Kilogram scale production of active and stable membrane proteins in vesicles
The core capabilities of Aquaporin A/S are 1) Fermentation and purification of membrane proteins at kilogram scale 2) Design of polymersomes leading to highly stable and active membrane protein formulation 3) Development and production of water filtration membranes based on this biomimetic technique Traditionally, membrane proteins are produced in research labs or assayed directly in living cells. Aquaporin's production process constitutes the largest scale membrane proteins have ever been produced at. Further, we have proven capabilities to formulate active and stable membrane proteins in large scale and at a cost orders of magnitude lower than conventional methods. This allows a new paradigme in applications of membrane proteins, simply due to availability and cost.
Accelerated wound healing using natural materials delivered subdermally in a nanoparticle dispersion
Research has established that birch bark triterpenes (TE) have notable wound healing properties. On a cellular level, TE enhances re-epithelialization and reformation of the epithelial barrier. Other research has shown improved epidermal tissue reconstitution, significantly enhancing cell proliferation in the basal layer and promoting the development of a well-formed stratum corneum. A recent research partnership has allowed us to create a novel, nano technology delivery system to deliver these triterpenes subdermally to improve efficacy. This technology was successfully demonstrated by Dr Eric Morrison in a Phase 1 NSF grant which showed the ability of this system to deliver ibuprofen subdermally through topical application. This delivery system can also be applied to other natural products and anti oxidant vitamins for wound and skin care applications
TiO2 nanowired delivery of DL-3-n-butylphthalide (DL-NBP) with neprilysin induces neuroprotection in Alzheimer's disease
TiO2 nanowired DL-3-n-butylphthalide (DL-NBP) reduces brain pathology in concussive head injury (CHI). Since, neprilysin (NPL) is the rate-limiting enzyme for amyloid-beta peptide (AβP) we demonstrate that DL-NBP with NPL if delivered together has superior neuroprotective effects in AD. DL-NBP alone in high doses was also able to significantly reduce AD pathology following intraventricular (i.c.v.) administration of AβP (1-40, 250 ng/10 µl), once daily) for 4 weeks. Co-administration of TiO2 nanowired DL-NBP (40 mg/kg, i.v. once daily for 7 days) with NPL (100 µl in 50 µl, i.c.v.) once daily for 1 week after 2 weeks of AβP infusion significantly reduced brain pathology and behavioural dysfunction. Increased NPL in hippocampus (400 pg/g) with significant decrease in the AβP deposition (45 pg/g from untreated control 75 pg/g; control 40±4 pg/g) was seen with this combined treatment. Interestingly, these changes were also evident when TiO2-DL-NBP was given alone at a high dose (60 mg/kg, i.v.) for 7 days or 10 days under identical conditions. These observations are the first to show that co-administration of TiO2-nanowired DL-NBP with NPL or NBP alone could reduce AD pathology if given for longer time periods or in high doses, not reported earlier.
Despite the burden on patients and the healthcare system, a cure for diabetes has not yet been elucidated. ECMedical’s curative solution in the form of pancreatic organoids is a first of its kind approach to functional organ replacement pushing the boundaries of biotechnological innovation on four separate fronts: 1) Biomaterial science, 2) Vascular network engineering, 3) Immunocompatible cell transplantation, and 4) Pioneering the FDA approval pathway for organ replacement technologies. 1. Biomaterial Science: Originally thought to solely provide structural support to the different organs and tissues of the body, the ECM is now recognized to constitute a reservoir of information in the form of molecular and mechanical cues that contribute to the maintenance of cellular homeostasis and the promotion of optimal tissue/organ function. 2. Vascular Tissue Engineering: ECMedical’s approach to vascular tissue engineering will permit the development of adequately-sized constructs capable of producing euglycemia in humans for the first time. 3. Immunocompatible cell transplantation: ECMedical’s solution to the problem of immunorejection is manufacturing universal donor cells via genetic engineering. 4. Regulatory approval: This project is innovative from a regulatory perspective because it would pave the way and set FDA precedent for organoids.
Wound healing has traditionally taken place either through primary or secondary intention, and in more complex cases through the assistance of complex and costly devices such as a Wound Vac, or via surgical intervention through various flaps or grafts. ECMedical’s innovative solution in the form of a bioactive wound dressing capable of promoting tissue regeneration is a first-of-its kind approach to functional tissue replacement for the treatment of pressure ulcers. This product is specifically designed to enhance healing in complicated clinical situations where factors such as age, smoking status, and comorbidities may be detrimental to the wound healing process.
Neuroprotection in Brain blast injury at hot environmentt by Nanowired delivery of cerebrolysin with mesenchymal stem cells
Blast brain injury (bBI) was inflicted in a shock tube where compressed air-and helium-induced membrane rupture causing pressure waves (100, 150 or 200 kPa) with velocity of ca. 400 to 450 m/sec. The animals survived 8 or 12 h after bBI. Identical bBI was induced in rats exposed to HE at 38°C for 2 h daily for 1 week. Our innovation showed a progressive BBB breakdown in the cerebral cortex, hippocampus, cerebellum, thalamus, hypothalamus and brain stem correlating well with blast overpressure strength. Regional cerebral blood flow (rCBF) reduced by -30 to -58 % associated with edema formation with 8 to 16 % higher volume swelling. Expansion of neuropil, sponginess and neuronal, glial and myelin damages are quite frequent in bBI that were 2-to 3-fold higher after identical bBI at HE. Nanodelivery of CBL (5 ml/kg, i.v.) or MSCs (106 cells, i.v.) either 30 min or 1 h after bBI significantly reduced brain pathology in normal animals however TiO2 nanodelivery of CBL (5 ml/kg, i.v.) together with MSCs (106 cells, i.v.) is needed for neuroprotection in bBI at HE. This innovation showed that nanowired CBL and MSCs is needed for neuroprotection after bBI in HE animals.
Self-assembly of Neural Stem Cells by Biocompatible Sculptured Extracellular Nanomatrices
The current leading and traditional products in the market of stem cell cultivation requires the addition of large amount of growth factors (i.e. divers kinds of polypeptides) and various organic compounds to increase cell division. Despite its capability in promoting cell development, the addition of growth facts also causes a high risk of stimulating growth of cancerous cells and tumors in vivo after transplantation. This is where our innovative product comes in. Our product, iSECnMs, can increase cell division and avoid the usage of additional growth factor for cultivating cells. As for the simply description of philosophy or mechanism, owing to the appropriate design and choice of materials & structure of the matrix, the physiological contact between NSCs and matrices resembles ‘physical massage’ in the Chinese medicine acupuncture technique that causes the cells to specific differentiate into functional cells that are in urgent demand in cell replacement therapy.
Autologous Neural Stem Cell Harvest (ANSc) Technology
Nowadays, there are over 100M people diagnosed with neurodegenerative diseases around the world, however, the neurodegenerative diseases remain incurable as the existing treatments. Our tailor-made magnetic nanoparticles used for extracting active adult neural stem cells in the ANSc Harvest technology. They attach to neural stem cells specifically by recognizing the markers expressed on the neural stem cells. They can also be tracked and real-time monitored by currently available bio-imaging devices, for example MRI. Once the micro-injection of the nanoparticles into the brain and applying magnetic field, the neural stem cells bound with the nanoparticles can be spinned off from the cell lining and extracted from the brain by a micro-syringe. The magnetically extracted neural stem cells are able to proliferate and differentiate in vitro and also give rise to viable cell population. After the extracted neural stem cells have been differentiated, they would be re-injected back to the disease region of the brain to replace the degenerated neural cells in the same patients for potential full recovery. ANSc Harvest Technology could be developed into personalized therapy for curing neurodegenerative diseases as patients utilize their own neural stem cells.
High throughput, benchtop instrument for analysis of protein structures in drug development.
The biological function of protein-based therapeutics are determined by the three dimensional structure of the molecule. Therefore, it is imperative to analyze the structure of protein therapeutics at several stages of the drug development process to ensure both safety and efficacy of the drug. Biopharmaceutical manufacturers are required to demonstrate the consistency of the protein structure to the FDA. Current techniques for protein structural characterization are slow, expensive and difficult to perform. We have developed a technology called Plasma Induced Modification to Biomolecules (PLIMB) that addresses the need of the industry for routine structural analysis of proteins. A way to quickly and efficiently analyze higher order structures of proteins on a benchtop scale, PLIMB will enable faster development of protein therapeutics, throughout the drug discovery process. Ultimately, PLIMB will enable fast, high resolution structural analysis of proteins, a capability which is highly sought after in the pharmaceutical industry.
A paper-based assay for quantitation and identification of glycan and oligosaccharide sugars
Viable synthetic human milk relies on the development of new methods of making the human milk oligosaccharides (HMO) that are necessary for supplementing formula. New synthesis methods will require monitoring and analysis of the glycan (oligosaccharide) products. Current analysis of glycans is limited in throughput and scope by the need for LC/MS technology, which can take roughly an hour for one sample, and requires expensive equipment, specialized columns and derivatizing agents, and expertise on the operation of the equipment. Higher throughput detection would make synthesis of glycans more viable via combinatorial chemical synthesis or metabolic engineering, both of which require the testing of many samples. Paper is a common material for low-cost point-of-use diagnostic where rapid throughput is desirable. A paper based glycan detection kit would therefore have utility for rapid detection for aiding glycan synthesis. Iowa State University researchers have developed a paper based assay that allows for colorimetric detection of glycans for rapid detection and quantification. The assay has enzymes imprinted onto the paper strips that selectively produce a color change when exposed to specific oligosaccharides linkages. The assay is quantifiable and can be completed in 5 minutes. The assay can be applied to high-throughput HMO detection.
Purple Bread: A New Superfood
According to current nutritional thinking, white bread is digested too fast, spikes blood sugar levels and is linked to obesity. In short, it's the enemy of healthy eaters. A Purple bread is invented by adding anthocyanin extract from black rice. Anthocyanin is a natural pigment that occurs in fruits and vegetables such as grapes, blueberries, and sweet potatoes, and is responsible for their vibrant hue. Studies have shown that anthocyanins can help prevent cardiovascular and neurological diseases and cancer, and play a role in controlling obesity and diabetes, as they can inhibit digestive enzymes and reduce glucose levels.
A nontoxic and novel way to prevent and treat cancer and inflammatory disease
We examined the expression of every single gene to find the most promising targets we can use to prevent and treat cancer. Our immediate efforts utilize our lead compound (called “MASL”) to target a unique receptor (called “PDPN”) on cancer cells. PDPN promotes cancer motility, invasion, and metastasis. We can use MASL to prevent and treat many forms of cancer. In addition, PDPN promotes inflammatory destruction leading to rheumatoid and osteoarthritis. MASL can be used to prevent and treat disease in addition to cancer.
Early Detection of Lung Cancer using Breath Test and VOC-Sensitive Aerosols
This proposed breath test device is small in size, noninvasivein nature, easy to use, and less expensive. These allow frequent testing and earlier detection 1. Generation of colorimetric chemical-sensitive aerosols. 2. Inhalation: Depending on the locations of suspected lung cancer, specified inhalation maneuver will be excised so that the inhaled aerosols reach the cancer site. This will be followed by a breath-holding so that the inhaled aerosols (cancer detectives) could have sufficient time to react with the VOCs. 3. Exhalation: The exhaled aerosol distributions will be captured by a filter, giving rise to an aerosol fingerprint. This fingerprint will be compared to the one with normal airways. A large difference between these two indicates carcinogens at the region of interest (ROI). 4. Aerosol samples can be further analyzed by gas chromatography and compared to the lung cancer database. Using the Numerical Inverse Method (NIM) developed in our lab, the site and stage of the lung cancer can retrieved. 5. The breath test can be performed periodically. A time series of aerosol fingerprints tells the development of the lung cancer, or the outcome of cancer treatments.
Diagnostic kit for real-time monitoring resistance in targeted cancer therapy
Targeted drug is common in cancer therapy. Immuno-oncology drugs only effect in 10-25％ cancers. CAR-T tumor immunotherapies only treat small number leukemias. Drug resistances may occur post-therapy: Some patients innately don't respond to a targeted drug in initial treatment; Tumor may develop resistance against an initial effective drug post-0.6-2 years. Drug resistances are often detected too late and cause mortality with CT scan 3 months post-therapy. About 60% drug resistances are not relative to current known oncogene, undetectable by liquid biopsy (ctDNA, CTC, exosome gene assay); their results often false-negative. Current liquid biopsy delay disease treatments, since presence of CTC or ctDNA indicate resistant tumors had metastasized earlier. By directly assaying serum biomarker downstream of oncoproteins that targeted drugs are acting, clinical trials showed our liquid biopsy IVD kit early (2-weeks post-therapy) and “truly” real-time monitor therapy resistance of most drugs and most cancer types, solve problems of CT, liquid biopsy in detecting resistance, help patients get correct drug in time. Our product can facilitate precision medicine; help CT and liquid biopsy get real-time monitoring resistance ability, save cancer treatment expenses.
Phospho-CSE1L Antibody-Drug Conjugates for Extensive Cancer Treatments
Typical antibody-drug conjugates (ADC) can only treat a specific cancer type. Also, as tumors are often heterogeneous, ADCs won’t be able to eradicate heterogeneous cancer cells and this renders cancer relapse. For example : kadcyla only treats Her2-positive breast tumor, recurrences are easy due to proliferation of HER2-nagative tumor cells. CSE1L (1) is highly expressed in most cancer types, (2) it exists as an microvesicle/exosome membrane protein that primarily accumulates in tumors. (3) CSE1L can be stimulated by oncogene to become phospho-CSE1L. Tumor cells often express abnormal oncogene thus have high phospho-CSE1L , while normal cells don’t express abnormal oncogene. Thus, phospho-CSE1L is a potential target for developing antibody-drug conjugates (ADC) that can specifically target tumor cells and extensive cancer treatments. Tumors release much more microvesicles/exosomes. The “CSE1L-ADC and phospho-CSE1L-ADC” featured in our patent (Claim: An anti-CSE1L carrier-conjugated antibody or an anti-phospho-CSE1L carrier-conjugated antibody for in vivo tumor targeting.) may be able to offer better therapy efficacy and enables development of ADCs cancer drugs, radiation cancer drugs, liposome cancer drugs for treating most cancer types, as well as targeted tumor contrast agents and so forth.
Tunable nanocomplexes for delivery of plasmid DNA into human and vertebrate primary cells
Historically, poly(D,L-lactide-co-plycolide) (PLGA) complexes with polyethylenimine (PEI) coated surfaces have been used as a DNA delivery mechanism in gene therapy. This allows for a complex that has relatively low cytotoxicity, binds DNA, and has a cationic surface to allow for easy entry into the target cell. However, these complexes still maintain an amount of cytotoxicity due to the nature of PEI. Smaller PEI molecules have low cytotoxic effects, but then have almost no transfection ability. The present invention uses lower molecular weight PEI along with an attached glycidyl hexadecyl ether to ultimately lower cytotoxic effects of the PEI-PLGA complex while increasing transfection ability. These PEI-PLGA complexes are successfully able to transfect genes into target cells, specifically macrophages, with decreased cytotoxicity and increased transfection ability.
Resveratrol nanoparticles for the prevention and treatment of obesity and its related metabolic disorders
This technology consists of a nanoparticle delivery system that specifically targets adipose stromal cells with resveratrol. Resveratrol delivery into adipose stromal cells causes them to differentiate into fat-burning beige adipose tissue (BAT) over the fat-storing WAT (white adipose tissue), aiding in weight loss and control of blood glucose and lipids. Obesity is a major public health problem affecting over 600 million adults and 100 million children worldwide. Obesity affects more than one third of adults and one sixth of children in the United States. Health risks associated with obesity include the development of cardiovascular disease, type 2 diabetes and some types of cancer.
New antibiotic drug candidates
New antibiotic drug candidates were discovered based on our biological research of bacterial transcription system. These first-in-class antimicrobial small molecules with new mechanism of action displayed excellent bactericidal effects against bacteria including antibiotic-resistant pathogens, unlike any current antibiotics on the market. These molecules target the important bacterial protein-protein interactions not existing in mammals, therefore demonstrated no toxicity to human cells, while the pre-clinical studies are ongoing. These new antibiotic drug candidates are expected to be developed to complement the current antibiotic drugs for treating bacteria caused infectious diseases, and contributing to the society and healthcare system threatened by antibiotic-resistant bacteria.
imple & Rapid Bacterial Detection Using Chimeric Phages
Rapidly identifying a bacterial infection in order to correctly target and kill bacteria has life-saving potential. Annually, millions of people are given the wrong antibiotic resulting in tens of thousands of deaths. This technology allows for the identification and potentially therapeutic targeting of multitudes of bacteria. The technology enables engineered-phages linked to a detection or cytotoxic platform to be designed against a vast array of bacteria. In order to generate a rapid colored result, the engineered-phages linked to gold or silver nanoparticles cause trigger the aggregation of the particles upon binding the targeted bacteria which in turn produces an easily measured visible color change. The color change is evident in 20 minutes or less making it substantially fasted that current technologies. The same approach can be applied killing bacteria if the engineered-phage is linked to a cytotoxin. This would allow for a specific and targeted antibacterial treatment. The flexibility in targeting, combined with the specificity of targeting and its speed make it unique and hugely versatile.
Polylactose Prebiotic Dietary Fiber
Polylactose, a novel dietary fiber, shows great promise as a prebiotic additive to human food products, as a supplement, an animal feed additive and potentially a therapeutic drug. Created by polymerizing lactose (a component of dairy whey), polylactose is highly fermentable, and has a profoundly positive effect on the colonic microflora. Current prebiotic ingredients, fructans (fructo-oligosaccharide and inulins) and galacto-oligosaccharide, offer health benefits such as weight reduction and glycemic control. Preclinical studies of polylactose showed significantly reduced body fat, lowered plasma leptin concentrations, improved blood glucose control and reduced fatty liver at a dietary concentration where other prebiotics were ineffective. While viscous dietary fibers (i.e., guar gum and hydroxypropyl methylcellulose) may offer these benefits, their slimy/gummy texture is unpleasant to most consumers. Polylactose powder, on the other hand, should be very easy to incorporate into foods and can be manufactured at a reasonable cost.
Liquid Crystal Detection of Endotoxins
A new test to detect endotoxin contamination of parenteral drugs. This test utilizes liquid crystals, as opposed to the current Limulus amebocyte lysate (LAL) test, which is derived from the blood of the horseshoe crab. Since the test is based on synthetic reagents, the test can be more replicable and more easily calibrated. The liquid crystal test is compatible with buffers and surfactants that would otherwise cause low endotoxin recovery (LER) issues. LER can lead to regulatory delays, including hold-times and post-marketing commitments The FDA has requested industry explore alternatives to the current LAL test.
Superior neuroprotection in concussive head injury at hot environment by nanowired delivery of cerebrolysin with neprilysin combined with antibodies to amyloid beta peptide
CHI was inflicted by dropping a weight of 114.6 g from 20 cm height on the exposed parietal skull bone in rats either acclimatized at RT (21±1°C) or at HE (34°C for 4 h per day for 2 weeks in biological oxygen demand incubator (BOD, relative humidity 45-47 %, wind speed 20-25 cm/sec). HE alone did not result in BBB breakdown, edema formation or changes in AbP or tau levels. However, CHI in HE resulted in 250 to 285 % higher breakdown of the BBB to Evans blue albumin and radioiodine (-I) and neuronal, glial and axonal damage following identical CHI at RT after 24 trauma. The AbP and tau in CHI at HE increased by 3- to 6-fold in the CSF (control AbP 0.23±0.04; CHI-RT 0.82±0.05; CHI-HE 2.34±0.12 ng/ml); (Control tau 20±2; CHI-RT 34±6; CHI-HE 76±8 pg/ml). Nanodelivery of cerebrolysin (2.5 ml/kg, i.v.) together with 50 µl 1:20 AbP antibodies i.c.v. 4 h after CHI resulted in significant reductions in AbP levels and brain pathology in CHI at HE. Our innovation show better therapeutic avenues in treating exacerbation of brain damage following CHI at HE.
Nanowired delivery of antibodies to tau and neuronal nitric oxide synthase with cerebrolysin reduced pathophysiology of Parkinson's disease after concussive had injury
PD like symptoms was produced in mice by administering 1-metyl-4-fenyl-1,2,3,6-tetrahydropyridin (MPTP, 20 mg/kg, i.p.) daily within 2-h intervals for 5 days in normal or following concussive head injury (CHI). CHI was inflicted by an impact of 0.224 N over the right parietal bone by dropping a weight of 114.6 g from 20 cm. On the 8th day brain pathology was examined. CHI exacerbated p-tau by 1.5 to 2.3 fold in the CSF and in the right and left hemispheres in PD as compared to uninjured PD group. CHI also enhanced greater expression of nNOS, neuronal or glial cell injuries in PD as compared to the uninjured PD group. TiO2 nanowired delivery of cerebrolysin (2.5 ml/kg, i.v.) together with monoclonal p-tau antibodies (phospho S396, 1:20, 30 µl, i.c.v.) with nNOS antibodies (EP1855Y, 1.20 40 µl, i.c.v.) into the left lateral cerebral ventricle 5 days after MPTP significantly reduced blood-brain barrier (BBB) disruption and edema formation in both hemispheres after CHI in PD as compared to CBL given alone. The p-tau levels and nNOS expression were also significantly reduced in the CSF and in brain after these combined treatments in PD following CHI indicating novel therapeutic measure in PD after CHI.