PMID- 23924466
OWN - NLM
STAT- MEDLINE
DCOM- 20140321
LR  - 20171116
IS  - 2542-5641 (Electronic)
IS  - 0366-6999 (Linking)
VI  - 126
IP  - 15
DP  - 2013
TI  - Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell 
      origin has little effect on recognition by serum thyroid peroxidase antibody.
PG  - 2907-11
AB  - BACKGROUND: Thyroid peroxidase (TPO) is an important autoantigen in Hashimoto's 
      thyroiditis (HT), and almost all epitopes are located in TPO ectodomain. The 
      glycosylation of TPO might contribute to breaking self-tolerance, therefore, 
      purified glycosylated recombinant TPO ectodomain is prerequisite of elucidating 
      its role in the pathogenesis of HT. The aim of our study was to investigate 
      whether the glycosylation has influence on the antigenic determinants of 
      recombinant TPO. METHODS: Bac-to-Bac baculovirus expression system was used to 
      generate recombinant human TPO ectodomain. The antigenicity was analyzed by 
      antigen specific enzyme-linked immunosorbant assays (ELISAs). The glycosylation 
      of recombinant human TPO ectodomain of High Five insect cell origin was detected 
      by lectin-ELISAs. RESULTS: TPO ectodomain was recovered from the culture media as 
      a soluble protein, and it was fused with a hexahistidine tag which allowed 
      purification by nickel-affinity chromatography. The recombinant TPO ectodomain 
      could be recognized by all the 54 HT patients and three TPO monoclonal 
      antibodies. Fucose, sialic acid and galactose were all detected on the 
      recombinant TPO ectodomain. Sera TPOAb binding decreased slightly after 
      non-specific deglycosylation of TPO by periodic acid. CONCLUSIONS: High Five 
      insect cells derived recombinant human TPO ectodomain had N-glycosylation sites, 
      which might have little effect on recognition by serum TPOAb.
FAU - Liu, Ming-ming
AU  - Liu MM
AD  - Department of Endocrinology, Peking University First Hospital, Beijing 100034, 
      China.
FAU - Li, Qing
AU  - Li Q
FAU - Zhao, Lan-lan
AU  - Zhao LL
FAU - Gao, Ying
AU  - Gao Y
FAU - Huang, You-yuan
AU  - Huang YY
FAU - Lu, Gui-zhi
AU  - Lu GZ
FAU - Gao, Yan-ming
AU  - Gao YM
FAU - Guo, Xiao-hui
AU  - Guo XH
FAU - Shi, Bing-yin
AU  - Shi BY
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - China
TA  - Chin Med J (Engl)
JT  - Chinese medical journal
JID - 7513795
RN  - 0 (Antibodies, Monoclonal)
RN  - 0 (Epitopes)
RN  - 0 (Recombinant Proteins)
RN  - EC 1.11.1.8 (Iodide Peroxidase)
SB  - IM
MH  - Animals
MH  - Antibodies, Monoclonal/immunology
MH  - Baculoviridae
MH  - Enzyme-Linked Immunosorbent Assay
MH  - *Epitopes
MH  - Glycosylation
MH  - Humans
MH  - Insecta/cytology
MH  - Iodide Peroxidase/*immunology
MH  - Recombinant Proteins
EDAT- 2013/08/09 06:00
MHDA- 2014/03/22 06:00
CRDT- 2013/08/09 06:00
PHST- 2013/08/09 06:00 [entrez]
PHST- 2013/08/09 06:00 [pubmed]
PHST- 2014/03/22 06:00 [medline]
PST - ppublish
SO  - Chin Med J (Engl). 2013;126(15):2907-11.