A metal-organic cage-derived cascade antioxidant nanozyme to mitigate renal ischemia-reperfusion injury.

In the field of contemporary medicine, inflammation has emerged as a significant concern in global public health. Among the current anti-inflammatory strategies, nanozymes possess distinctive advantages and demonstrate unexpected efficacy in combating inflammation. However, the indeterminate structures and limited enzyme-like activity exhibited by most developed nanozymes impede their clinical translation and therapeutic effectiveness. In this paper, we developed a nanozyme derived from a well-defined metal-organic cage (MOC). The oxidized MOC (MOC-O), containing pyridine nitrogen oxide moieties, exhibited effective cascade superoxide dismutase (SOD) and catalase (CAT)-like activities for scavenging reactive oxygen species (ROS). This ROS scavenging ability was confirmed through flow cytometry analysis using DCFH-DA in a hypoxia/reoxygenation (H/R) model, where MOC-O significantly alleviated oxidative stress. Furthermore, the administration of MOC-O resulted in preserved renal function during renal ischemia-reperfusion (I/R) injury due to downregulated oxidative stress levels and reduced cell apoptosis.

Recent advances in carbon monoxide-releasing nanomaterials.

As an endogenous signaling molecule, carbon monoxide (CO) has emerged as an increasingly promising option regarding as gas therapy due to its positive pharmacological effects in various diseases. Owing to the gaseous nature and potential toxicity, it is particularly important to modulate the CO release dosages and targeted locations to elucidate the biological mechanisms of CO and facilitate its clinical applications. Based on these, diverse CO-releasing molecules (CORMs) have been developed for controlled release of CO in biological systems. However, practical applications of these CORMs are limited by several disadvantages including low stability, poor solubility, weak releasing controllability, random diffusion, and potential toxicity. In light of rapid developments and diverse advantages of nanomedicine, abundant nanomaterials releasing CO in controlled ways have been developed for therapeutic purposes across various diseases. Due to their nanoscale sizes, diversified compositions and modified surfaces, vast CO-releasing nanomaterials (CORNMs) have been constructed and exhibited controlled CO release in specific locations under various stimuli with better pharmacokinetics and pharmacodynamics. In this review, we present the recent progress in CORNMs according to their compositions. Following a concise introduction to CO therapy, CORMs and CORNMs, the representative research progress of CORNMs constructed from organic nanostructures, hybrid nanomaterials, inorganic nanomaterials, and nanocomposites is elaborated. The basic properties of these CORNMs, such as active components, CO releasing mechanisms, detection methods, and therapeutic applications, are discussed in detail and listed in a table. Finally, we explore and discuss the prospects and challenges associated with utilizing nanomaterials for biological CO release.

Chiral metal-organic cages decorated with binaphthalene moieties.

The construction of chiral nanoobjects with atomically precise nanostructures has attracted much more attention in the past decades. However, this field is still in its early stages. We designed and synthesized a series of chiral ligands containing the binaphthalene moiety and isophthalate module. Then, four chiral metal-organic cages (MOCs) were obtained through the coordination between isophthalate modules and copper ions. These chiral MOCs exhibit discrete, uniform and stable structures, good solubility and photoluminescence behaviors.

Bilateral visual loss after selective artery embolization in the neck for hemorrhage of malignant tumor.

INTRODUCTION AND IMPORTANCE: Currently, selective arterial embolization (SAE) has been widely applied for the treatment of many diseases due to its minimal invasiveness. But the complications caused by SAE can be serious. CASE PRESENTATION: Here, we report a case of a patient who experienced bilateral blindness 4 h after selective arterial embolization (SAE). A 67-year-old man, with a 13-year history of nasopharyngeal carcinoma, was admitted to our hospital for nasopharyngeal carcinoma hemorrhage and was scheduled for SAE. The patient did not have any thromboembolic complications. His had a platelet count of 43 × 109/L (range 150-400 × 109/L) and a prothrombin time (PT) of 9.3 s. The surgery was completed under local anesthesia. However, 4 h after the surgery, the patient complained of visual loss. We performed a fundoscopy examination, which showed bilateral ophthalmic artery embolism. CLINICAL DISCUSSION: Bilateral ophthalmic artery embolism is fatal to vision. When this occurs, it would be difficult to save the eyes. So, the relevant selection of the optimal properties of the used PVA and coil embolization materials is important during SAE. CONCLUSION: It is important to improve the existing understanding of the involvement various vessels during embolization of head and neck tumors. Furthermore, special and paramount attention is to be paid to the specific pre-operative angio-architecture, particular patient condition, and the prudent choice of embolic material to prevent the occurrence of ectopic embolization.

DRPADC: A novel drug repositioning algorithm predicting adaptive drugs for COVID-19.

Given that the usual process of developing a new vaccine or drug for COVID-19 demands significant time and funds, drug repositioning has emerged as a promising therapeutic strategy. We propose a method named DRPADC to predict novel drug-disease associations effectively from the original sparse drug-disease association adjacency matrix. Specifically, DRPADC processes the original association matrix with the WKNKN algorithm to reduce its sparsity. Furthermore, multiple types of similarity information are fused by a CKA-MKL algorithm. Finally, a compressed sensing algorithm is used to predict the potential drug-disease (virus) association scores. Experimental results show that DRPADC has superior performance than several competitive methods in terms of AUC values and case studies. DRPADC achieved the AUC value of 0.941, 0.955 and 0.876 in Fdataset, Cdataset and HDVD dataset, respectively. In addition, the conducted case studies of COVID-19 show that DRPADC can predict drug candidates accurately.

Catechol-Coordinated Framework Film-based Micro-Supercapacitors with AC Line Filtering Performance.

Coordination polymer frameworks (CPFs) have broad applications due to their excellent features, including stable structure, intrinsic porosity, and others. However, preparation of thin-film CPFs for energy storage and conversion remains a challenge because of poor compatibility between conductive substrates and CPFs and crucial conditions for thin-film preparation. In this work, a CPF film was prepared by the coordination of the anisotropic four-armed ligand and CuII at the liquid-liquid interface. Such film-based micro-supercapacitors (MSCs) are fabricated through high-energy scribing and electrolytes soaking. As-fabricated MSCs displayed high volumetric specific capacitance of 121.45 F cm-3 . Besides, the volumetric energy density of MSCs reached 52.6 mWh cm-3 , which exceeds the electrochemical performance of most reported CPF-based MSCs. Especially, the device exhibited alternating current (AC) line filtering performance (-84.2° at 120 Hz) and a short resistance capacitance (RC) constant of 0.08 ms. This work not only provides a new CPF for MSCs with AC line filtering performance but also paves the way for thin-film CPFs preparation with versatile applications.

Noble Metal Nanomaterials for NIR-Triggered Photothermal Therapy in Cancer.

It is of great significance to develop anticancer therapeutic agents or technologies with high degree of specificity and patient compliance, while low toxicity. The emerging photothermal therapy (PTT) has become a new and powerful therapeutic technology due to its noninvasiveness, high specificity, low side effects to normal tissues and strong anticancer efficacy. Noble metal nanomaterials possess strong surface plasmon resonance (SPR) effect and synthetic tunability, which make them facile and effective PTT agents with superior optical and photothermal characteristics, such as high absorption cross-section, incomparable optical-thermal conversion efficiency in the near infrared (NIR) region, as well as the potential of bioimaging. By incorporating with various functional reagents such as antibodies, peptides, biocompatible polymers, chemo-drug and immune factors, noble metal nanomaterials have presented strong potential in multifunctional cancer therapy. Herein, the recent development regarding the application of noble metal nanomaterials for NIR-triggered PTT in cancer treatment is summarized. A variety of studies with good therapeutic effects against cancer from impressive photothermal efficacy of noble metal nanomaterials are concluded. Intelligent nanoplatforms through ingenious fabrication showing potential of multifunctional PTT, combined with chemo-therapy, immunotherapy, photodynamic therapy (PDT), as well as simultaneous imaging modality are also demonstrated.

Nanofabrication within unimolecular nanoreactors.

Nanoparticles (NPs) have been a research focus over the last three decades owing to their unique properties and extensive applications. It is crucial to precisely control the features of NPs including topology, architecture, composition, size, surface and assembly because these features will affect their properties and then applications. Ingenious nanofabrication strategies have been developed to precisely control these features of NPs, especially for templated nanofabrication within predesigned nanoreactors. Compared with conventional nanoreactors (hard templates and supramolecular nanoreactors), unimolecular nanoreactors exhibit (1) covalently stable nanostructures uninfluenced by environmental variations, (2) extensively regulated features of the structure including topology, composition, size, surface and valence due to the rapid development of polymer chemistry, and (3) effective encapsulation of abundant guests with or without strong interaction to achieve the function of loading, delivery and conversion of guests. Thus, unimolecular nanoreactors have shown fascinating prospects as templates for nanofabrication. Various NPs with expected topologies (sphere, rod, tube, branch, and ring), architectures (compact, hollow, core-shell, and necklace-like), compositions (metal, metal oxide, semiconductor, doping, alloy, silica, and composite), sizes (generally 1-100 nm), surface properties (hydrophilic, hydrophobic, reactivity, valence and responsivity) and assemblies (oligomer, chain, and aggregate) can be fabricated easily within reasonably designed unimolecular nanoreactors in a programmable way. In this review, we provide a brief introduction of the properties and types of unimolecular nanoreactors, a condensed summary of representative methodologies of nanofabrication within various unimolecular nanoreactors and a predicted outlook of the potential further developments of this charming nanofabrication approach.

Methotrexate-Mn2+ based nanoscale coordination polymers as a theranostic nanoplatform for MRI guided chemotherapy.

Theranostic nanoplatforms based on magnetic resonance imaging (MRI) technology have drawn much attention due to their inherent advantages such as non-invasiveness and high spatial resolution, but their construction is usually complicated and the compositions are uncontrollable. Herein, a simple and controllable theranostic nanoplatform by replacing a chelating agent with a chemotherapeutic drug directly was proposed. In detail, we elaborately synthesized Mn2+ and methotrexate (MTX) based nanoscale coordination polymers (NCPs) coated with poly(ethylene glycol) (PEG), MTX-Mn@PEG, in which Mn2+ ions served as MRI contrast agents (CAs), MTX was used as a chemotherapy drug and PEG provided a stable environment for nanoparticles, respectively. The obtained MTX-Mn@PEG NCPs showed a relatively uniform size, a pH-responsive feature and long blood circulation to enrich in the tumor location through the enhanced permeability and retention (EPR) effect. We determined the composition and coordination mechanism of MTX-Mn@PEG through thermogravimetric analysis (TGA), Fourier transform infrared (FI-TR) spectroscopy, powder X-ray diffraction (XRD) and so on. We determined the composition and coordination mechanism of MTX-Mn@PEG with the molar ratio of MTX to Mn2+ as 1 : 1. Then we confirmed that the antitumor ability was mediated by MTX through MTT and cell apoptosis assay in vitro. In vivo MR imaging and antitumor assays demonstrated the excellent effect of the NCPs in diagnosis and therapy of tumors. Overall, these novel MTX-Mn@PEG NCPs provide a theranostic nanoplatform for cancer treatment.

PLAC1-specific TCR-engineered T cells mediate antigen-specific antitumor effects in breast cancer.

Placenta-specific 1 (PLAC1), a novel cancer-testis antigen (CTA), is expressed in a number of different human malignancies. It is frequently produced in breast cancer, serving a function in tumorigenesis. Adoptive immunotherapy using T cell receptor (TCR)-engineered T cells against CTA mediates objective tumor regression; however, to the best of our knowledge, targeting PLAC1 using engineered T cells has not yet been attempted. In the present study, the cDNAs encoding TCRα- and ß-chains specific for human leukocyte antigen (HLA)-A*0201-restricted PLAC1 were cloned from a cytotoxic T-lymphocyte, generated by in vitro by the stimulation of CD8+ T cells using autologous HLA-A2+ dendritic cells loaded with a PLAC1-specific peptide (p28-36, VLCSIDWFM). The TCRα/ß-chains were linked by a 2A peptide linker (TCRα-Thosea asigna virus-TCRß), and the constructs were cloned into the lentiviral vector, followed by transduction into human cytotoxic (CD8+) T cells. The efficiency of transduction was up to 25.16%, as detected by PLAC1 multimers. TCR-transduced CD8+ T cells, co-cultured with human non-metastatic breast cancer MCF-7 cells (PLAC1+, HLA-A2+) and triple-negative breast cancer MDAMB-231 cells (PLAC1+, HLA-A2+), produced interferon γ and tumor necrosis factor α, suggesting TCR activation. Furthermore, the PLAC1 TCR-transduced CD8+ T cells efficiently and specifically identified and annihilated the HLA-A2+/PLAC1+ breast cancer cell lines in a lactate dehydrogenase activity assay. Western blot analysis demonstrated that TCR transduction stimulated the production of mitogen-activated protein kinase signaling molecules, extracellular signal-regulated kinases 1/2 and nuclear factor-κB, through phosphoinositide 3-kinase γ-mediated phosphorylation of protein kinase B in CD8+ T cells. Xenograft mouse assays revealed that PLAC1 TCR-transduced CD8+T cells significantly delayed the tumor progression in mice-bearing breast cancer compared with normal saline or negative control-transduced groups. In conclusion, a novel HLA-A2-restricted and PLAC1-specific TCR was identified. The present study demonstrated PLAC1 to be a potential target for breast cancer treatment; and the usage of PLAC1-specific TCR-engineered T cells may be a novel strategy for PLAC1-positive breast cancer treatment.

Successful Coupling of a Bis-Amidoxime Uranophile with a Hydrophilic Backbone for Selective Uranium Sequestration.

The amidoxime group (-RNH2NOH) has long been used to extract uranium from seawater on account of its high affinity toward uranium. The development of tunable sorbent materials for uranium sequestration remains a research priority as well as a significant challenge. Herein, we report the design, synthesis, and uranium sorption properties of bis-amidoxime-functionalized polymeric materials (BAP 1-3). Bifunctional amidoxime monomers were copolymerized with an acrylamide cross-linker to obtain bis-amidoxime incorporation as high as 2 mmol g-1 after five synthetic steps. The resulting sorbents were able to uptake nearly 600 mg of uranium per gram of polymer after 37 days of contact with a seawater simulant containing 8 ppm uranium. Moreover, the polymeric materials exhibited low vanadium uptake with a maximum capacity of 128 mg of vanadium per gram of polymer. This computationally predicted and experimentally realized selectivity of uranium over vanadium, nearly 5 to 1 w/w, is one of the highest reported to date and represents an advancement in the rational design of sorbent materials with high uptake capacity and selectivity.

Construction of Polyarylenes with Various Structural Features via Bergman Cyclization Polymerization.

Synthetic polymer chemistry is a fundamental part of polymer science, and highly efficient polymerization reactions are essential for the synthesis of high-performance polymers. Development of new synthetic methods for emerging polymer science is of great importance in this regard. Bergman cyclization is a chemical process in which highly reactive aryl diradicals form from enediyne precursors, having a strong impact in a number of fields including pharmaceutics, synthetic chemistry, and materials science. Diradical intermediates stemming from enediynes can cause DNA cleavage under physiological conditions, leading to the strong cytotoxicity of many naturally occurring enediyne antibiotics. Meanwhile, diradical intermediates can quickly couple with each other to construct polyarylenes, providing a novel method to synthesize these conjugated polymers with the advantages of facile and catalyst-free operation, high efficiency, and tailored structure. Moreover, conjugated polymers generated by Bergman cyclization exhibit many remarkable properties, such as excellent thermal stability and good solubility and processability, enabling their further processing into carbon-rich materials. This review presents a brief overview of the trajectory of Bergman cyclization in polymer science, followed by an introduction to research advances, mainly from our group, in developing polymerization methods based on Bergman cyclization, taking advantages of its catalyst-free, byproduct-free, in situ polymerization mechanism to synthesize new polymeric materials with various structures and morphologies. These synthetic strategies include fabrication of rod-like polymers with polyester, dendrimer, and chiral imide side chains, functionalization of carbon nanomaterials by surface-grafting conjugated polymers, formation of nanoparticles by intramolecular collapse of single polymer chains, and construction of carbon nanomembranes on the external and internal surface of inorganic nanomaterials. These polymers with novel structural features have been used in a variety of fields, such as energy transformation, energy storage, catalyst support, and fluorescent detection. Finally, the outlook for future developments of Bergman cyclization in polymer science is presented.

From natural cotton thread to sewable energy dense supercapacitors.

Considering cost and flexibility, cotton thread is an ideal material for the fabrication of wearable and portable electronics. However, the capacitance of cotton thread based supercapacitors remains extremely low (below 50 mF cm-2) due to the insufficient capacitive utilization of active materials. In this work, ordered mesoporous carbon (OMC) membranes are rationally coupled with chemical vapour deposition derived graphene (CVD gr), to form a highly conductive carbon coating around cotton yarn. In this material design, OMC membranes act as hydrophilic nanoporous "ion reservoirs" to accumulate sufficient cations from a gel electrolyte, while CVD gr endows the composite thread low liner resistance (3.7 Ω cm-1) and high mechanical strength. Using a butyl-3-methylimidazolium chloride modified gel as an ionic conducting electrolyte, the efficiency in capacitive utilization of coated MnO2 microparticles has been doubled, delivering an areal capacitance of 1.1 F cm2 with a volumetric energy of 2.7 mWh cm-3. Such a supercapacitor thread is lightweight, sewable and durable in bending fatigue tests, and can be fabricated through a facile dip-coating method. Impressively, this device can power a photodetector based on TiO2 nanowires without applying any external bias voltage, which opens up a new opportunity for development of wearable and self-powered nanodevices in the near future.

Anti-VEGFR2-interferon-α2 regulates the tumor microenvironment and exhibits potent antitumor efficacy against colorectal cancer.

Interferon-α (IFNα) has multiple antitumor effects including direct antitumor toxicity and the ability to potently stimulate both innate and adaptive immunity. However, its clinical applications in the treatment of malignancies have been limited because of short half-life and serious adverse reactions when attempting to deliver therapeutically effective doses. To address these issues, we fused IFNα2a to the anti-vascular endothelial growth factor and receptor 2 (VEGFR2) antibody JZA00 with the goal of targeting it to the tumor microenvironment where it can stimulate the antitumor immune response. The fusion protein, JZA01, is effective against colorectal cancer by inhibiting angiogenesis, exhibiting direct cytotoxicity, and activating the antitumor immune response. Although JZA01 exhibited reduced IFNα2 activity in vitro compared with native IFNα2, VEGFR2 targeting permitted efficient antiproliferative, proapoptotic, antiangiogenesis, and immune-stimulating effects against the colorectal tumors HCT-116 and SW620. JZA01 showed in vivo efficacy in NOD-SCID mice-bearing established HCT-116 tumors. In conclusion, this study describes an antitumor immunotherapy that is highly promising for the treatment of colorectal cancer.

A Novel Fusion Antibody Exhibits Antiangiogenic Activity and Stimulates NK Cell-mediated Immune Surveillance Through Fused NKG2D Ligand.

A single-chain variable fragment (scFv) targeting vascular endothelial growth factor receptor 2 was previously generated from a phage display library in our laboratory. However, it has shortened half-life and lacks Fc fragment for effector cell recognition. To address these challenges, a ligand of NK-cell receptor NKG2D was fused to the scFv and created a fusion protein scFv-major histocompatibility complex class I-related chain A (MICA), which is expected to recognize tumor cells through the scFv moiety and stimulate NK cells through the MICA. The fusion protein demonstrated specific binding to both vascular endothelial growth factor receptor 2 and NKG2D in protein-based and cell-based assays. In addition, it demonstrated antiangiogenic activities including restraining the proliferation, migration, transwell invasion, and tube formation of human umbilical vein endothelial cells. Furthermore, the fusion protein exhibited significant cytotoxicity on K562, MDA-MB-435, and B16F10 cells and triggered NK92 cell-mediated cytotoxicity on MDA-MB-435 cells by stimulating the release of significant cytokines. The fusion protein targeting strategy, therefore, provides a means to engage lymphocyte effector cells against tumor specific antigen overexpressing tumor cells.

A high performance flexible all solid state supercapacitor based on the MnO2 sphere coated macro/mesoporous Ni/C electrode and ionic conducting electrolyte.

A high contact resistance between the active materials and the current collector, a low ionic conductivity of the gel electrolyte, and an impenetrable electrode structure are the three major barriers which greatly limit the capacitance of MnO2 in solid state supercapacitors. As a potential solution to these problems, in this work we report a novel electrode for solid state supercapacitors, based on a ternary system composed of hierarchical MnO2 spheres as the active material, macroporous Ni foam as gel penetrable skeletons and an ordered mesoporous carbon (OMC) membrane as the charge-transport accelerating layer. By employing butyl-3-methylimidazolium chloride (BMIMCl) modified gels as the ionic conducting electrolyte, the utilization efficiency of MnO2 on the specific capacitance was enhanced up to 88% of the theoretical value, delivering a volumetric capacitance of 81 F cm(-3), which is the highest value among MnO2 based solid state supercapacitors. Moreover, such a flexible device exhibits exceptional volumetric energy and power density (6.6 Wh L(-1) and 549 W L(-1), based on the whole device volume) combined with a small capacity loss of 8.5% after 6000 cycles under twisting. These encouraging findings unambiguously overcome the energy bottleneck of MnO2 in solid state supercapacitors, and open up a new application of macro/mesoporous materials in flexible devices.

VEGFR2-targeted fusion antibody improved NK cell-mediated immunosurveillance against K562 cells.

MHC class I polypeptide-related sequence A (MICA), which is normally expressed on cancer cells, activates NK cells via NK group 2-member D pathway. However, some cancer cells escape NK-mediated immune surveillance by shedding membrane MICA causing immune suppression. To address this issue, we designed an antibody-MICA fusion targeting tumor-specific antigen (vascular endothelial growth factor receptor 2, VEGFR2) based on our patented antibody (mAb04) against VEGFR2. In vitro results demonstrate that the fusion antibody retains both the antineoplastic and the immunomodulatory activity of mAb04. Further, we revealed that it enhanced NK-mediated immunosurveillance against K562 cells through increasing degranulation and cytokine production of NK cells. The overall data suggest our new fusion protein provides a promising approach for cancer-targeted immunotherapy and has prospects for potential application of chronic myeloid leukemia.

Nanoscale Metal-Organic Frameworks for Ratiometric Oxygen Sensing in Live Cells.

We report the design of a phosphorescence/fluorescence dual-emissive nanoscale metal-organic framework (NMOF), R-UiO, as an intracellular oxygen (O2) sensor. R-UiO contains a Pt(II)-porphyrin ligand as an O2-sensitive probe and a Rhodamine-B isothiocyanate ligand as an O2-insensitive reference probe. It exhibits good crystallinity, high stability, and excellent ratiometric luminescence response to O2 partial pressure. In vitro experiments confirmed the applicability of R-UiO as an intracellular O2 biosensor. This work is the first report of a NMOF-based intracellular oxygen sensor and should inspire the design of ratiometric NMOF sensors for other important analytes in biological systems.

VEGFR2 targeted antibody fused with MICA stimulates NKG2D mediated immunosurveillance and exhibits potent anti-tumor activity against breast cancer.

Binding of MHC class I-related chain molecules A and B (MICA/B) to the natural killer (NK) cell receptor NK group 2, member D (NKG2D) is thought critical for activating NK-mediated immunosurveillance. Angiogenesis is important for tumor growth and interfering with angiogenesis using the fully human IgG1 anti-VEGFR2 (vascular endothelial growth factor receptor 2) antibody (mAb04) can be effective in treating malignancy. In an effort to make mAb04 more effective we have generated a novel antibody fusion protein (mAb04-MICA) consisting of mAb04 and MICA. We found that mAb04-MICA maintained the anti-angiogenic and antineoplastic activities of mAb04, and also enhanced immunosurveillance activated by the NKG2D pathway. Moreover, in human breast tumor-bearing nude mice, mAb04-MICA demonstrated superior anti-tumor efficacy compared to combination therapy of mAb04 + Docetaxel or Avastin + Docetaxel, highlighting the immunostimulatory effect of MICA. In conclusion, mAb04-MICA provided new inspiration for anti-tumor treatment and had prospects for clinical application.

A novel bispecific diabody targeting both vascular endothelial growth factor receptor 2 and epidermal growth factor receptor for enhanced antitumor activity.

Epidermal growth factor receptor (EGFR) and vascular endothelial growth factor receptor 2 (VEGFR2) are receptor tyrosine kinases known to play critical roles in the development and progression of tumors. Based on the cross-talk between EGFR and VEGFR2 signal pathways, we designed and produced a bispecific diabody (bDAb) targeting both EGFR and VEGFR2 simultaneously. The bispecific molecule (EK-02) demonstrated that it could bind to HUVEC (VEGFR2 high-expressing) and A431 (EGFR overexpressing) cells. Additionally, similar to the parental antibodies, it was able to inhibit proliferation and migration, and induced apoptosis in these cells (HUVECs and A431), demonstrating that it had retained the functional properties of its parental antibodies. Furthermore, the efficacy of EK-02 was evaluated using the human colon adenocarcinoma cell line HT29 (VEGFR2 and EGFR coexpressing). In vitro assay showed that EK-02 could bind to HT29 cells, restrain cell growth and migration, and induce apoptosis with enhanced efficacy compared to both parental antibodies. Further, it inhibited the neovascularization and tumor formation on an HT29 cell bearing chicken chorioallantoic membrane (CAM) tumor model in vivo. In conclusion, these data suggest that the novel bDAb (EK-02) has antiangiogenesis and antitumor capacity both in vitro and in vivo, and can possibly be used as cotargeted therapy for the treatment of EGFR and VEGFR2 overexpressing tumors. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:294-302, 2016.