Led by Nathan Alves, PhD, the research conducted within the Alves Lab focuses largely on the development of translational technologies, treatments and techniques that can be used to have a positive impact on people’s lives. The interests in this lab are highly interdisciplinary, as researchers apply engineering principles and designs to create translational technologies for clinical implementation. A chemical and bimolecular engineer, Dr. Alves leads a lab that is uniquely situated to access relevant patient samples as researchers develop novel methods of treating and diagnosing disease.
The Alves Lab is located in the Medical Sciences Building on the IU School of Medicine—Indianapolis campus and spans 1350 square feet that are split across three rooms, each representing a diverse and state-of-the-art translational research center. This space includes a biomarker, biochemistry and small, dander-free molecule/protein purification room; an animal surgery/housing, exercise testing, echocardiography; and a Langendorff room.
Specialized equipment in the Alves Lab includes an Oroboros Oxygraph-2k to provide a unique, high-resolution approach to monitoring cellular and mitochondrial respiratory function and two Haemonetics TEG Analyzer 5000 thromboelastography systems (four channels in total) that enable complete mapping of coagulation, fibrinolysis and platelet function using appropriate excitatory and inhibitory additives. The lab has molecular devices M5, SpectraMax M5 microplate/cuvette reader used for fluorescence intensity (top and bottom reads), enzyme kinetics, luminescence, fluorescence polarization, time resolved fluorescence and absorbance for biochemical assays with current version of SoftMax® as well as a Thermo Scientific Dionex UltiMate 3000 uHPLC System for purification of peptides, small molecules and nucleic acids in the milligram scale of target product.
Additional equipment includes a Burrell wrist-action shaker Model-95 and Butchi R-100 Rotate Evaporator System used to facilitate chemical synthesis and compound isolation processing. Molecular Devices ImageXpress Micro IXM XLS Multimode Microscope for bright field, fluorescence (solid state white-light engine), interchangeable filter cubes (5 locations), interchangeable objectives (4 locations), high resolution 4.66 megapixel scientific CMOS camera, linear encoded voice coil driven X, Y, Z stage with <100 nm resolution are also available in the Alves Lab.
Often blood clots are beneficial in that they’re the body’s appropriate response to injury and the prevention of bleeding by sealing off the wound in a fibrous clot. In some circumstances, a person may develop a blood clot that significantly blocks blood flow, which greatly increases vascular resistance and produces significant strain on the heart. Such is the case with pulmonary embolism (PE).
Patients presenting to the Emergency Department with a PE diagnosis get risk stratified and scanned to determine the best route of clinical intervention. Severe PE can require aggressive therapeutic interventions, such as infusion of tissue plasminogen activator (tPA), as heart strain can transition to heart failure very rapidly. Administering tPA results in the activation of endogenous plasminogen to plasmin, the protein responsible for digesting the insoluble fibrin backbone present in blood clots, but possess significant systemic bleeding risks. Researchers in this lab seek to develop a safer direct fibrinolysis treatment with reduced bleeding risk compared to tPA by infusing active plasmin via lipid-based nanoparticles to protect and target the plasmin to the clot of interest.
Having an appropriate model system to test a pharmaceuticals can be just as important as the therapy itself. As researchers seek to develop direct fibrinolytic agents, they also seek to develop physiologically relevant ex vivo clot lysis assays under flowing conditions. Blood is a complex and intricate mixture of cells, proteins, soluble and insoluble factors that, outside of the body, has an exceedingly short stability. This makes it difficult to perform blood coagulation and lysis assays because fresh blood is necessary from volunteers on an as-needed basis, directly before an assay is conducted.
To further confound results, each volunteer, and even the same volunteer on different days, will exhibit different blood compositions, making comparing lysis and coagulation therapeutic interventions very difficult. The goal of the synthetic human blood clot project is to develop a method for producing a fully human ex vivo blood clot on demand without the need for a fresh blood draw. The clot would then be placed into various flow chambers to mimic in vivo flowing conditions to match a variety of clinically relevant clot orientations.
Antibodies are often utilized across a variety of different industries, including use as pharmaceutical agents to treat disease, medical diagnostic tools to capture and quantify molecules of interest and for many purification applications. In many circumstances, it’s necessary to modify the antibodies to endow them with unnatural capabilities, as the naked antibody is not sufficient to meet the needs of each unique therapeutic or diagnostic application. Common antibody tags are fluorescent probes, affinity molecules, cytotoxic drugs and biotin—just to name a few.
Often, antibody modification techniques are not site-specific and result in inactivation of antibodies as well as increased off-target cross reactivity. Researchers in the Alves Lab utilize a unique site-specific photocrosslinking technique for antibody modification to development designer multifunctional pharmaceutical agents to treat and diagnose disease. Taking advantage of the conserved NBS site present in the antibody Fab region, investigators can site-specifically modify nearly any off the shelf antibody for use in nearly any therapeutic or diagnostic application in which antibody tagging is necessary. They also utilize the NBS for oriented antibody immobilization for enhanced medical diagnostics to meet the ever-increasing demand for faster, earlier and more selective detection of disease biomarkers as we enter the age of personalized medicine.