Recent advances in self‐immobilizing fluorescent probes for in vivo imaging

Smart Molecules, Journal Year: 2024, Volume and Issue: 2(3)

Published: Aug. 23, 2024

Abstract In situ precise detection of bioactive molecules with high sensitivity and spatiotemporal resolution is essential for studying physiological events disease diagnosis. The utilization versatile fluorescent probes in fluorescence imaging offers a powerful tool vivo biomarkers closely associated pathological conditions. However, the dynamic behavior leading to rapid clearance small molecule from regions interest severely compromises their potential imaging. Notably, self‐immobilizing that selectively recognize diseased tissues while improving retention enrichment enable accurate high‐fidelity this review, we aim summarize strategies employed recent advances performance precision using techniques. Lastly, discuss prospects challenges promote further development application more delicate probes.

Language: Английский

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Cellular and Biophysical Applications of Genetic Code Expansion

Chemical Reviews, Journal Year: 2024, Volume and Issue: 124(11), P. 7465 - 7530

Published: May 16, 2024

Despite their diverse functions, proteins are inherently constructed from a limited set of building blocks. These compositional constraints pose significant challenges to protein research and its practical applications. Strategically manipulating the cellular synthesis system incorporate novel blocks has emerged as critical approach for overcoming these in application. In past two decades, field genetic code expansion (GCE) achieved advancements, enabling integration numerous functionalities into across variety organisms. This technological evolution paved way extensive application multiple domains, including imaging, introduction probes research, analysis protein–protein interactions, spatiotemporal control function, exploration proteome changes induced by external stimuli, endowed with functions. this comprehensive Review, we aim provide an overview biophysical applications that have employed GCE technology over decades.

Language: Английский

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Thermally Triggered Triazolinedione–Tyrosine Bioconjugation with Improved Chemo- and Site-Selectivity

Journal of the American Chemical Society, Journal Year: 2024, Volume and Issue: 146(18), P. 12672 - 12680

Published: April 29, 2024

A bioconjugation strategy is reported that allows the derivatization of tyrosine side chains through triazolinedione-based "Y-clicking". Blocked triazolinedione reagents were developed that, in contrast to classical reagents, can be purified before use, stored for a long time, and allow functionalization with wider range cargoes labels. These are bench-stable at room temperature but steadily release highly reactive triazolinediones upon heating 40 °C buffered media physiological pH, showing sharp response over 0 range. This conceptually interesting strategy, which complementary existing photo- or electrochemical bioorthogonal bond-forming methods, not only avoids synthesis handling difficulties these click-like also markedly improves selectivity profile conjugation reaction itself. It oxidative damage "off-target" tryptophan labeling, it even site-selectivity discriminating between different on same protein polypeptide chains. In this research article, we describe stepwise development from their short modular small-molecule model studies proof-of-principle chemistry peptides proteins.

Language: Английский

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Peptide and Protein Cysteine Modification Enabled by Hydrosulfuration of Ynamide

ACS Central Science, Journal Year: 2024, Volume and Issue: 10(9), P. 1742 - 1754

Published: Aug. 21, 2024

Efficient functionalization of peptides and proteins has widespread applications in chemical biology drug discovery. However, the chemoselective site-selective modification remains a daunting task. Herein, highly efficient chemo-, regio-, stereoselective hydrosulfuration ynamide was identified as an method for precise by uniquely targeting thiol group cysteine (Cys) residues. This novel could be facilely operated aqueous buffer fully compatible with wide range proteins, including small model large full-length antibodies, without compromising their integrity functions. Importantly, this reaction provides Z-isomer corresponding conjugates exclusively superior stability, offering approach to peptide protein therapeutics. The potential application further exemplified Cys-bioconjugation variety ynamide-bearing functional molecules such molecule drugs, fluorescent/affinity tags, PEG polymers. It also proved redox proteomic analysis through Cys-alkenylation. Overall, study bioorthogonal tool Cys-specific functionalization, which will find broad synthesis peptide/protein conjugates.

Language: Английский

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Exploring Protein Bioconjugation: A Redox-Based Strategy for Tryptophan Targeting

Research, Journal Year: 2024, Volume and Issue: 7

Published: Jan. 1, 2024

Amino acid bioconjugation technology has emerged as a pivotal tool for linking small-molecule fragments with proteins, antibodies, and even cells. The study in

Language: Английский

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3-Acyl-4-Pyranone as a Lysine Residue-Selective Bioconjugation Reagent for Peptide and Protein Modification

Bioconjugate Chemistry, Journal Year: 2024, Volume and Issue: 35(3), P. 286 - 299

Published: March 7, 2024

Chemoselective protein modification plays extremely important roles in various biological, medical, and pharmaceutical investigations. Mimicking the mechanism of chemoselective reaction between natural azaphilones primary amines, this work successfully simplified azaphilone scaffold into much simpler 3-acyl-4-pyranones. Examinations confirmed that these slim-size mimics perfectly kept unique reactivity for selective conjugation with amines including lysine residues peptides proteins. The newly developed pyranone tool presents remarkably increased aqueous solubility compatible second-order rate constant by comparison original azaphilone. Additional advantages also include ease biorthogonal combinative use a copper-catalyzed azide–alkyne Click reaction, which was conveniently applied to decorate lysozyme neutral-, positive- negative-charged functionalities parallel. Moderate-degree positively charged quaternary ammoniums revealed increase enzymatic activities.

Language: Английский

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Assembling Branched and Macrocyclic Peptides on Proteins

Chemical Communications, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 1, 2025

A small molecular adapter enables site-specific peptide–protein conjugation via cysteine modification and cyanopyridine–aminothiol reactions, yielding complex branched or cyclic architectures for potential protein therapeutics.

Language: Английский

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Antibody‐Nanoparticle Conjugates in Therapy: Combining the Best of Two Worlds

Small, Journal Year: 2025, Volume and Issue: unknown

Published: March 6, 2025

Monoclonal antibodies (mAbs) and antibody fragments have revolutionized medicine as highly specific binding agents inhibitors. At the same time, several types of nanomaterials, including liposomes, lipid nanoparticles (NPs), polymersomes, metal oxide NPs, protein nanostructures, are increasingly utilized explored for therapeutic potential due to their versatility, chemical physical properties, tunability. However, nanomaterials alone often lack specificity, leading relatively low efficacy and/or high toxicity. To address this problem, a rapidly emerging area is antibody-nanomaterial conjugates (ANCs), which combine precise targeting specificity with effector functionality nanomaterial. In review, we give brief introduction mAbs major conjugation techniques, describe classes being studied potential, review literature on ANCs each class. Special focus given applications addressing blood-brain barrier, delivering nucleic acids, light-activated ANCs. While many disease targets related cancer, also under development autoimmune, neurological, infectious diseases. important challenges remain, poised become next-generation technology.

Language: Английский

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Phosphine-promoted Thiol-Specific Bioconjugation with Allylic Acetates

Research Square (Research Square), Journal Year: 2025, Volume and Issue: unknown

Published: March 24, 2025

Thiol-specific modification of proteins through conjugation with small molecules represents a critical advancement in biological research and therapeutic development, particularly the context antibody-drug conjugates (ADCs) for targeted cancer therapy. Despite widespread use maleimide-based linkers, their stability under physiological conditions remains major limitation, often compromising efficacy. In this investigation, we developed novel efficient thiol-specific bioconjugation strategy that employs allylic acetate, activated by recyclable solid phosphine catalyst. This approach achieves high yields demonstrates robust bio-compatible, room-temperature conditions. Notably, it sets new record fastest cysteine-conjugation reaction rate reported to date, constant k₂ = 1.49 × 10⁶ M^{− 1}s^{− 1}. The method expands substrate scope introduces sustainable, environmentally friendly ADC linker design, offering significant creation stable, biocompatible, therapeutically effective compounds.

Language: Английский

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Orthogonal Site-Specific Dual Bioconjugation of Aryl and Alkyl Thiols

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

Published: May 5, 2025

We introduce aryl thiols as nucleophiles for site-specific protein and antibody bioconjugation, which allows the orthogonal labeling of native cysteines double modification strategies. In a high-yielding synthesis, we aromatic thiol substituents in two amino acids (4-SH-L-Phe 3-SH-L-Tyr), can be site-specifically incorporated into C-terminus using enzyme tubulin tyrosine ligase (TTL, Tub-tag labeling). particular, found that 3-SH-L-tyrosine shows excellent water solubility incorporation rates, similar to previously described Tyr-derivatives. 2D NMR experiments revealed pKa value 5.5 modality 3-SH-L-tyrosine, matches pH-dependent reactivity profile toward thiol-selective ethynyl-triazolyl-phosphinate (ETP) electrophiles. Most importantly, addition glutathione had no significant effect on reaction between ETPs at pH 7.0 below, supporting alkyl thiols. utilized these findings develop an dual bioconjugation protocol proteins, featuring TTL-ligation incorporate arylthiol-containing acid derivative, followed by thiolate functionalization subsequent conjugation 8.3. This strategy was used generate highly fluorescent photostabilized nanobody fully functionalized antibody-drug conjugate carrying different cytotoxic payloads, displays potent cytotoxicity cells target antigen strong bystander effect.

Language: Английский

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