The CSIR Biosciences researchers are developing a platform for robust techniques for glyco-profiling of therapeutic glycoproteins. The extracellular lipase (Lip2) (Aloulou, A et al;2007) was used as a model in establishing and evaluating glycan profiling techniques. The gene encoding Lip2 was cloned as a C-terminally His-tagged protein, expressed in Yarrowia lipolytica (Madzak, C et al;2004) and the glycan composition of the purified protein was analysed by HPLC and MALDITOF. The HPLC techniques was also used to profile the glycan composition of EPO expressed in Yarrowia lipolytica yeast.
Reference:
Kahari, D, Koorsen, G, Stoychev, S et al. 2008. Glycoprofiling of N-linked glycans of erythropoietin therapeutic protein expressed in Yarrowia lipolytica. Science Real and Relevant: 2nd CSIR Biennial Conference, CSIR International Convention Centre Pretoria, 17 & 18 November 2008, pp 1
Kahari, D., Koorsen, G., Stoychev, S. H., Chipeta, Z., Labuschagne, M., Van Zyl, P., & Nthangeni, B. (2008). Glycoprofiling of N-linked glycans of erythropoietin therapeutic protein expressed in Yarrowia lipolytica. CSIR. http://hdl.handle.net/10204/2866
Kahari, D, G Koorsen, Stoyan H Stoychev, Z Chipeta, M Labuschagne, P Van Zyl, and B Nthangeni. "Glycoprofiling of N-linked glycans of erythropoietin therapeutic protein expressed in Yarrowia lipolytica." (2008): http://hdl.handle.net/10204/2866
Kahari D, Koorsen G, Stoychev SH, Chipeta Z, Labuschagne M, Van Zyl P, et al, Glycoprofiling of N-linked glycans of erythropoietin therapeutic protein expressed in Yarrowia lipolytica; CSIR; 2008. http://hdl.handle.net/10204/2866 .