Aptamers are valuable alternative ligands for affinity separations. Here, we describe the aptamer-based affinity separation of His-tagged proteins using an aptamer directed against the His-tag. The immobilization of the aptamer to magnetic beads is described as well as the aptamer-based purification and proper methods for the characterization of the process. Moreover, indications for the transfer of the process to other aptamers are given.
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http://dx.doi.org/10.1007/978-1-4939-2447-9_7 | DOI Listing |
Drug Test Anal
January 2025
European Monitoring Center for Emerging Doping Agents, German Sport University Cologne, Cologne, Germany.
A cost minimized immunoaffinity protocol was developed, which allows the direct purification of ERAs (urinary and recombinant human EPO, Darbepoetin, EPO-Fc, CERA) from human urine. The method applies magnetic beads and needs no covalent immobilization of the capture antibody. It requires only 10 mL of urine, 1 μg of anti-EPO antibody, and 25 μL of bead slurry.
View Article and Find Full Text PDFJ Proteome Res
January 2025
Discovery Research, AbbVie, Inc., 1 North Waukegan Rd., North Chicago, Illinois 60064, United States.
Affinity capture (AC) combined with mass spectrometry (MS)-based proteomics is highly utilized throughout the drug discovery pipeline to determine small-molecule target selectivity and engagement. However, the tedious sample preparation steps and time-consuming MS acquisition process have limited its use in a high-throughput format. Here, we report an automated workflow employing biotinylated probes and streptavidin magnetic beads for small-molecule target enrichment in the 96-well plate format, ending with direct sampling from EvoSep Solid Phase Extraction tips for liquid chromatography (LC)-tandem mass spectrometry (MS/MS) analysis.
View Article and Find Full Text PDFAnal Chem
January 2025
State Key Laboratory of Food Science and Resources, Jiangnan University, Wuxi 214122, China.
Timely and accurate detection of trace mycotoxins in agricultural products and food is significant for ensuring food safety and public health. Herein, a deep learning-assisted and entropy-driven catalysis (EDC)-Argonaute powered fluorescence single-particle aptasensing platform was developed for ultrasensitive detection of fumonisin B (FB) using single-stranded DNA modified with biotin and red fluorescence-encoded microspheres as a signal probe and streptavidin-conjugated magnetic beads as separation carriers. The binding of aptamer with FB releases the trigger sequence to mediate EDC cycle to produce numerous 5'-phosphorylated output sequences, which can be used as the guide DNA to activate downstream Argonaute (Ago) for cleaving the signal probe, resulting in increased number of fluorescence microspheres remaining in the final reaction supernatant after magnetic separation.
View Article and Find Full Text PDFEar Nose Throat J
January 2025
Department of Otolaryngology, People's Hospital of Jingshan, Jingshan Union Hospital of Huazhong University of Science and Technology, Jingmen City, Hubei Province, People's Republic of China.
This case involved a 21-year-old male patient who was admitted due to having a lump behind the left ear that had been present for 2 years and had gradually increased in size for over a year. This was accompanied by palpable hard masses on the same side of the neck. Laboratory tests indicated an elevated eosinophil count, and magnetic resonance imaging confirmed the "string-of-beads" sign in the left cervical lymph nodes.
View Article and Find Full Text PDFTalanta
January 2025
Key Laboratory of Chemical Biology and Traditional Chinese Medicine Research (Ministry of Education of China), College of Chemistry and Chemical Engineering, Hunan Normal University, Changsha, 410081, PR China. Electronic address:
E-selectin (CD62E) is an adhesion molecule expressed on the surface of endothelial cells (ECs) and its level increases significantly upon the stimulation of ECs by inflammatory factors. Quantitative analysis of CD62E is of great importance to early diagnosis and treatment of vascular diseases and hypertension. A new method for the determination of CD62E was developed using a portable pH meter in this work.
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