Wakarchuk Lab

GlycoNet Project

Wakarchuk lab researchers are engineering E. coli cells to complete human-like glycosylation functions on human therapeutic proteins
Dr. Wakarchuk is the chair and a professor in the Department of Chemistry and Biology of Ryerson University in Toronto, ON, Canada

A national research network harnessing the power of glycomics (carbohydrates) to deliver solutions to important health issues & improve the quality of life of Canadians
Includes more than 110 researchers in 28 institutions across Canada
Five Research themes:
- Antimicrobials
- Rare Genetic Diseases
- Diabetes and Obesity
- Chronic Disease
- Therapeutic Proteins and Vaccines

It is the most prevalent and diverse protein modification
Over 70% of human proteins undergo this process
It is the addition of a carbohydrate unit or chain to a specific location on a protein
E. coli are not capable of glycosylation
Sialylation is a type of glycosylation involving the addition of sialic acid, a sugar

http://www.creativebiomart.net/Protein-Glycosylation-Service.htm

Glycosylation of therapeutic proteins mimics natural human structures
- Leads to reduced side effects in the patient
Sialylation improves the circulatory half-life of therapeutic proteins in the body
- They last longer which leads to reduced dose frequency in the patient
Achieving glycosylation (and sialylation) of therapeutic proteins in E. coli will reduce production costs making them a more accessible form of treatment

Steps

Target protein is isolated from a human cell and the gene encoding it is inserted into a plasmid
A plasmid containing human-like glycosylation machinery is made
Both engineered recombinant plasmids are inserted into E. coli by the process of transformation
Transformed bacteria with both plasmids are isolated and grown in broth
E. coli cells are lysed and the glycosylated human therapeutic protein is isolated
Therapeutic protein undergoes testing to determine dosage and half-life