Combinatorial High-Throughput Screening of Complex Polymeric Enzyme Immobilization Supports

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(13), P. 9112 - 9123

Published: March 19, 2024

Recent advances have demonstrated the promise of complex multicomponent polymeric supports to enable supra-biological enzyme performance. However, discovery such has been limited by time-consuming, low-throughput synthesis and screening. Here, we describe a novel combinatorial high-throughput platform that enables rapid screening heterogeneous copolymer brushes as immobilization supports, named support (CHESS). Using 384-well plate format, synthesized arrays three-component polymer in microwells using photoactivated surface-initiated polymerization immobilized enzymes situ. The utility CHESS identify optimal under thermally chemically denaturing conditions was usingBacillus subtilisLipase A (LipA). identification with compositions validated immobilizing LipA on polymer-brush-modified biocatalyst particles. We further could be used predict composition priori for previously unexplored enzyme, alkaline phosphatase (AlkP). Our findings demonstrate represents predictable reliable dramatically accelerating search chemical facilitates biocompatible stabilizing materials.

Language: Английский

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Pretargeted Multimodal Tumor Imaging by Enzymatic Self-Immobilization Labeling and Bioorthogonal Reaction

Journal of the American Chemical Society, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 12, 2025

Covalent modification of cell membranes has shown promise for tumor imaging and therapy. However, existing membrane labeling techniques face challenges such as slow kinetics poor selectivity cancer cells, leading to off-target effects suboptimal in vivo efficacy. Here, we present an enzyme-triggered self-immobilization strategy, termed E-SIM, which enables rapid selective with bioorthogonal trans-cycloctene (TCO) handles vivo. E-SIM utilizes P-TCO, alkaline phosphatase (ALP) responsive quinone methide (QM) precursor a TCO group, facilitating the conjugation high-density onto via proximity labeling. These groups then react efficiently tetrazine (Tz)-bearing reporters fast reaction, resulting significant enrichment various sizes modalities on membranes. We demonstrate efficacy reaction pretargeted multimodality tumors Notably, achieve efficient installation Tz-modified Renilla luciferase cells vivo, thereby offering highly sensitive bioluminescence signals detecting guiding surgical removal small human HepG2 liver peritoneal metastases. represents robust tool precise complex environments, feasible multimodal applications.

Language: Английский

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Design of nanostructured lipid carriers and solid lipid nanoparticles for enhanced cellular uptake

International Journal of Pharmaceutics, Journal Year: 2022, Volume and Issue: 624, P. 122014 - 122014

Published: July 16, 2022

In this study PEG-free and zeta potential changing lipid-based nanocarriers providing enhanced cellular uptake were developed. Nanostructured lipid carriers (NLC), consisting of paraffin wax, caprylic/ capric triglyceride, cetyltrimethylammoniumchloride either soy lecithin or polyglycerol-4 laurate solid nanoparticles (SLN) with the same composition but without liquid content All formulations exposed a positive surface charge then coated polyphosphate Graham's salt. Phosphate release from these was evaluated by incubation intestinal alkaline phosphatase as well on Caco-2 monolayer potentials measured. Additionally, studies performed. Within 5 h, remarkable amount phosphate released all incubated phosphatase. Enzymatically induced led to shift up Δ 26 mV. Results change confirmed cells. Cellular cells showed an 5.6-times higher compared inhibited According results, coating is powerful tool obtain for uptake.

Language: Английский

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Emerging procedures and solvents in biological sample pre-treatment

Advances in Sample Preparation, Journal Year: 2023, Volume and Issue: 6, P. 100066 - 100066

Published: April 9, 2023

The treatment of biological samples, especially from complex matrices, has consistently challenged analytical operators. classic problems to be faced for any analysis, regardless the origin sample, such as example contamination and loss analyte, in are particularly emphasized. In particular, main cause error is due degradation analyte which several cases interaction. Many factors can influence stability drugs, chief among them physicochemical properties drug, characteristics matrix, tendency conjugation/deconjugation, sample collection procedure, container (e.g., oxidation, adsorption), use preservatives or other additives. problem severe toxicological forensic fields, analyzes considered "non-repeatable." this review, we will explore all major pre-extraction phase chemical-analytical aspect pharmacotoxicological fields.

Language: Английский

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Emerging technologies to increase gastrointestinal transit times of drug delivery systems

Journal of Controlled Release, Journal Year: 2022, Volume and Issue: 346, P. 289 - 299

Published: April 28, 2022

Apart from already established technologies to increase gastrointestinal transit times, including devices rapidly increasing in size once they have reached the stomach order retard passage through pylorus, formulations that float on gastric fluids and mucoadhesive drug delivery systems adhering mucosa, there are new emerging might be game changing. They include mucus permeating nanocarriers able diffuse deeply into gel layer of intestinal mucosa remaining for a prolonged time period (i), charge-converting shift their zeta potential negative positive within providing strong ionic bonds with anionic glycoproteins (ii) thiolated cyclodextrins form even covalent cysteine-rich subdomains (iii). Within this review we will provide an overview about these critically discuss shortcomings.

Language: Английский

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Biosensing Intestinal Alkaline Phosphatase by Pregnancy Test Strips Based on Target-Triggered CRISPR-Cas12a Activity to Monitor Intestinal Inflammation

Analytical Chemistry, Journal Year: 2023, Volume and Issue: 95(37), P. 14111 - 14118

Published: Sept. 5, 2023

With an increasing incidence worldwide, inflammatory bowel disease (IBD) is a chronic affecting the gastrointestinal tract, which impairs life quality of patients. Therefore, it great significance to construct sensitive, simple, and convenient biosensor analyze IBD-associated biomarkers for auxiliary diagnosis IBD. Intestinal alkaline phosphatase (IAP), expressed by intestinal epithelium, endogenous protein that thought play vital role in maintaining homeostasis considered potential biomarker Here, IAP detection method was developed using pregnancy test strips dephosphorylation. Initially, double-stranded DNA (dsDNA) designed respond acted as activator Cas12a. In presence IAP, dsDNA not digested lambda exonuclease (λ exo), hybridized Cas12a-crRNA duplex resulted activation trans-cleavage Further, activated Cas12a cleaved single-strand (ssDNA) linker MBs-ssDNA-hCG probe, triggering release hCG. magnetic separation, released hCG could be quantitatively detected strips. levels were analyzed feces from colitis healthy mice The results showed level (3.89 ± 1.92 U/L) much lower than (39.64 24.93 U/L), indicating correlation between inflammation. Taken together, user-friendly assay based on constructed monitor used diagnostic approach IBD clinical scene.

Language: Английский

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Host–Guest Adduct as a Stimuli‐Responsive Prodrug: Enzyme‐Triggered Self‐Assembly Process of a Short Peptide Within Mitochondria to Induce Cell Apoptosis

Advanced Healthcare Materials, Journal Year: 2024, Volume and Issue: 14(4)

Published: Nov. 6, 2024

To address the issue of nonspecific biodistribution a chemotherapeutic drug, stable [2]pseudorotaxane complexes (PK@CAOPP and PR@CAOPP) are used to demonstrate proof concept. Cationic -PPh3 + moiety in CAOPP allows specific localization PK@CAOPP/ PR@CAOPP mitochondrial membrane (MM). Electrostatic interaction between cationic LysinePK or ArgininePR negatively charged phosphoesterCAOPP functionality favours strong adduct formation. The ALP-induced hydrolytic cleavage phosphoester cancer cells triggers dephosphorylation releases PK/ PR from PK@CAOPP/PR@CAOPP. PK PR, derived Phe-Phe dipeptide, formed fibril-like molecular aggregates MM induce dysfunction, depolarization, ROS generation apoptotic MCF7 cell death. Such phenomena were not observed ALP-negative HEK293 normal cells. These propositions confirmed through control studies using NBDK PE, other guest molecules. Smaller size inclusion short peptides (PK PR) within hydrophobic interior CAOPP, attributed their stability blood serum. Thus, we have demonstrated use supramolecular adducts as potential therapeutic option for treating without affecting healthy efficacy was also established with an in-vivo tumour xenograft model Balb/c nude mice.

Language: Английский

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Responsive shape-shifting nanoarchitectonics and its application in tumor diagnosis and therapy

Journal of Controlled Release, Journal Year: 2022, Volume and Issue: 352, P. 600 - 618

Published: Nov. 3, 2022

Language: Английский

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Fosfatriclaben, a prodrug of triclabendazole: Preparation, stability, and fasciolicidal activity of three new intramuscular formulations

Veterinary Parasitology, Journal Year: 2024, Volume and Issue: 327, P. 110113 - 110113

Published: Jan. 12, 2024

Language: Английский

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Unraveling Caco-2 Cells through Functional and Transcriptomic Assessments.

Regulatory Toxicology and Pharmacology, Journal Year: 2025, Volume and Issue: 156, P. 105771 - 105771

Published: Jan. 5, 2025

Language: Английский

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