Seltsam Axel, Grueger Daniela, Blasczyk Rainer, Flegel Willy A
German Red Cross Blood Donor Service NSTOB, Institute Springe, Springe, Germany.
Transfusion. 2009 Oct;49(10):2090-6. doi: 10.1111/j.1537-2995.2009.02255.x. Epub 2009 Jun 23.
Identification of antibodies against high-prevalence Scianna (Sc; ERMAP) antigens, like Sc1 and Sc5, is difficult and may incur delays in blood procurement and costs. The detection of additional clinically significant alloantibodies is hampered in the presence of anti-Scianna. Soluble recombinant Scianna protein is demonstrated to facilitate antibody diagnostics in both cases.
Soluble recombinant Scianna protein (Sc:1,-2,3,-4,5,6,7) was produced comprising the antigenic extracellular domain fused to a V5-His tag. The protein was isolated from eukaryotic cell culture supernatants of stably transfected HEK293 cells. Seven serum samples with anti-Sc1, anti-Sc2, and anti-Sc5 and 30 serum samples with antibodies to other blood group antigens were evaluated in hemagglutination inhibition assays. Antisera with mixed antibody specificities and autoantibodies were also tested.
Soluble Scianna protein inhibited specifically antibodies to the high-prevalence Scianna antigens Sc1 and Sc5. No antibodies were neutralized that were directed to the low-prevalence Sc2 antigen or to a large representative set of antigens from other blood group systems. Clinically relevant antibodies could be identified despite being masked by anti-Sc1 and anti-Sc5. A mixture of Scianna and JMH proteins allowed detecting a common antibody despite the presence of antibodies to high-prevalence antigens of the Scianna or JMH blood group systems.
Antibody detection systems comprising soluble recombinant Scianna protein provide an easy single-step method for detection and identification of antibodies to high-prevalence Scianna antigens. Reagents with Scianna and other recombinant blood group proteins and mixtures of such proteins would be useful routine reagents in immunohematology.
鉴定针对高流行率的斯恰纳(Sc;ERMAP)抗原(如Sc1和Sc5)的抗体很困难,可能会导致血液采集延迟和成本增加。在存在抗斯恰纳抗体的情况下,检测其他具有临床意义的同种抗体受到阻碍。可溶性重组斯恰纳蛋白被证明在这两种情况下都有助于抗体诊断。
制备了可溶性重组斯恰纳蛋白(Sc:1、-2、3、-4、5、6、7),其包含与V5-组氨酸标签融合的抗原性细胞外结构域。该蛋白从稳定转染的HEK293细胞的真核细胞培养上清液中分离得到。在血凝抑制试验中评估了7份含有抗Sc1、抗Sc2和抗Sc5的血清样本以及30份含有针对其他血型抗原抗体的血清样本。还检测了具有混合抗体特异性的抗血清和自身抗体。
可溶性斯恰纳蛋白特异性抑制针对高流行率斯恰纳抗原Sc1和Sc5的抗体。未中和针对低流行率Sc2抗原或来自其他血型系统的大量代表性抗原集的抗体。尽管被抗Sc1和抗Sc5掩盖,但仍可鉴定出临床相关抗体。斯恰纳蛋白和JMH蛋白的混合物能够检测到一种常见抗体,尽管存在针对斯恰纳或JMH血型系统高流行率抗原的抗体。
包含可溶性重组斯恰纳蛋白的抗体检测系统为检测和鉴定针对高流行率斯恰纳抗原的抗体提供了一种简便的单步方法。含有斯恰纳蛋白和其他重组血型蛋白以及此类蛋白混合物的试剂将是免疫血液学中有用的常规试剂。