Microfluidic nanochips for rapid exosome isolation and multiplex biomarker profiling from a single blood drop: Toward sample-to-answer liquid biopsy

Abstract

Background: Exosomes and other small extracellular vesicles carry molecular cargo that can reflect disease status, therapeutic response, and relapse dynamics. Currently, most exosome-analysis workflows are centralized, require milliliter-sized blood samples, involve extended isolation procedures, and depend on separate downstream assays. Single-drop microfluidic nanochips, which can process finger-prick blood volumes of approximately 1050 µL within a closed cartridge, offer a promising route toward integrated sample-to-answer liquid biopsy platforms.

Main Body: These systems may integrate plasma handling, exosome enrichment, cargo access, multiplex biomarker detection, and algorithmic classification. This review highlights key on-chip enrichment techniques, including immunoaffinity, size selection, electrokinetic, acoustofluidic, and hybrid methods. It also discusses nano-biointerface designs that improve vesicle capture efficiency while reducing fouling and shear-induced vesicle damage. In addition, multiplex sensing architectures for optical, electrochemical, and plasmonic transduction are explored, with particular attention to challenges associated with whole-blood measurements, including anticoagulation, sample variability, platelet activation, viscosity, and hemolysis.

Conclusions: Single-drop exosome nanochips have the potential to transform liquid biopsy by enabling rapid, low-volume, integrated analysis of extracellular vesicles. However, successful clinical translation will require clear validation requirements, standardized reporting expectations, and defined use-case roadmaps for screening, disease monitoring, and therapeutic-response assessment. Establishing realistic performance standards and robust engineering parameters will be essential for making single-lab exosome nanochip platforms reproducible and clinically applicable.

Keywords

exosomes / extracellular vesicles / liquid biopsy / microfluidics / multiplex biosensing / nanochips / point-of-care diagnostics

References

[1] Aheget H, Mazini L, Martin F, Belqat B, Marchal JA, Benabdellah K. Exosomes: their role in pathogenesis, diagnosis and treatment of diseases. Cancers. 2020; 13: 84.

[2] Almohaimeed HM, Almars AI, Tounsi WA, et al. Exosomes in autoimmunity: mediators of autoantigen presentation and targets for therapeutic intervention in autoimmune diseases. J Clin Immunol. 2025; 46: 3.

[3] Lee M-J, Shin M, Lee S, Liu S, Lee S-N, Choi J-W. Nanomaterial-based total analysis systems for isolation and detection of exosomal biomarkers in cancer diagnosis. Mater Today Bio. 2026; 37: 102866.

[4] Liangsupree T, Multia E, Riekkola M-L. Recent advances in modern extracellular vesicle isolation and separation techniques. J Chromatogr A. 2026; 1767: 466602.

[5] Chen J, Li P, Zhang T et al. Review on strategies and technologies for exosome isolation and purification. Front Bioeng Biotechnol. 2022; 9: 811971.

[6] Ramnauth N, Neubarth E, Makler-Disatham A, et al. Development of a microfluidic device for exosome isolation in point-of-care settings. Sensors. 2023; 23: 8292.

[7] Saghezi BV, Mansouri K, Hallaj R. Microfluidic systems in cancer diagnosis: advancing exosome and CTC detection from laboratory research to clinical application. Nano Sel. 2025; 6: e202400108.

[8] Werner R, Crosbie R, Dorney M, et al. Implementation of an ISO 15189 accredited next generation sequencing service for cell-free total nucleic acid (cfTNA) analysis to facilitate driver mutation reporting in blood: the experience of a clinical diagnostic laboratory. J Clin Pathol. 2024; 78: 783-791.

[9] Zhang P, Zhou X, He M, et al. Ultrasensitive detection of circulating exosomes with a 3D-nanopatterned microfluidic chip. Nat Biomed Eng. 2019; 3: 438-451.

[10] Raju D, Bathini S, Badilescu S, Ghosh A, Packirisamy M. Microfluidic platforms for the isolation and detection of exosomes: a brief review. Micromachines. 2022; 13: 730.

[11] Yasamineh S, Nikben N, Hamed Ahmed M, Abdul Kareem R, Kadhim Al-Aridhy A, Hosseini Hooshiar M. Increasing the sensitivity and accuracy of detecting exosomes as biomarkers for cancer monitoring using optical nanobiosensors. Cancer Cell Int. 2024; 24: 189.

[12] Bowling F, Badrick T. Methods for determining clinical utility. Clin Biochem. 2023; 121–122: 110674.

[13] Lan X, Li D, Yu Y, Prabhu SN, Liu G. Extracellular vesicles isolation: a focus on magnetic beads-assisted platforms. Front Bioeng Biotechnol. 2025; 13: 1646385.

[14] Lee J, Zhu Y, Tseng H-R. Bioorthogonal click chemistry-enabled enrichment of extracellular vesicles for integrated molecular and functional liquid biopsy§. Acc Chem Res. 2026; 59: 1813-1828.

[15] Baillargeon KR, Mace CR. Microsampling tools for collecting, processing, and storing blood at the point-of-care. Bioeng Transl Med. 2022; 8: e10476.

[16] Phillips W, Willms E, Hill AF. Understanding extracellular vesicle and nanoparticle heterogeneity: novel methods and considerations. Proteomics. 2021; 21: 2000118.

[17] Clegg L, Jensen RB, Bæk R, et al. Direct profiling of extracellular vesicle surface markers in plasma: a proof-of-concept study. Biol Cell. 2025; 117: e70047.

[18] Mathieu M, Névo N, Jouve M, et al. Specificities of exosome versus small ectosome secretion revealed by live intracellular tracking of CD63 and CD9. Nat Commun. 2021; 12: 4389.

[19] Zhang Y, Meng X, Greening DW, et al. Unveiling heterogeneity: innovations and challenges in single-vesicle analysis for clinical translation. J Extracell Vesicles. 2025; 14: e70209.

[20] Ljungström M, Oltra E. Methods for Extracellular vesicle isolation: relevance for encapsulated mirnas in disease diagnosis and treatment. Genes. 2025; 16: 330.

[21] Nguyen PHD, Le AH, Pek JSQ, et al. Extracellular vesicles and lipoproteins—smart messengers of blood cells in the circulation. J Extracell Biol. 2022; 1: e49.

[22] Aljohani MA. Exosomes in clinical laboratory: from biomarker discovery to diagnostic implementation. Medicina (Mex). 2025; 61: 1930.

[23] Burla B, Arita M, Arita M, et al. MS-based lipidomics of human blood plasma: a community-initiated position paper to develop accepted guidelines1. J Lipid Res. 2018; 59: 2001-2017.

[24] Woodland B, Farrell LA, Brockbals L, et al. Sample preparation for multi-omics analysis: considerations and guidance for identifying the ideal workflow. Proteomics. 2025; 25: 76-101.

[25] Parmentier L, Vermeersch E, Van Vlierberghe S. Recapitulating the bone extracellular matrix through 3D bioprinting using various crosslinking chemistries. Front Bioeng Biotechnol. 2025; 13: 1506122.

[26] Khan OA, Rai D, Crabtree B, et al. Three-dimensional assessment of finger individuation reveals finger- and joint-specific selective motor control deficits in children with cerebral palsy. J NeuroEngineering Rehabil. 2026; 23: 73.

[27] Sanfeliu-Cerdán N, Krieg M. The mechanobiology of biomolecular condensates. Biophys Rev. 2025; 6: 011310.

[28] Trębacz P, Pawlik M, Kurkowska A, Wilk K, Piątek A, Czopowicz M. Low-temperature plasma activation of biomaterials and its stability over time and post-sterilisation effects. Materials. 2026; 19: 643.

[29] Buschmann D, Mussack V, Byrd JB. Separation, characterization, and standardization of extracellular vesicles for drug delivery applications. Adv Drug Deliv Rev. 2021; 174: 348-368.

[30] Rahmani H, Larachi F, Taghavi SM. Modeling of shear flows over superhydrophobic surfaces: from Newtonian to non-Newtonian fluids. ACS Eng. Au. 2024; 4: 166-192.

[31] Willms E, Cabañas C, Mäger I, Wood MJA, Vader P. Extracellular vesicle heterogeneity: subpopulations, isolation techniques, and diverse functions in cancer progression. Front Immunol. 2018; 9: 738.

[32] Wang W, Qiao S, Kong X, Zhang G, Cai Z. The role of exosomes in immunopathology and potential therapeutic implications. Cell Mol Immunol. 2025; 22: 975-995.

[33] Li C, Ni Y-Q, Xu H, et al. Roles and mechanisms of exosomal non-coding RNAs in human health and diseases. Signal Transduct Target Ther. 2021; 6: 383.

[34] Ghadami S, Dellinger K. The lipid composition of extracellular vesicles: applications in diagnostics and therapeutic delivery. Front Mol Biosci. 2023; 10: 1198044.

[35] Li X, Wang Q, Wang R. Roles of Exosome Genomic DNA in Colorectal Cancer. Front Pharmacol. 2022; 13: 923232.

[36] Fu S, Wang Y, Xia X, Zheng JC. Exosome engineering: current progress in cargo loading and targeted delivery. NanoImpact. 2020; 20: 100261.

[37] Pattanayak P, Singh SK, Gulati M, et al. Microfluidic chips: recent advances, critical strategies in design, applications and future perspectives. Microfluid Nanofluidics. 2021; 25: 99.

[38] Alhalaili B, Popescu IN, Rusanescu CO, Vidu R. Microfluidic devices and microfluidics-integrated electrochemical and optical (bio) sensors for pollution analysis: a review. Sustainability. 2022; 14: 12844.

[39] Jensen MJ, Peel A, Horne R, et al. Antifouling and mechanical properties of photografted zwitterionic hydrogel thin-film coatings depend on the cross-link density. ACS Biomater Sci Eng. 2021; 7: 4494-4502.

[40] Hanson EK, Wang C-W, Minkoff L, Whelan RJ. Strategies for mitigating commercial sensor chip variability with experimental design controls. Sensors. 2023; 23: 6703.

[41] Chakrapani G, Vijay Kumar V, Zare M. Next-generation biomaterials: design strategies, clinical translation, and the rise of intelligent therapeutic platforms. ACS Omega. 10: 46272-46293.

[42] De Sousa KP, Rossi I, Abdullahi M, Ramirez MI, Stratton D, Inal JM. Isolation and characterization of extracellular vesicles and future directions in diagnosis and therapy. Wiley Interdiscip Rev Nanomed Nanobiotechnol. 2023; 15: e1835.

[43] Li L, Willard B, Rachdaoui N, et al. Plasma proteome dynamics: analysis of lipoproteins and acute phase response proteins with 2H₂O metabolic labeling. Mol Cell Proteomics MCP. 2012; 11: M111.014209.

[44] Cruchinho P, López-Franco MD, Capelas ML, et al. Translation, cross-cultural adaptation, and validation of measurement instruments: a practical guideline for novice researchers. J Multidiscip Healthc. 2024; 17: 2701-2728.

[45] Sauer-Budge AF, Brookfield SJ, Janzen R, et al. A novel device for collecting and dispensing fingerstick blood for point of care testing. PLoS ONE. 2017; 12: e0183625.

[46] Lina. Microfluidic Prototyping for Scalable Manufacturing | Micromolds. https://www.micromolds.eu/post/microfluidic-prototyping-for-scalable-manufacturing

[47] Dinc R, Ardic N. Artificial intelligence for exosomal biomarker discovery for cardiovascular diseases: multi-omics integration, reproducibility, and translational prospects. Cells. 2026; 15: 304.

[48] Kim Y, Park J, Choi J, et al. Physiological barriers to nucleic acid therapeutics and engineering strategies for lipid nanoparticle design, optimization, and clinical translation. Pharmaceutics. 2025; 17: 1309.

[49] Sonbhadra S, Mehak , Pandey LM. Biogenesis, isolation, and detection of exosomes and their potential in therapeutics and diagnostics. Biosensors. 2023; 13: 802.

[50] Solovicová V, Ďatková A, Bertók T, et al. Advances in magnetic affinity-based isolation/detection of exosomes for robust diagnostics. Mikrochim Acta. 2025; 192: 206.

[51] Feingold KR. Introduction to Lipids and Lipoproteins. In: KR Feingold, RA Adler, SF Ahmed, et al. (eds.) Endotext. MDText.com, Inc., (2000)

[52] Onukwugha N-E, Kang Y-T, Nagrath S. Emerging micro-nanotechnologies for extracellular vesicles in immuno-oncology: from target specific isolations to immunomodulation. Lab Chip. 2022; 22: 3314-3339.

[53] Afzal MA, Brickey KP, Gomez ED, Zydney AL. Quantitative analysis of protein fouling in virus removal filtration membranes through electron tomography. Membranes. 2025; 15: 369.

[54] Kutluk H, Viefhues M, Constantinou I. Integrated Microfluidics for Single-Cell Separation and On-Chip Analysis: novel Applications and Recent Advances. Small Sci. 2024; 4: 2300206.

[55] Costa CM, Cardoso VF, Martins P, et al. Smart and multifunctional materials based on electroactive poly (vinylidene fluoride): recent advances and opportunities in sensors, actuators, energy, environmental, and biomedical applications. Chem Rev. 2023; 123: 11392-11487.

[56] Mansor MA, Jamrus MA, Lok CK, Ahmad MR, Petrů M, Koloor SSR. Microfluidic device for both active and passive cell separation techniques: a review. Sens Actuators Rep. 2025; 9: 100277.

[57] Mikkelsen NS, Ravendran S, Broksø AD, et al. Orthogonal CRISPR systems for targeted integration and multiplex base editing enable nonviral engineering of allogeneic CAR-T cells. Mol Ther. 2025; 33: 6082-6100.

[58] Mondal TK, Bangaru AVB, Williams SJ. A review on AC-dielectrophoresis of nanoparticles. Micromachines. 2025; 16: 453.

[59] Li P, Huang TJ. Applications of acoustofluidics in bioanalytical chemistry. Anal Chem. 2019; 91: 757-767.

[60] Deivasigamani R, Mohd Maidin NN, Abdul Nasir NS, et al. A correlation of conductivity medium and bioparticle viability on dielectrophoresis-based biomedical applications. Electrophoresis. 2023; 44: 573-620.

[61] Lan Z, Chen R, Zou D, Zhao C. Microfluidic nanoparticle separation for precision medicine. Adv Sci. 2024; 12: 2411278.

[62] Zhao Y, Zhang T, Zhao G, et al. Biological and bioinspired vesicles for wound healing: insights, advances and challenges. Int J Nanomedicine. 2025; 20: 8497-8528.

[63] Wang X, Fu C, Zhang C, Qiu Z, Wang B. A Comprehensive review of wetting transition mechanism on the surfaces of microstructures from theory and testing methods. Materials. 2022; 15: 4747.

[64] Hoffman MSF, McKeage JW, Xu J, Ruddy BP, Nielsen PMF, Taberner AJ. Minimally invasive capillary blood sampling methods. Expert Rev Med Devices. 2023; 20: 5-16.

[65] Tran HL, Zheng W, Issadore DA, et al.: Extracellular vesicles for clinical diagnostics: from bulk measurements to single-vesicle analysis. ACS Nano. 19: 28021-28109.

[66] Zhao Z, Wijerathne H, Godwin AK, Soper SA. Isolation and analysis methods of extracellular vesicles (EVs). Extracell Vesicles Circ Nucleic Acids. 2021; 2: 80-103.

[67] Filipović L, Spasojević M, Prodanović R, et al. Affinity-based isolation of extracellular vesicles by means of single-domain antibodies bound to macroporous methacrylate-based copolymer. New Biotechnol. 2022; 69: 36-48.

[68] Leggio L, Paternò G, Vivarelli S, et al. Label-free approaches for extracellular vesicle detection. iScience. 2023; 26: 108105.

[69] Henriksson A, Neubauer P, Birkholz M. Dielectrophoresis: an approach to increase sensitivity, reduce response time and to suppress nonspecific binding in biosensors? Biosensors. 2022; 12: 784.

[70] Mello GAB, Benjamin SR, Lima F, Dutra RF. Recent advances in electrochemical sensors for the detection of anti-inflammatory and antibiotic drugs: a comprehensive review. Biosensors. 2025; 15: 676.

[71] Zhao Z, Yang Y, Zeng Y, He M. A microfluidic ExoSearch chip for multiplexed exosome detection towards blood-based ovarian cancer diagnosis. Lab Chip. 2016; 16: 489-496.

[72] Jeong S, Park J, Pathania D, Castro CM, Weissleder R, Lee H. Integrated magneto-electrochemical sensor for exosome analysis. ACS Nano. 2016; 10: 1802-1809.

[73] Poncelet L, Morton KJ, Shiu M, et al. EV-Blade: an automated centrifugal-pneumatic cartridge for size- and affinity-based exosome isolation from whole blood. Lab Chip. 2026; 26: 770-782.

[74] Fan Y, Wang X, Ren J, Lin F, Wu J. Recent advances in acoustofluidic separation technology in biology. Microsyst Nanoeng. 2022; 8: 94.

[75] Ozcelik A, Cevik O. Microfluidic methods used in exosome isolation. Biocell. 2023; 47: 959-964.

[76] Wu Y, Wang Y, Lu Y, et al. Microfluidic technology for the isolation and analysis of exosomes. Micromachines. 2022; 13: 1571.

[77] Huang S, Ji X, Jackson KK et al. Rapid separation of blood plasma exosomes from low-density lipoproteins via a hydrophobic interaction chromatography method on a polyester capillary-channeled polymer fiber phase. Anal Chim Acta. 2021; 1167: 338578.

[78] Quek C, Bellingham SA, Jung C-H, et al. Defining the purity of exosomes required for diagnostic profiling of small RNA suitable for biomarker discovery. RNA Biol. 2016; 14: 245-258.

[79] Bian J, Gobalasingham N, Purchel A, Lin J. The power of field-flow fractionation in characterization of nanoparticles in drug delivery. Molecules. 2023; 28: 4169.

[80] Jeon S, Kim Y, Shin S. Selective lipoprotein removal enables high-purity EV isolation from plasma via aptamer-based mesh filtration. Small. 2026; 22: e14724.

[81] Hassanzadeh-Barforoushi A, Sango X, Johnston EL, Haylock D, Wang Y. Microfluidic devices for manufacture of therapeutic extracellular vesicles: advances and opportunities. J Extracell Vesicles. 2025; 14: e70132.

[82] Chen J, Zheng M, Xiao Q, et al. Recent advances in microfluidic-based extracellular vesicle analysis. Micromachines. 2024; 15: 630.

[83] Dai Y, Cui Y, Li J, Li P, Huang X. Integrated microfluidic platforms for extracellular vesicles: separation, detection, and clinical translation. APL Bioeng. 2025; 9: 031502.

[84] Wang J, Ma P, Kim DH, Liu B-F, Demirci U. Towards microfluidic-based exosome isolation and detection for tumor therapy. Nano Today. 2021; 37: 101066.

[85] Meggiolaro A, Moccia V, Brun P, et al. Microfluidic strategies for extracellular vesicle isolation: towards clinical applications. Biosensors. 2022; 13: 50.

[86] Lee SM, Balakrishnan HK, Doeven EH, Yuan D, Guijt RM. Chemical trends in sample preparation for nucleic acid amplification testing (NAAT): a review. Biosensors. 2023; 13: 980.

[87] Mahran R, Vello N, Komulainen A, Malakoutikhah M, Härmä H, Kopra K. Isothermal chemical denaturation assay for monitoring protein stability and inhibitor interactions. Sci Rep. 2023; 13: 20066.

[88] Shen L, Pang S, Zhong M, et al. A comprehensive review of ultrasonic assisted extraction (UAE) for bioactive components: principles, advantages, equipment, and combined technologies. Ultrason Sonochem. 2023; 101: 106646.

[89] Jiang Y, Rex DAB, Schuster D, et al. Comprehensive overview of bottom-up proteomics using mass spectrometry. ACS Meas Sci Au. 2024; 4: 338-417.

[90] Dohnalová H, Seifert M, Matoušková E, et al. Temperature-dependent twist of double-stranded rna probed by magnetic tweezer experiments and molecular dynamics simulations. J Phys Chem B. 2024; 128: 664-675.

[91] Cheng Y-H, Chande C, Li Z, Menon NH, Kaaliveetil S, Basuray S. Optimization of Electrolytes with Redox Reagents to Improve the Impedimetric Signal for Use with a Low-Cost Analyzer. Biosensors. 2023; 13: 999.

[92] Chicea ED, Chicea R, Teacoe DA, et al. Advancements in sonication-based extraction techniques for ovarian follicular fluid analysis: implications for infertility diagnostics and assisted reproductive technologies. Int J Mol Sci. 2025; 26: 10368.

[93] Jain K, Salamat-Miller N, Taylor K. Freeze–thaw characterization process to minimize aggregation and enable drug product manufacturing of protein based therapeutics. Sci Rep. 2021; 11: 11332.

[94] Schimek A, Ng JKM, Hubbuch J. Navigating the purification process: maintaining the integrity of replication-competent enveloped viruses. Vaccines. 2025; 13: 444.

[95] Yang H, Ren X, Zhao Y, Xu T, Xiao J, Chen H. Enhancing alkaline protease stability through enzyme-catalyzed crosslinking and its application in detergents. Processes. 2024; 12: 624.

[96] Cui X, Ruan Q, Zhuo X, et al. Photothermal nanomaterials: a powerful light-to-heat converter. Chem Rev. 2023; 123: 6891-6952.

[97] Șoldănescu I, Lobiuc A, Caliman-Sturdza OA, Covasa M, Mangul S, Dimian M. The Potential of Nanopore Technologies in Peptide and Protein Sensing for Biomarker Detection. Biosensors. 2025; 15: 540.

[98] Jiawei S, Zhi C, Kewei T, Xiaoping L. Magnetic bead-based adsorption strategy for exosome isolation. Front Bioeng Biotechnol. 2022; 10: 942077.

[99] Kruk T, Bzowska M, Hinz A, Szuwarzyński M, Szczepanowicz K. Control of specific/nonspecific protein adsorption: functionalization of polyelectrolyte multilayer films as a potential coating for biosensors. Materials. 2021; 14: 7629.

[100] Vargas R, Abbott L, Bower D, Frahm N, Shaffer M, Yu W-H. Gene signature discovery and systematic validation across diverse clinical cohorts for TB prognosis and response to treatment. PLOS Comput Biol. 2023; 19: e1010770.

[101] Earm K, Earm YE, Noble D. Extracellular vesicles as targeted communicators in complementary medical treatments. Int J Mol Sci. 2025; 26: 5896.

[102] Duché G, Sanderson JM. The chemical reactivity of membrane lipids. Chem Rev. 2024; 124: 3284-3330.

[103] Mak GC, Lau SS, Wong KK, et al. Analytical sensitivity and clinical sensitivity of the three rapid antigen detection kits for detection of SARS-CoV-2 virus. J Clin Virol. 2020; 133: 104684.

[104] O'Sullivan S, Ali Z, Jiang X, et al. Developments in transduction, connectivity and AI/machine learning for point-of-care testing. Sensors. 2019; 19: 1917.

[105] Gimondi S, Ferreira H, Reis RL, Neves NM. Microfluidic devices: a tool for nanoparticle synthesis and performance evaluation. ACS Nano. 2023; 17: 14205-14228.

[106] Li Y, Dong J, Liu J, Han S. Thermodynamic analysis of wetting transitions on micro/nanopillared superhydrophobic surfaces. Coatings. 2025; 15: 140.

[107] Sánchez-Trasviña C, Flores-Gatica M, Enriquez-Ochoa D, Rito-Palomares M, Mayolo-Deloisa K. Purification of modified therapeutic proteins available on the market: an analysis of chromatography-based strategies. Front Bioeng Biotechnol. 2021; 9: 717326.

[108] Protti M, Mercolini L, Mandrioli R. Review: the role of automation in improving the performance and throughput of microsample bioanalysis. Anal Chim Acta. 2025; 1359: 344018.

[109] Zhang Y, Gao S, Li H, et al. Design principles of nanosensors for multiplex detection of contaminants in food. Small. 2025; 21: 2412271.

[110] Tian D, Yang J, Zhang L, Yang H, Wang S, Su X. Spatial fluorescence barcode by transiently luminescent DNA beads. Nat Commun. 2025; 17: 656.

[111] Camarca A, Varriale A, Capo A, et al. Emergent biosensing technologies based on fluorescence spectroscopy and surface plasmon resonance. Sensors. 2021; 21: 906.

[112] Berezin MY, Achilefu S. Fluorescence lifetime measurements and biological imaging. Chem Rev. 2010; 110: 2641-2684.

[113] Kim EJ, Kim H, Park E, et al. Paper-based multiplex surface-enhanced raman scattering detection using polymerase chain reaction probe codification. Anal Chem. 2021; 93: 3677-3685.

[114] Pedersen T, Gurevich L, Magnusson NE. Aspects of electrochemical biosensors using affinity assays. Biosensors. 2025; 15: 166.

[115] Fan H, Ye D, Gao X, Luo Y, Wang L. Nucleic acid-based field-effect transistor biosensors. Biosensors. 2026; 16: 95.

[116] Butt MA. Surface plasmon resonance-based biodetection systems: principles, progress and applications—a comprehensive review. Biosensors. 2025; 15: 35.

[117] Kan CW, Tobos CI, Rissin DM, et al. Digital enzyme-linked immunosorbent assays with sub-attomolar detection limits based on low numbers of capture beads combined with high efficiency bead analysis. Lab Chip. 2020; 20: 2122-2135.

[118] Fang X, Yang Y, Wang H, Xu H. Bead-based microfluidic platforms for multiplex and ultrasensitive immunoassays in clinical diagnosis and treatment. Mechanobiol Med. 2024; 2: 100063.

[119] Stoeva D, Gencheva D, Radoslavov G, Hristov P, Yordanova R, Beev G. Novel DNA barcoding and multiplex PCR strategy for the molecular identification and mycotoxin gene detection of Fusarium spp. in maize from Bulgaria. Methods Protoc. 2025; 8: 78.

[120] Sousa AM, Ferreira D, Rodrigues LR, Pereira MO. Aptamer-based therapy for fighting biofilm-associated infections. J Controlled Release. 2024; 367: 522-539.

[121] Liu H, Yin H, Xiu L, et al. One-Pot Isothermal nucleic acid amplification assisted CRISPR/Cas detection technology: challenges, strategies, and perspectives. Adv Sci. 2025; 12: e06716.

[122] Al Mahmud A, Ahmed MB, Abbey B, Kabir A, Hessel V, Moni MA. Electrochemical nucleic acid biosensors: advancing blood-based diagnostics toward intelligent healthcare in resource-limited settings. TrAC Trends Anal Chem. 2026; 202: 118959.

[123] Tatarova Z, Blumberg DC, Korkola JE, et al. A multiplex implantable microdevice assay identifies synergistic combinations of cancer immunotherapies and conventional drugs. Nat Biotechnol. 2022; 40: 1823-1833.

[124] Soler M, Lechuga LM. Label-free photonic biosensors: key technologies for precision diagnostics. ChemistryEurope. 2025; 3: e202400106.

[125] Couacault P, Avella D, Londoño-Osorio S, et al. Targeted and untargeted metabolomics and lipidomics in dried blood microsampling: recent applications and perspectives. Anal Sci Adv. 2024; 5: e2400002.

[126] Saadh MJ, Saeed TN, Al-Hussainy AF, et al. Revolutionizing cancer diagnostics: the promise of exosome-based biosensors. Anal Biochem. 2025; 705: 115912.

[127] Chin LK, Son T, Hong J-S, et al. Plasmonic sensors for extracellular vesicle analysis: from scientific development to translational research. ACS Nano. 2020; 14: 14528-14548.

[128] Guo R, Yin F, Sun Y, et al. Ultrasensitive simultaneous detection of multiplex disease-related nucleic acids using double-enhanced surface-enhanced raman scattering nanosensors. ACS Appl Mater Interfaces. 2018; 10: 25770-25778.

[129] Siddiqui SM, Welch NL, Nguyen TG, et al. Bead-based approaches for increased sensitivity and multiplexing of CRISPR diagnostics. Nat Biomed Eng. 2026; 10: 939-951.

[130] Jeon H-B, Song D-Y, Park YJ, Kim D-M. Enhancing ELISA sensitivity: from surface engineering to synthetic biology. Biosensors. 2025; 15: 434.

[131] Morris MC, Veile R, Friend LA, et al. Effects of whole blood leukoreduction on platelet function and hemostatic parameters. Transfus Med Oxf Engl. 2019; 29: 351-357.

[132] Liu X, Xu W, Jiang H, et al. Paper-based microfluidic devices: a powerful strategy for rapid detection. Micromachines. 2025; 17: 64.

[133] Ning Y, Li S, Ong MEH, et al. A novel interpretable machine learning system to generate clinical risk scores: an application for predicting early mortality or unplanned readmission in a retrospective cohort study. PLOS Digit Health. 2022; 1: e0000062.

[134] Goodwin L, Faber J, Holmberg KH, et al. Whole blood stability in bioanalytical method validation: updated recommendations from the European Bioanalysis Forum. Bioanalysis. 2025; 17: 767-768.

[135] Choi S-J, Choi J, Kim J, et al. Advanced technologies in extracellular vesicle biosensing: platforms, standardization, and clinical translation. Molecules. 2026; 31: 227.

[136] Watterson WJ, Tanyeri M, Watson AR, et al. Droplet-based high-throughput cultivation for accurate screening of antibiotic resistant gut microbes. eLife. 2020; 9: e56998.

[137] Marcelletti JF, Evans CL, Saxena M, Lopez AE. Calculations for adjusting endogenous biomarker levels during analytical recovery assessments for ligand-binding assay bioanalytical method validation. AAPS J. 2015; 17: 939-947.

[138] Nam Y, Lee JH, Kim SM, et al. Periodic comparability verification and within-laboratory harmonization of clinical chemistry laboratory results at a large healthcare center with multiple instruments. Ann Lab Med. 2022; 42: 150-159.

[139] Théry C, Witwer KW, Aikawa E, Alcaraz MJ, Zuba-Surma EK. Minimal information for studies of extracellular vesicles 2018 (MISEV2018): a position statement of the International Society for Extracellular Vesicles and update of the MISEV2014 guidelines. J Extracell Vesicles. 2018; 7: 1535750.

[140] Fredericks CM, Kunihiro JKI, Zheng H, Waghu NR, Kamkar M. Chemical enhancements and advanced manufacturing methods of poly (lactic acid) for tissue engineering applications. Polymer. 2024; 313: 127691.

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