Blue Sky Biotech had a commanding presence at the Society for Biomolecular Sciences (SBS) annual meeting in Phoenix this year. Here are some highlights to which we would like to draw your attention.
- Several talks, including one by Kathleen Seyb of Brigham and Women’s Hospital and another by William LaMarr of BIOCIUS Life Sciences
Blue Sky Biotech, Inc. is a supplier of biological services and products in the pharmaceutical R&D market whose aim is to alleviate labor-intensive tasks that take scientists away from key experimentation, novel theory exploration, and other optimal uses of their time. Blue Sky continuously develops new and innovative research services while rapidly developing its own products and technologies. The vision of the company is to increase the speed-to-market from “idea” to “drug” by improving the overall efficiency of the drug discovery process and by staying focused on the delivery of time as it relates to molecular biology, tissue culture, fermentation, protein/antibody expression, and cell biology research services and products.
Both Blue Sky BioServices and Blue Sky BioProducts had exhibits located in the main hall, two booths in #402. An interactive display of our custom service and product offerings was available as well as reprints of presentations. The reprints will be made available on our website. Contact us for information.
As part of our exhibition at SBS, Blue Sky BioProducts highlighted our new, novel SmartScreen™ assay kits. Based on the patented TDA 2.0™ technology, these kits enable quantization of membrane-associated enzyme activity and facilitate drug discovery. Extensive information regarding the technology and the product line will be available at the booth and is already available online at www.blueskybiotech.com.
Talks
Further highlighting this unique technology, were several presentations. A talk by Kathleen Seyb of Brigham and Women’s Hospital was given on , titled “Mimicking Membrane-Associated Kinase Activity in an In Vitro Enzymatic Assay for HTS: Template-Directed Assembly”. This talk highlighted the utility of our template-directed assembly, TDA 2.0™, which is featured in our SmartScreen™ kits. The laboratory of Marcie Glicksman at Harvard was interested in screening the membrane-associated kinase Lyn, an important target in the pathogenesis of lymphoma. Measurement of Lyn activity in vitro proved difficult in solution and HTS was impractical. However, by employing TDA 2.0™, the activity of Lyn was robust, making the screen feasible. The results were presented.
Blue Sky distributed an application note in partnership with BIOCIUS Life Sciences, describing label-free high-throughput mass spectrometry assays for the Sirtuin class of histone deacetylases as highlighted in the talk by William LaMarr.
Posters
The following posters were also presented in the Advances in Assay Technology section on Wednesday, April 14 between and , as well as at the reception on Wednesday evening from to . The posters were on display throughout the conference, and reprints and discussion were available at the Blue Sky booth (402).
- Poster B218:
Mimicking Membrane-Associated Kinase Activity in an In Vitro Enzymatic Assay for HTS: Template-Directed Assembly of Lyn Kinase Using TDA 2.0™
Kathleen I. Seyb, Susan Austin, Colin Coombs, Ed Esposito, Erin Sutherland, Anthony Shrout, Marcie A. Glicksman, and Scott Gridley
- Membrane-associated kinases continue to be attractive targets for drug discovery despite being a difficult class of proteins to assay. In most instances the soluble kinase domain is employed because of the ease in expression, purification, and handling. However, many of these fragments exhibit low activity due to the removal of the cell membrane from the natural environment. The loss of full function is likely due to improper orientation and the inability of the enzyme fragments to cluster and interact with neighboring proteins, which is now thought to contribute significantly to activity, function, regulation, and pharmacology. Here we describe the development of a novel enzymatic assay for the membrane-associated Lyn kinase utilizing the template-directed assembly reagent TDA 2.0™—synthetic membrane lipospheres that serve to mimic the membrane surface and restore the native context of the enzyme. In the presence of TDA 2.0™, there is a dramatic increase in Lyn kinase activity compared to assays performed in the absence of TDA 2.0™. TDA 2.0™ allows for a reduction of protein concentration 100-fold, enabling the development and optimization of an HTS-compatible assay that was previously not feasible. TDA 2.0™ represents an enabling technology platform which will allow interrogation of membrane proteins in a more biologically relevant context.
Blue Sky BioProducts Download - Poster B218(PDF): Mimicking Membrane-Associated Kinase Activity in an In Vitro Enzymatic Assay for HTS: Template-Directed Assembly of Lyn Kinase Using TDA 2.0™ - Poster B216:
Analysis of TrkB activity in complex with TDA 2.0™: an HTS-compatible assay with altered relevant substrate preference
Ed Esposito, Erin Sutherland, Anthony Shrout, Scott Gridley, and Paul Wengender
Blue Sky BioProducts, 60 Prescott Street, Worcester, MA 01605
- Membrane-associated proteins derive significant structural, conformational, and relational organization from being constrained on the fluid two-dimensional surface of the membrane. The formation of higher-order complexes of membrane-associated enzymes through template-directed assembly with TDA 2.0™, synthetic membrane lipospheres (which serve to mimic cell membranes) is proposed to contribute significantly to activity, function, regulation, and pharmacology. Here we describe the development of a novel in vitro assay for the membrane-associated kinase, TrkB. Assay of TrkB activity in the presence of TDA 2.0™ reveals a several-fold increase in activity, indicative of complex formation and activation. Additionally, a screen of a library of peptide substrates reveals altered substrate selection in the presence of TDA 2.0™. Further, in contrast to data obtained in the absence of TDA 2.0™, the preferred substrates for TrkB in the presence of TDA 2.0™ include many known biologically relevant substrates. TDA 2.0™ represents an enabling technology platform which will allow interrogation of membrane proteins in a more biologically relevant context.
Blue Sky BioProducts Download - Poster B216(PDF): Analysis of TrkB activity in complex with TDA 2.0™: an HTS-compatible assay with altered relevant substrate preference - Poster B217:
Catalytic Activity of Receptor Tyrosine Kinase Cytoplasmic Domains Strongly Dependent on Template-Directed Assembly
Ed Esposito, Erin Sutherland, Anthony Shrout, Scott Gridley, and Rajiv Gangurde
Blue Sky BioProducts, 60 Prescott Street, Worcester, MA 01605
- The receptor tyrosine kinase (RTK) family of proteins is responsible for sensing the presence of extracellular hormones, cytokines, and growth factors through a cell-surface localized receptor domain and for transducing a signal through an intracellular tyrosine kinase domain to elicit cellular responses. Certain disease states, such as transformed growth (cancer) and insulin insensitivity (diabetes, obesity) arise when RTK signaling becomes dysregulated or inactivated, making this family an important target in drug discovery. The RTKs function as multimers and utilize the membrane to provide a context for the assembly of complex signaling networks. The use of recombinant, soluble, truncated tyrosine kinase domains in early-stage HTS efforts has yielded some questionable results including false positives, false negatives, and poor selectivity data. Our patented reagent, TDA 2.0™, directs the assembly of functional protein complexes on a homogeneous, soluble, nanometer-scale template, enabling recombinant proteins to self-assemble in vitro and more faithfully represent cellular biology. We expressed full-length cytoplasmic domains containing the tyrosine kinases of two closely related representatives of the RTK family, the insulin receptor (IR) and insulin-like growth factor receptor (IGF1R). Our results indicate that these proteins exhibit trans-phosphorylation consistent with dimerization on TDA 2.0™. Furthermore, tyrosine kinase activity is enhanced and substrate selectivity is altered compared to assays conducted in the absence of TDA 2.0™. In summary, TDA 2.0™ provides a context-sensitive environment allowing assembly and activity assessment of complex signaling networks with a higher degree of biological relevance and discrimination of specific inhibitors with higher fidelity.
Blue Sky BioProducts Download - Poster B217(PDF): Catalytic Activity of Receptor Tyrosine Kinase Cytoplasmic Domains Strongly Dependent on Template-Directed Assembly
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SBS Conference & Exhibition