This presentation will introduce you to the HSC technology, it’s power, and how it can be used to achieve your research and development goals.
The solublization and stabilization of formulated protein solutions and the crystallization of proteins are inextricably linked to the thermodynamics of protein interactions. The HSC technology is used to quantitatively measure protein-protein interactions. These interactions are calculated as the second virial coefficient, or B value, which is the sum of all potential forces between two proteins. -Positive B values indicate net repulsion -whereas negative B values indicate net attraction. Additives can be introduced to solutions, which can alter the B value so that the a protein displays mild attraction conducive to crystallization. Alternatively, additives can be used to enhance repulsion thereby increasing the protein’s solubility and physical stability.
This image shows the HSC Instrument used to calculate the B values of proteins in solution. It has two components. On the left is the high-throughput instrument where the target protein and additives are analyzed and on the right is the computer used to capture the raw data required to calculate the B values. In approximately 3 weeks, all of this information is compiled and stored for subsequent processing by the artificial neural network.
This illustration depicts how the HSC technology derives the B values from the interactions of the customer’s target protein. A small amount of the customer’s protein is bound to the matrix of a column within the HSC Instrument. Proteins in solution with one of 40 additives at either a high or low concentration are then injected into the column and allowed to pass over the bound protein. A UV detector is used to establish the flow through speed. The flow through speed, in addition to a number of molecular parameters, is an input to an equation that allows the high-throughput system to automatically calculate a B value. However, generally speaking, a slower flow through speed indicates greater protein-protein interaction and thus a more negative B value. Conversely, a faster flow through speed indicates less interaction and thus a more positive B value.
For ease of understanding and to best serve our customers, we offer our services as three screening options. Once the protein has been received and its condition confirmed, the initial screen is performed. During the initial screen, the HSC Instrument is used by our highly experienced staff to determine the B values of your protein in combination with 40 different additives in 80 conditions. The discovery screen then expands upon the conditions of the 9 most promising additives identified from the initial screen. The data derived from the initial and discovery screens combined with data derived from calorimetry assays are analyzed by an artificial neural network as part of the formulation screen.
The artificial neural network, depicted here, provides a powerful companion analysis and predictive capability to the high-throughput component of the HSC technology. It uses the data generated from the 161 assays performed in the initial and discovery screens combined with the algorithms’ predictive components to generate in silico B values for more than 12,000 formulations. It is important to point out that this entire process, including the initial, discovery, and formulation screens takes about one month to complete.
In this example, performed with an inherently insoluble protein, osteoprotegerin, the maximum original concentration of 1 mg/ml was ultimately increased to 50 mg/ml. The data generated from initial and discovery screens performed on osteoprotegerin resulted in a formulation containing two additional additives that allowed the protein to be concentrated 10-fold without any aggregation. Using the data from the initial and discovery screens, combined with the artificial neural network, we then determined a completely new formulation. We were able to deliver this new formulation providing an additional 5 fold increase, allowing the customer to progress to the next stage of development. We were able to deliver…
The HSC technology will enhance and speed research, development, and production of protein-based therapeutics, vaccines, and drugs by minimizing protein aggregation and maximizing solubility and physical stability. Additionally, the HSC technology can be used to rapidly screen crystallization conditions to identify those most likely to yield diffraction quality crystals. The power and the value of the HSC technology is ultimately derived from its ability to save months to years of time and hundreds of thousands of dollars through faster formulation of solutions for proteins.
Thank you for your time. For questions, comments and additional information on the initial, discovery, and formulation screens, please feel free to email us at: firstname.lastname@example.org or call us at 205-420-2055.
HSC Technology - Solutions For Proteins
Solutions for proteins Information provided in this presentation is proprietary and shall not be distributed without the prior written consent of Soluble Therapeutics, LLC
HSC TM Technology <ul><li>Quantitative measure of protein-protein interaction. </li></ul><ul><li>B value - Sum of all potential forces between two proteins in all orientations and distances: Ionic, Dipole, Hydrophobic, Van der Waals. </li></ul>Correlated to crystallization and solubility. B Value Measurements Proteins in Crystal Conditions
HSC TM Technology Rapid screening of the influence of additives on protein-protein interaction.
Case Study: Improving the solubility of Osteoprotegerin using the HSC Technology 1 11 50 Formulation Contents Solubility (mg/ml) Original 20mM HEPES pH 7, 10mM NaCl, 1mM TCEP HSC (Initial and Discovery) 50mM HEPES pH 7, 0.2M NaCl, 10% Glycerol, 0.15M Trehalose, 1mM TCEP HSC + ANN (Formulation) 0.1M MES pH 6.1, 0.8M LiCl, 0.1M Glycine, 15% MPD
Benefits of the HSC TM Technology <ul><li>Rapid determination of protein formulations in support of therapeutic, vaccine, and drug discovery </li></ul><ul><ul><ul><ul><li>Minimize protein aggregation </li></ul></ul></ul></ul><ul><ul><ul><ul><li>Maximize solubility and stability </li></ul></ul></ul></ul><ul><li>Rapid screening of potential crystallization conditions for diffraction quality crystals </li></ul><ul><li>Faster formulation - Complete screen in about a month </li></ul><ul><li>Cost savings of tens of thousands of dollars per protein </li></ul>
Contacts Email: [email_address] Phone: 205-420-2055 Information provided in this presentation is proprietary and shall not be distributed without the prior written consent of Soluble Therapeutics, LLC