Research

The ability of antibodies to recognize target molecules (antigens) with high affinity and specificity is central to their widespread use in diagnostic and therapeutic applications. The binding activity of antibodies is encoded in up to six of their solvent-exposed peptide loops that directly contact antigens. Antibodies are generated by randomly varying the sequences of their antigen-binding loops and selecting rare variants that are complementary to target antigens. Due to the daunting number of possible antibody sequences with variation only within their antigen-binding loops (>10^30 variants), it seems unlikely that the needles (antibodies with desired binding activity) in the haystack (all possible antibody variants) can be predicted instead of being selected. We are challenging this conventional wisdom by reducing the seemingly intractable problem of designing multiple antibody loops to cooperatively bind antigens to a tractable one in which we design individual antibody loops with binding activity.

Based on this paradigm, we have developed a strategy for designing antibodies specific for aggregated proteins associated with several conformational disorders (e.g., Parkinson’s and prion diseases) that is inspired by the molecular interactions governing protein aggregation. We find that grafting small hydrophobic peptides from the Aβ42 peptide associated with Alzheimer’s disease into the complementarity determining regions of a domain antibody generates antibody variants that recognize Aβ oligomers and fibrils with nanomolar affinity. We refer to these antibodies as gammabodies for Grafted AMyloid-Motif AntiBODIES. Interestingly, these grafted antibodies have predictable binding sites within aggregated Aβ conformers since they bind via homotypic interactions between identical peptide motifs. We expect that our antibody design strategy is not limited to Aβ and can be used to readily generate gammabodies against other toxic misfolded proteins.

Selected publications:

Ladiwala, A.R.A., Bhattacharya, M., Perchiacca, J.M., Cao, P., Raleigh, D.P., Abedini, A., Schmidt, A.M., Varkey, J., Langen, R., Tessier, P.M., "Rational design of potent domain antibody inhibitors of amyloid fibril assembly", P. Natl. Acad. Sci. U. S. A., ePub Nov 15 (2012). [link]

Perchiacca, J.P., Ladiwala, A.R.A, Bhattacharya, M.B., Tessier, P.M., "Structure-based design of conformation- and sequence-specific antibodies against amyloid β", P. Natl. Acad. Sci. U. S. A, 109, 84 (2012). [link]

Funding: NSF (CAREER Award), NSF (BBBE), Pew Charitable Trust (Pew Scholar Award in Biomedical Science), American Health Assistance Foundation

Lab news:

Tessier named an
Assistant Editor of the Journal of Biological Chemistry (April 2013)

Mark Julian (graduate student in Tessier lab) awarded an NSF Graduate Fellowship (March 2013)

Tessier wins Rensselaer's School of Engineering Teaching Excellence Award (March 2013)

Lab reports a high-throughput method for assaying antibody stability during antibody selection in Molecular Pharmaceutics (Feb 2013) [link]

Tessier granted early tenure and will be promoted to Associate Professor in July 2013 (Dec 2012)

Lab reports a new method for designing potent antibody inhibitors of amyloid formation in PNAS (Oct 2012) [link to PNAS] [link to news story]

Lab awarded grant from NY State to develop Alzheimer’s antibodies (Sept 2012) [link]

Kathryn Tiller (graduate student in Tessier lab) awarded an NSF Graduate Fellowship (Sept 2012)

Lab awarded an NSF grant to develop methods for designing conformation-specific antibodies (Sept 2012) [link]

Lab reports a novel approach for engineering aggregation-resistant domain antibodies in Protein Engineering, Design & Selection (June 2012) [link]

Tessier presents the Allan P. Colburn Memorial Lecture at the University of Delaware (May 2012) [link]

Lab discovers key structural features of toxic oligomers of the Alzheimer's beta-amyloid peptide (April 2012) [link]

Tessier wins Rensselaer's Early Career Award (April 2012) [link]

Tessier wins Rensselaer's School of Engineering Research Excellence Award (April 2012) [link]

Ali Reza Ladiwala (graduate student in Tessier lab) wins Rensselaer's Karen and Lester Gerhardt Prize in Science and Engineering (April 2012)

Lab discovers that polyphenolic disaccharides are unusually effective at preventing protein aggregation (February 2012) [link]

Lab reports a motif-grafting strategy for designing conformation-specific antibodies against beta-amyloid oligomers and fibrils in PNAS (October 2011) [link to PNAS] [link to news story]

Lab reports a high-throughput, nanoparticle-based screening method for measuring monoclonal antibody self-association in Biophysical Journal (August 2011) [link]

Lab identifies structural mechanisms used by infectious prions to cross species barriers (August 2011) [link]

Lab discovers that aromatic compounds conjugated with sugars potently disaggregate toxic oligomers of beta-amyloid associated with Alzheimer's disease (June 2011) [link]

Lab identifies novel mutations that prevent antibody aggregation (May 2011) [link]

Lab awarded grant from the American Health Assistance Foundation to investigate structural differences between toxic and non-toxic oligomers of beta-amyloid associated with Alzheimer's disease (March 2011) [link]

Genetic Engineering News featured high-throughput protein self-interaction research in Tessier lab (February 2011) [link]

Lab reports three pathways used by aromatic small molecules to selectively remodel toxic soluble oligomers and fibrils of beta-amyloid in the Journal of Biological Chemistry (November 2010) [link]

Tessier wins Pew Scholars Award in Biomedical Sciences (June 2010) [link]

Lab reports that resveratrol selectively remodels toxic soluble oligomers and fibrils of beta-amyloid in the Journal of Biological Chemistry (May 2010) [link]

Tessier wins NSF CAREER Award (March 2010) [link]

Tessier & Lindquist publish a review in Nature Structural & Molecular Biology on prion structure, conformational variants and species barriers (June 2009) [link]

Lab publishes new approach for measuring weak protein interactions in Biotechnology & Bioengineering (June 2009) [link]

Lab awarded NIH grant to study the structural basis of species-specific infectivities of prion strain variants (May 2009)

Tessier awarded Alzheimer’s Association New Investigator Research Grant (July 2008) [link]

Lab publishes JACS paper on self-interaction nanoparticle spectroscopy (Feb 2008) [link]

I. Design of antibodies specific for misfolded (aggregated) proteins

Protein aggregation is the seminal event in conformational disorders such as Alzheimer’s and prion diseases. It is critical to identify therapeutic molecules ranging from small aromatic molecules to large antibodies to inhibit and/or reverse toxic protein aggregation. We have identified several aromatic compounds that potently neutralize toxic protein aggregates either by disaggregating them or converting them to non-toxic aggregates. We are also developing novel strategies for designing peptides and antibodies for selectively and potently inhibiting toxic protein aggregation. Our long-term goal is to develop these and related molecules as therapeutic candidates for treating protein aggregation disorders.

Selected publications:

Ladiwala, A.R, Litt, J., Kane, R.S., Aucoin, D.S., Smith, S.O., Ranjan, S, Davis, J., Van Nostrand, W.E., Tessier, P.M., "Conformational differences between two amyloid β oligomers of similar size and dissimilar toxicity", J. Biol. Chem., 287, 24765 (2012). [link]

Ladiwala, A.R, Mora-Pale, M., Lin, J.C., Bale, S.S., Fishman, Z.S., Dordick, J.S., Tessier, P.M., "Polyphenolic glycosides and aglycones utilize opposing pathways to selectively remodel and inactivate toxic oligomers of amyloid β", ChemBioChem, 12, 1749 (2011). [link]

Ladiwala, A.R., Dordick J.S., Tessier, P.M., “Aromatic small molecules remodel toxic soluble oligomers of amyloid ß through three independent pathways”, J. Biol. Chem., 286, 3209 (2011). [link]

Ladiwala, A.R., Lin, J.C., Bale, S.S., Marcelino-Cruz, A.M., Bhattacharya, M., Dordick, J.S., Tessier, P.M., “Resveratrol selectively remodels soluble oligomers and fibrils of amyloid Aß into off-pathway conformers”, J. Biol. Chem., 285, 24228 (2010). [link]

Funding: Alzheimer’s Association, American Health Assistance Foundation, New York Capital Alliance

II. Inhibition of protein aggregation

High-affinity antibodies are critical for numerous detection and therapeutic applications, yet their utility is limited by their propensity to aggregate. Therefore, it is critical to determine the sequence and structural features that differentiate aggregation-resistant antibodies from aggregation-prone ones to improve antibody activity. Our hypothesis is that the complementarity-determining regions (CDRs) – which commonly contain solvent-exposed hydrophobic residues to mediate high binding affinity – contribute disproportionately to the aggregation propensity of antibodies. Consistent with this hypothesis, we have found that the aggregation behavior of several human antibodies is governed primarily by their CDRs, and this poor solubility can often be localized to a single CDR loop. Moreover, we have found that charged mutations within or near hydrophobic CDRs greatly increase antibody solubility. We are currently using these findings to guide the design and optimization of high-affinity antibodies that are extremely resistant to aggregation.

Selected publications:

Perchiacca, J.M., Ladiwala, A.R.A., Bhattacharya, M., Tessier, P.M., "Aggregation-resistant domain antibodies engineered with charged mutations near the edges of the complementarity-determining regions", Protein Eng. Des. Sel., 12, 591 (2012). [link]

Perchiacca, J.M., Bhattacharya, M., Tessier, P.M., "Mutational analysis of domain antibodies reveals aggregation hotspots within and near the complementarity determining regions", Proteins, 79, 2637 (2011). [link]

Perchiacca, J.M., Tessier, P.M., "Engineering aggregation-resistant antibodies", Ann. Rev. Chem. Biomol. Eng., 3, 263 (2012). [link]

Funding: NSF CAREER Award

III. Engineering aggregation-resistant antibodies

Monoclonal antibodies are typically monomeric and non-viscous at low concentrations, yet they display highly variable associative and viscous behavior at elevated concentrations. Although measurements of antibody self-association are critical for understanding this complex behavior, traditional biophysical methods are not capable of characterizing such concentration-dependent self-association in a high-throughput manner. We are developing a nanoparticle-based method (self-interaction nanoparticle spectroscopy or SINS) capable of rapidly measuring self-interactions between monoclonal antibodies. We find that gold nanoparticles conjugated with antibodies at low protein concentrations display self-association behavior (as measured by their optical properties that depend on interparticle separation distance) that is well correlated with light scattering measurements obtained at orders of magnitude higher antibody concentrations. We are currently using SINS to optimize the selection and formulation of aggregation-resistant antibodies.

Selected publications:

Sule, S.V., Dickinson, C.G., Lu, J., Chow, C.-K., Tessier, P.M., “Rapid analysis of antibody self-association in complex mixtures using immunogold conjugates”, Mol. Pharm., ePub Feb 5 (2013). [link]

Sule, S.V., Sukumar, M., Weiss, W.F., Marcelino-Cruz, A.M., Sample, T., Tessier, P.M., "High-throughput analysis of concentration-dependent antibody self-association", Biophys. J., 101, 1749 (2011). [link]

IV. High-throughput screening of antibody stability