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 patients suffering from venous thromboembolism (VTE), the broader category of pulmonary embolism (PE) and deep vein thrombosis (DVT).

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 initiates 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 leveraging multivalency and nanoparticles to protect and target the plasmin to the clot of interest. 

Having an appropriate model system to test a pharmaceutical 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 digestion 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 that match a variety of clinically relevant clot orientations.  Therapeutics are then infused into the system and clot targeting, digestion, and pressure fluctuations are monitored in real time.  

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.

Common antibody modification techniques are not typically 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.