Humanizing N-linked glycosylation in Escherichia coli

Award Information
Agency:
Department of Health and Human Services
Branch
n/a
Amount:
$115,988.00
Award Year:
2009
Program:
SBIR
Phase:
Phase I
Contract:
1R43GM088905-01
Agency Tracking Number:
GM088905
Solicitation Year:
n/a
Solicitation Topic Code:
n/a
Solicitation Number:
n/a
Small Business Information
GLYCOBIA, INC.
GLYCOBIA, INC., 216 WAIT AVE, ITHACA, NY, 14850
Hubzone Owned:
N
Minority Owned:
N
Woman Owned:
N
Duns:
824602135
Principal Investigator:
ADAM FISHER
() -
Business Contact:
ADAM FISHER
() -
Research Institution:
n/a
Abstract
DESCRIPTION (provided by applicant): Protein-based therapeutics currently represent one in every four new drugs approved by the FDA and command a market in excess of 30 billion. The vast majority of therapeutic proteins require additional post-translation al modifications to attain their full biological and therapeutic function. One of the most important of these modifications, N-linked protein glycosylation, is predicted to affect more than half of all eukaryotic protein species and is often essential for proper folding, pharmacokinetic stability, systemic half-life, immunogenicity and overall efficacy for a large number of human therapeutic proteins. Since most bacteria do not glycosylate their own proteins, expression of many therapeutically relevant glyc oproteins, including antibodies, is relegated to mammalian cells. Unfortunately, mammalian cell culture suffers from a number of drawbacks including low volumetric productivity, product heterogeneity, high media cost, retroviral contamination and the relat ively long time required to generate stable cell lines. Recently, however, it has been reported that the Campylobacter jejuni asparagine-linked (Nlinked) protein glycosylation system can be functionally transferred into Escherichia coli, giving the recipie nt E. coli cells the ability to glycosylate proteins. Although the bacterial N-glycan is structurally different from its eukaryotic counterparts, it stands to reason that such glyco-enabled E. coli could be engineered to perform sequential glycosylation re actions that mimic the early processing of N-glycans in humans and other higher mammals. Such an accomplishment would open the door for performing complex human-like protein glycosylation in bacteria. Thus, the long-term goal of this research project is to humanize the N-linked protein glycosylation process in E. coli for the routine production of authentic human glycoproteins in this simple host organism. Towards this goal, the objective of this particular application is to begin the early stages of hum anizing the bacterial glycosylation system via glycan engineering (specific aim 1) and site-specific transfer of novel glycans onto target proteins (specific aim 2). The specific hypothesis behind the proposed research is that reconstitution of a eukaryot ic N-glycosylation pathway in E. coli will result in N-glycoproteins with structurally homogeneous human-like glycans. These studies are significant because they should (i) provide the necessary genetic tools to help elucidate the crucial role of glycosyla tion in a myriad of biological phenomena and (ii) enable the biotechnological synthesis of novel glycoconjugates and potential immunostimulating agents for research, industrial and therapeutic applications. PUBLIC HEALTH RELEVANCE: N-linked glycosyl ation is predicted to affect more than half of all eukaryotic protein species and is often essential for proper folding, pharmacokinetic stability, systemic half-life, immunogenicity and overall efficacy for a large number of therapeutically relevant prote ins (e.g., human erythropoietin). Since most bacteria do not glycosylate their own proteins, expression of many therapeutically relevant glycoproteins, including antibodies, is relegated to mammalian cells despite the fact that mammalian cell culture suffe rs from a number of well-documented drawbacks. Therefore, the focus of these studies is the development of glyco-engineered bacteria that are capable of producing authentic human glycoproteins.

* information listed above is at the time of submission.

Agency Micro-sites


SBA logo

Department of Agriculture logo

Department of Commerce logo

Department of Defense logo

Department of Education logo

Department of Energy logo

Department of Health and Human Services logo

Department of Homeland Security logo

Department of Transportation logo

Enviromental Protection Agency logo

National Aeronautics and Space Administration logo

National Science Foundation logo
US Flag An Official Website of the United States Government