Todd L. Lowary Organic and Carbohydrate Web site: http://www.chem.ualberta.ca/~lowary/ Email: tlowary@ualberta.ca Phone: 780-492-1861

Research keywords: Organic synthesis, carbohydrate chemistry, glycoimmunochemistry, carbohydrate biochemistry. R&D capabilities: Synthesis of organic compounds particularly those containing carbohydrates Techniques and instrumentation: NMR spectroscopy, computational methods Examples of industrial collaborations and commercialization: • Carbohydrate Arrays as Diagnostics Two different carbohydrate arrays have been developed. One is for use in monitoring the immune response following an ABO-incompatible organ transplant and is currently being tested by clinicians. A second is being developed as a tool for probing the role of carbohydrates in the progression of tuberculosis and has applications in novel diagnostics for this disease. • Custom synthesis of oligosaccharides and other carbohydrate-containing molecules The laboratory has nearly two decades experience making complex carbohydrates, which have a range of applications including vaccine, assay and diagnostic development. In conjunction with the Alberta Glycomics Centre and Canadian Glycomics Network, we seek partnerships where our expertise can be used to solve important industrial challenges. Licensing opportunities: representative patents and invention reports: 1) Patent title: Methods and Systems for Inducing Immunogenic Tolerance to Non-Self

Antigens, International PCT Application CA2009/001814, filed on May 12, 2009. Potential applications: ABO-incompatible organ transplantation. 2) Patent title: Method and System for ABO Compatible Blood Type Matching. PCT Application

CA2012/050602. Filed August 30, 2012. Potential applications: ABO-incompatible organ transplantation, blood typing.

Todd L. Lowary

17

CS-35 CS-40 906.4321 Figure 2. Screening of 55-member glycan array against three anti-LAM antibodies; CS-35, CS-40 and 906.4321. Each series of triplicate spots represents a different structurally-defined glycan; those binding to the protein appear as green spots. The array provides a rapid (less than one day) method for determining the specificity of antibodies, as well as other proteins, that bind to MCW glycans. Projec t 2. Deve lopment o f nove l approaches for the synthes is o f campylobacter ce l l wal l g lycans to fa-c i l i tate biosynthet i c and structure/funct ion studies .

Context Campylobacter jejuni is an important food-borne pathogen that can cause severe enteritis and is a leading

cause of diarrheal disease.13 Infections by C. jejuni are also linked to Guillain–Barré syndrome, a neurologi-cal disorder that results in paralysis of varying severity and which is sometimes fatal. 14 The surface of all campylobacters is functionalized with capsular polysaccharide (CPS), the production of which is required for virulence.15 For the past several years we have been interested in the function and biosynthesis of cam-pylobacter CPS and have initiated a program dedicated to synthesizing various fragments of them. These synthetic investigations will provide compounds that can be used in biosynthetic or structure/function studies.16 Information derived from such investigations could lead ultimately to novel therapeutics to treat-ing food-borne diseases or strategies for preventing or mitigating the effect of these infections.

Campylobacter CPSs vary from strain-to-strain and possess a dizzying array of structural diversity (see Figure 3 for representative examples). In particular, they are characterized by unusual furanose and/or hep-tose residues, often decorated with methyl and/or methyl phosphoramidate (MeOPN) moieties.17 This rich array of structures will serve as a platform upon which significant new chemistry can be developed and re-fined. This project will thus: 1) serve as an important testing ground, and motivation, for developing new synthetic methods and approaches, and 2) provide a library of compounds for our collaborative biosyn-thetic studies with C. Szymanski characterizing the C. jejuni glycosyltransferases and MeOPN-transferases being expressed in her lab at the University of Alberta.

OHO O

HO O

O

NHAcOH

OPO

H3COH2N

OOO HO

NHO

O

HOOH

HO

H3COHO

HO OH

HS:2(Campylobacter jejuni)

HS:1(Campylobacter jejuni)

O

O

OO

OHPO

O

O

OOH

O

HO

HO

OH

OO

HO HO

OHO

PO

H3COH2N

PO

H3COH2N

OOO

HO

O

HO O

OH

O

HO

OH

HS:41 (Campylobacter jejuni)

OHHO

OHO

HO

HO

OHO OO

OOHOAcHN

OHO

O OH

HO

O

HOP

O

O

ATCC35221 (Campylobacter lari) Figure 3. Examples of CPS repeating unit structure from of various campyloacter strains. Methodology

To date, we have focused on the HS:2 serotype, and we have developed new methods for synthesizing heptoses18 and for introducing MeOPN motifs into glycans.19 We will use the considerable expertise we have developed in this area to expand the scope of the program. In particular, we will synthesize the re-peating units of all known C. jejuni CPSs, as well as those from the related organisms C. lari and C. coli. 20

Judiciously chosen fragments of these glycans will also be synthesized. The major challenges in synthesiz-ing these compounds are: 1) preparation of the unusual sugar residues (usually heptoses) and 2) installation of the MeOPN groups. We propose the de novo synthesis of the unusual heptose residues from a known compound 6 (see example in Figure 4A).21 An approach we have developed for introducing MeOPN groups, using a P(V) reagent, will be used.19 We will also explore introducing these groups via a P(III) spe-

Screening of a monoclonal antibody against a tuberculosis carbohydrate array