15 results on '"Paul, Sudhir"'
Search Results
2. Anti-Amyloid-β Single-Chain Antibody Brain Delivery Via AAV Reduces Amyloid Load But May Increase Cerebral Hemorrhages in an Alzheimer's Disease Mouse Model.
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Kou, Jinghong, Kim, HongDuck, Pattanayak, Abhinandan, Song, Min, Lim, Jeong-Eun, Taguchi, Hiroaki, Paul, Sudhir, Cirrito, John R., Ponnazhagan, Selvarangan, and Fukuchi, Ken-ichiro
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AMYLOID beta-protein ,ALZHEIMER'S disease ,IMMUNOTHERAPY ,CEREBRAL hemorrhage ,IMMUNOGLOBULINS - Abstract
Accumulation of amyloid-β protein (Aβ) in the brain is thought to be a causal event in Alzheimer's disease (AD). Immunotherapy targeting Aβ holds great promise for reducing Aβ in the brain. Here, we evaluated the efficacy and safety of anti-Aβ single-chain antibody (scFv59) delivery via recombinant adeno-associated virus (rAAV) on reducing Aβ deposits in an AD mouse model (TgAβPPswe/PS1dE9). First, delivery of scFv59 to the brain was optimized by injecting rAAV serotypes 1, 2, and 5 into the right lateral ventricle. Symmetrical high expression of scFv59 was found throughout the hippocampus and partly in the neocortex in both hemispheres via rAAV1 or rAAV5, while scFv59 expression via rAAV2 was mostly limited to one hemisphere. rAAV1, however, induced apoptosis and microglial activation but rAAV5 did not. Therefore, rAAV5 was selected for therapeutic scFv59 delivery in TgAβPPswe/PS1dE9 mice. rAAV5 was similarly injected into the ventricle of 10-month-old TgAβPPswe/PS1dE9 mice and 5 months later its efficacy and safety were evaluated. Immunoreactive Aβ deposits reduced in the hippocampus. Aβ
42 levels in cerebrospinal fluid (CSF) tended to increase and the Aβ40 : 42 ratio decreased in CSF, suggesting that Aβ42 was relocated from the parenchyma to CSF. Hemorrhages associated with a focal increase in blood vessel amyloid were found in the brain. While immunotherapy has great potential for clearing cerebral Aβ, caution for cerebrovascular effects should be exercised when rAAV-mediated anti-Aβ immunotherapy is applied. [ABSTRACT FROM AUTHOR]- Published
- 2011
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3. Theory of proteolytic antibody occurrence
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Paul, Sudhir, Nishiyama, Yasuhiro, Planque, Stephanie, and Taguchi, Hiroaki
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NATURAL immunity , *LABORATORY animals , *IMMUNOGLOBULINS , *PHYSIOLOGY - Abstract
Abstract: Antibodies (Abs) with proteolytic and other catalytic activities have been characterized in the blood and mucosal secretions of humans and experimental animals. The catalytic activity can be traced to nucleophilic sites of innate origin located in Ab germline variable regions. Discoveries of the natural chemical reactivity of Abs were initially met with bewilderment, as the notion had taken hold that catalytic activities can be introduced into Abs by artificial means, but somatically operative selection pressures are designed only to adapt non-covalent Ab binding to antigen ground states. Unsurprisingly, initial efforts to engineer Abs with catalytic activity were oriented towards improving the non-covalent binding at the atoms immediately within the transition state reaction center. Slowly, however, dogmatic approaches to Ab catalysis have given way to the realization that efficient and specific catalytic Abs can be prepared by improving the natural nucleophilic reactivity combined with non-covalent recognition of epitope regions remote from the reaction center. The field remains beset, however, with controversy. This article attempts to provide a rational basis for natural Ab catalysis, in the hope that understanding this phenomenon will stimulate medical and basic science advances in the field. [Copyright &y& Elsevier]
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- 2006
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4. Specific HIV gp120-cleaving Antibodies Induced by Covalently Reactive Analog of gp120.
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Paul, Sudhir, Planque, Stephanie, Yong-Xin Zhou, Taguchi, Hiroaki, Bhatia, Gita, Karle, Sangeeta, Hanson, Carl, and Nishiyama, Yasuhiro
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IMMUNOGLOBULINS , *NUCLEOPHILIC reactions , *VIRAL proteins , *HIV - Abstract
Examines the natural nucleophilic activity of antibodies for the purpose of specific cleavage of the human immunodeficiency virus-1 coat protein gp120. Enzyme-linked immunoabsorbent assay; Nucleophilic reactivity; Proteolysis.
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- 2003
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5. Metal-dependent amyloid β-degrading catalytic antibody construct.
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Nishiyama, Yasuhiro, Taguchi, Hiroaki, Hara, Mariko, Planque, Stephanie A., Mitsuda, Yukie, and Paul, Sudhir
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AMYLOID beta-protein , *METAL catalysts , *IMMUNOGLOBULINS , *CHELATION , *HYDROLYSIS , *NUCLEOPHILIC reactions , *CONFORMATIONAL analysis - Abstract
Highlights: [•] Chelation of bound metal inhibited Aβ hydrolysis by a catalytic antibody fragment. [•] The metal accelerated a reaction step after initial antibody nucleophilic attack. [•] The hydrolytic but not chelator-inactivated antibody dissolved Aβ aggregates. [•] Zn-induced conformational transitions were associated with restored catalysis. [•] The antibody requires bound metal for efficient Aβ hydrolysis and dissolution. [ABSTRACT FROM AUTHOR]
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- 2014
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6. Constitutive Production of Catalytic Antibodies to a Staphylococcus aureus Virulence Factor and Effect of Infection.
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Brown, Eric L., Nishiyama, Yasuhiro, Dunkle, Jesse W., Aggarwal, Shreya, Planque, Stephanie, Watanabe, Kenji, Csencsits-Smith, Keri, Bowden, M. Gabriela, Kaplan, Sheldon L., and Paul, Sudhir
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SYNDECANS , *STAPHYLOCOCCUS aureus , *MICROBIAL virulence , *FIBRINOGEN , *IMMUNOGLOBULINS , *B cells - Abstract
Antibodies that recognize microbial B lymphocyte superantigenic epitopes are produced constitutively with no requirement for adaptive immune maturation. We report cleavage of the Staphylococcus aureus virulence factor extracellular fibrinogen-binding protein (Efb) by catalytic antibodies produced with no exposure to the bacterium and reduction of the catalytic antibody activity following infection. IgG catalytic antibodies that specifically hydrolyzed Efb via a nucleophilic catalytic mechanism were found in the blood of healthy humans and aseptic mice free of S. aureus infection. IgG hydrolyzed peptide bonds on the C-terminal side of basic amino acids, including a bond located within the C3b-binding domain of Efb. Efb digested with the IgG lost its ability to bind C3b and inhibit complement-dependent antibody-mediated red blood cell lysis. In addition to catalysis, the IgG expressed saturable Efb binding activity. IgG from S. aureus-infected mice displayed reduced Efb cleaving activity and increased Efb binding activity compared with uninfected controls, suggesting differing effects of the infection on the antibody subsets responsible for the two activities. IgG from children hospitalized for S. aureus infection also displayed reduced Efb cleavage compared with healthy children. These data suggest a potential defense function for constitutively produced catalytic antibodies to a putative superantigenic site of Efb, but an adaptive catalytic response appears to be proscribed. [ABSTRACT FROM AUTHOR]
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- 2012
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7. Antigen-specific Proteolysis by Hybrid Antibodies Containing Promiscuous Proteolytic Light Chains Paired with an Antigen-binding Heavy Chain.
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Sapparapu, Gopal, Planque, Stephanie A., Nishiyama, Yasuhiro, Foung, Steven K., and Paul, Sudhir
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PROTEOLYSIS , *ANTIGENS , *IMMUNOGLOBULINS , *PROTEOLYTIC enzymes , *IMMUNE recognition , *HYDROLYSIS - Abstract
The antigen recognition site of antibodies consists of the heavy and light chain variable domains (VL and VH domains). VL domains catalyze peptide bond hydrolysis independent of VH domains (Mei, S., Mody, B., Ekiund, S. H., and Paul, S. (1991) I. Biol. Chem. 266, 15571-15574). VH domains bind antigens noncovalently independent of VL domains (Ward, E. S., Güssow, D., Griffiths, A. D., Jones, P. T., and Winter, G. (1989) Nature 341, 544-546). We describe specific hydrolysis of fusion proteins of the hepatitis C virus E2 protein with glutathione S-transferase (GST-E2) or FLAG peptide (FLAG-E2) by antibodies containing the VH domain of an anti-E2 IgG paired with promiscuously catalytic VL domains. The hybrid IgG hydrolyzed the E2 fusion proteins more rapidly than the unpaired light chain. An active site-directed inhibitor of serine proteases inhibited the proteolytic activity of the hybrid IgG, indicating a serine protease mechanism. The hybrid IgG displayed noncovalent E2 binding in enzyme-linked immunosorbent assay tests. Immunoblotting studies suggested hydrolysis of FLAG-E2 at a bond within E2 located ∼11 kDa from the N terminus. GST-E2 was hydrolyzed by the hybrid lgG at bonds in the GST tag. The differing cleavage pattern of FLAG-E2 and GST-E2 can be explained by the split-site model of catalysis, in which conformational differences in the E2 fusion protein substrates position alternate peptide bonds in register with the antibody catalytic subsite despite a common noncovalent binding mechanism. These studies provide proof-of-principle that the catalytic activity of a light chain can be rendered antigen-specific by pairing with a noncovalently binding heavy chain subunit. [ABSTRACT FROM AUTHOR]
- Published
- 2009
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8. Exceptional Amyloid β Peptide Hydrolyzing Activity of Nonphysiological Immunoglobulin Variable Domain ScaffoIds.
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Taguchi, Hiroaki, Planque, Stephanie, Sapparapu, Gopal, Boivin, Stephane, Hara, Mariko, Nishiyama, Yasuhiro, and Paul, Sudhir
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ALZHEIMER'S disease treatment , *NUCLEOPHILIC reactions , *PEPTIDES , *IMMUNOGLOBULINS , *HYDROLYSIS , *IMMUNOTHERAPY , *CLONING - Abstract
Nucleophilic sites in the paired variable domains of the light and heavy chains (V[subL] and V[subH] domains) of Ig can catalyze peptide bond hydrolysis. Amyloid β (Aβ)-binding Igs are under consideration for immunotherapy of Alzheimer disease. We searched for Aβ-hydrolyzing human IgV domains (IgVs) in a library containing a majority of single chain Fv clones mimicking physiological V[subL]-V[subH]-combining sites and minority IgV populations with nonphysiological structures generated by cloning errors. Random screening and covalent selection of phage-displayed IgVs with an electrophilic Aβ analog identified rare IgVs that hydrolyzed Aβ mainly at His[sup14]-Gln[sup15]. Inhibition of IgV catalysis and irreversible binding by an electrophilic hapten suggested a nucleophilic catalytic mechanism. Structural analysis indicated that the catalytic IgVs are nonphysiological structures, a two domain heterodimeric V[subL] (IgV[subL2]-t) and single domain V[subL] clones with aberrant polypeptide tags (IgV[subL]-t'). The IgVs hydrolyzed Aβ at rates superior to naturally occurring Igs by 3-4 orders of magnitude. Forced pairing of the single domain V[subL] with V[subH] or V[subL] domains resulted in reduced Aβ hydrolysis, suggesting catalysis by the unpaired V[subL] domain. Angstrom level amino acid displacements evident in molecular models of the two domain and unpaired V[subL] domain clones explain alterations of catalytic activity. In view of their superior catalytic activity, the V[subL] domain IgVs may help attain clearance of medically important antigens more efficiently than natural Igs. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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9. Catalytic antibodies to HIV: Physiological role and potential clinical utility
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Planque, Stephanie, Nishiyama, Yasuhiro, Taguchi, Hiroaki, Salas, Maria, Hanson, Carl, and Paul, Sudhir
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IMMUNOGLOBULINS , *HIV , *HYDROLYSIS , *BACTERIAL proteins , *MICROBIAL proteins , *BLOOD proteins , *PLASMA cells , *AMINO acids - Abstract
Abstract: Immunoglobulins (Igs) in uninfected humans recognize residues 421–433 located in the B cell superantigenic site (SAg) of the HIV envelope protein gp120 and catalyze its hydrolysis by a serine protease-like mechanism. The catalytic activity is encoded by germline Ig variable (V) region genes, and is expressed at robust levels by IgMs and IgAs but poorly by IgGs. Mucosal IgAs are highly catalytic and neutralize HIV, suggesting that they constitute a first line of defense against HIV. Lupus patients produce the Igs at enhanced levels. Homology of the 421–433 region with an endogenous retroviral sequence and a bacterial protein may provide clues about the antigen driving anti-SAg synthesis in lupus patients and uninfected subjects. The potency and breadth of HIV neutralization revives hopes of clinical application of catalytic anti-421–433 Igs as immunotherapeutic and topical microbicide reagents. Adaptive improvement of anti-SAg catalytic Igs in HIV infected subjects is not customary. Further study of the properties of the naturally occurring anti-SAg catalytic Igs should provide valuable guidance in designing a prophylactic vaccine that amplifies protective catalytic immunity to HIV. [Copyright &y& Elsevier]
- Published
- 2008
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10. Covalent Inactivation of Factor VIII Antibodies from Hemophilia A Patients by an Electrophilic FVIII AnaIog.
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Plariquet, Stephanie, Escobart, Miguel A., Smith, Ken C., Taguchi, Hiroaki, Nishiyama, Yasuhiro, Donnachie, Elizabeth, Pratt, Kathleen P., and Paul, Sudhir
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HEMOPHILIA , *ANTIGENS , *IMMUNOGLOBULINS , *ENZYMES , *SCISSION (Chemistry) - Abstract
The antigen-binding sites of antibodies (Abs) can express enzyme-like nucleophiles that react covalently with electrophilic compounds. We examined the irreversible and specific inactivation of antibodies (Abs) to Factor VIII (FVIII) responsible for failure of FVIII replacement therapy in hemophilia A (HA) patients. Electrophilic analogs of FVIII (E-FVIII) and its C2 domain (E-C2) were prepared by placing the strongly electrophilic phosphonate groups at surface-exposed Lys side chains of diverse antigenic epitopes. IgG Abs to FVIII from HA patients formed stable immune complexes with E-FVIII and E-C2 that were refractory to dissociation by SDS treatment and boiling, procedures that dissociate noncovalent Ab-antigen complexes. The rate-limiting step in the reaction was formation of the initial noncovalent complexes. Conversion of the initial complexes to the irreversible state occurred rapidly. The antigenic epitopes of E-FVIII were largely intact, and most of the Abs were consumed covalently. E-FVIII expressed poor FVIII cofactor activity in clotting factor assays. Nonspecific interference by E-FVIII in clotting factor function was not evident. Treatment with E-FVIII, and to a lesser extent E-C2, irreversibly relieved the FVIII inhibitory effect of HA IgG in clotting factor assays. Small FVIII peptides did not display useful reactivity, highlighting the diverse epitope specificities of the Abs and the conformational character of FVIII epitopes. E-FVIII is a prototype reagent able to attain irreversible and specific inactivation of pathogenic Abs. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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11. Catalytic antibodies to amyloid β peptide in defense against Alzheimer disease
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Taguchi, Hiroaki, Planque, Stephanie, Nishiyama, Yasuhiro, Szabo, Paul, Weksler, Marc E., Friedland, Robert P., and Paul, Sudhir
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IMMUNOGLOBULINS , *AMYLOID , *PEPTIDES , *CLINICAL trials , *IMMUNOTHERAPY - Abstract
Abstract: Immunoglobulins (Igs) that bind amyloid β peptide (Aβ) are under clinical trials for immunotherapy of Alzheimer disease (AD). We have identified IgMs and recombinant Ig fragments that hydrolyze Aβ. Hydrolysis of peripheral Aβ by the IgMs may induce increased Aβ release from the brain. The catalytic IgMs are increased in AD patients, presumably reflecting a protective autoimmune response. Reduced Aβ aggregation and neurotoxicity attributable to the catalytic function were evident. These findings provide a foundation for development of catalytic Igs for AD immunotherapy. [Copyright &y& Elsevier]
- Published
- 2008
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12. Towards Covalent Vaccination: IMPROVED POLYCLONAL HIV NEUTRALIZING ANTIBODY RESPONSE INDUCED BY AN ELECTROPHILIC gp 120 V3 PEPTIDE ANALOG.
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Nishiyama, Yasuhiro, Mitsuda, Yukie, Taguchi, Hiroaki, Planque, Stephanie, Salas, Maria, Hanson, Carl V., and Paul, Sudhir
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VACCINATION , *HIV , *ANTIGENS , *IMMUNOGLOBULINS , *BACTERIA , *NUCLEOPHILIC reactions - Abstract
Rare monoclonal antibodies (Abs) can form irreversible complexes with antigens by enzyme-like covalent nucleophile-electrophile pairing. To determine the feasibility of applying irreversible antigen inactivation by Abs as the basis of vaccination against microbes, we studied the polyclonal nucleophilic Ab response induced by the electrophilic analog of a synthetic peptide corresponding to the principal neutralizing determinant (PND) of human immunodeficiency virus type-1 (HIV) gp120 located in the V3 domain. Abs from mice immunized with the PND analog containing electrophilic phosphonates (E-PND) neutralized a homologous HIV strain (MN) ∼50-fold more potently than control Abs from mice immunized with PND. The IgG fractions displayed binding to intact HIV particles. HIV complexes formed by anti-E-PND IgG dissociated noticeably more slowly than the complexes formed by anti-PND IgG. The slower dissociation kinetics are predicted to maintain long-lasting blockade of host cell receptor recognition by gp120. Pre-treatment of the anti-PND IgG with a haptenic electrophilic phosphonate compound resulted in more rapid dissociation of the HIV-IgG complexes, consistent with the hypothesis that enhanced Ab nucleophilic reactivity induced by electrophilic immunization imparts irreversible character to the complexes. These results suggest that electrophilic immunization induces a sufficiently robust nucleophilic Ab response to enhance the anti-microbial efficacy of candidate polypeptide vaccines. [ABSTRACT FROM AUTHOR]
- Published
- 2007
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13. Lupus-Derived Antiprothrombin Autoantibodies from a V Gene Phage Display Library Are Specific for the Kringle 2 Domain of Prothrombin.
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Le, Anhquyen, Dasgupta, Swapan, Paul, Stephanie, Paul, Sudhir, and Thiagarajan, Perumal
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LUPUS erythematosus , *BLOOD coagulation factor IX , *AUTOANTIBODIES , *PROTHROMBIN , *GENES , *IMMUNOGLOBULINS - Abstract
Autoantibodies to prothrombin are common in patients with systemic lupus erythematosus. Although theft presence is a risk factor for thrombosis, neither their origin nor their precise role in inducing the procoagulant state is known. We have developed a phage-display antibody library from patients with systemic lupus erythematosus with antiprothrombin antibodies, and we have selected two single-chain Fv antibody fragments (ScFvs) by panning on a prothrombin-coated surface. In prothrombin activation assays using purified components, these antibodies promoted prothrombin activation. These ScFvs, termed AN78 and AN129, bound to immobilized prothrombin in a concentration-dependent specific manner but not to other anionic phospholipid binding proteins such as β2-glycoprotein I or annexin V. Phosphatidylserine-bound prothrombin, but not soluble prothrombin, inhibited the binding suggesting that the epitope is available only on immobilized prothrombin. To localize the epitope, prothrombin was treated with thrombin or factor Xa and various prothrombin activation fragments were subsequently isolated and tested in ELISA with the ScFvs. Both AN78 and AN129 bound to prethrombin I (the fragment lacking the Gla domain and the first kringle domain), to fragment 1.2 (containing Gla and the two kringle domains only) and to fragment 2 but not to thrombin, thus localizing the cognate epitope to the kringle 2 domain in prothrombin. Analysis of the cDNA sequences of these antibodies show clustered mutational patterns in the complementarity determining region, suggesting that variable domains are the products of antigen-driven B cell clonal maturation. [ABSTRACT FROM AUTHOR]
- Published
- 2004
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14. Toward Selective Covalent Inactivation of Pathogenic Antibodies.
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Nishiyama, Yasuhiro, Bhatia, Gita, Bangale, Yogesh, Planque, Stephanie, Mitsuda, Yukie, Taguchi, Hiroaki, Karle, Sangeeta, and Paul, Sudhir
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IMMUNOGLOBULINS , *VASOACTIVE intestinal peptide , *NEUROPEPTIDES , *HYDROLYSIS , *PHOSPHONATES , *BINDING sites - Abstract
We report the selective inactivation of proteolytic antibodies (Abs) to an autoantigen, the neuropeptide vasoactive intestinal peptide (VIP), by a covalently reactive analog (CRA) of VIP containing an electrophilic phosphonate diester at the Lys20 residue. The VIP-CRA was bound irreversibly by a monoclonal Ab that catalyzes the hydrolysis of VIP. The reaction with the VIP-CRA proceeded more rapidly than with a hapten CRA devoid of the VIP sequence. The covalent binding occurred preferentially at the light chain subunit of the Ab. Covalent VIP-CRA binding was inhibited by VIP devoid of the phosphonate diester group. These results indicate the importance of noncovalent VIP recognition in guiding Ab nucleophilic attack on the phosphonate group. Consistent with the covalent binding data, the VIP-CRA inhibited catalysis by the recombinant light chain of this Ab with potency greater than the hapten-CRA. Catalytic hydrolysis of VIP by a polyclonal VIPase autoantibody preparation that cleaves multiple peptide bonds located between residues 7 and 22 essentially was inhibited completely by the VIP-CRA, suggesting that the electrophilic phosphonate at Lys20 enjoys sufficient conformational freedom to react covalently with Abs that cleave different peptide bonds in VIP. These results suggest a novel route to antigen-specific covalent targeting of pathogenic Abs. [ABSTRACT FROM AUTHOR]
- Published
- 2004
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15. Broadly Distributed Chemical Reactivity of Natural Antibodies Expressed in Coordination with Specific Antigen Binding Activity.
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Planque, Stephanie, Taguchi, Hiroaki, Burr, Gary, Bhatia, Gita, Karle, Sangeeta, Yong-Xin Zhou, Nishiyama, Yasuhiro, and Paul, Sudhir
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IMMUNOGLOBULINS , *REACTIVITY (Chemistry) - Abstract
Examines the chemical reactivity of natural antibodies expressed in coordination with the specific antigen binding activity. Enzyme-linked immunoabsorbent assay; Covalently reactive antigen analog; Proteolysis assay.
- Published
- 2003
- Full Text
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