Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan.
Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan.
Anal Biochem. 2024 Nov;694:115601. doi: 10.1016/j.ab.2024.115601. Epub 2024 Jul 5.
The measurement of free hemoglobin (free Hb) in blood is crucial for assessing the risk of organ damage in patients with hemolytic diseases. However, the colorimetric method, commonly used in clinical practice, does not distinguish between free Hb and the hemoglobin-haptoglobin complex (Hb-Hp) in the blood, instead reflecting the total Hb level. Although size-exclusion high-performance liquid chromatography (SEC-HPLC) can specifically measure free Hb, its clinical use is limited by long assay times. Here, we developed a novel assay method for the rapid quantification of free Hb in serum, distinguishing it from Hb-Hp, using a latex agglutination immunoturbidimetric assay (LATIA). This method could be used to measure free Hb in sera in the range of 1-100 μg/mL in approximately 15 min using an automatic biochemistry analyzer. Using Hb-spiked serum samples from healthy adults, there was a high correlation with Hb levels determined using the newly developed method and SEC-HPLC, indicating a high specificity for free Hb. This novel assay can be used to monitor levels of free Hb in patients with various hemolytic diseases and to design therapeutic strategies based on measured values. However, further studies are required to assess its clinical performance.
游离血红蛋白(free Hb)的测量对于评估溶血性疾病患者的器官损伤风险至关重要。然而,临床实践中常用的比色法不能区分血液中的游离血红蛋白和血红蛋白-触珠蛋白复合物(Hb-Hp),而是反映总血红蛋白水平。虽然大小排阻高效液相色谱法(SEC-HPLC)可以特异性地测量游离血红蛋白,但由于测定时间长,其临床应用受到限制。在这里,我们开发了一种新的检测方法,使用胶乳凝集免疫比浊法(LATIA)快速定量血清中的游离血红蛋白,将其与 Hb-Hp 区分开来。该方法可在大约 15 分钟内使用自动生化分析仪在 1-100μg/mL 的范围内测量血清中的游离血红蛋白。使用来自健康成年人的 Hb 加标血清样本,与使用新开发方法和 SEC-HPLC 测定的 Hb 水平高度相关,表明对游离 Hb 具有高度特异性。这种新的检测方法可用于监测各种溶血性疾病患者的游离血红蛋白水平,并根据测量值设计治疗策略。然而,还需要进一步的研究来评估其临床性能。