title: Surface display of glycosylated Tyrosinase related protein-2 (TRP-2) tumour antigen on Lactococcus lactis.
abstract: BACKGROUND
  The exploitation of the surface display system of food and commensal lactic acid bacteria (LAB) for bacterial, viral, or protozoan antigen delivery has received strong interest recently. The Generally Regarded as Safe (GRAS) status of the Lactococcus lactis coupled with a non-recombinant strategy of in-trans surface display, provide a safe platform for therapeutic drug and vaccine development. However, production of therapeutic proteins fused with cell-wall anchoring motifs is predominantly limited to prokaryotic expression systems. This presents a major disadvantage in the surface display system particularly when glycosylation has been recently identified to significantly enhance epitope presentation. In this study, the glycosylated murine Tyrosinase related protein-2 (TRP-2) with the ability to anchor onto the L. lactis cell wall was produced in suspension adapted Chinese Hamster Ovary (CHO-S) cells by expressing TRP-2 fused with cell wall anchoring LysM motif (cA) at the C-terminus.
  
  
  RESULTS
  A total amount of 33 μg of partially purified TRP-2-cA from ~6.0 g in wet weight of CHO-S cells was purified by His-tag affinity chromatography. The purified TRP-2-cA protein was shown to be N-glycosylated and successfully anchored to the L. lactis cell wall.
  
  
  CONCLUSIONS
  Thus cell surface presentation of glycosylated mammalian antigens may now permit development of novel and inexpensive vaccine platforms.
authors.affiliation: Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia. gs31997@mutiara.upm.edu.my.
authors.forename: Jeevanathan
authors.initials: J
authors.lastname: Kalyanasundram
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor Darul Ehsan, Malaysia. suetlin@upm.edu.my.
authors.forename: Suet Lin
authors.initials: SL
authors.lastname: Chia
authors.affiliation: Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia. adelene_song@yahoo.com.
authors.forename: Adelene Ai-Lian
authors.initials: AA
authors.lastname: Song
authors.affiliation: Department of Cell and Molecular Biology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia. raha@upm.edu.my.|Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia. raha@upm.edu.my.
authors.forename: Abdul Rahim
authors.initials: AR
authors.lastname: Raha
authors.affiliation: Cancer and Inflammation Program, Center for Cancer Research, National Cancer Institute, Frederick, MD, USA. YoungHow@mail.nih.gov.
authors.forename: Howard A
authors.initials: HA
authors.lastname: Young
authors.affiliation: Department of Microbiology, Faculty of Biotechnology and Biomolecular Sciences, Universiti Putra Malaysia, 43400 UPM, Serdang, Selangor Darul Ehsan, Malaysia. kyusoff@upm.edu.my.|Institute of Bioscience, Universiti Putra Malaysia, Serdang, Selangor Darul Ehsan, Malaysia. kyusoff@upm.edu.my.
authors.forename: Khatijah
authors.initials: K
authors.lastname: Yusoff
mesh_terms: D000818:Animals; D017951:Antigen Presentation / Q000235:genetics* / Q000502:physiology; D016466:CHO Cells; D002473:Cell Wall / Q000378:metabolism*; D002846:Chromatography, Affinity; D006224:Cricetinae; D003412:Cricetulus; D017931:DNA Primers; D019746:Intramolecular Oxidoreductases / Q000378:metabolism*; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism*; D051379:Mice; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000235:genetics
chemical_list: D017931:DNA Primers; D019746:Intramolecular Oxidoreductases; C042501:dopachrome isomerase