Aggregation-Induced Emission Luminogen-Based Dual-Mode Enzyme-Linked Immunosorbent Assay for Ultrasensitive Detection of Cancer Biomarkers in a Broad Concentration Range DOI
Jianlei Shen, Bo Situ, Xianchao Du

et al.

ACS Sensors, Journal Year: 2022, Volume and Issue: 7(3), P. 766 - 774

Published: Feb. 18, 2022

The enzyme-linked immunosorbent assay (ELISA) is one of the most commonly used methods for measuring antibodies and antigens in biological samples. However, developing new ELISAs with high detection sensitivity broad dynamic ranges without resorting to complicated signal processing equipment setups remains a challenge. In this work, we report strategy simultaneously improve broaden range by replacing chromogenic reagents traditional an aggregation-induced emission luminogen (AIEgen). developed AIE–ELISA could generate complementary absorbance fluorescence signals linear 1.6–25,000 pg/mL. application dual-mode prostate-specific antigen (PSA) realized limit 1.3 pg/mL (3.78 × 10–14 M) improvement approximately 2 orders magnitude compared single-mode ELISA, which enabled it discriminate minor PSA difference patient's serum. simpler experimental operation, faster enzyme response speed, better photostability AIEgen than showed that our holds great potential fields immunoassay, immunohistochemistry, immunocytochemistry.

Language: Английский

Ultrasensitive and Highly Specific Lateral Flow Assays for Point-of-Care Diagnosis DOI Creative Commons
Yilin Liu, Li Zhan, Zhenpeng Qin

et al.

ACS Nano, Journal Year: 2021, Volume and Issue: 15(3), P. 3593 - 3611

Published: Feb. 19, 2021

Lateral flow assays (LFAs) are paper-based point-of-care (POC) diagnostic tools that widely used because of their low cost, ease use, and rapid format. Unfortunately, traditional commercial LFAs have significantly poorer sensitivities (μM) specificities than standard laboratory tests (enzyme-linked immunosorbent assay, ELISA: pM-fM; polymerase chain reaction, PCR: aM), thus limiting impact in disease control. In this Perspective, we review the evolving efforts to increase sensitivity specificity LFAs. Recent work improve through assay improvement includes optimization kinetics signal amplification by either reader systems or additional reagents. Together, these produced with ELISA-level (pM-fM). addition, sample preamplification can be applied both nucleic acids (direct amplification) other analytes (indirect prior LFA testing, which lead PCR-level (aM) sensitivity. However, strategies also detection time complexity, inhibits large-scale POC use Perspectives achieve future (<30 min), ultrasensitive (PCR-level), "sample-to-answer" diagnostics provided. case specificity, recent research focused on high-affinity molecules reduce nonspecific binding. Furthermore, novel highly specific molecules, such as CRISPR/Cas systems, integrated into diagnosis produce not only but diagnostics. summary, continuing improvements, may soon offer performance at is competitive techniques while retaining a

Language: Английский

Citations

424
The emerging landscape of single-molecule protein sequencing technologies DOI Open Access
Javier A. Alfaro,

Peggy R. Bohländer,

Mingjie Dai

et al.

Nature Methods, Journal Year: 2021, Volume and Issue: 18(6), P. 604 - 617

Published: June 1, 2021

Language: Английский

Citations

290
Rolling Circle Replication for Biosensing, Bioimaging, and Biomedicine DOI
Shuzhen Yue, Yuwei Li,

Zhenjie Qiao

et al.

Trends in biotechnology, Journal Year: 2021, Volume and Issue: 39(11), P. 1160 - 1172

Published: March 12, 2021

Language: Английский

Citations

172
The Human Proteoform Project: Defining the human proteome DOI Creative Commons
Lloyd M. Smith, Jeffrey N. Agar, Julia Chamot‐Rooke

et al.

Science Advances, Journal Year: 2021, Volume and Issue: 7(46)

Published: Nov. 12, 2021

Proteins are the primary effectors of function in biology, and thus, complete knowledge their structure properties is fundamental to deciphering basic translational research. The chemical diversity proteins expressed many proteoforms, which result from combinations genetic polymorphisms, RNA splice variants, posttranslational modifications. This foundational for biological complexes networks that control biology yet remains largely unknown. We propose here an ambitious initiative define human proteome, is, generate a definitive reference set proteoforms produced genome. Several examples power importance proteoform-level disease-based research presented along with call improved technologies two-pronged strategy Human Proteoform Project.

Language: Английский

Citations

170
Emerging ELISA derived technologies for in vitro diagnostics DOI
Ping Peng, Chang Liu, Zedong Li

et al.

TrAC Trends in Analytical Chemistry, Journal Year: 2022, Volume and Issue: 152, P. 116605 - 116605

Published: March 25, 2022

Language: Английский

Citations

133
Framework for rapid comparison of extracellular vesicle isolation methods DOI Creative Commons
Dmitry Ter‐Ovanesyan, Maia Norman, Roey Lazarovits

et al.

eLife, Journal Year: 2021, Volume and Issue: 10

Published: Nov. 16, 2021

Extracellular vesicles (EVs) are released by all cells into biofluids and hold great promise as reservoirs of disease biomarkers. One the main challenges in studying EVs is a lack methods to quantify that sensitive enough can differentiate from similarly sized lipoproteins protein aggregates. We demonstrate use ultrasensitive, single-molecule array (Simoa) assays for quantification using three widely expressed transmembrane proteins: tetraspanins CD9, CD63, CD81. Using Simoa measure these EV markers, well albumin contamination, we were able compare relative efficiency purity several commonly used isolation plasma cerebrospinal fluid (CSF): ultracentrifugation, precipitation, size exclusion chromatography (SEC). further assays, on one platform, improve SEC CSF. Our results highlight utility quantifying proteins provide rapid framework comparing improving biofluids.

Language: Английский

Citations

109
High-Throughput, High-Multiplex Digital Protein Detection with Attomolar Sensitivity DOI

Connie Wu,

Tyler Dougan, David R. Walt

et al.

ACS Nano, Journal Year: 2022, Volume and Issue: 16(1), P. 1025 - 1035

Published: Jan. 14, 2022

A major challenge in many clinical diagnostic applications is the measurement of low-abundance proteins and other biomolecules biological fluids. Digital technologies such as digital enzyme-linked immunosorbent assay (ELISA) have enabled 1000-fold increases sensitivity over conventional protein detection methods. However, current ELISA still possess insufficient sensitivities for rare biomarkers require specialized instrumentation or time-consuming workflows that limited their widespread implementation. To address these challenges, we developed a more sensitive streamlined platform, Molecular On-bead Signal Amplification Individual Counting (MOSAIC), which attains low attomolar limits detection, with an order magnitude enhancement MOSAIC uses rapid, automatable flow cytometric readout vastly throughput easily integrated into existing laboratory infrastructure. As provides high sampling efficiencies target molecules, bead number can readily be tuned to enhance signal-to-background precision. Furthermore, solution-based signal expands analytes simultaneously measured higher-order multiplexing femtomolar below, compared microwell- droplet-based proof principle, apply toward improving detectability cytokines saliva ultrasensitive multiplexed measurements eight plasma saliva. The sensitivity, throughput, broad abilities provide highly accessible versatile capabilities potentially accelerate biomarker discovery testing diverse disease applications.

Language: Английский

Citations

94
Future of Digital Assays to Resolve Clinical Heterogeneity of Single Extracellular Vesicles DOI
Renee‐Tyler T. Morales, Jina Ko

ACS Nano, Journal Year: 2022, Volume and Issue: 16(8), P. 11619 - 11645

Published: July 29, 2022

Extracellular vesicles (EVs) are complex lipid membrane vehicles with variable expressions of molecular cargo, composed diverse subpopulations that participate in the intercellular signaling biological responses disease. EV-based liquid biopsies demonstrate invaluable clinical potential for overhauling current practices disease management. Yet, EV heterogeneity is a major needle-in-a-haystack challenge to translate their use into practice. In this review, existing digital assays will be discussed analyze EVs at single vesicle resolution, and future opportunities optimize throughput, multiplexing, sensitivity highlighted. Furthermore, review outline challenges impact translation technologies diagnostics treatment monitoring.

Language: Английский

Citations

86
Emerging biosensing and transducing techniques for potential applications in point-of-care diagnostics DOI Creative Commons
Junjie Qin, Wei Wang, Liqian Gao

et al.

Chemical Science, Journal Year: 2022, Volume and Issue: 13(10), P. 2857 - 2876

Published: Jan. 1, 2022

POC diagnostics are driven by the rapid advances in CRISPR, electrochemical and optical biosensors. Related emerging strategies described discussed from perspective of facilitating practical application biosensors testing.

Language: Английский

Citations

80
Computer Vision-Based Artificial Intelligence-Mediated Encoding-Decoding for Multiplexed Microfluidic Digital Immunoassay DOI

Weiqi Zhao,

Yang Zhou, Y. X. Feng

et al.

ACS Nano, Journal Year: 2023, Volume and Issue: 17(14), P. 13700 - 13714

Published: July 17, 2023

Digital immunoassays with multiplexed capacity, ultrahigh sensitivity, and broad affordability are urgently required in clinical diagnosis, food safety, environmental monitoring. In this work, a multidimensional digital immunoassay has been developed through microparticle-based encoding artificial intelligence-based decoding, enabling detection high sensitivity convenient operation. The information encoded the features of microspheres, including their size, number, color, allows for simultaneous identification accurate quantification multiple targets. Computer vision-based intelligence can analyze microscopy images decoding output results visually. Moreover, optical imaging be well integrated microfluidic platform, allowing encoding-decoding computer intelligence. This simultaneously inflammatory markers antibiotics within 30 min range from pg/mL to μg/mL, which holds great promise as an intelligent bioassay next-generation biosensing.

Language: Английский

Citations

45