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  1. Article ; Online: High throughput purification of monoclonal recombinant antibodies using a Protein-A coated membrane plate system.

    Leal-Lopes, Camila / D'Angelo, Sara / Erasmus, M Frank / Teixeira, Andre A R / Temples, Graham / Zhou, Jinxiang / Bradbury, Andrew R M / Ferrara, Fortunato

    New biotechnology

    2023  Volume 77, Page(s) 111–119

    Abstract: The therapeutic use of monoclonal antibodies (mAbs) ranges from cancer treatment to immune-mediated conditions, covering infectious and cardiovascular disorders, among others. The development of improved methods for therapeutic antibody discovery has ... ...

    Abstract The therapeutic use of monoclonal antibodies (mAbs) ranges from cancer treatment to immune-mediated conditions, covering infectious and cardiovascular disorders, among others. The development of improved methods for therapeutic antibody discovery has accelerated the identification of numerous mAbs: a discovery campaign can be deeply mined, resulting in hundreds, even thousands, of potential antibody leads for a given target of interest. High throughput mAb expression and purification methods are required for the rapid validation of those leads. In this work, we describe the implementation of a Protein-A coated membrane plate system, the Purexa™ AHT membrane plate, for robust preparative purification of hundreds of recombinant mAbs, without the need for automation. The high efficiency (>80%) recovery generated sufficient mAb for downstream screening analyses such as ELISA and surface plasmon resonance (SPR). This new system allows the functional validation of hundreds of lead antibodies from discovery campaigns in a timely manner regardless of operational size.
    MeSH term(s) Antibodies, Monoclonal ; Recombinant Proteins ; Staphylococcal Protein A ; Surface Plasmon Resonance ; Enzyme-Linked Immunosorbent Assay
    Chemical Substances Antibodies, Monoclonal ; Recombinant Proteins ; Staphylococcal Protein A
    Language English
    Publishing date 2023-08-28
    Publishing country Netherlands
    Document type Journal Article
    ZDB-ID 2400836-9
    ISSN 1876-4347 ; 1876-4347
    ISSN (online) 1876-4347
    ISSN 1876-4347
    DOI 10.1016/j.nbt.2023.08.003
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Schistosoma mansoni vaccine candidates identified by unbiased phage display screening in self-cured rhesus macaques.

    Woellner-Santos, Daisy / Tahira, Ana C / Malvezzi, João V M / Mesel, Vinicius / Morales-Vicente, David A / Trentini, Monalisa M / Marques-Neto, Lázaro M / Matos, Isaac A / Kanno, Alex I / Pereira, Adriana S A / Teixeira, André A R / Giordano, Ricardo J / Leite, Luciana C C / Pereira, Carlos A B / DeMarco, Ricardo / Amaral, Murilo S / Verjovski-Almeida, Sergio

    NPJ vaccines

    2024  Volume 9, Issue 1, Page(s) 5

    Abstract: Schistosomiasis, a challenging neglected tropical disease, affects millions of people worldwide. Developing a prophylactic vaccine against Schistosoma mansoni has been hindered by the parasite's biological complexity. In this study, we utilized the ... ...

    Abstract Schistosomiasis, a challenging neglected tropical disease, affects millions of people worldwide. Developing a prophylactic vaccine against Schistosoma mansoni has been hindered by the parasite's biological complexity. In this study, we utilized the innovative phage-display immunoprecipitation followed by a sequencing approach (PhIP-Seq) to screen the immune response of 10 infected rhesus macaques during self-cure and challenge-resistant phases, identifying vaccine candidates. Our high-throughput S. mansoni synthetic DNA phage-display library encoded 99.6% of 119,747 58-mer peptides, providing comprehensive coverage of the parasite's proteome. Library screening with rhesus macaques' antibodies, from the early phase of establishment of parasite infection, identified significantly enriched epitopes of parasite extracellular proteins known to be expressed in the digestive tract, shifting towards intracellular proteins during the late phase of parasite clearance. Immunization of mice with a selected pool of PhIP-Seq-enriched phage-displayed peptides from MEG proteins, cathepsins B, and asparaginyl endopeptidase significantly reduced worm burden in a vaccination assay. These findings enhance our understanding of parasite-host immune responses and provide promising prospects for developing an effective schistosomiasis vaccine.
    Language English
    Publishing date 2024-01-04
    Publishing country England
    Document type Journal Article
    ISSN 2059-0105
    ISSN (online) 2059-0105
    DOI 10.1038/s41541-023-00803-x
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Simultaneous affinity maturation and developability enhancement using natural liability-free CDRs.

    Teixeira, Andre A R / D'Angelo, Sara / Erasmus, M Frank / Leal-Lopes, Camila / Ferrara, Fortunato / Spector, Laura P / Naranjo, Leslie / Molina, Esteban / Max, Tamara / DeAguero, Ashley / Perea, Katherine / Stewart, Shaun / Buonpane, Rebecca A / Nastri, Horacio G / Bradbury, Andrew R M

    mAbs

    2022  Volume 14, Issue 1, Page(s) 2115200

    Abstract: Abbreviations: CDR: complementarity determining region; FACS: fluorescence-activated cell sorting; ... ...

    Abstract Abbreviations: CDR: complementarity determining region; FACS: fluorescence-activated cell sorting; k
    MeSH term(s) Antibody Affinity ; Complementarity Determining Regions ; Single-Chain Antibodies ; Surface Plasmon Resonance
    Chemical Substances Complementarity Determining Regions ; Single-Chain Antibodies
    Language English
    Publishing date 2022-09-22
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2537838-7
    ISSN 1942-0870 ; 1942-0870
    ISSN (online) 1942-0870
    ISSN 1942-0870
    DOI 10.1080/19420862.2022.2115200
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: A ligand motif enables differential vascular targeting of endothelial junctions between brain and retina.

    Tang, Fenny H F / Staquicini, Fernanda I / Teixeira, André A R / Michaloski, Jussara S / Namiyama, Gislene M / Taniwaki, Noemi N / Setubal, João C / da Silva, Aline M / Sidman, Richard L / Pasqualini, Renata / Arap, Wadih / Giordano, Ricardo J

    Proceedings of the National Academy of Sciences of the United States of America

    2019  Volume 116, Issue 6, Page(s) 2300–2305

    Abstract: Endothelial heterogeneity has important implications in health and disease. Molecular markers selectively expressed in the vasculature of different organs and tissues are currently being explored in targeted therapies with promising results in ... ...

    Abstract Endothelial heterogeneity has important implications in health and disease. Molecular markers selectively expressed in the vasculature of different organs and tissues are currently being explored in targeted therapies with promising results in preclinical and clinical studies. Noteworthy is the role that combinatorial approaches such as phage display have had in identifying such markers by using phage as nanoparticles and surrogates for billions of different peptides, screening noninvasively the vascular lumen for binding sites. Here, we show that a new peptide motif that emerged from such combinatorial screening of the vasculature binds selectively to blood vessels in the brain in vivo but not to vessels in other organs. Peptides containing a conserved motif in which amino acids Phenylalanine-Arginine-Tryptophan (FRW) predominate could be visualized by transmission electron microscopy bound to the junctions between endothelial cells in all areas of the brain, including the optic nerve, but not in other barrier-containing tissues, such as intestines and testis. Remarkably, peptides containing the motif do not bind to vessels in the retina, implying an important molecular difference between these two vascular barriers. Furthermore, the peptide allows for in vivo imaging, demonstrating that new tools for studying and imaging the brain are likely to emerge from this motif.
    MeSH term(s) Amino Acid Motifs ; Amino Acid Sequence ; Animals ; Brain/blood supply ; Brain/metabolism ; Cell Surface Display Techniques ; Cerebrovascular Circulation/drug effects ; Endothelium, Vascular/drug effects ; Endothelium, Vascular/metabolism ; Endothelium, Vascular/ultrastructure ; Female ; Fluorescent Antibody Technique ; Ligands ; Mice ; Peptides/chemistry ; Peptides/metabolism ; Protein Binding ; Retinal Vessels/drug effects ; Retinal Vessels/metabolism
    Chemical Substances Ligands ; Peptides
    Language English
    Publishing date 2019-01-22
    Publishing country United States
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 209104-5
    ISSN 1091-6490 ; 0027-8424
    ISSN (online) 1091-6490
    ISSN 0027-8424
    DOI 10.1073/pnas.1809483116
    Database MEDical Literature Analysis and Retrieval System OnLINE

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