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

Autor: Yui M; Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan., Nagatake Y; Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan., Takehara S; Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan., Ito M; Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan. Electronic address: mitsuki_ito@eiken.co.jp., Watanabe K; Research and Development Division, Eiken Chemical Co., Ltd., Shimotsuga-gun, Tochigi, Japan.
Jazyk: angličtina
Zdroj: Analytical biochemistry [Anal Biochem] 2024 Nov; Vol. 694, pp. 115601. Date of Electronic Publication: 2024 Jul 05.
DOI: 10.1016/j.ab.2024.115601
Abstrakt: 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.
Competing Interests: Declaration of competing interest The authors declare the following financial interests/personal relationships which may be considered as potential competing interests: M. Y., Y. N., M. I., S. T. and K. W. are employees of Eiken Chemical Co., Ltd. No other potential competing interests relevant to this article were reported. This study did not receive any specific grants from funding agencies in the public, commercial, or non-profit sectors.
(Copyright © 2024 The Authors. Published by Elsevier Inc. All rights reserved.)
Databáze: MEDLINE
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