An Enzymatic Biosensor for the Detection of D-2-Hydroxyglutaric Acid in Serum and Urine

Biosensors

卷:12

Fuping Lu¹  Bo Wu¹  Zehua Li²  Zepeng Kang²  Chunling Ma²  Zhiguang Zhu²  Haiyan Song² 

[1] Key Laboratory of Industrial Fermentation Microbiology, Ministry of Education, Tianjin University of Science and Technology, No.9, 13th Avenue, Tianjin Economic and Technological Development Area, Tianjin 300457, China;

[2] Tianjin Institute of Industrial Biotechnology, Chinese Academy of Sciences, 32 West 7th Avenue, Tianjin Airport Economic Area, Tianjin 300308, China;

关键词: D-2-hydroxyglutaric acid;    MXene;    biosensor;    screen-printed electrode;    D-2-hydroxyglutarate dehydrogenase;   

DOI  :  10.3390/bios12020066

来源: DOAJ

【 摘 要 】

D-2-hydroxyglutaric acid (D2HG) is overproduced as a result of the D-2-hydroxyglutaric aciduria and relevant cancers, caused by gene mutation. Accurate analysis of D2HG could help rapid diagnosis of these diseases and allow for timely treatment. In this work, a D-2-hydroxyglutarate dehydrogenase from Ralstonia solanacearum (RsD2HGDH) is cloned and recombinantly expressed. This enzyme features the direct electron transfer to chemical electron mediators (such as methylene blue (MB)) in the absence of additional coenzymes. Therefore, NAD⁺, a natural electron acceptor for the commercial D2HGDH and usually known for being unstable and difficult for immobilization can be avoided in the preparation of biosensors. The RsD2HGDH and MB are co-immobilized on a two-dimensional material, Ti₃C₂ MXene, followed by drop-coating on the gold screen-printed electrode (AuSPE) to construct a compact and portable biosensor. The D2HG in samples can be catalyzed by RsD2HGDH, where the current change is measured by chronoamperometry at −0.23 V. The biosensor shows a D2HG detection range of 0.5 to 120 µM (R² = 0.9974) with a sensitivity of 22.26 μA mM⁻¹ cm⁻² and a detection limit of 0.1 µM (S/N = 3). The biosensor retains 72.52% performance of its incipient state after 30 days of storage. The samples of D2HG-containing fetal bovine serum and artificial urine were analyzed with the recovery of 99.56% to 106.83% and 97.30% to 102.47% further indicating the great application potential of our portable D2HG biosensor.

【 授权许可】

Unknown