[Mannosylation of ligand binding domain 2 of HER-2/neu].
OBJECTIVE
To establish a new method for protein mannosylation, which will facilitate the study of antigen presentation mediated by mannose receptor.


METHODS
Recombinant HER-2/neu ligand binding domain 2 (LBD2) was purified through DEAE sepharose FF column and cobalt-based affinity resins. LBD2 protein was mannosylated in-vitro,and then analyzed by MALDI-TOF-MS and resorcinol-sulfuric acid method.


RESULTS
The purity of LBD2 obtained was 90%. Analysis of MALDI-TOF-MS and resorcinol-sulfuric acid method suggested LBD2 was mannosylated.


CONCLUSIONS
LBD2 neoglycoprotein has been obtained, which lays the foundation for the study of antigen uptake and presentation mediated by mannose receptor and the development of tumor vaccine.
Xu
Ming
M
Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Beijing 100850, China.
Guo
Ning
N
Wang
Hong-xia
HX
Li
Ai-ling
AL
Hu
Mei-ruo
MR
Shen
Bei-fen
BF
D001665:Binding Sites; D015153:Blotting, Western; D008024:Ligands; D008358:Mannose / Q000378:metabolism*; D008970:Molecular Weight; D018719:Erb-b2 Receptor Tyrosine Kinases / Q000737:chemistry* / Q000378:metabolism; D011994:Recombinant Proteins / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D008024:Ligands; D008358:Mannose; D018719:Erb-b2 Receptor Tyrosine Kinases; D011994:Recombinant Proteins