The discovery that certain diseases have specific miRNA signatures which correspond to disease progression opens a new biomarker category. The detection of these small non-coding RNAs is performed routinely using body fluids or tissues with real-time PCR, next-generation sequencing, or amplification-based miRNA assays. Antibody-based detection systems allow an easy onset handling compared to PCR or sequencing and can be considered as alternative methods to support miRNA diagnostic in the future. In this study, we describe the generation of a camelid heavy-chain-only antibody specifically recognizing miRNAs to establish an antibody-based detection method. The generation of nucleic acid-specific binders is a challenge. We selected camelid binders via phage display, expressed them as VHH as well as full-length antibodies, and characterized the binding to several miRNAs from a signature specific for dilated cardiomyopathy. The described workflow can be used to create miRNA-specific binders and establish antibody-based detection methods to provide an additional way to analyze disease-specific miRNA signatures.
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http://dx.doi.org/10.3390/ijms23116275 | DOI Listing |
Anal Chem
January 2025
The MOE Key Laboratory of Spectrochemical Analysis and Instrumentation, State Key Laboratory of Physical Chemistry of Solid Surfaces, Department of Chemistry, College of Chemistry and Chemical Engineering, Xiamen University, Xiamen 361005, China.
CD28 is a costimulatory receptor that provides the second signal necessary for T-cell activation and is associated with diseases, including rheumatoid arthritis, asthma, and cancer. Targeting CD28 is crucial for both functional bioanalysis and therapeutic development. Molecular probes, particularly fluorescent probes, can enhance our understanding of CD28's cellular roles.
View Article and Find Full Text PDFVet Med Sci
January 2025
Department of Microbiology, Faculty of Veterinary and Animal Science, Hajee Mohammad Danesh Science and Technology University, Dinajpur, Bangladesh.
Background: Brucellosis is a zoonotic disease caused by Brucella spp., affecting various animals and humans, leading to significant economic and public health impacts. Traditional diagnostic methods, mainly serological, often fail to detect seronegative carriers, which continue to spread the infection.
View Article and Find Full Text PDFAnal Chem
January 2025
Guangdong Provincial Key Laboratory of Food Quality and Safety, South China Agricultural University, Guangzhou 510642, China.
The rapid, sensitive, and accurate detection of paralytic shellfish toxins (PSTs), such as saxitoxin (STX), is critical for protecting human health due to the frequent occurrence of toxic red tides. In this work, to address the low affinity of traditional mouse monoclonal antibodies (m-mAbs), rabbit monoclonal antibodies (r-mAbs) against STX were produced by a single B-cell sorting culture and a cross-selection strategy. The r-mAbs showed 100-fold improvement in sensitivity (IC = 0.
View Article and Find Full Text PDFJ Microbiol Methods
December 2024
Applied Microbiology Research Center, Biomedicine Technologies Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran. Electronic address:
Alpha-toxin of Staphylococcus aureus belongs to the pore-forming toxin (PFT) family, which can lyse red and white blood cells. In addition to the existence of the hla gene in the majority of S. aureus strains (about 95 %), higher expression exhibits enhanced pathogenicity to the bacteria.
View Article and Find Full Text PDFSci Rep
December 2024
Department of Civil, Environmental and Construction Engineering, University of Central Florida, Orlando, FL, USA.
Harmful algal blooms (HABs) can release cyanotoxins such as microcystins (MCs), especially, microcystin-leucine-arginine (MC-LR) which is one of the commonest and most toxic, into our water bodies and can lead to several acute or chronic diseases such as liver diseases and respiratory irritation in humans. There is an increasing need for rapid and simple detection of MC-LR in water bodies for early warning of HABs. In this study, we developed an innovative on-site screening electrochemical impedance spectroscopy (EIS) biosensor with a simplified calibration curve that can rapidly detect blooms for early action in similar water bodies.
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