[ { "id": "authors:xq7xw-5cp13", "collection": "authors", "collection_id": "xq7xw-5cp13", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20190729-100027029", "type": "article", "title": "Inhibition of heme sequestration of histidine-rich protein 2 using multiple epitope-targeted peptides", "author": [ { "family_name": "Liang", "given_name": "JingXin", "orcid": "0000-0001-6600-8409", "clpid": "Liang-JingXin" }, { "family_name": "Bunck", "given_name": "David N.", "orcid": "0000-0002-6246-9846", "clpid": "Bunck-D-N" }, { "family_name": "Mishra", "given_name": "Anvita", "clpid": "Mishra-A" }, { "family_name": "Hong", "given_name": "Sunga", "clpid": "Hong-Sunga" }, { "family_name": "Idso", "given_name": "Matthew N.", "clpid": "Idso-M-N" }, { "family_name": "Heath", "given_name": "James R.", "orcid": "0000-0001-5356-4385", "clpid": "Heath-J-R" } ], "abstract": "Plasmodium falciparum is the most lethal species of malaria. In infected human red blood cells, P. falciparum digests hemoglobin as a nutrient source, liberating cytotoxic free heme in the process. Sequestration and subsequent conversion of this byproduct into hemozoin, an inert biocrystalline heme aggregate, plays a key role in parasite survival. Hemozoin has been a longstanding target of antimalarials such as chloroquine (CQ), which inhibit the biocrystallization of free heme. In this study, we explore heme\u2010binding interactions with histidine\u2010rich\u2010protein 2 (HRP2), a known malarial biomarker and purported player in free heme sequestration. HRP2 is notoriously challenging to target due to its highly repetitious sequence and irregular secondary structure. We started with three protein\u2010catalyzed capture agents (PCCs) developed against epitopes of HRP2, inclusive of heme\u2010binding motifs, and explored their ability to inhibit heme:HRP2 complex formation. Cocktails of the individual PCCs exhibit an inhibitory potency similar to CQ, while a covalently linked structure built from two separate PCCs provided considerably increased inhibition relative to CQ. Epitope\u2010targeted disruption of heme:HRP2 binding is a novel approach towards disrupting P. falciparum\u2010related hemozoin formation.", "doi": "10.1002/psc.3203", "issn": "1075-2617", "publisher": "Wiley", "publication": "Journal of Peptide Science", "publication_date": "2019-09", "series_number": "9", "volume": "25", "issue": "9", "pages": "Art. No. e3203" }, { "id": "authors:65yf9-kty62", "collection": "authors", "collection_id": "65yf9-kty62", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20180706-134625112", "type": "article", "title": "An Allosteric Inhibitor of KRas Identified Using a Barcoded Rapid Assay Microchip Platform", "author": [ { "family_name": "McCarthy", "given_name": "Amy Michelle", "orcid": "0000-0003-3456-0383", "clpid": "McCarthy-Amy-Michelle" }, { "family_name": "Kim", "given_name": "Jungwoo", "orcid": "0000-0002-5215-2044", "clpid": "Kim-Jungwoo" }, { "family_name": "Museth", "given_name": "A. Katrine", "clpid": "Museth-A-Katrine" }, { "family_name": "Henning", "given_name": "Ryan K.", "clpid": "Henning-Ryan-K" }, { "family_name": "Heath", "given_name": "John E.", "clpid": "Heath-John-E" }, { "family_name": "Winson", "given_name": "Emma", "clpid": "Winson-Emma" }, { "family_name": "Oh", "given_name": "Joseph J.", "clpid": "Oh-Joseph-J" }, { "family_name": "Liang", "given_name": "Jingxin", "orcid": "0000-0001-6600-8409", "clpid": "Liang-JingXin" }, { "family_name": "Hong", "given_name": "Sunga", "clpid": "Hong-Sunga" }, { "family_name": "Heath", "given_name": "James R.", "orcid": "0000-0001-5356-4385", "clpid": "Heath-J-R" } ], "abstract": "Protein catalyzed capture agents (PCCs) are synthetic antibody surrogates that can target a wide variety of biologically relevant proteins. As a step toward developing a high-throughput PCC pipeline, we report on the preparation of a barcoded rapid assay platform for the analysis of hits from PCC library screens. The platform is constructed by first surface patterning a micrometer scale barcode composed of orthogonal ssDNA strands onto a glass slide. The slide is then partitioned into microwells, each of which contains multiple copies of the full barcode. Biotinylated candidate PCCs from a click screen are assembled onto the barcode stripes using a complementary ssDNA-encoded cysteine-modified streptavidin library. This platform was employed to evaluate candidate PCC ligands identified from an epitope targeted in situ click screen against the two conserved allosteric switch regions of the Kirsten rat sarcoma (KRas) protein. A single microchip was utilized for the simultaneous evaluation of 15 PCC candidate fractions under more than a dozen different assay conditions. The platform also permitted more than a 10-fold savings in time and a more than 100-fold reduction in biological and chemical reagents relative to traditional multiwell plate assays. The best ligand was shown to exhibit an in vitro inhibition constant (IC_(50)) of \u223c24 \u03bcM.", "doi": "10.1021/acs.analchem.8b00706", "pmcid": "PMC6734936", "issn": "0003-2700", "publisher": "American Chemical Society", "publication": "Analytical Chemistry", "publication_date": "2018-08-07", "series_number": "15", "volume": "90", "issue": "15", "pages": "8824-8830" }, { "id": "authors:8gghn-9pz16", "collection": "authors", "collection_id": "8gghn-9pz16", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20160606-134847419", "type": "article", "title": "Detection of a Geographically Diverse Malarial Biomarker via Multi-Epitope Targeted Screening", "author": [ { "family_name": "Liang", "given_name": "JingXin", "orcid": "0000-0001-6600-8409", "clpid": "Liang-JingXin" }, { "family_name": "Nag", "given_name": "Arundhati", "clpid": "Nag-A" }, { "family_name": "Das", "given_name": "Samir", "clpid": "Das-S" }, { "family_name": "Bunck", "given_name": "David", "orcid": "0000-0002-6246-9846", "clpid": "Bunck-D-N" }, { "family_name": "McCarthy", "given_name": "Amy", "clpid": "McCarthy-A" }, { "family_name": "Mishra", "given_name": "Anvita", "clpid": "Mishra-A" }, { "family_name": "Heath", "given_name": "John E.", "clpid": "Heath-J-E" }, { "family_name": "Villalonga", "given_name": "Belen", "clpid": "Villalonga-B" }, { "family_name": "Heath", "given_name": "James R.", "orcid": "0000-0001-5356-4385", "clpid": "Heath-J-R" } ], "abstract": "We report on the development of high affinity macrocyclic peptides as ligands for the capture and detection of PfHRP2, a protein without a defined tertiary structure. PfHRP2 is a key biomarker of the fatal falciparum strain of malaria. This intrinsically disordered protein has repeated epitopes that have inconstant occurrence over geographic regions. Thus, current antibody based diagnostic tests that target a single epitope do not account for the sequence diversity of PfHRP2 and exhibit variable performance. Ligands selected via high throughput epitope-targeted in situ click screening with combinatorial macrocylic peptide libraries yield binders with high affinity and selectivity for their protein targets. These macrocyclic peptide binders offer greater biochemical and thermal stability than antibodies. We provide a general strategy for the amplification of sensitivity in disease detection through the development of ligands that target multiple epitopes in a single and unstructured protein biomarker. We apply our macrocyclic peptide ligands towards the development of an antibody free diagnostic test for PfHRP2.", "doi": "10.1016/j.bpj.2015.11.2774", "issn": "0006-3495", "publisher": "Biophysical Society", "publication": "Biophysical Journal", "publication_date": "2016-02-16", "series_number": "3", "volume": "110", "issue": "3", "pages": "519A" }, { "id": "authors:4cd3j-mm213", "collection": "authors", "collection_id": "4cd3j-mm213", "cite_using_url": "https://resolver.caltech.edu/CaltechAUTHORS:20150921-111839917", "type": "article", "title": "A General Synthetic Approach for Designing Epitope Targeted Macrocyclic Peptide Ligands", "author": [ { "family_name": "Das", "given_name": "Samir", "clpid": "Das-Samir" }, { "family_name": "Nag", "given_name": "Arundhati", "orcid": "0000-0002-1328-1695", "clpid": "Nag-Arundhati" }, { "family_name": "Liang", "given_name": "JingXin", "orcid": "0000-0001-6600-8409", "clpid": "Liang-JingXin" }, { "family_name": "Bunck", "given_name": "David N.", "orcid": "0000-0002-6246-9846", "clpid": "Bunck-David-N" }, { "family_name": "Umeda", "given_name": "Aiko", "clpid": "Umeda-Aiko" }, { "family_name": "Farrow", "given_name": "Blake", "orcid": "0000-0002-1007-7111", "clpid": "Farrow-Blake" }, { "family_name": "Coppock", "given_name": "Matthew B.", "orcid": "0000-0001-9305-3846", "clpid": "Coppock-Matthew-B" }, { "family_name": "Sarkes", "given_name": "Deborah A.", "clpid": "Sarkes-Deborah-A" }, { "family_name": "Finch", "given_name": "Amethist S.", "orcid": "0000-0002-4650-6301", "clpid": "Finch-Amethist-S" }, { "family_name": "Agnew", "given_name": "Heather D.", "orcid": "0000-0003-0010-3248", "clpid": "Agnew-Heather-D" }, { "family_name": "Pitram", "given_name": "Suresh", "clpid": "Pitram-Suresh" }, { "family_name": "Lai", "given_name": "Bert", "clpid": "Lai-Bert" }, { "family_name": "Yu", "given_name": "Mary Beth", "clpid": "Yu-Mary-Beth" }, { "family_name": "Museth", "given_name": "A. Katrine", "clpid": "Museth-A-Katrine" }, { "family_name": "Deyle", "given_name": "Kaycie M.", "clpid": "Deyle-Kaycie-M" }, { "family_name": "Lepe", "given_name": "Bianca", "clpid": "Lepe-Bianca" }, { "family_name": "Rodriguez-Rivera", "given_name": "Frances P.", "orcid": "0000-0003-0651-8639", "clpid": "Rodriguez-Rivera-Frances-P" }, { "family_name": "McCarthy", "given_name": "Amy", "orcid": "0000-0003-3456-0383", "clpid": "McCarthy-Amy-Michelle" }, { "family_name": "Alvarez-Villalonga", "given_name": "Belen", "clpid": "Alvarez-Villalonga-Belen" }, { "family_name": "Chen", "given_name": "Ann", "clpid": "Chen-Ann" }, { "family_name": "Heath", "given_name": "John", "orcid": "0000-0002-5004-2693", "clpid": "Heath-John-E" }, { "family_name": "Stratis-Cullum", "given_name": "Dimitra N.", "orcid": "0000-0002-4832-6945", "clpid": "Stratis-Cullum-Dimitra-N" }, { "family_name": "Heath", "given_name": "James R.", "orcid": "0000-0001-5356-4385", "clpid": "Heath-J-R" } ], "abstract": "We describe a general synthetic strategy for developing high-affinity peptide binders against specific epitopes of challenging protein biomarkers. The epitope of interest is synthesized as a polypeptide, with a detection biotin tag and a strategically placed azide (or alkyne) presenting amino acid. This synthetic epitope (SynEp) is incubated with a library of complementary alkyne or azide presenting peptides. Library elements that bind the SynEp in the correct orientation undergo the Huisgen cycloaddition, and are covalently linked to the SynEp. Hit peptides are tested against the full-length protein to identify the best binder. We describe development of epitope-targeted linear or macrocycle peptide ligands against 12 different diagnostic or therapeutic analytes. The general epitope targeting capability for these low molecular weight synthetic ligands enables a range of therapeutic and diagnostic applications, similar to those of monoclonal antibodies.", "doi": "10.1002/anie.201505243", "pmcid": "PMC4890538", "issn": "1433-7851", "publisher": "Wiley", "publication": "Angewandte Chemie International Edition", "publication_date": "2015-11-02", "series_number": "45", "volume": "54", "issue": "45", "pages": "13219-13224" } ]