10 results on '"Manchulenko, K"'
Search Results
2. Pharmacology of AMG 181, a human anti-α4β7antibody that specifically alters trafficking of gut-homing T cells
- Author
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Pan, WJ, primary, Hsu, H, additional, Rees, WA, additional, Lear, SP, additional, Lee, F, additional, Foltz, IN, additional, Rathanaswami, P, additional, Manchulenko, K, additional, Chan, BM, additional, Zhang, M, additional, Xia, XZ, additional, Patel, SK, additional, Prince, PJ, additional, Doherty, DR, additional, Sheckler, CM, additional, Reynhardt, KO, additional, Krill, CD, additional, Harder, BJ, additional, Wisler, JA, additional, Brandvig, JL, additional, Lynch, JL, additional, Anderson, AA, additional, Wienkers, LC, additional, and Borie, DC, additional
- Published
- 2013
- Full Text
- View/download PDF
3. Pharmacology of AMG 181, a human anti-α4 β7 antibody that specifically alters trafficking of gut-homing T cells.
- Author
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Pan, Wj, Hsu, H, Rees, Wa, Lear, Sp, Lee, F, Foltz, In, Rathanaswami, P, Manchulenko, K, Chan, Bm, Zhang, M, Xia, Xz, Patel, Sk, Prince, Pj, Doherty, Dr, Sheckler, Cm, Reynhardt, Ko, Krill, Cd, Harder, Bj, Wisler, Ja, and Brandvig, Jl
- Abstract
Background and Purpose: AMG 181 is a human anti-α4 β7 antibody currently in phase 1 and 2 trials in subjects with inflammatory bowel diseases. AMG 181 specifically targets the α4 β7 integrin heterodimer, blocking its interaction with mucosal addressin cell adhesion molecule-1 (MAdCAM-1), the principal ligand that mediates α4 β7 T cell gut-homing.Experimental Approach: We studied the in vitro pharmacology of AMG 181, and the pharmacokinetics and pharmacodynamics of AMG 181 after single or weekly i.v. or s.c. administration in cynomolgus monkeys for up to 13 weeks.Key Results: AMG 181 bound to α4 β7 , but not α4 β1 or αE β7 , and potently inhibited α4 β7 binding to MAdCAM-1 (but not vascular cell adhesion molecule-1) and thus inhibited T cell adhesion. Following single i.v. administration, AMG 181 Cmax was dose proportional from 0.01 to 80 mg·kg(-1) , while AUC increased more than dose proportionally. Following s.c. administration, dose-proportional exposure was observed with single dose ranging from 5 to 80 mg·kg(-1) and after 13 weekly doses at levels between 20 and 80 mg·kg(-1) . AMG 181 accumulated two- to threefold after 13 weekly 80 mg·kg(-1) i.v. or s.c. doses. AMG 181 had an s.c. bioavailability of 80%. The linear elimination half-life was 12 days, with a volume of distribution close to the intravascular plasma space. The mean trend for the magnitude and duration of AMG 181 exposure, immunogenicity, α4 β7 receptor occupancy and elevation in gut-homing CD4+ central memory T cell count displayed apparent correlations.Conclusions and Implications: AMG 181 has in vitro pharmacology, and pharmacokinetic/pharmacodynamic and safety characteristics in cynomolgus monkeys that are suitable for further investigation in humans. [ABSTRACT FROM AUTHOR]- Published
- 2013
- Full Text
- View/download PDF
4. Pharmacology of AMG 181, a human anti-α4β7 antibody that specifically alters trafficking of gut-homing T cells.
- Author
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Pan, WJ, Hsu, H, Rees, WA, Lear, SP, Lee, F, Foltz, IN, Rathanaswami, P, Manchulenko, K, Chan, BM, Zhang, M, Xia, XZ, Patel, SK, Prince, PJ, Doherty, DR, Sheckler, CM, Reynhardt, KO, Krill, CD, Harder, BJ, Wisler, JA, and Brandvig, JL
- Subjects
INTEGRINS ,IMMUNOGLOBULINS ,T cells ,INFLAMMATORY bowel diseases ,CROHN'S disease ,ULCERATIVE colitis - Abstract
Background and Purpose AMG 181 is a human anti-α
4 β7 antibody currently in phase 1 and 2 trials in subjects with inflammatory bowel diseases. AMG 181 specifically targets the α4 β7 integrin heterodimer, blocking its interaction with mucosal addressin cell adhesion molecule-1 ( MAdCAM-1), the principal ligand that mediates α4 β7 T cell gut-homing. Experimental Approach We studied the in vitro pharmacology of AMG 181, and the pharmacokinetics and pharmacodynamics of AMG 181 after single or weekly i.v. or s.c. administration in cynomolgus monkeys for up to 13 weeks. Key Results AMG 181 bound to α4 β7 , but not α4 β1 or αE β7 , and potently inhibited α4 β7 binding to MAdCAM-1 (but not vascular cell adhesion molecule-1) and thus inhibited T cell adhesion. Following single i.v. administration, AMG 181 Cmax was dose proportional from 0.01 to 80 mg·kg−1 , while AUC increased more than dose proportionally. Following s.c. administration, dose-proportional exposure was observed with single dose ranging from 5 to 80 mg·kg−1 and after 13 weekly doses at levels between 20 and 80 mg·kg−1 . AMG 181 accumulated two- to threefold after 13 weekly 80 mg·kg−1 i.v. or s.c. doses. AMG 181 had an s.c. bioavailability of 80%. The linear elimination half-life was 12 days, with a volume of distribution close to the intravascular plasma space. The mean trend for the magnitude and duration of AMG 181 exposure, immunogenicity, α4 β7 receptor occupancy and elevation in gut-homing CD4+ central memory T cell count displayed apparent correlations. Conclusions and Implications AMG 181 has in vitro pharmacology, and pharmacokinetic/pharmacodynamic and safety characteristics in cynomolgus monkeys that are suitable for further investigation in humans. [ABSTRACT FROM AUTHOR]- Published
- 2013
- Full Text
- View/download PDF
5. Impact of antibody subclass and disulfide isoform differences on the biological activity of CD200R and βklotho agonist antibodies.
- Author
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Grujic O, Stevens J, Chou RY, Weiszmann JV, Sekirov L, Thomson C, Badh A, Grauer S, Chan B, Graham K, Manchulenko K, Dillon TM, Li Y, and Foltz IN
- Subjects
- Animals, CHO Cells, Cricetulus, Disulfides chemistry, Disulfides immunology, Epitope Mapping methods, Klotho Proteins, Orexin Receptors, Protein Isoforms chemistry, Protein Isoforms immunology, Structure-Activity Relationship, Antibodies, Monoclonal chemistry, Antibodies, Monoclonal immunology, Antigens, Surface chemistry, Antigens, Surface immunology, Membrane Proteins chemistry, Membrane Proteins immunology, Receptors, Cell Surface chemistry, Receptors, Cell Surface immunology
- Abstract
Agonism of cell surface receptors by monoclonal antibodies is dependent not only on its ability to bind the target, but also to deliver a biological signal through receptors to the cell. Immunoglobulin G2 antibodies (IgG2s) are made up of a mixture of distinct isoforms (IgG2-A, -B and A/B), which differ by the disulfide connectivity at the hinge region. When evaluating panels of agonistic antibodies against CD200 receptor (CD200R) or βklotho receptor (βklotho), we noticed striking activity differences of IgG1 or IgG2 antibodies with the same variable domains. For the CD200R antibody, the IgG2 antibody demonstrated higher activity than the IgG1 or IgG4 antibody. More significantly, for βklotho, agonist antibodies with higher biological activity as either IgG2 or IgG1 were identified. In both cases, ion exchange chromatography was able to isolate the bioactivity to the IgG2-B isoform from the IgG2 parental mixture. The subclass-related increase in agonist activity was not correlated with antibody aggregation or binding affinity, but was driven by enhanced avidity for the CD200R antibody. These results add to the growing body of evidence that show that conformational differences in the antibody hinge region can have a dramatic impact on the antibody activity and must be considered when screening and engineering therapeutic antibody candidates. The results also demonstrate that the IgG1 (IgG2-A like) or the IgG2-B form may provide the most active form of agonist antibodies for different antibodies and targets., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
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6. A novel antibody engineering strategy for making monovalent bispecific heterodimeric IgG antibodies by electrostatic steering mechanism.
- Author
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Liu Z, Leng EC, Gunasekaran K, Pentony M, Shen M, Howard M, Stoops J, Manchulenko K, Razinkov V, Liu H, Fanslow W, Hu Z, Sun N, Hasegawa H, Clark R, Foltz IN, and Yan W
- Subjects
- Amino Acids chemistry, Animals, Antibody-Dependent Cell Cytotoxicity, CHO Cells, Cell Line, Cricetulus, Dimerization, Enzyme-Linked Immunosorbent Assay, Humans, Immunoglobulin G immunology, Mice, Protein Engineering, Surface Plasmon Resonance, Immunoglobulin G chemistry, Static Electricity
- Abstract
Producing pure and well behaved bispecific antibodies (bsAbs) on a large scale for preclinical and clinical testing is a challenging task. Here, we describe a new strategy for making monovalent bispecific heterodimeric IgG antibodies in mammalian cells. We applied an electrostatic steering mechanism to engineer antibody light chain-heavy chain (LC-HC) interface residues in such a way that each LC strongly favors its cognate HC when two different HCs and two different LCs are co-expressed in the same cell to assemble a functional bispecific antibody. We produced heterodimeric IgGs from transiently and stably transfected mammalian cells. The engineered heterodimeric IgG molecules maintain the overall IgG structure with correct LC-HC pairings, bind to two different antigens with comparable affinity when compared with their parental antibodies, and retain the functionality of parental antibodies in biological assays. In addition, the bispecific heterodimeric IgG derived from anti-HER2 and anti-EGF receptor (EGFR) antibody was shown to induce a higher level of receptor internalization than the combination of two parental antibodies. Mouse xenograft BxPC-3, Panc-1, and Calu-3 human tumor models showed that the heterodimeric IgGs strongly inhibited tumor growth. The described approach can be used to generate tools from two pre-existent antibodies and explore the potential of bispecific antibodies. The asymmetrically engineered Fc variants for antibody-dependent cellular cytotoxicity enhancement could be embedded in monovalent bispecific heterodimeric IgG to make best-in-class therapeutic antibodies., (© 2015 by The American Society for Biochemistry and Molecular Biology, Inc.)
- Published
- 2015
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7. Remediating agitation-induced antibody aggregation by eradicating exposed hydrophobic motifs.
- Author
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Clark RH, Latypov RF, De Imus C, Carter J, Wilson Z, Manchulenko K, Brown ME, and Ketchem RR
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- Algorithms, Amino Acid Motifs, Antibodies, Monoclonal immunology, Antibodies, Monoclonal pharmacology, Antibody Affinity immunology, Cells, Cultured, Computational Biology methods, HEK293 Cells, Humans, Hydrophobic and Hydrophilic Interactions, Immunoglobulin G immunology, Immunoglobulin G pharmacology, Interferon-gamma immunology, Interferon-gamma metabolism, Killer Cells, Natural drug effects, Killer Cells, Natural immunology, Killer Cells, Natural metabolism, Models, Molecular, Protein Binding immunology, Protein Stability, Protein Structure, Tertiary, Stress, Mechanical, Antibodies, Monoclonal chemistry, Immunoglobulin G chemistry, Protein Aggregates, Protein Multimerization
- Abstract
Therapeutic antibodies must encompass drug product suitable attributes to be commercially marketed. An undesirable antibody characteristic is the propensity to aggregate. Although there are computational algorithms that predict the propensity of a protein to aggregate from sequence information alone, few consider the relevance of the native structure. The Spatial Aggregation Propensity (SAP) algorithm developed by Chennamsetty et. al. incorporates structural and sequence information to identify motifs that contribute to protein aggregation. We have utilized the algorithm to design variants of a highly aggregation prone IgG2. All variants were tested in a variety of high-throughput, small-scale assays to assess the utility of the method described herein. Many variants exhibited improved aggregation stability whether induced by agitation or thermal stress while still retaining bioactivity.
- Published
- 2014
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8. MEDI0639: a novel therapeutic antibody targeting Dll4 modulates endothelial cell function and angiogenesis in vivo.
- Author
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Jenkins DW, Ross S, Veldman-Jones M, Foltz IN, Clavette BC, Manchulenko K, Eberlein C, Kendrew J, Petteruti P, Cho S, Damschroder M, Peng L, Baker D, Smith NR, Weir HM, Blakey DC, Bedian V, and Barry ST
- Subjects
- Animals, Antibodies, Monoclonal administration & dosage, Antibodies, Monoclonal metabolism, Cell Line, Epitope Mapping, Humans, Intracellular Signaling Peptides and Proteins metabolism, Membrane Proteins metabolism, Mice, Mice, SCID, Neovascularization, Pathologic, Protein Binding, Receptors, Notch metabolism, Signal Transduction drug effects, Antibodies, Monoclonal pharmacology, Endothelial Cells drug effects, Endothelial Cells metabolism, Intracellular Signaling Peptides and Proteins antagonists & inhibitors, Membrane Proteins antagonists & inhibitors, Neovascularization, Physiologic drug effects
- Abstract
The Notch signaling pathway has been implicated in cell fate determination and differentiation in many tissues. Accumulating evidence points toward a pivotal role in blood vessel formation, and the importance of the Delta-like ligand (Dll) 4-Notch1 ligand-receptor interaction has been shown in both physiological and tumor angiogenesis. Disruption of this interaction leads to a reduction in tumor growth as a result of an increase in nonfunctional vasculature leading to poor perfusion of the tumor. MEDI0639 is an investigational human therapeutic antibody that targets Dll4 to inhibit the interaction between Dll4 and Notch1. The antibody cross-reacts to cynomolgus monkey but not mouse species orthologues. In vitro MEDI0639 inhibits the binding of Notch1 to Dll4, interacting via a novel epitope that has not been previously described. Binding to this epitope translates into MEDI0639 reversing Notch1-mediated suppression of human umbilical vein endothelial cell growth in vitro. MEDI0639 administration resulted in stimulation of tubule formation in a three-dimensional (3D) endothelial cell outgrowth assay, a phenotype driven by disruption of the Dll4-Notch signaling axis. In contrast, in a two-dimensional endothelial cell-fibroblast coculture model, MEDI0639 is a potent inhibitor of tubule formation. In vivo, MEDI0639 shows activity in a human endothelial cell angiogenesis assay promoting human vessel formation and reducing the number of vessels with smooth muscle actin-positive mural cells coverage. Collectively, the data show that MEDI0639 is a potent modulator of Dll4-Notch signaling pathway.
- Published
- 2012
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9. Kinetic analysis of unpurified native antigens available in very low quantities and concentrations.
- Author
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Rathanaswami P, Richmond K, Manchulenko K, and Foltz IN
- Subjects
- Animals, Antibodies, Monoclonal analysis, Cells, Cultured, Humans, Interleukin-13 analysis, Kinetics, T-Lymphocytes immunology, Antibodies, Monoclonal immunology, Antibody Affinity, Interleukin-13 immunology
- Abstract
Affinity measurements of antigen-antibody interactions are generally performed using known concentrations of purified or recombinant materials. In addition, many technologies that measure affinity require the interacting components to be present in at least microgram quantities. Specifically, if the antigen is either available only in low quantities or unable to be purified, or if the quantity is unknown, then the measurement of affinity can be very difficult. Using the Kinetic Exclusion Assay (KinExA) technology, here we describe a method that overcomes the requirement for large amounts of purified and known quantities of antigen. We used this method to precisely measure the affinity of fully human anti-human interleukin 13 (IL13) monoclonal antibodies to IL13 produced in native form from primary T cells derived from a variety of species, including human. These antigens were available only in the limited quantities present in the conditioned cell culture medium, and the affinity was measured directly without further purification., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2011
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10. Enhancing antibody Fc heterodimer formation through electrostatic steering effects: applications to bispecific molecules and monovalent IgG.
- Author
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Gunasekaran K, Pentony M, Shen M, Garrett L, Forte C, Woodward A, Ng SB, Born T, Retter M, Manchulenko K, Sweet H, Foltz IN, Wittekind M, and Yan W
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- Animals, Dimerization, Female, Humans, Kinetics, Leukocytes, Mononuclear cytology, Mass Spectrometry methods, Mice, Mice, Inbred C57BL, Mice, Inbred NOD, Neoplasm Transplantation, Protein Conformation, Protein Structure, Tertiary, Tumor Necrosis Factor-alpha chemistry, Antibodies, Bispecific chemistry, Immunoglobulin G chemistry
- Abstract
Naturally occurring IgG antibodies are bivalent and monospecific. Bispecific antibodies having binding specificities for two different antigens can be produced using recombinant technologies and are projected to have broad clinical applications. However, co-expression of multiple light and heavy chains often leads to contaminants and pose purification challenges. In this work, we have modified the CH3 domain interface of the antibody Fc region with selected mutations so that the engineered Fc proteins preferentially form heterodimers. These novel mutations create altered charge polarity across the Fc dimer interface such that coexpression of electrostatically matched Fc chains support favorable attractive interactions thereby promoting desired Fc heterodimer formation, whereas unfavorable repulsive charge interactions suppress unwanted Fc homodimer formation. This new Fc heterodimer format was used to produce bispecific single chain antibody fusions and monovalent IgGs with minimal homodimer contaminants. The strategy proposed here demonstrates the feasibility of robust production of novel Fc-based heterodimeric molecules and hence broadens the scope of bispecific molecules for therapeutic applications.
- Published
- 2010
- Full Text
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