Controllable Contact-Destructive Hydrogel Actuators.

Constructing hydrogels with spatially heterogeneous structures are crucial for unlocking novel applications. To this end, selectively removing a specific portion of hydrogels by facile and intricate destructive strategies is worth exploring. Herein, a "contact-destructive" hydrogel actuator is presented, composed of a dynamic hydrogel network doped with hydrophilic polyethylene glycol (PEG). The destructive behavior of the hydrogel actuator is attributed to the surface tension-induced spreading effect and the enhanced water absorption due to the additive PEG. Parameters that act on these mechanisms are used to control the destruction of the hydrogel. During the destructive process, the hydrogel actuator exhibits locomotion routes predetermined by the graphic pattern with the aid of 3D printing. Additionally, such self-destructive behavior can be terminated by UV light irradiation when PEG is replaced with poly(ethylene glycol) diacrylate (PEGDA). Significantly, diverse applications including controllable 3D structures collapse, self-erasing, and on-demand cell release, are realized with such self-destructive hydrogel. These results demonstrate that the present hydrogel has great values in soft robotics, anti-counterfeiting, controlled drug delivery, and other related fields.

Self-Healing Dynamic Hydrogel Microparticles with Structural Color for Wound Management.

Chronic diabetic wounds confront a significant medical challenge because of increasing prevalence and difficult-healing circumstances. It is vital to develop multifunctional hydrogel dressings, with well-designed morphology and structure to enhance flexibility and effectiveness in wound management. To achieve these, we propose a self-healing hydrogel dressing based on structural color microspheres for wound management. The microsphere comprised a photothermal-responsive inverse opal framework, which was constructed by hyaluronic acid methacryloyl, silk fibroin methacryloyl and black phosphorus quantum dots (BPQDs), and was further re-filled with a dynamic hydrogel. The dynamic hydrogel filler was formed by Knoevenagel condensation reaction between cyanoacetate and benzaldehyde-functionalized dextran (DEX-CA and DEX-BA). Notably, the composite microspheres can be applied arbitrarily, and they can adhere together upon near-infrared irradiation by leveraging the BPQDs-mediated photothermal effect and the thermoreversible stiffness change of dynamic hydrogel. Additionally, eumenitin and vascular endothelial growth factor were co-loaded in the microspheres and their release behavior can be regulated by the same mechanism. Moreover, effective monitoring of the drug release process can be achieved through visual color variations. The microsphere system has demonstrated desired capabilities of controllable drug release and efficient wound management. These characteristics suggest broad prospects for the proposed composite microspheres in clinical applications.

Sprayable Multifunctional Black Phosphorus Hydrogel with On-Demand Removability for Joint Skin Wound Healing.

Wound healing remains a critical challenge in regenerative engineering. Great efforts are devoted to develop functional patches for wound healing. Herein, a novel sprayable black phosphorus (BP)-based multifunctional hydrogel with on-demand removability is presented as a joints' skin wound dressing. The hydrogel is facilely prepared by mixing dopamine-modified oxidized hyaluronic acid, cyanoacetategroup-functionalized dextran containing black phosphorus, and the catalyst histidine. The catechol-containing dopamine can not only enhance tissue adhesiveness, but also endow the hydrogel with antioxidant capacity. In addition, benefiting from the photothermal conversion ability of the BP and thermally reversible performance of the formed C═C double bonds between aldehyde groups and cyanoacetate groups, the resulting hydrogel displays excellent antibacterial performance and on-demand dissolving ability under NIR irradiation. Moreover, by loading vascular endothelial growth factor into the hydrogel, the promoted migration and angiogenesis effects of endothelial cells can also be achieved. Based on these features, it is demonstrated that such sprayable BP hydrogels can effectively facilitate joint wounds healing by accelerating angiogenesis, alleviating inflammation, and improving wound microenvironment. Thus, it is believed that this NIR-responsive removable BP hydrogel dressing will put forward an innovative concept in designing wound dressings.

Designing self-healing hydrogels for biomedical applications.

Self-healing hydrogels have emerged as the most promising alternatives to conventional brittle hydrogels used in the biomedical field due to the features of long-term stability and durability. However, the incompatibility between the fast self-healing property and enough mechanical strength of hydrogels remains a challenge. Therefore, hydrogels that possess not only mechanical toughness but also autonomous self-healing capacity are sought after. This review presents a comprehensive summary of the latest self-healing mechanisms. Specifically, we review various systems based on dynamic bonds, ranging from dynamic covalent bonds to non-covalent bonds. Additionally, this review presents different characterization methods for self-healing hydrogels, and also highlights their potential applications in the biomedical field, such as tissue engineering, drug delivery, cell therapy, and wound dressing. Furthermore, this review aims to provide valuable guidance for constructing diverse self-healing hydrogels with tailored functions.

Immersion ultrasound improves the repeatability of shear-wave elastography for measuring median nerve elasticity.

OBJECTIVE: To study the factors that influence the measurement of median nerve elasticity, to provide a more reproducible test for the assessment of median nerve elasticity using shear wave elasticity (SWE), and to reduce operator empirical dependence. To compare the repeatability of the median nerve elasticity measurement using immersion SWE with that using contact SWE, analyze the factors affecting SWE measurement, and provide a more repeatable method for doctors without SWE operation experience. METHODS: Two doctors without SWE operation experience measured the median nerve mean elastic modulus (Emean) at the same position and at different time points on the right wrist of 58 healthy volunteers using immersion and contact ultrasound methods. The intraobserver and interobserver repeatability of measurements was assessed using the interclass correlation coefficient (ICC), while the repeatability was assessed using the Bland-Altman diagram. RESULTS: The intraobserver and interobserver repeatability of the median nerve elasticity measured via contact SWE by inexperienced operators were classified as good, with ICCs of 0.633 (95% CI 0.380-0.783) and 0.552 (95% CI 0.243-0.735), respectively. The intraobserver and interobserver repeatability of the median nerve elasticity measured by immersion SWE were very good, with ICCs of 0.975 (95% CI 0.958-0.985) and 0.942 (95% CI 0.902-0.966), respectively. The intraobserver and interobserver Bland-Altman diagram of median nerve elasticity measured by immersion SWE showed that 98% of the points fell within the 95% limits of agreement. The intraobserver and interobserver Bland-Altman diagram of median nerve elasticity measured by contact SWE showed that 94% of the points fell within the 95% limits of agreement. CONCLUSION: Immersion ultrasound can improve the repeatability of median nerve elasticity measurements by inexperienced operators.

A BrLINE1-RUP insertion in BrCER2 alters cuticular wax biosynthesis in Chinese cabbage (Brassica rapa L. ssp. pekinensis).

Glossiness is an important quality-related trait of Chinese cabbage, which is a leafy vegetable crop in the family Brassicaceae. The glossy trait is caused by abnormal cuticular wax accumulation. In this study, on the basis of a bulked segregant analysis coupled with next-generation sequencing (BSA-seq) and fine-mapping, the most likely candidate gene responsible for the glossy phenotype of Chinese cabbage was identified. It was subsequently named Brcer2 because it is homologous to AtCER2 (At4g24510). A bioinformatics analysis indicated a long interspersed nuclear element 1 (LINE-1) transposable element (named BrLINE1-RUP) was inserted into the first exon of Brcer2 in HN19-G via an insertion-mediated deletion mechanism, which introduced a premature termination codon. Gene expression analysis showed that the InDel mutation of BrCER2 reduced the transcriptional expression levels of Brcer2 in HN19-G. An analysis of cuticular waxes suggested that a loss-of-function mutation to BrCER2 in Chinese cabbage leads to a severe decrease in the abundance of very-long-chain-fatty-acids (> C28), resulting in the production of a cauline leaf, inflorescence stem, flower, and pistil with a glossy phenotype. These findings imply the insertion of the LINE-1 transposable element BrLINE1-RUP into BrCER2 can modulate the waxy traits of Chinese cabbage plants.

Biomimetic Alveoli System with Vivid Mechanical Response and Cell-Cell Interface.

Alveolar microenvironmental models are important for studying the basic biology of the alveolus, therapeutic trials, and drug testing. However, a few systems can fully reproduce the in vivo alveolar microenvironment including dynamic stretching and the cell-cell interface. Here, a novel biomimetic alveolus-on-a-chip microsystem is presented suitable for visualizing physiological breathing for simulating the 3D architecture and function of human pulmonary alveoli. This biomimetic microsystem contains an inverse opal structured polyurethane membrane that achieves real-time observation of mechanical stretching. In this microsystem, the alveolar-capillary barrier is created by alveolar type 2 (ATII) cells cocultured with vascular endothelial cells (ECs) on this membrane. Based on this microsystem, the phenomena of flattening and the tendency of differentiation in ATII cells are observed. The synergistic effects of mechanical stretching and ECs on the proliferation of ATII cells are also observed during the repair process following lung injury. These features indicate the potential of this novel biomimetic microsystem for exploring the mechanisms of lung diseases, which can provide future guidance concerning drug targets for clinical therapies.

Psychrophilic Yeasts: Insights into Their Adaptability to Extremely Cold Environments.

Psychrophilic yeasts are distributed widely on Earth and have developed adaptation strategies to overcome the effect of low temperatures. They can adapt to low temperatures better than bacteriophyta. However, to date, their whole-genome sequences have been limited to the analysis of single strains of psychrophilic yeasts, which cannot be used to reveal their possible psychrophilic mechanisms to adapt to low temperatures accurately and comprehensively. This study aimed to compare different sources of psychrophilic yeasts at the genomic level and investigate their cold-adaptability mechanisms in a comprehensive manner. Nine genomes of known psychrophilic yeasts and three representative genomes of mesophilic yeasts were collected and annotated. Comparative genomic analysis was performed to compare the differences in their signaling pathways, metabolic regulations, evolution, and psychrophilic genes. The results showed that fatty acid desaturase coding genes are universal and diverse in psychophilic yeasts, and different numbers of these genes exist (delta 6, delta 9, delta 12, and delta 15) in the genomes of various psychrophilic yeasts. Therefore, they can synthesize polyunsaturated fatty acids (PUFAs) in a variety of ways and may be able to enhance the fluidity of cell membranes at low temperatures by synthesizing C18:3 or C18:4 PUFAs, thereby ensuring their ability to adapt to low-temperature environments. However, mesophilic yeasts have lost most of these genes. In this study, psychrophilic yeasts could adapt to low temperatures primarily by synthesizing PUFAs and diverse antifreeze proteins. A comparison of more psychrophilic yeasts' genomes will be useful for the study of their psychrophilic mechanisms, given the presence of additional potential psychrophilic-related genes in the genomes of psychrophilic yeasts. This study provides a reference for the study of the psychrophilic mechanisms of psychrophilic yeasts.

Multifunctional GO Hybrid Hydrogel Scaffolds for Wound Healing.

Hydrogel dressings have received extensive attention for the skin wound repair, while it is still a challenge to develop a smart hydrogel for adapting the dynamic wound healing process. Herein, we develop a novel graphene oxide (GO) hybrid hydrogel scaffold with adjustable mechanical properties, controllable drug release, and antibacterial behavior for promoting wound healing. The scaffold was prepared by injecting benzaldehyde and cyanoacetate group-functionalized dextran solution containing GO into a collection pool of histidine. As the GO possesses obvious photothermal behavior, the hybrid hydrogel scaffold exhibited an obvious stiffness decrease and effectively promoted cargo release owing to the breaking of the thermosensitive C=C double bond at a high temperature under NIR light. In addition, NIR-assisted photothermal antibacterial performance of the scaffold could be also achieved with the local temperature rising after irradiation. Therefore, it is demonstrated that the GO hybrid hydrogel scaffold with vascular endothelial growth factor (VEGF) encapsulation can achieve the adjustable mechanical properties, photothermal antibacterial, and angiogenesis during the wound healing process. These features indicated that the proposed GO hybrid hydrogel scaffold is potentially valuable for promoting wound healing and other biomedical application.

Hierarchical Microparticles Delivering Oxaliplatin and NLG919 Nanoprodrugs for Local Chemo-immunotherapy.

Chemo-immunotherapy shows promising antitumor therapeutic outcomes for many primary cancers. Research in this area has been focusing on developing an ideal formula that enables the potent efficacy of chemo-immunotherapy in combating various cancers with reduced systemic toxicity. Herein, we present novel hierarchical hydrogel microparticles (MDDP) delivering oxaliplatin and NLG919 nanoprodrugs for local chemo-immunotherapy with desired features. The oxaliplatin prodrug and NLG919 were efficiently loaded in the dual-drug polymeric nanoparticles (DDP NPs), which were further encapsulated into a MDDP by using microfluidic technology. When delivered to the tumor site, the DDP NPs will be sustainedly released from the MDDP and retained locally to reduce systemic toxicity. After being endocytosed by cancer cells, the cytotoxic oxaliplatin and NLG919 could be successfully triggered to release from DDP NPs in a chain-shattering manner, leading to the immunogenic cell death (ICD) of tumor cells and the suppression of intratumoral immunosuppressive Tregs, respectively. With the assistance of an immune modulator, the chemotherapeutics-induced ICD could trigger robust systemic antitumor immune responses, presenting superior synergistic antitumor efficacies. Thus, the hierarchical microparticles could substantially inhibit the growth of mouse subcutaneous colorectal tumors, breast tumors, and colorectal tumors with large initial sizes via synergized chemo-immunotherapy, showing great potential in the practical clinical application of oncotherapy.

Histidine-Triggered GO Hybrid Hydrogels for Microfluidic 3D Printing.

Graphene oxide (GO) hydrogels have provided tremendous opportunities in designing and fabricating complex constructs for diverse applications, while their 3D printing without photocuring is still a challenging task due to their low viscosity, uncontrollable gelation, and low interfacial tension. Here, we report a histidine-assisted printing strategy to prepare GO hybrid hydrogels through the microfluidic 3D printing technique. We found that the GO additive could significantly hamper the Knoevenagel condensation (KC) reaction between benzaldehyde and cyanoacetate group-functionalized polymers to form a hydrogel, while these GO mixed solutions were rapidly solidified into a hydrogel when histidine was added. This fascinating phenomenon enabled us to prepare low-viscosity GO mixed polymer solutions as printable inks and generate hydrogel microfibers in histidine solutions. The hydrogel fibers could support cell survival and be further constructed into complex 3D structures through microfluidic 3D printing techniques. Moreover, due to the addition of GO, the microfibers exhibited excellent electrical conductivity and could sense the motion changes and convert these stimuli as electrical resistance signals. This strategy adds an option for the design and application of 3D printable aqueous GO inks in many fields.

A Multistage Cooperative Nanoplatform Enables Intracellular Co-Delivery of Proteins and Chemotherapeutics for Cancer Therapy.

Combining intracellularly active proteins with chemotherapeutics represents a promising strategy for synergistic cancer therapy. However, the lack of nanocarrier systems for delivery into cancer cells and controlled intracellular release of both physicochemically very distinct cargos significantly impedes the biomedical translation of this combination strategy in cancer therapy. Here, a well-designed triblock copolymer, mPEG-b-PGCA-b-PGTA, is reported for application in a multistage cooperative drug delivery nanoplatform that accomplishes effective intracellular co-delivery of hydrophilic ribonuclease A (RNase A) and hydrophobic doxorubicin (DOX). RNase A bioreversibly modified with phenylboronic acid groups via a ROS-cleavable carbamate linker is incorporated into the triblock copolymer nanoparticles with high efficiency through a pH-reversible phenylboronic acid-catechol linkage. The reversible covalent conjugations between RNase A and the triblock copolymer endow the nanoparticles with high stability under normal physiological conditions. Upon cellular internalization, the cooperative release of DOX and RNase A from the triblock copolymer nanoparticles is triggered at multiple stages by endosomal acidic environment and subsequent DOX-enhanced intracellular ROS environment. This leads to enhanced synergistic anticancer effects as demonstrated both in vitro and in vivo. Given the versatility of dynamic covalent conjugations, this work provides a universal and stable platform for intracellular co-delivery of various combinations of proteins and chemotherapeutics.

Enhanced nanoparticle accumulation by tumor-acidity-activatable release of sildenafil to induce vasodilation.

Inefficient nanoparticle accumulation in solid tumors hinders the clinical translation of cancer nanomedicines. Herein, we proposed that sildenafil, a vasodilator ampholyte, could be used to promote nanoparticle accumulation by inducing vasodilation after its tumor acidity-triggered release from the nanocarriers. To confirm this, sildenafil was first encapsulated in a cisplatin-incorporated polymeric micelle. The dense PEG shell of the micelle reduced its endocytosis by cancer cells, which in return resulted in accumulative extracellular release of protonated sildenafil in the acidic tumor microenvironment. The released sildenafil was found to be more effective in enlarging the tumor blood vessels than could be achieved without sildenafil. As a result, we demonstrated considerable improvement in the intratumoral accumulation of the sildenafil-cisplatin co-loaded nanoparticle and its enhanced cancer therapeutic efficacy over the control group. Given the generality of a dense PEG shell and a hydrophobic part in most clinically developed nanomedicines, this work implies the great potential of sildenafil as a simple and universal adjuvant to selectively promote the intratumoral accumulation of nanomedicines, thus improving their clinical translation.

Constructing Thermally Reversible Dynamic Hydrogels via Catalysis-Free Knoevenagel Condensation.

Thermally reversible dynamic covalent bonds (TRDCBs) have attracted great interest for building polymers with self-healing and adaptable properties in bulk. However, none of the developed TRDCBs can be used in aqueous media for the fabrication of thermally reversible dynamic hydrogels due to the requirement of high temperature to initiate the retro-reaction or the susceptibility to hydrolysis. Herein, we report a thermally reversible dynamic covalent C═C double bond that was formed by catalysis-free Knoevenagel condensation (CKC) between benzaldehyde and cyanoacetate end-functionalized polymers in aqueous solution. The as-formed TRDCB shows typical thermal reversibility in the aqueous media under mild temperatures (4-70 °C). Constructing hydrogels with this TRDCB led to the formation of thermally reversible dynamic hydrogels with intriguing self-healing, injectable, thermosensitive, and thermoplastic properties. Overall, this work not only broadens the application of TRDCBs in aqueous media but also provides a thermally reversible dynamic hydrogel for potential use in various biomedical fields.

Cell penetrating peptide-modified nanoparticles for tumor targeted imaging and synergistic effect of sonodynamic/HIFU therapy.

BACKGROUND: Theranostics based on multifunctional nanoparticles (NPs) is a promising field that combines therapeutic and diagnostic functionalities into a single nanoparticle system. However, the major challenges that lie ahead are how to achieve accurate early diagnosis and how to develop efficient and noninvasive treatment. Sonodynamic therapy (SDT) utilizing ultrasound combined with a sonosensitizer represents a novel noninvasive modality for cancer therapy. Different ultrasound frequencies have been used for SDT, nevertheless, whether the effect of SDT can enhance synergistic HIFU ablation remains to be investigated. MATERIALS AND METHODS: We prepared a nanosystem for codelivery of a sonosensitizer (methylene blue, MB) and a magnetic resonance contrast agent (gadodiamide, Gd-DTPA-BMA) based on hydrophilic biodegradable polymeric NPs composed of poly (lactic-co-glycolic acid) (PLGA). To enhance accumulation and penetration of the NPs at the tumor site, the surface of PLGA NPs was decorated with a tumor-homing and penetrating peptide-F3 and polyethylene glycol (PEG). The physicochemical, imaging and therapeutic properties of F3-PLGA@MB/Gd and drug safety were thoroughly evaluated both in vitro and in vivo. F3-PLGA@MB/Gd was evaluated by both photoacoustic and resonance imaging. RESULTS: F3-PLGA@MB/Gd NPs exhibited higher cellular association than non-targeted NPs and showed a more preferential enrichment at the tumor site. Furthermore, with good drug safety, the apoptosis triggered by ultrasound in the F3-PLGA@MB/Gd group was greater than that in the contrast group. CONCLUSION: F3-PLGA@MB/Gd can work as a highly efficient theranostic agent, and the incorporation of targeted multimodal and combined therapy could be an encouraging strategy for cancer treatment.

Enhancement of HIFU ablation by sonosensitizer-loading liquid fluorocarbon nanoparticles with pre-targeting in a mouse model.

High intensity focused ultrasound (HIFU) is a noninvasive thermal ablation technique for the treatment of benign and malignant solid masses. To improve the efficacy of HIFU ablation, we developed poly (lactide-co-glycolide) (PLGA) nanoparticles encapsulating perfluoropentane (PFP) and hematoporphyrin monomethyl ether (HMME) as synergistic agents (HMME+PFP/PLGA). Two-step biotin-avidin pre-targeting technique was applied for the HIFU ablation. We further modified the nanoparticles with streptavidin (HMME+PFP/PLGA-SA). HMME+PFP/PLGA-SA were highly dispersed with spherical morphology (477.8 ± 81.8 nm in diameter). The encapsulation efficiency of HMME and PFP were 46.6 ± 3.3% and 40.1 ± 2.6%, respectively. The binding efficiency of nanoparticles to streptavidin was 95.5 ± 2.5%. The targeting ability of the HMME+PFP/PLGA-SA nanoparticles was tested by parallel plate flow chamber in vitro. In the pre-targeting group (HMME+PFP/PLGA-SA), a large number of nanoparticles bound to the peripheral and surface of the cell. In the HIFU ablation experiment in vivo, compared with the other groups, the largest gray-scale changes and coagulation necrosis areas were observed in the pre-targeting (HMME+PFP/PLGA-SA) group, with the lowest energy efficiency factor value. Moreover, the microvessel density and proliferation index declined, while the apoptotic index increased, in the tumor tissue surrounding the coagulation necrosis area in the pre-targeting group. Meanwhile, the survival time of the tumor-bearing nude mice in the pre-targeting group was significantly longer than that in the HIFU treatment group. These results suggest that HMME+PFP/PLGA-SA have high potential to act as synergistic agents in HIFU ablation.

In Vitro and In Vivo Investigation of High-Intensity Focused Ultrasound (HIFU) Hat-Type Ablation Mode.

BACKGROUND The aim of this study was to investigate the feasibility of the application of high-intensity focused ultrasound (HIFU) hat-type ablation mode in in vitro and in vivo models, and to compare the ablation effects of different parameter combinations. MATERIAL AND METHODS HIFU hat-type ablation was performed in isolated bovine liver tissue and in the liver tissue in living rabbits, and the coagulative necrosis for different parameter combinations (plane angles and irradiation order) was investigated. We also analyzed and compared the ablation effects of traditional ablation and hat-type ablation modes. Coagulative necrosis morphology was detected with TTC staining, and the coagulative necrosis volume and energy efficiency factor (EEF) were calculated and compared. RESULTS Coagulative necrosis was observed in all the ablated groups, and the coagulative necrosis volume was much larger than the irradiation area. The coagulative necrosis induced by the hat-type ablation was more regular and controllable than the traditional ablation. The angles between the ablation planes determined the coagulative necrosis morphology, but did not affect the coagulative necrosis volume. Moreover, the irradiation order significantly influenced the coagulative necrosis. Importantly, under certain conditions, hat-type ablation achieved higher efficiency compared with the traditional ablation mode. CONCLUSIONS Compared with the traditional ablation mode, HIFU hat-type ablation effectively shortened the irradiation time, reduced the over-accumulation of energy, and increased the HIFU ablation efficiency.

HematoPorphyrin Monomethyl Ether polymer contrast agent for ultrasound/photoacoustic dual-modality imaging-guided synergistic high intensity focused ultrasound (HIFU) therapy.

This study is to prepare a hematoporphyrin monomethyl ether (HMME)-loaded poly(lactic-co-glycolic acid) (PLGA) microcapsules (HMME/PLGA), which could not only function as efficient contrast agent for ultrasound (US)/photoacoustic (PA) imaging, but also as a synergistic agent for high intensity focused ultrasound (HIFU) ablation. Sonosensitizer HMME nanoparticles were integrated into PLGA microcapsules with the double emulsion evaporation method. After characterization, the cell-killing and cell proliferation-inhibiting effects of HMME/PLGA microcapsules on ovarian cancer SKOV3 cells were assessed. The US/PA imaging-enhancing effects and synergistic effects on HIFU were evaluated both in vitro and in vivo. HMME/PLGA microcapsules were highly dispersed with well-defined spherical morphology (357 ± 0.72 nm in diameter, PDI = 0.932). Encapsulation efficiency and drug-loading efficiency were 58.33 ± 0.95% and 4.73 ± 0.15%, respectively. The HMME/PLGA microcapsules remarkably killed the SKOV3 cells and inhibited the cell proliferation, significantly enhanced the US/PA imaging results and greatly enhanced the HIFU ablation effects on ovarian cancer in nude mice by the HMME-mediated sono-dynamic chemistry therapy (SDT). HMME/PLGA microcapsules represent a potential multifunctional contrast agent for HIFU diagnosis and treatment, which might provide a novel strategy for the highly efficient imaging-guided non-invasive HIFU synergistic therapy for cancers by SDT in clinic.

The evaluation of a novel approach for the profiling and identification of N-linked glycan with a procainamide tag by HPLC with fluorescent and mass spectrometric detection.

Procainamide was investigated as a multifunctional oligosaccharide label for glycan profiling and identification in a HPLC-FL/ESI-QTOF system. Addition of this aromatic amine to glycans through reductive amination improves fluorescence detection and ESI ionization efficiency. Both procainamide and 2-AB derivatives of N-linked glycans released from three glycoproteins (Human IgG, Mouse IgG, and RNase B) were quantitatively profiled with HPLC-FL and identified with ESI-QTOF. The procainamide derivatives produced FL glycan profiles comparable to the 2-AB derivatives, but with a few extra minor peaks, which suggests better labeling efficiency for procainamide derivatives for minor peaks. The procainamide derivatives also improve ESI ionization efficiency by 10-50 times over the respective 2-AB derivatives and the ESI-QTOF method sensitivity is at the low picomole to high femtomole level. Using the procainamide tag, all N-linked glycans released from three tested glycoproteins can be quantitatively detected with HPLC-FL and identified with ESI-QTOF at the same time. Monosaccharide sequence confirmation was also demonstrated in this study.