Kanto-Koshinetsu Block Blood Center, Japanese Red Cross Society, Tokyo, Japan.
Fujikura Kasei Co., Ltd, Tokyo, Japan.
Vox Sang. 2023 Dec;118(12):1109-1114. doi: 10.1111/vox.13543. Epub 2023 Oct 5.
In Japan, the prevalence of haptoglobin deficiency is approximately 1 in 4000. Haptoglobin-deficient individuals may produce anti-haptoglobin from allo-immunization, leading to serious transfusion reactions. Therefore, implementation of a consistent supply of haptoglobin-deficient fresh frozen plasma is crucial. We developed a novel reagent to facilitate large-scale identification of haptoglobin-deficient individuals as potential donors of plasma products.
We established mouse monoclonal anti-haptoglobin-producing cell lines (three clones) using the hybridoma method by immunizing mice with the haptoglobin protein. Purified antibodies were conjugated with carboxylate-modified polystyrene latex beads and used for haptoglobin measurements by the latex agglutination method using an automatic analyser (LABOSPECT008). Samples with low protein concentrations were re-examined by enzyme-linked immunosorbent assay to confirm the results. Additionally, the haptoglobin gene was amplified by polymerase chain reaction to confirm the haptoglobin deletion allele (Hp ).
From February to October 2022, 7476 blood donor samples were screened. Two haptoglobin-deficient and 21 low-haptoglobin-expressing individuals were identified. Two haptoglobin-deficient donors were found homozygous for Hp , and 19 (90%) of the 21 low-haptoglobin-expressing individuals were heterozygous for Hp , which includes the first reported case of heterozygous Hp /Hp .
We developed a new reagent for the detection of haptoglobin deficiency, which is automatable and inexpensive and appears useful for large-scale screening of blood donors.
在日本，血色素结合蛋白缺乏症的患病率约为 1/4000。血色素结合蛋白缺乏症患者可能会因同种免疫产生抗血色素结合蛋白，从而导致严重的输血反应。因此，确保血色素结合蛋白缺乏的新鲜冷冻血浆的稳定供应至关重要。我们开发了一种新的试剂，以方便大规模鉴定血色素结合蛋白缺乏的个体，作为血浆制品的潜在供体。
我们使用杂交瘤方法，用血色素蛋白免疫小鼠，建立了生产抗血色素结合蛋白的小鼠单克隆抗体产生细胞系（三个克隆）。纯化的抗体与羧基化修饰的聚苯乙烯乳胶珠偶联，并用于使用自动分析仪（LABOSPECT008）通过乳胶凝集法进行血色素结合蛋白测量。对于蛋白浓度低的样本，通过酶联免疫吸附试验重新检查以确认结果。此外，通过聚合酶链反应扩增血色素结合蛋白基因，以确认血色素结合蛋白缺失等位基因（Hp ）。
2022 年 2 月至 10 月，筛选了 7476 份献血者样本。鉴定出 2 例血色素结合蛋白缺乏症和 21 例低血色素结合蛋白表达个体。2 例血色素结合蛋白缺乏症供者均为 Hp 纯合子，21 例低血色素结合蛋白表达个体中 19 例（90%）为 Hp 杂合子，其中包括首例报道的 Hp /Hp 杂合子。
我们开发了一种新的血色素结合蛋白缺乏检测试剂，该试剂具有自动化和低成本的特点，似乎非常适合大规模筛选献血者。
