopentaose, 2-deoxy-[18F]-fluoro-cellotriose, 2- deoxy-[18F]-fluoro-cellotetraose, 2-deoxy-[18F]-fluoro-cellopentaose, and 2-deoxy-[18F]-fluoro- cellohexaose.

33. A pharmaceutical composition comprising an effective amount of the [18F]-labeled oligosaccharide of claim 32 and a pharmaceutically acceptable excipient.

34. The pharmaceutical composition of claim 33, further comprising an effective amount of an ^–glycosidase inhibitor.

35. The pharmaceutical composition of claim 34, wherein the ^–glycosidase inhibitor is voglibose, acarbose, miglitol or a combination thereof.

36. A method of detecting a disease in a subject in need thereof, the method comprising administering to the subject an effective amount of the [18F]-labeled oligosaccharide of claim 32, and detecting the [18F]-labeled oligosaccharide in said subject, thereby detecting the disease.

37. The method of claim 36, wherein the detecting comprises positron emission tomography–computed tomography (PET-CT) or positron emission tomography– magnetic resonance imaging (PET-MRI).

38. The method of claim 36, wherein the disease is a microbe infection.

39. The method of claim 38, wherein the microbe infection is a bacterial infection.

40. The method of claim 39, wherein the bacterial infection is a gram-positive bacterial infection.

41. The method of claim 39, wherein the bacterial infection is a gram-negative bacterial infection.

42. The method of claim 39, wherein the bacterial infection is caused by one or more of Staphylococcus epidermis, Acinetobacter baumannii, Streptococcus pneumonia, Enterobacter cloacae, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella typhimurium, Proteus mirabilis, Escherichia coli, Klebsiella pneumoniae Staphylococcus aureus, Staphylococcus epidermidis, or Enterococcus faecalis.

43. The method of claim 38, wherein the microbe infection is a fungal infection.

44. The method of claim 43, wherein the fungal is caused by one or more of Mycobacterium tuberculosis, Candida auris, Candida albicans, Aspergillus sp, Pneumocystis jirovecii, Criptococcus neoformans, or Taloromyces sp.

45. The method of any one of claims 36 to 44, wherein the microbe infection is a live microbe infection.

46. The method of claim 36, wherein the disease is myositis, vertebral discitis- osteomyelitis, pneumonia, or cancer.

47. The method of claim 36, wherein the method further comprises administering to the subject an effective amount of an ^–glycosidase inhibitor.

48. The method of claim 47, wherein the ^–glycosidase inhibitor is voglibose, acarbose, miglitol or a combination thereof.

49. The method of claim 36, wherein the method further comprises treating the disease by administering to the subject one or more of an anti-inflammatory agent, an antibiotic, an antifungal agent, and an anti-cancer agent.

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