Detection of anti-topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system.
Detection of anti-topoisomerase I antibodies using a full length human topoisomerase I recombinant protein purified from a baculovirus expression system.
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使用从杆状病毒表达系统纯化的全长人拓扑异构酶 I 重组蛋白检测抗拓扑异构酶 I 抗体。
DOI：
10.1111/j.1365-2249.1995.tb03655.x
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发表时间：
1995
影响因子：
4.6
通讯作者：
McHugh,NJ
中科院分区：
文献类型：
--
作者：
Whyte,J;Earnshaw,WC;Champoux,JJ;Parker,LH;Stewart,L;Hall,ND;McHugh,NJ
拓扑异构酶I（topo I）是一种主要的系统性硬化症（SSc）相关自身抗原。将在重组杆状病毒转移载体中编码全长人topo I的cDNA构建体用于感染培养物中的昆虫细胞，从中纯化重组蛋白。使用重组蛋白的ELISA评价340例血清，包括134例SSc患者的血清，其中33例通过免疫扩散检测到抗拓扑I抗体。获得了高产率的预期分子量和催化活性的纯拓扑异构体I。重组拓扑酶I ELISA检测抗拓扑酶I抗体的敏感性为92%，特异性为98%，这些抗体几乎只存在于SSc患者中。因此，证实了在真核系统中表达人自身抗原用于诊断目的的潜在优势。
Topoisomerase I (topo I) is a major systemic sclerosis (SSc)-associated autoantigen. A cDNA construct encoding full length human topo I in a recombinant baculovirus transfer vector was used to infect insect cells in culture from which recombinant protein was purified. An ELISA using recombinant protein was evaluated in 340 sera including sera from 134 patients with SSc, of whom 33 had anti-topo I antibodies detected by immunodiffusion. A high yield of pure topo I of expected molecular mass and catalytic activity was obtained. The recombinant topo I ELISA was 92% sensitive and 98% specific in detecting anti-topo I antibodies which were present almost exclusively in patients with SSc. Therefore, the potential advantages of expressing human autoantigens in eukaryotic systems for diagnostic purposes were confirmed.