Bioconjugate Chemistry最新文献

{"title":"Bivalent Hapten Display Strategies for Conjugate Vaccines Targeting Opioid Mixtures Containing Fentanyl.","authors":"Carly Baehr, Rajwana Jahan, Ann Gebo, Jennifer Vigliaturo, Daihyun Song, Md Toufiqur Rahman, Davide Tronconi, Aaron Khaimraj, Robert Seaman, Courtney Marecki, Caroline M Kim, Stefano Persano, Scott P Runyon, Marco Pravetoni","doi":"10.1021/acs.bioconjchem.4c00548","DOIUrl":"10.1021/acs.bioconjchem.4c00548","url":null,"abstract":"<p><p>Increasingly, street mixtures of opioids are reported to contain combinations of synthetic opioids, such as fentanyl with fentanyl analogues or counterfeit oxycodone pills containing fentanyl. While antiopioid immunotherapeutics have been investigated as a possible approach to address the opioid epidemic, the efficacy of vaccines and antibodies is limited to specific target opioids, based on the chemical structure of the haptens used in vaccines. Hence, there is a need for rational design of antiopioid conjugate vaccines that simultaneously target multiple opioids. Here, four novel haptens were synthesized, which were designed to elicit antibodies capable of binding to fentanyl other target opioids, including carfentanil, alfentanil, or oxycodone. Haptens were conjugated to CRM carrier protein and formulated with an aluminum salt adjuvant, and vaccines containing bivalent haptens were compared to admixtures of individual conjugate vaccines targeting the two opioids separately. Rats were immunized with monovalent, admixed, or novel bivalent vaccines, and the blockade of opioid effects was assessed against the individual drugs and their mixtures. Opioid-specific antibody titer was measured, and in vivo effects of vaccines were assessed in terms of preventing opioid-induced antinociception and respiratory depression and opioid distribution to the brain. While the bivalent vaccines reduced the effects of some target opioids, the admixed vaccine formulations were more effective against fentanyl/carfentanil and fentanyl/alfentanil mixtures. The bivalent fentanyl/oxycodone vaccine was as effective as the monovalent vaccines against a single drug challenge. These results inform the design of future vaccines against opioids and other drugs, particularly in the context of vaccines against polysubstance use that require optimization of response against multiple drugs of interest.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"676-687"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12090901/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143646486","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Rational Design of Site-Specific Fatty Acid Derivatives to Extend the Half-Life of Fibroblast Growth Factor 21.","authors":"Chengcheng Wang, Yapeng Wang, Yuanzhen Dong, Yu Duan, Ying Zhang, Hao Huang, Zhiru Xu, Jianguang Lu, Chunyong Ding, Zhengyan Cai, Dianwen Ju, Jun Feng","doi":"10.1021/acs.bioconjchem.4c00549","DOIUrl":"10.1021/acs.bioconjchem.4c00549","url":null,"abstract":"<p><p>Fibroblast growth factor 21 (FGF21) is a crucial regulator of glucose and lipid metabolism, showing significant therapeutic promise for metabolic disorders. However, its clinical application is limited by poor pharmacokinetics. One potential strategy to improve its half-life is to facilitate albumin binding through fatty acid derivation. Despite this promise, achieving site-specific modifications of FGF21 while preserving its biological activity has been challenging. In this study, we applied a rational design approach to create site-specific fatty acid derivatives of FGF21, guided by the structure of the FGF21-receptor complex. This strategy successfully enhances albumin binding without interfering with receptor interactions. The modified FGF21 derivatives exhibited dramatically extended half-lives in mice, increasing from 0.73 h to 11.36 and 13.36 h, respectively. Furthermore, these analogues showed superior biological activity in the presence of albumin, outperforming the C-terminal-derived variant zalfermin. This rational design approach not only improves the pharmacokinetic profile of FGF21 but also provides a framework for enhancing the therapeutic potential of other small proteins.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"688-696"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143629977","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Binary Copolymer Blending Enhances pDNA Delivery Performance and Colloidal Shelf Stability of Quinine-Based Polyplexes.","authors":"Punarbasu Roy, Nicholas W Kreofsky, Cristiam F Santa Chalarca, Theresa M Reineke","doi":"10.1021/acs.bioconjchem.5c00040","DOIUrl":"10.1021/acs.bioconjchem.5c00040","url":null,"abstract":"<p><p>Successful gene therapies require the efficient delivery of the therapeutic nucleic acids in the target cells, which is a major bottleneck. Our group has demonstrated that quinine-based polymers are effective and promising carriers for delivering nucleic acids, such as plasmid DNA (pDNA). However, the inherent hydrophobicity of quinine-based polymers makes the polymer-pDNA complexes (polyplexes) colloidally unstable leading to aggregation, which is relevant in clinical scenarios as larger particles (diameter >1000 nm) tend to perform poorly when administered systemically in vivo. Herein, we overcome the hydrophobicity-induced aggregation by using two types of quinine-based polymer systems to form polyplexes via a facile blending approach. We balanced desirable properties using quinine-based copolymers (HQ-<i>X</i>) as the pDNA binding component along with a quinine-based diblock copolymer (PHQ), having a polyethylene glycol chain, to provide colloidal stability to the particles. Using 5 polymer pairs, 5 mixing ratios, and 3 mixing sequences, we screened 66 formulations out of which 37 resulted in nonaggregating small polyplexes (diameter <300 nm) with colloidal stability tested up to 7 days at 4 °C. Furthermore, 18 out of these 37 colloidally stable formulations showed transfection performance better than or comparable to the commercial control, jetPEI. Our results clearly indicated that while the three mixing sequences generate polyplexes of similar characteristics, the best balance of transfection efficiency, toxicity, and colloidal stability is achieved at moderate PHQ % in the mixture when colloidal stability does not compromise payload binding. Our results showcase that polymer blending, in a manner similar to lipids, is an effective and parallel approach to achieving desirable polyplex characteristics, such as particle size, colloidal stability, and performance.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"770-781"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603009","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Linker Design Principles for the Precision Targeting of Oncogenic G-Quadruplex DNA with G4-Ligand-Conjugated Oligonucleotides.","authors":"Alva Abrahamsson, Andreas Berner, Justyna Golebiewska-Pikula, Namrata Chaudhari, Emelie Keskitalo, Cecilia Lindgren, Marcin K Chmielewski, Sjoerd Wanrooij, Erik Chorell","doi":"10.1021/acs.bioconjchem.5c00008","DOIUrl":"10.1021/acs.bioconjchem.5c00008","url":null,"abstract":"<p><p>G-quadruplex (G4) DNA structures are noncanonical secondary structures found in key regulatory regions of the genome, including oncogenic promoters and telomeres. Small molecules, known as G4 ligands, capable of stabilizing G4s hold promise as chemical probes and therapeutic agents. Nevertheless, achieving precise specificity for individual G4 structures within the human genome remains a significant challenge. To address this, we expand upon G4-ligand-conjugated oligonucleotides (GL-Os), a modular platform combining the stabilizing properties of G4-ligands with the sequence specificity of guide DNA oligonucleotides. Central to this strategy is the linker that bridges the G4 ligand and the guide oligonucleotide. In this study, we develop multiple conjugation strategies for the GL-Os that enabled a systematic investigation of the linker in both chemical composition and length, enabling a thorough assessment of their impact on targeting oncogenic G4 DNA. Biophysical, biochemical, and computational evaluations revealed GL-Os with optimized linkers that exhibited enhanced binding to target G4s, even under thermal or structural stress. Notably, longer linkers broadened the range of targetable sequences without introducing steric hindrance, thereby enhancing the platform's applicability across diverse genomic contexts. These findings establish GL-Os as a robust and versatile tool for the selective targeting of individual G4s. By facilitating precise investigations of G4 biology, this work provides a foundation for advancing G4-targeted therapeutic strategies and exploring their role in disease contexts.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"724-736"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12006964/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143668579","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Advances in the Treatment of Spinal Cord Injury with Nanozymes.","authors":"Zuohong Chen, Yili Wang, Shaofang Zhang, Huanhuan Qiao, Shuquan Zhang, Hao Wang, Xiao-Dong Zhang","doi":"10.1021/acs.bioconjchem.5c00100","DOIUrl":"10.1021/acs.bioconjchem.5c00100","url":null,"abstract":"<p><p>Spinal cord injury (SCI) with increasing incidence can lead to severe disability. The pathological process involves complex mechanisms such as oxidative stress, inflammation, and neuron apoptosis. Current treatment strategies focusing on the relief of oxidative stress and inflammation have achieved good effects, while many problems and challenges remain such as the side effect and short half-life of the therapeutic agents. Nanozymes exhibiting good biocatalytic activities can sustainably scavenge free radicals, inhibit neuroinflammation, and protect the neurons. With high stability in physiological conditions and cost-effectiveness, the nanozymes provide a new strategy for SCI treatment. In this Review, we outline the advances of nanozymes and their enzyme-mimicking activities and highlight the progress in the intervention of SCI-adopting nanozymes. We also propose future directions and clinical translation for the nanozyme strategy against SCI.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"627-651"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750135","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Introducing TAPY as a Versatile Alternative to TPP for Selective Mitochondrial Targeting in Cancer Cells.","authors":"Jean C Neto, Federico Lucantoni, Leydy V González, Eva Falomir, Juan F Miravet, Francisco Galindo","doi":"10.1021/acs.bioconjchem.4c00554","DOIUrl":"10.1021/acs.bioconjchem.4c00554","url":null,"abstract":"<p><p>The understanding of diseases such as cancer and Alzheimer's, along with natural aging processes, heavily relies on the study of mitochondrial function. Optical techniques like fluorescence imaging microscopy are pivotal for this purpose, enabling precise mapping of subcellular structures, including mitochondria. In this study, we explored TAPY (triarylpyridinium) cations, a novel family of mitochondrial carriers resembling the well-known triphenylphosphonium cation (TPP). Six TAPY-bodipy (BDP) dyads were prepared and chemically characterized. Confocal Laser Scanning Microscopy (CLSM) studies demonstrated that the systems were delivered selectively to the mitochondria of cancer cells (MCF-7, A549, HT-29). Remarkably, these dyads did not target the mitochondria of normal cells (HEK-293, HMEC-1), suggesting their potential use in distinguishing cancerous cells from healthy ones. A model compound comprised of the same bodipy cargo but attached to TPP was also synthesized and tested. Notably, in preliminary comparative assays with MCF-7 cells, the dyad TAPY(OMe)-BDP outperformed the TPP derivative in mitochondrial imaging, achieving twice the final fluorescence intensity. The potential chemical diversity achievable with TAPY cations is considerable, with many derivatives being accessible starting from readily available commercial products. This implies that, based on the strategy outlined in this study, carefully optimized TAPY derivatives for targeted mitochondrial delivery could potentially be developed in the future as alternatives or complements to TPP, with the present work acting as a proof of concept.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"697-706"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12129258/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750148","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Recent Advances in Aggregation-Induced Emission Bioconjugates.","authors":"Guiquan Zhang, Daming Zhou, Rong Hu, Anjun Qin, Ben Zhong Tang","doi":"10.1021/acs.bioconjchem.5c00036","DOIUrl":"10.1021/acs.bioconjchem.5c00036","url":null,"abstract":"<p><p>Fluorescence imaging technology is playing increasing roles in modern personalized and precision medicine. Thanks to their excellent photophysical properties, organic luminogens featuring aggregation-induced emission (AIE) characteristics (AIEgens) have attracted considerable attention over the past two decades. Because of their superior biocompatibility, ease of processing and functionalization, excellent water solubility, high responsiveness, and exceptional signal-to-noise ratio (SNR) for biotargets, AIE bioconjugates, formed by covalently linking AIEgens with biomolecules, have emerged as an ideal candidate for bioapplications. In this review, we summarize the progress in preparation, properties, and application of AIE bioconjugates in the last five years. Moreover, the challenges and opportunities of AIE bioconjugates are also briefly discussed.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"609-626"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143750151","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Photochemical-Promoted Cross-Coupling Reaction of Alkyl Boronate Esters with DNA-Conjugated Aryl Bromides for DNA-Encoded Library Synthesis.","authors":"Baiyang Mu, Yiwei Zhang, Xudong Wang, Rui Jin, Weiwei Lu, Nidhal Selmi, Sixiu Liu, Avinash Bhat, Sara Pahlén, Giulia Bergonzini, Zhiqiang Duan, Yinan Song, Xiaojie Lu","doi":"10.1021/acs.bioconjchem.4c00581","DOIUrl":"10.1021/acs.bioconjchem.4c00581","url":null,"abstract":"<p><p>The C(sp²)-C(sp³) cross-coupling reaction is an effective way to increase the C(sp³) content in compound collections for drug discovery, enhancing molecular diversity and offering a unique chemistry starting point. In this study, we report a mild, DNA-compatible, and off-DNA-inert photochemical cross-coupling reaction inspired by the amino radical transfer strategy. This method demonstrates broad substrate scopes for DNA-encoded library (DEL) constructions, utilizing commonly available structures on DNA and diverse alkyl boronate ester building blocks, which have not been widely applied in the current DEL chemical space.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"718-723"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143727012","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Improved Pendant-Bearing Glucose-Neopentyl Glycols for Membrane Protein Stability.","authors":"Taeyeol Youn, Ganghee Kim, Parameswaran Hariharan, Xianglan Li, Waqar Ahmed, Bernadette Byrne, Xiangyu Liu, Lan Guan, Pil Seok Chae","doi":"10.1021/acs.bioconjchem.4c00556","DOIUrl":"10.1021/acs.bioconjchem.4c00556","url":null,"abstract":"<p><p>Membrane proteins are biologically and pharmaceutically significant, and determining their 3D structures requires a membrane-mimetic system to maintain protein stability. Detergent micelles are widely used as membrane mimetics; however, their dynamic structures often lead to the denaturation and aggregation of encapsulated membrane proteins. To address the limitations of classical detergents in stabilizing membrane proteins, we previously reported a class of glucose-neopentyl glycols (GNGs) and their pendant-bearing versions (P-GNGs), several of which proved more effective than DDM in stabilizing membrane proteins. In this study, we synthesized additional GNG derivatives by varying the lengths of the pendant (P-GNGs), and by introducing hemifluorinated pendants to the GNG scaffold (fluorinated pendant-bearing GNGs or FP-GNGs). The synthetic flexibility of the GNG chemical architecture allowed us to create a diverse range of derivatives, essential for the effective optimization of detergent properties. When tested with two model membrane proteins (a transporter and a G-protein coupled receptor (GPCR)), most of the new (F)P-GNGs demonstrated superior stabilization of these membrane proteins compared to DDM, the original GNG (OGNG)), and a previously developed P-GNG (i.e., GNG-3,14). Notably, several P-GNGs synthesized in this study were as effective as or even better than lauryl maltose neopentyl glycol (LMNG) in stabilizing a human GPCR, beta2 adrenergic receptor (β2AR). Enhanced protein stability was particularly observed for the P-GNGs with a butyl (C4) or pentyl (C5) pendant, indicating that these pendant sizes are optimal for membrane protein stability. The volumes of these pendants appear to minimize the empty spaces in the micelle interiors, thereby enhancing detergent-detergent interactions in micelles complexed with the membrane proteins. Additionally, we identified one FP-GNG that was more efficient at extracting the transporter and more effective at stabilizing the GPCR than DDM. Thus, the current study demonstrates that both chain length and number of fluorine atoms in the pendants of the P-GNGs were crucial determinants for membrane protein stability. We not only developed a few (F)P-GNGs that are significantly more effective than maltoside detergents (LMNG/DDM) for protein extraction and stability but we also provided an effective strategy for detergent design through the utilization of partially fluorinated pendants of varying length.</p>","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"707-717"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12110327/pdf/","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143655463","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Indium-111-Labeled Single-Domain Antibody for <i>In Vivo</i> CXCR4 Imaging Using Single-Photon Emission Computed Tomography.","authors":"Muriel Aline Spahn, Stephanie Mareike Anbuhl, Kaat Luyten, Tom Van Loy, Matti F Pronker, Christopher Cawthorne, Christophe M Deroose, Dominique Schols, Raimond Heukers, Guy Bormans, Frederik Cleeren","doi":"10.1021/acs.bioconjchem.5c00024","DOIUrl":"10.1021/acs.bioconjchem.5c00024","url":null,"abstract":"&lt;p&gt;&lt;p&gt;C-X-C chemokine receptor type 4 (CXCR4) is highly expressed in a range of pathologies, including cancers like multiple myeloma and non-Hodgkin lymphoma, inflammatory diseases such as rheumatoid arthritis, and viral infections like HIV. Currently, the most advanced radiotracer for CXCR4 imaging in clinics is [&lt;sup&gt;68&lt;/sup&gt;Ga]PentixaFor. However, its structure is prone to modifications, complicating the development of a specific CXCR4 fluorine-18-labeled tracer with good pharmacokinetic properties. This study aimed to screen multiple CXCR4-targeting variable domains of heavy-chain-only antibody (VHH or single-domain antibody (sdAb)) constructs to identify the most promising sdAb as a vector molecule for the future development of a CXCR4 fluorine-18 tracer. We have generated five CXCR4-specific sdAb constructs with a cysteine-containing C-terminal tag (C-Direct tag) (VUN400-C-Direct, VUN401-C-Direct, VUN410-C-Direct, VUN411-C-Direct, and VUN415-C-Direct) and one probe (VUN400-C) without. The reduced sdAbs were coupled to maleimide-DOTAGA for &lt;sup&gt;111&lt;/sup&gt;In-labeling. Their binding affinity against human CXCR4 (hCXCR4) was assessed by using a previously described BRET-based displacement assay. The &lt;i&gt;in vivo&lt;/i&gt; profile was assessed using naive mice. Based on the plasma stability (60 min post injection (p.i.)), we selected VUN400-C-Direct and its derivative VUN400-C for further evaluation. These compounds ([&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C-Direct and [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C) were tested in mice bearing xenografts derived from U87.CD4, U87.CXCR4, and U87.CD4.CXCR4 cells through &lt;i&gt;ex vivo&lt;/i&gt; biodistribution studies and SPECT/CT imaging. The six sdAb constructs were labeled with a high radiochemical conversion (75-97%) and purity (&gt;95%). In radioactive binding assays using U87.CD4.CXCR4 cells, [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C-Direct and [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN401-C-Direct displayed the highest cellular uptake, achieving 10.4 ± 1.6% and 11.5 ± 1.1%, respectively. In naive mice, [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C-Direct showed the most favorable biodistribution profile, with low uptake across all organs except the kidneys (Standardized Uptake Value (SUV) &gt; 50, &lt;i&gt;n&lt;/i&gt; = 3, 60 min p.i.), but average plasma stability (40.6 ± 9.4%, &lt;i&gt;n&lt;/i&gt; = 3, 60 min p.i.). In a xenografted tumor model, [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C-Direct showed only minor uptake (SUV&lt;sub&gt;U87.CXCR4&lt;/sub&gt; 0.71 ± 0.002, &lt;i&gt;n&lt;/i&gt; = 3, 60 min p.i.). [&lt;sup&gt;111&lt;/sup&gt;In]In-DOTAGA-VUN400-C demonstrated nearly identical plasma stability (41.08 ± 5.45%, &lt;i&gt;n&lt;/i&gt; = 4) but showed high and specific uptake in the CXCR4-expressing xenografted tumor (SUV&lt;sub&gt;U87.CD4.CXCR4&lt;/sub&gt; 3.75 ± 1.08 vs SUV&lt;sub&gt;U87.CD4&lt;/sub&gt; = 0.64 ± 0.19, &lt;i&gt;n&lt;/i&gt; = 5, 60 min p.i.), which could be blocked by coinjection of AMD3100 (5 mg/kg) (SUV&lt;sub&gt;U87.CD4.CXCR4&lt;/sub&gt; 0.55 ± 0.32 vs SUV&lt;sub&gt;U87.CD4&lt;/sub&gt; = 0.39 ± 0.07, &lt;i&gt;n&lt;/i&gt; = 2, 60 min p.i.). In conclusion, all six sdAbs exhibite","PeriodicalId":29,"journal":{"name":"Bioconjugate Chemistry","volume":" ","pages":"737-747"},"PeriodicalIF":4.0,"publicationDate":"2025-04-16","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"143603017","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"化学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}