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. 2024 Jan 15;25(2):1044.
doi: 10.3390/ijms25021044.

New Advances in Rapid Pretreatment for Small Dense LDL Cholesterol Measurement Using Shear Horizontal Surface Acoustic Wave (SH-SAW) Technology

Tai-Hua Chou  1 , Chia-Hsuan Cheng  2   3 , Chi-Jen Lo  4   5 , Guang-Huar Young  1   6 , Szu-Heng Liu  1   3 , Robert Y-L Wang  1   6   7   8
Affiliations

New Advances in Rapid Pretreatment for Small Dense LDL Cholesterol Measurement Using Shear Horizontal Surface Acoustic Wave (SH-SAW) Technology

Tai-Hua Chou et al. Int J Mol Sci. .

Abstract

Atherosclerosis is an inflammatory disease of the arteries associated with alterations in lipid and other metabolism and is a major cause of cardiovascular disease (CVD). LDL consists of several subclasses with different sizes, densities, and physicochemical compositions. Small dense LDL (sd-LDL) is a subclass of LDL. There is growing evidence that sd-LDL-C is associated with CVD risk, metabolic dysregulation, and several pathophysiological processes. In this study, we present a straightforward membrane device filtration method that can be performed with simple laboratory methods to directly determine sd-LDL in serum without the need for specialized equipment. The method consists of three steps: first, the precipitation of lipoproteins with magnesium harpin; second, the collection of effluent from a 100 nm filter; and third, the quantification of sd-LDL-ApoB in the effluent with an SH-SAW biosensor. There was a good correlation between ApoB values obtained using the centrifugation (y = 1.0411x + 12.96, r = 0.82, n = 20) and filtration (y = 1.0633x + 15.13, r = 0.88, n = 20) methods and commercially available sd-LDL-C assay values. In addition to the filtrate method, there was also a close correlation between sd-LDL-C and ELISA assay values (y = 1.0483x - 4489, r = 0.88, n = 20). The filtration treatment method also showed a high correlation with LDL subfractions and NMR spectra ApoB measurements (y = 2.4846x + 4.637, r = 0.89, n = 20). The presence of sd-LDL-ApoB in the effluent was also confirmed by ELISA assay. These results suggest that this filtration method is a simple and promising pretreatment for use with the SH-SAW biosensor as a rapid in vitro diagnostic (IVD) method for predicting sd-LDL concentrations. Overall, we propose a very sensitive and specific SH-SAW biosensor with the ApoB antibody in its sensitive region to monitor sd-LDL levels by employing a simple delay-time phase shifted SH-SAW device. In conclusion, based on the demonstration of our study, the SH-SAW biosensor could be a strong candidate for the future measurement of sd-LDL.

Keywords: apolipoprotein B; shear horizontal surface acoustic wave SH-SAW; small dense LDL.

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Conflict of interest statement

Chia-Hsuan Cheng is an employee of tst Biomedical Electronics Co., Ltd., Taiwan. He is currently a PhD student at the Graduate School of Science and Technology, Shizuoka University Japan. Szu-Heng Liu is the director of a new business division in tst Biomedical Electronics Co., Ltd., Taiwan. He is also an adjunct assistant professor in Chang Gung University, Taiwan. The authors declare no conflicts of interest. The tst Biomedical Electronics company worked together with Robert YL Wang in implementing Taiwanese government programs to promote innovative research and development in small and medium enterprises through which the SH-SAW technology was developed and manufactured by tst Biomedical Electronics company.

Figures

Figure 1
Figure 1
Phase shifts of the SH-SAW biosensor at different concentrations of ApoB samples and the 4PL curve of the SH-SAW biosensor. The purchased apolipoprotein B calibrator was diluted to different concentrations (24–148 mg/dL) and the samples were diluted to 20× and blank (PBS). Then, 5 μL sample drops were measured in the reaction area of the SAW chip and repeated three times. (a) Real-time curve of the measurements; (b) the 4PL fitting curve of the 30-s phase shift.
Figure 2
Figure 2
Phase shift measurements were performed on different samples using SH-SAW biosensors. Fresh serum was prepared for ApoB measurements. No precipitant was added to the serum. Samples, including centrifuged supernatant, 300 kDa filtered effluent, 100 nm filter effluent, and 200 nm filtered effluent, were spiked with precipitating heparin-Mg2+ precipitation reagents. After the incubation step, the supernatant samples were centrifuged and the ultrafiltrate was collected. The effluent samples were filtered through 300 kDa, 100 nm, and 200 nm filters and the effluent were collected. (a) Measured real-time curves; (b) SH-SAW ApoB measurements of different pretreatment serum samples.
Figure 3
Figure 3
Comparative study between commercial sd-LD-C assay and SH-SAW ApoB biosensor measurements of different pre-treatment samples. (a) Correlation between commercial sd-LDL assay results and SH-SAW ApoB biosensor measurements of heparin-Mg2+ precipitated centrifuged supernatant (N = 20). (b) Correlation between commercial sd-LDL-C assay results and SH-SAW ApoB biosensor measurement on 100 nm pore size filtered effluent samples (N = 20).
Figure 4
Figure 4
Correlation between the results of the commercial measurement of the sd-LDL-C assay and sd-LDL-ApoB measurement on 100 nm pore size filtered effluent samples using the ELISA ApoB-100 assay (N = 20).
Figure 5
Figure 5
Correlation between 100 nm pore size filtration performed by the SH-SAW biochip in the sd-LDL-ApoB measurement of 20 individual plasma samples and flesh plasma performed by NMR spectra of the LDL subfractions, Apo-B, LDL-6 (N = 20). Correlation between 100 nm pore size filtration performed by the SH-SAW biochip in the sd-LDL-ApoB measurement of 20 individual plasma samples and fresh plasma performed by NMR spectra of the LDL subfractions, Apo-B, LDL-6 (N = 20).
Figure 6
Figure 6
Correlation between the results of sd-LDL-ApoB concentrations measured in 100 nm pore size filtered effluent from 20 plasma samples using the SH-SAW biosensors and the ELISA ApoB-100 assay kit.
Figure 7
Figure 7
Schematic diagram of the centrifugation and filtration procedures for the detection ApoB in plasma.
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References

    1. Di Cesare M., Khang Y.-H., Asaria P., Blakely T., Cowan M.J., Farzadfar F., Guerrero R., Ikeda N., Kyobutungi C., Msyamboza K.P. Inequalities in non-communicable diseases and effective responses. Lancet. 2013;381:585–597. doi: 10.1016/S0140-6736(12)61851-0. - DOI - PubMed
    1. Jin J.L., Zhang H.W., Cao Y.X., Liu H.H., Hua Q., Li Y.F., Zhang Y., Wu N.Q., Zhu C.G., Xu R.X., et al. Association of small dense low-density lipoprotein with cardiovascular outcome in patients with coronary artery disease and diabetes: A prospective, observational cohort study. Cardiovasc. Diabetol. 2020;19:45. doi: 10.1186/s12933-020-01015-6. - DOI - PMC - PubMed
    1. Holvoet P., Vanhaecke J., Janssens S., Van de Werf F., Collen D. Oxidized LDL and malondialdehyde-modified LDL in patients with acute coronary syndromes and stable coronary artery disease. Circulation. 1998;98:1487–1494. doi: 10.1161/01.CIR.98.15.1487. - DOI - PubMed
    1. Barquera S., Pedroza-Tobias A., Medina C., Hernandez-Barrera L., Bibbins-Domingo K., Lozano R., Moran A.E. Global Overview of the Epidemiology of Atherosclerotic Cardiovascular Disease. Arch. Med. Res. 2015;46:328–338. doi: 10.1016/j.arcmed.2015.06.006. - DOI - PubMed
    1. Tanaga K., Bujo H., Inoue M., Mikami K., Kotani K., Takahashi K., Kanno T., Saito Y. Increased circulating malondialdehyde-modified LDL levels in patients with coronary artery diseases and their association with peak sizes of LDL particles. Arterioscler. Thromb. Vasc. Biol. 2002;22:662–666. doi: 10.1161/01.ATV.0000012351.63938.84. - DOI - PubMed

Grants and funding

This study was supported in part by Chang Gung memorial hospital research fund (CMRPD1M0421-2 and CMRPD1M0851).

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