We have developed a rapid and sensitive thin film assay for in-process monitoring of target protein purification. This novel biosensor method provides rapid (5-min) visual evaluation of column purification fractions. The method can be used to monitor the efficiency of purification and potential loss of protein if the column binding capacity is exceeded. The eluted fractions containing the highest yield of target protein can be quickly identified, pooled, and processed. This convenient platform, known as the SILAS product, is a thin-film detection technology in which specific molecular interactions are transduced into visible color changes based on changes in the optical thickness of layers on a silicon surface. The results are interpreted without instrumentation. Proteins eluted from a purification column are adsorbed to the assay surface, and the ligand of interest (target) can be identified with specific binding reagents. Here we demonstrate two protein purification applications for the SILAS technology product: monitoring antibody elution from a Protein G column and evaluating the efficiency of purification of a glutathione-S-transferase (GST)-tagged recombinant protein through each step of the purification process.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.2144/02324pt05 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Gut microbiota modulation via fecal microbiota transplantation mitigates hyperoxaluria and calcium oxalate crystal depositions induced by high oxalate diet.
Gut Microbes
December 2025
Department of Urology, The Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, Guangdong, China.
Hyperoxaluria, including primary and secondary hyperoxaluria, is a disorder characterized by increased urinary oxalate excretion and could lead to recurrent calcium oxalate kidney stones, nephrocalcinosis and eventually end stage renal disease. For secondary hyperoxaluria, high dietary oxalate (HDOx) or its precursors intake is a key reason. Recently, accumulated studies highlight the important role of gut microbiota in the regulation of oxalate homeostasis.
View Article and Find Full Text PDFMouse-derived Synaptosomes Trypsin Cleavage Assay to Characterize Synaptic Protein Sub-localization.
Bio Protoc
January 2025
Department of Structural Interactomics, Leibniz-Forschungsinstitut für Molekulare Pharmakologie (FMP), Berlin, Germany.
Neurons communicate through neurotransmission at highly specialized junctions called synapses. Each neuron forms numerous synaptic connections, consisting of presynaptic and postsynaptic terminals. Upon the arrival of an action potential, neurotransmitters are released from the presynaptic site and diffuse across the synaptic cleft to bind specialized receptors at the postsynaptic terminal.
View Article and Find Full Text PDFScreening, identification, and mechanism of novel antioxidant peptides in walnut meal under aerobic stress.
Food Chem
April 2025
Engineering Research Center of Bio-process, Ministry of Education/Key Laboratory for Agricultural Products Processing of Anhui Province/School of Food and Biological Engineering, Hefei University of Technology, Hefei 230601, China. Electronic address:
Walnut (Juglans regia L.) meal, being the primary by-product of walnut oil processing, is rich in high-quality proteins and of significant potential for development and utilization. The study used multi-stage gradient purification, liquid-quantity chromatography, and computerized virtual screening to isolate and characterize antioxidant peptides from walnut meal.
View Article and Find Full Text PDFPreclinical evaluation and preliminary clinical study of Ga-NODAGA-NM-01 for PET imaging of PD-L1 expression.
Cancer Imaging
January 2025
Department of Nuclear Medicine, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Hongkou District, No. 100, Haining Road, Shanghai, 200080, China.
Background: Programmed cell death 1/programmed death ligand-1 (PD-L1)-based immune checkpoint blockade is an effective treatment approach for non-small-cell lung cancer (NSCLC). However, immunohistochemistry does not accurately or dynamically reflect PD-L1 expression owing to its spatiotemporal heterogeneity. Herein, we assessed the feasibility of using a Ga-labeled anti-PD-L1 nanobody, Ga-NODAGA-NM-01, for PET imaging of PD-L1.
View Article and Find Full Text PDFAssociation of LPCAT1-rs8352 genetic variant with susceptibility and severity of pediatric bronchial asthma: a case-control study.
BMC Pediatr
January 2025
Department of Pediatrics, Faculty of Medicine, Assiut University, Egypt, 71515, Assiut, Egypt.
Background: This study aimed to investigate the possible association of LPCAT1-rs8352 genetic variant (single nucleotide change C to G) with the onset and severity of pediatric asthma. Additionally, the study examined the influence of LPCAT1-rs8352 genotypes on asthma-related biomarkers including blood eosinophils count (BEC), eosinophil cationic protein (ECP), high-sensitivity C-reactive protein (hs-CRP), and immunoglobulin E (IgE) and on lung function [forced expiratory volume in one second (FEV1) and the forced vital capacity (FVC)].
Patients And Methods: The study included ninety-six participant grouped into two groups: G1 (46 asthmatics) and G2 (50 healthy controls).
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!