Calpastatin autoantibodies: detection, epitope mapping, and development of a specific peptide ELISA.
Calpastatin autoantibodies: detection, epitope mapping, and development of a specific peptide ELISA.
复制标题
钙蛋白酶抑制剂自身抗体：检测、表位作图和特定肽 ELISA 的开发。
作者：
U. Schlosser;K. Lackner;C. Scheckenhofer;G. Schmitz
在一名有关节炎和血栓形成病史的53岁女性患者中检测到抗钙蛋白酶抑制蛋白（CAST）的自身抗体。通过筛选表达cDNA文库、阳性克隆的序列分析和随后的针对重组抗原的Western印迹来确定自身抗体的特异性。由于Western印迹缺乏令人满意的重现性，建立了抗CAST抗体的ELISA。24份Western blot阳性血清识别的主要抗原表位位于C端27个氨基酸内。因此，ELISA基于代表这些氨基酸的合成肽。用阳性参考血清的系列稀释液校准测定。低滴度和高滴度样本的批内精密度较高，CV约为4%。低和中等滴度血清的批间精密度（CV）为5.6-8.2%[10-60个任意单位（Au），其中阳性参考血清的1：100稀释度= 100 Au，1：1000稀释度= 10 Au等]，其随着滴度的增加而增加（> 60 Au）。在205名健康献血者中，平均值+3 SD（对数转换后）为30 Au;在138名住院患者中，2.9%的患者观察到较高值。新开发的ELISA将是一个有用的工具，为进一步的临床研究的协会抗CAST抗体的疾病，因为它允许快速和可重复的分析患者血清。
Autoantibodies against calpastatin (CAST) were detected in a 53-year-old female patient with a history of arthritis and thrombosis. The specificity of the autoantibodies was determined by screening expression cDNA libraries, sequence analysis of positive clones, and subsequent Western blotting against recombinant antigen. Because the Western blot lacked satisfactory reproducibility, an ELISA for anti-CAST antibodies was established. The major epitope recognized by 24 Western blot-positive sera was located within the C-terminal 27 amino acids. The ELISA was therefore based on a synthetic peptide representing these amino acids. The assay was calibrated with serial dilutions of a positive reference serum. Intraassay precision is high with a CV of approximately 4% for low- and high-titer samples. Interassay precision (CV) was 5.6-8.2% for sera with low and intermediate titers [10-60 arbitrary units (AU), where 1:100 dilution of the positive reference serum = 100 AU, 1:1000 dilution = 10 AU, etc.], which increases with higher titers ( > 60 AU). Among 205 healthy blood donors, the mean + 3 SD (after logarithmic transformation) was 30 AU; higher values were seen in 2.9% of 138 hospitalized patients. The newly developed ELISA will be a useful tool for further clinical studies on the association of anti-CAST antibodies to disease, because it permits rapid and reproducible analysis of patient sera.
通过不育患者血清鉴定的人类睾丸 cDNA：免疫避孕开发的分子生物学方法。
DOI：
10.1071/rd9940297
发表时间：
1994
期刊：
Reproduction, fertility, and development
影响因子：
--
作者：
Liang,ZG;O'Hern,PA;Yavetz,B;Yavetz,H;Goldberg,E
通讯作者：
Goldberg,E
通过由激肽原和 S-(3-硝基-2-吡啶硫基)-半胱氨酸的硫醇蛋白酶抑制序列衍生的合成肽抑制钙蛋白酶来调节凝血酶诱导的血小板聚集。
DOI：
10.1111/j.1432-1033.1993.tb17916.x
发表时间：
1993
期刊：
European journal of biochemistry
影响因子：
--
作者：
Puri,RN;Matsueda,R;Umeyama,H;Bradford,HN;Colman,RW
通讯作者：
Colman,RW