Dunstan R A, Simpson M B, Rosse W F
Am J Clin Pathol. 1985 Jan;83(1):90-4. doi: 10.1093/ajcp/83.1.90.
One- and two-stage radioligand assays were used to determine if human platelets possess the Lea antigen. Goat IgG anti-Lea antibody was purified by multiple adsorptions with Le(a-b-) human red blood cells, followed by affinity chromatography with synthetic Lea substance and labeling with 125I. Human IgG anti-Lea antibody was used either in a two stage radioassay with 125I-labeled mouse monoclonal IgG anti-human IgG as the second antibody or, alternatively, purified by Staph protein A chromatography, labeled with 125I, and used in a one-stage radioassay. Platelets from donors of appropriate red blood cell phenotypes were incubated with the antisera, centrifuged through phthalate esters, and assayed in a gamma scintillation counter. Dose response and saturation curve analysis demonstrate the presence of Lewis a antigen on platelets from Lea+ donors. Furthermore, platelets from an Le(a-b-) donor incubated in Le (a+b-) plasma adsorb Lea antigen in a similar manner to red blood cells. The clinical significance of these antigens in platelet transfusion remains undefined.
采用单阶段和双阶段放射性配体测定法来确定人类血小板是否具有Lea抗原。山羊抗Lea IgG抗体通过与Le(a-b-)人红细胞多次吸附进行纯化,随后用合成Lea物质进行亲和层析并用125I标记。人抗Lea IgG抗体要么在双阶段放射测定法中使用,以125I标记的小鼠抗人IgG单克隆抗体作为第二抗体,要么通过葡萄球菌蛋白A层析纯化,用125I标记,并用于单阶段放射测定法。将具有合适红细胞表型的供体的血小板与抗血清孵育,通过邻苯二甲酸酯离心,并在γ闪烁计数器中进行测定。剂量反应和饱和曲线分析表明Lea+供体的血小板上存在Lewis a抗原。此外,在Le(a+b-)血浆中孵育的Le(a-b-)供体的血小板以与红细胞类似的方式吸附Lea抗原。这些抗原在血小板输注中的临床意义尚不清楚。