The Wittrup Lab develops protein engineering technology and applies it to the discovery of new biopharmaceuticals. In particular, yeast surface display is used for the directed evolution of protein expression stability, affinity, and specificity. A particular focus is on the development of anti-cancer drugs, with quantitative studies of cellular-level pharmacokinetics and pharmacodynamics.
Engineers now have the tools to design biological products and processes at the molecular level. Proteins are of particular therapeutic interest, because proteins mediate most biochemical processes both inside and outside cells. The ability to manipulate the strength and specificity of protein binding events provides tremendous leverage for the development of novel biopharmaceuticals. We are developing powerful new tools for protein engineering, and applying them both to particular disease targets and to bettering our understanding of protein structure/function relationships. In the absence of predictive capabilities for protein design, a directed evolution or combinatorial library screening strategy can be effectively applied to alter protein properties in a desired fashion. We apply quantitative engineering analyses of the relevant kinetic and statistical processes to develop optimal search strategies on the protein ìfitness landscape.î In particular, we have developed a method for protein display on the surface of yeast cells that, for example, enabled engineering of a noncovalent protein-ligand bond with a dissociation half-time over one week. We are engineering potential protein biopharmaceuticals in areas where molecular understanding of disease pathology is sufficient to hypothesize particular objective functions to target. For example, antibodies can be used to target cell-killing modalities to cancerous cells, given sufficiently strong and specific binding properties. Growth factors that carry signals between cells do so via particular binding events that, if manipulated to alter intracellular trafficking or signalling outcomes, could alter immune responses in precisely defined ways. Finally, viral and nonviral vectors for gene therapy could be targeted to specific cells and tissues via alteration of an exchangeable antibody recognition module. Altered proteins developed in this work can also provide a potential vehicle for new insights into the mechanisms of protein-ligand binding. We are performing biophysical analyses of the kinetic, thermodynamic, and structural aspects of engineered protein function in order to contribute to an improved understanding of protein binding processes.
Rao BM, Driver I, Lauffenburger DA, Wittrup KD. High-affinity CD25-binding IL-2 mutants potently stimulate persistent T cell growth. Biochemistry. 44(31):10696-701, 2005.
Rao, BM, Lauffenburger, DA, Wittrup KD. Integrating cell-level kinetic modeling into the design of engineered protein therapeutics. Nature Biotechnology, 23(2):191-4, 2005.
Colby DW, Chu Y, Cassady JP, Duennwald M, Zazulak H, Webster JM, Messer A, Lindquist S, Ingram VM, Wittrup KD. Potent inhibition of huntingtin aggregation and cytotoxicity by a disulfide bond-free single-domain intracellular antibody. Proc. Natl. Acad. Sci. USA 101(51): 17616-17621, 2004.
Mohr L, Yeung A, Aloman C, Wittrup D, Wands JR. Antibody-directed therapy for human hepatocellular carcinoma. Gastroenterology. 127(5 Suppl 2):S225-31, 2004.
Midelfort, K.S., Hernandez, H.H., Lippow, S.M., Tidor, B., Drennan, C.L., Wittrup, K.D. Substantial Energetic Improvement with Minimal Structural Perturbation in a High Affinity Mutant Antibody. J. Mol. Biol., 343(3):685-701, 2004.
Rao, B.M., Driver, I., Lauffenburger D.A., Wittrup K.D. IL-2 variants engineered for increased IL 2R± affinity exhibit increased potency arising from a cell surface ligand reservoir effect. Molecular Pharmacology 66(4):864-9, 2004.
Chao, G., Cochran, J.R., Wittrup, K.D. Fine Epitope Mapping of anti-Epidermal Growth Factor Receptor Antibodies through Random Mutagenesis and Yeast Surface Display. J. Mol. Biol. 342(2): 539-50, 2004.
Colby, D.W., Garg, P., Holden, T., Chao, G., Webster, J.M., Messer, A., Ingram, V.M., Wittrup, K.D. Development of a Human Light Chain Variable Domain (VL) Intracellular Antibody Specific for the Amino Terminus of Huntingtin via Yeast Surface Display. J. Mol. Biol. 342(3): 901-12, 2004.
Johns TG, Adams TE, Cochran JR, Hall NE, Hoyne PA, Olsen MJ, Kim YS, Rothacker J, Nice EC, Walker F, Ritter G, Jungbluth AA, Old LJ, Ward CW, Burgess AW, Wittrup KD, Scott AM. Identification of the Epitope for the Epidermal Growth Factor Receptor-specific Monoclonal Antibody 806 Reveals That It Preferentially Recognizes an Untethered Form of the Receptor. J Bio. Chem. 279(29):30375-30384, 2004.
Coughlan, C.M., Walker J.L., Cochran J.C., Wittrup K.D., Brodsky, J.L. Degradation of mutated bovine pancreatic trypsin inhibitor (BPTI) in the yeast vacuole suggests post-endoplasmic reticulum protein quality control. J. Bio.. Chem. 279(15):15289-97,2004.
Graff, C.P., Chester K., Begent R., Wittrup, K.D. Directed Evolution of an Anti-Carcinoembryonic Antigen scFv with a Four-Day Monovalent Dissociation Half-time at 37 °C. Protein Engineering, Design & Selection, 17:293, 2004.
room 66-552
phone (617) 253-4578
email wittrup@mit.edu
