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Glycosylation of recombinant human thyroid peroxidase ectodomain of insect cell origin has little effect on recognition by serum thyroid peroxidase antibody

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Chinese Medical Journal 2013年第15期126卷 2907-2911页

作者： LILT Ming-ming LI Qing ZHAO Lan-lan GAO Ying HUANG You-yuan LU Gui-zhi GAO Yan-ming GUO Xiao-hui SHI Bing-yin Peking Univ Hosp 1 Dept Endocrinol Beijing 100034 Peoples R China Chinese Acad Sci Inst Microbiol CAS Key Lab Pathogen Microbiol & Immunol Beijing 100101 Peoples R China Xi An Jiao Tong Univ Sch Med Affiliated Hosp 1 Dept Endocrinol Xian 710061 Shaanxi Peoples R China

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, pudfied 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.

关键词： thyroid peroxidase baculovirus expression vector system sialic acid galactose fucose

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Isolation and Characterization of Novel Insecticidal cryl-Type Genes from Bacillus thuringiensis K1 Strains

Isolation and Characterization of Novel Insecticidal cryl-Ty...

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“环境与健康”学术研讨会

作者：李明顺 Jae Young Choi Jong Yul Roh Yeon Ho Je State Key Laboratory of Agricultural Microbiology Huazhong Agricultural UniversityWuhanChina College of Agriculture and Life Sciences Seoul National UniversitySeoulKorea

To identify novel cryl-type insecticidal genes,56 lepidopteran-specific Bacillus thuringiensis (Bt) strains were isolated from *** oligonucleotide primers, K5un2 and K3un2,were designed and used to amplify all known c... 详细信息

关键词： Bacillus thuringiensis Kl RFLP cryl-type gene inverse PCR insecticidal activity baculovirus expression vector system

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