Method for quantifying free hemoglobin, distinct from the hemoglobin-haptoglobin complex, in human serum

IF 2.6 4区生物学 Q2 BIOCHEMICAL RESEARCH METHODS Analytical biochemistry Pub Date : 2024-07-05 DOI:10.1016/j.ab.2024.115601

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Abstract

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.

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人血清中游离血红蛋白（有别于血红蛋白-aptoglobin 复合物）的定量方法。

测量血液中的游离血红蛋白（游离 Hb）对于评估溶血性疾病患者器官受损的风险至关重要。然而，临床上常用的比色法并不能区分血液中的游离血红蛋白和血红蛋白-aptoglobin 复合物（Hb-Hp），而只能反映总血红蛋白水平。虽然尺寸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，但其临床应用受到检测时间长的限制。在此，我们采用乳胶凝集免疫比浊法（LATIA）开发了一种新的检测方法，用于快速定量血清中的游离 Hb，并将其与 Hb-Hp 区分开来。使用该方法，可在约 15 分钟内利用自动生化分析仪测定血清中 1 至 100 μg /mL 范围内的游离血红蛋白。使用新开发的方法和 SEC-HPLC 测定的游离血红蛋白水平与健康成人血清样本中的游离血红蛋白水平高度相关，表明该方法对游离血红蛋白具有高度特异性。这种新型检测方法可用于监测各种溶血性疾病患者的游离 Hb 水平，并根据测量值设计治疗策略。不过，还需要进一步的研究来评估其临床性能。

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来源期刊

Analytical biochemistry 生物-分析化学

CiteScore

5.70

自引率

0.00%

发文量

283

审稿时长

44 days

期刊介绍： The journal''s title Analytical Biochemistry: Methods in the Biological Sciences declares its broad scope: methods for the basic biological sciences that include biochemistry, molecular genetics, cell biology, proteomics, immunology, bioinformatics and wherever the frontiers of research take the field. The emphasis is on methods from the strictly analytical to the more preparative that would include novel approaches to protein purification as well as improvements in cell and organ culture. The actual techniques are equally inclusive ranging from aptamers to zymology. The journal has been particularly active in: -Analytical techniques for biological molecules- Aptamer selection and utilization- Biosensors- Chromatography- Cloning, sequencing and mutagenesis- Electrochemical methods- Electrophoresis- Enzyme characterization methods- Immunological approaches- Mass spectrometry of proteins and nucleic acids- Metabolomics- Nano level techniques- Optical spectroscopy in all its forms. The journal is reluctant to include most drug and strictly clinical studies as there are more suitable publication platforms for these types of papers.