|Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody|
|Liu Ming-ming1; Li Qing2; Zhao Lan-lan1; Gao Ying1; Huang You-yuan1; Lu Gui-zhi1; Gao Yan-ming1; Guo Xiao-hui1; Shi Bing-yin3|
|关键词||thyroid peroxidase baculovirus expression vector system sialic acid galactose fucose|
|刊名||CHINESE MEDICAL JOURNAL|
|WOS标题词||Science & Technology|
|类目[WOS]||Medicine, General & Internal|
|研究领域[WOS]||General & Internal Medicine|
|关键词[WOS]||HIGH-LEVEL EXPRESSION ; HASHIMOTOS-THYROIDITIS ; BACULOVIRUS SYSTEM ; DISEASE ; AUTOANTIBODIES ; PURIFICATION ; SUBCLASSES|
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.
|项目编号||7082096 ; 2011011 ; BMU20110266 ; 201002002 ; Z111107058811079|
|资助机构||Beijing Natural Science Foundation ; Beijing Nova Program ; Program for New Century Excellent Talents in University ; Sector Funds of Ministry of Health ; Beijing Special Clinical Application Fund|
|作者单位||1.Peking Univ, Hosp 1, Dept Endocrinol, Beijing 100034, Peoples R China|
2.Chinese Acad Sci, Inst Microbiol, CAS Key Lab Pathogen Microbiol & Immunol, Beijing 100101, Peoples R China
3.Xi An Jiao Tong Univ, Sch Med, Affiliated Hosp 1, Dept Endocrinol, Xian 710061, Shaanxi, Peoples R China
|Liu Ming-ming,Li Qing,Zhao Lan-lan,et al. Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody[J]. CHINESE MEDICAL JOURNAL,2013,126(15):2907-2911.|
|APA||Liu Ming-ming.,Li Qing.,Zhao Lan-lan.,Gao Ying.,Huang You-yuan.,...&Shi Bing-yin.(2013).Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody.CHINESE MEDICAL JOURNAL,126(15),2907-2911.|
|MLA||Liu Ming-ming,et al."Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody".CHINESE MEDICAL JOURNAL 126.15(2013):2907-2911.|