Production and characterization of bispecific single-chain antibody fragments.

Abstract:

:We report the construction, expression and purification of a bispecific single-chain Fv antibody fragment produced in Escherichia coli. The protein possesses a dual specificity: the single-chain FvB1 portion is directed to the Idiotype of BCL1 lymphoma cells, the single-chain Fv2C11 moiety binds to the CD3 marker on T cells. The two domains are joined by a flexible peptide linker. Using Immobilized Metal Affinity Chromatography, the recombinant protein was purified from bacterial insoluble membrane fractions. After refolding of the bispecific protein, it was affinity-purified. As demonstrated by flow cytometry, both binding sites are retained in the refolded protein. Retargeted cytotoxicity and T cell proliferation assays further prove the biological activity and specificity of the bispecific single-chain Fv. Thus, these bispecific molecules show a potential anti-tumor activity.

journal_name

Mol Immunol

journal_title

Molecular immunology

authors

De Jonge J,Brissinck J,Heirman C,Demanet C,Leo O,Moser M,Thielemans K

doi

10.1016/0161-5890(95)00089-5

subject

Has Abstract

pub_date

1995-12-01 00:00:00

pages

1405-12

issue

17-18

eissn

0161-5890

issn

1872-9142

journal_volume

32

pub_type

杂志文章
  • C1q binding by a high affinity anti-fluorescein murine monoclonal IgM antibody and monomeric subunits.

    abstract::Comparative interactions of purified rabbit C1q with 18-2-3, a high affinity (2-3 X 10(10) M-1) anti-fluorescein (anti-F1) murine monoclonal IgM antibody (pentamer) and constitutive monomeric subunits (IgMs) were studied. Using a solid phase radioimmunoassay (SPRIA), based on immobilized polyvalent antigen, it was sho...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(88)90076-4

    authors: Swanson SM,Dombrink-Kurtzman MA,Voss EW Jr

    更新日期:1988-06-01 00:00:00

  • Variation in the ordered structure of complexes between CD154 and anti-CD154 monoclonal antibodies.

    abstract::The cell surface co-stimulatory protein CD154 (CD40L) is a target for monoclonal antibody (mAb) inhibitors of T-cell mediated immune diseases. This protein, like most other members of the TNF ligand family, forms homotrimeric complexes on the cell surface and in solution, with a three-fold axis of symmetry. We find th...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/s0161-5890(02)00045-7

    authors: Ferrant JL,Wilson CA,Benjamin CD,Hess DM,Hsu YM,Karpusas M,Roux KH,Taylor FR

    更新日期:2002-09-01 00:00:00

  • The TNFα converting enzyme (TACE) from ayu (Plecoglossus altivelis) exhibits TNFα shedding activity.

    abstract::TNFα converting enzyme (TACE) is responsible for converting membrane-anchored TNFα to its soluble form in mammalian. However, the function and characteristics of TACE in teleosts is unclear. In this study, we report the cloning of a cDNA sequence of the PaTACE from ayu, Plecoglossus altivelis. PaTACE encodes an 865-aa...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2014.10.010

    authors: Lu XJ,Chen Q,Yang GJ,Chen J

    更新日期:2015-02-01 00:00:00

  • A conserved WW domain-like motif regulates invariant chain-dependent cell-surface transport of the NKG2D ligand ULBP2.

    abstract::Malignant cells expressing NKG2D ligands on their cell surface can be directly sensed and killed by NKG2D-bearing lymphocytes. To ensure this immune recognition, accumulating evidence suggests that NKG2D ligands are trafficed via alternative pathways to the cell surface. We have previously shown that the NKG2D ligand ...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2015.04.022

    authors: Uhlenbrock F,van Andel E,Andresen L,Skov S

    更新日期:2015-08-01 00:00:00

  • Monitoring the formation of soluble immune complexes composed of idiotype and anti-idiotype antibodies by electron microscopy.

    abstract::We have previously used immunoelectron microscopy (IEM) to generate a three-dimensional map of idiotypic (Id) and isotypic epitopes on the Fab arms of HGAC 39 (Roux et al., 1987, Proc. natn. Acad. Sci. U.S.A. 84, 4984-4988), a mouse IgG3 monoclonal antibody (Mab). In this report, we analyse the geometry of the various...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(94)90167-8

    authors: Roux KH,Greenspan NS

    更新日期:1994-06-01 00:00:00

  • Complement deficiencies and dysregulation: Pathophysiological consequences, modern analysis, and clinical management.

    abstract::Complement defects are associated with an enhanced risk of a broad spectrum of infectious as well as systemic or local inflammatory and thrombotic disorders. Inherited complement deficiencies have been described for virtually all complement components but can be mimicked by autoantibodies, interfering with the activit...

    journal_title:Molecular immunology

    pub_type: 杂志文章,评审

    doi:10.1016/j.molimm.2019.08.002

    authors: Schröder-Braunstein J,Kirschfink M

    更新日期:2019-10-01 00:00:00

  • Molecular interactions of the CTLA-4 cytoplasmic region with the phosphoinositide 3-kinase SH2 domains.

    abstract::During T-cell regulation, T-cell receptors and CD28 lead to signaling activation, while T-lymphocyte antigen 4 (CTLA-4) is known to lead to downregulation, similar to programmed cell death-1 (PD-1). In the cytoplasmic tails of CD28 and CTLA-4, phosphoinositide 3-kinase (PI3K) binds to the consensus sequence including ...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2020.12.002

    authors: Iiyama M,Numoto N,Ogawa S,Kuroda M,Morii H,Abe R,Ito N,Oda M

    更新日期:2020-12-29 00:00:00

  • High molecular weight components containing N-linked oligosaccharides of Ascaris suum extract inhibit the dendritic cells activation through DC-SIGN and MR.

    abstract::Helminths, as well as their secretory/excretory products, induce a tolerogenic immune microenvironment. High molecular weight components (PI) from Ascaris suum extract down-modulate the immune response against ovalbumin (OVA). The PI exerts direct effect on dendritic cells (DCs) independent of TLR 2, 4 and MyD88 molec...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2017.03.015

    authors: Favoretto BC,Casabuono AAC,Portes-Junior JA,Jacysyn JF,Couto AS,Faquim-Mauro EL

    更新日期:2017-07-01 00:00:00

  • Inducible turnover of optineurin regulates T cell activation.

    abstract::Optineurin (Optn) is an adaptor protein with homology to NF-κB essential modulator (NEMO), the regulatory subunit of the IκB kinase (IKK) complex. Dysregulation of Optn has been linked to neurodegenerative, autoimmune and bone diseases. Optn shares a high degree of homology with NEMO, but is not part of the same high-...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2017.01.027

    authors: Montecalvo A,Watkins SC,Orange J,Kane LP

    更新日期:2017-05-01 00:00:00

  • NF-kappaB is essential for induction of pro-inflammatory cytokine genes by filarial parasitic sheath proteins.

    abstract::The transcription factor NF-kappaB plays critical roles in immune and inflammatory responses. Here we show that filarial parasitic sheath proteins cause activation of NF-kappaB in the airway epithelial HEp-2 cell line. This activation was transient and saturable, and involved degradation of the cytoplasmic inhibitor p...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/s0161-5890(00)00039-0

    authors: Narayanan K,Balakrishnan A,Miyamoto S

    更新日期:2000-02-01 00:00:00

  • Comparative molecular modelling of the Fas-ligand and other members of the TNF family.

    abstract::A number of proteins with significant similarity to the tumour necrosis factor (TNF) have been identified over the last years. Upon interaction with their cognate receptor (members of the TNF-receptor family), all members of this protein family induce either cell death or proliferation/differentiation of the receptor-...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(95)00016-8

    authors: Peitsch MC,Tschopp J

    更新日期:1995-07-01 00:00:00

  • Chemokine-mediated inflammation: Identification of a possible regulatory role for CCR2.

    abstract::The chemokine receptor CCR2 binds four pro-inflammatory monocyte chemoattractant proteins, designated MCP1/CCL2, MCP2/CCL8, MCP3/CCL7 and MCP4/CCL13. This study demonstrates the important biology of this receptor during the response to the chemokine milieu. Competitive chemotaxis and calcium flux assays were performed...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2006.09.033

    authors: O'Boyle G,Brain JG,Kirby JA,Ali S

    更新日期:2007-03-01 00:00:00

  • Immunoglobulin G binding activity of Brucella abortus.

    abstract::A Brucella abortus protein with a molecular weight of 50 kDa has been shown to bind bovine immunoglobulin G from healthy, brucellosis-free animals. The Brucella immunoglobulin G binding molecule appears to be a protein, since it is susceptible to proteolysis. The protein is presumed to be located on the cell surface, ...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(91)90084-w

    authors: Bricker BJ,Tabatabai LB,Mayfield JE

    更新日期:1991-01-01 00:00:00

  • Identification of peptidic inhibitors of the alternative complement pathway based on Staphylococcus aureus SCIN proteins.

    abstract::The complement system plays a central role in a number of human inflammatory diseases, and there is a significant need for development of complement-directed therapies. The discovery of an arsenal of anti-complement proteins secreted by the pathogen Staphylococcus aureus brought with it the potential for harnessing th...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2015.05.012

    authors: Summers BJ,Garcia BL,Woehl JL,Ramyar KX,Yao X,Geisbrecht BV

    更新日期:2015-10-01 00:00:00

  • Purification and antimicrobial function of ubiquitin isolated from the gill of Pacific oyster, Crassostrea gigas.

    abstract::An antimicrobial polypeptide was purified from an acidified gill extract of Pacific oyster (Crassostrea gigas) by C(18) reversed-phase HPLC. The purified polypeptide had a molecular weight of 8471Da containing 74 amino acid residues. Comparison of the obtained N-terminal sequences with those of others revealed that it...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2012.07.003

    authors: Seo JK,Lee MJ,Go HJ,Kim GD,Jeong HD,Nam BH,Park NG

    更新日期:2013-01-01 00:00:00

  • Effects of surfactant protein D on growth, adhesion and epithelial invasion of intestinal Gram-negative bacteria.

    abstract::Surfactant protein D (SP-D) interacts with various different microorganisms and plays an important role in pulmonary innate immunity. SP-D expression has also been detected in extrapulmonary tissues, including the gastro-intestinal tract. However, its function in the intestine is unknown and may differ considerably fr...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2007.03.013

    authors: Hogenkamp A,Herías MV,Tooten PC,Veldhuizen EJ,Haagsman HP

    更新日期:2007-07-01 00:00:00

  • Specificity and degeneracy: T cell recognition in CNS autoimmunity.

    abstract::T cells play a crucial role in the pathogenesis of most autoimmune disorders. However, target antigens and pathomechanisms leading to human autoimmune diseases are still largely unknown. Cross-recognition of T cells between self and foreign antigens has been considered as a driving force in generating autoimmunity. He...

    journal_title:Molecular immunology

    pub_type: 杂志文章,评审

    doi:10.1016/j.molimm.2003.11.012

    authors: Zhou D,Hemmer B

    更新日期:2004-02-01 00:00:00

  • Regulation and functions of NLRP3 inflammasome during influenza virus infection.

    abstract::The NLRP3 inflammasome constitutes a major antiviral host defense mechanism during influenza virus infection. Inflammasome assembly in virus-infected cells facilitates autocatalytic processing of pro-caspase-1 and subsequent cleavage and secretion of proinflammatory cytokines IL-1β and IL-18. The NLRP3 inflammasome is...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2017.01.023

    authors: Kuriakose T,Kanneganti TD

    更新日期:2017-06-01 00:00:00

  • Structural comparison of murine T-cell (B151K12)-derived T-cell-replacing factor (IL-5) with rIL-5: dimer formation is essential for the expression of biological activity.

    abstract::T-cell-replacing factor (TRF)/IL-5 is a T-cell-derived glycoprotein which has pleiotropic activity on lymphoid and myeloid cells. IL-5 polypeptide translated into Xenopus oocytes are heterogeneous in molecular size (40,000 to 60,000 under nonreducing conditions) and yields a monomeric form (Mr of 25,000 to 30,000) und...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(90)90158-v

    authors: Takahashi T,Yamaguchi N,Mita S,Yamaguchi Y,Suda T,Tominaga A,Kikuchi Y,Miura Y,Takatsu K

    更新日期:1990-09-01 00:00:00

  • LMIR5 extracellular domain activates myeloid cells through toll-like receptor 4.

    abstract::LMIR5/CD300b is an activating immunoglobulin-like receptor whose extracellular domain (LMIR5-Fc) is constitutively released from immune cells. The release of LMIR5-Fc is augmented upon stimulation with TLR agonists. LMIR5-Fc is reported to possess inflammatory activity and amplify LPS-induced lethal inflammation; howe...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2014.06.012

    authors: Phongsisay V,Iizasa E,Hara H,Yamasaki S

    更新日期:2014-11-01 00:00:00

  • Cloning, sequence analysis and confirmation of derived gene sequences for three epitope-mapped monoclonal antibodies against human phagocyte flavocytochrome b.

    abstract::The integral membrane protein flavocytochrome b (Cyt b) is the catalytic core of the NADPH oxidase complex, a multicomponent enzyme system that initiates a cascade of reactive oxygen species that play a critical role in innate immunity and vascular physiology. Epitope-mapped, monoclonal antibodies (mAb) that recognize...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2005.10.022

    authors: Taylor RM,Maaty WS,Lord CI,Hamilton T,Burritt JB,Bothner B,Jesaitis AJ

    更新日期:2007-01-01 00:00:00

  • A retro-inverso peptide analogue of influenza virus hemagglutinin B-cell epitope 91-108 induces a strong mucosal and systemic immune response and confers protection in mice after intranasal immunization.

    abstract::In this study, a novel approach for the development of a peptide-based vaccine has been tested. We investigated the possibility of replacing an all-L amino acid peptide sequence corresponding to the protective B-cell epitope hemagglutinin (HA) 91-108 from influenza HA with a retro-inverso analogue encompassing this se...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/s0161-5890(02)00095-0

    authors: Ben-Yedidia T,Beignon AS,Partidos CD,Muller S,Arnon R

    更新日期:2002-10-01 00:00:00

  • Amino acid sequence of a light chain variable region of a human rheumatoid factor of the Wa idiotypic group, in part predicted by its reactivity with antipeptide antibodies.

    abstract::Antipeptide antisera were raised to the second and third complementarity-determining regions of the light chain derived from a human monoclonal IgM (Sie) which had antigammaglobulin activity and belonged to the Wa cross-reactive idiotypic group of human rheumatoid factors, two of whose members (Sie, Wo1) had been prev...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(86)90049-0

    authors: Newkirk M,Chen PP,Carson D,Posnett D,Capra JD

    更新日期:1986-03-01 00:00:00

  • Complement regulates inhalation tolerance at the dendritic cell/T cell interface.

    abstract::Pulmonary exposure to innocuous aeroallergens is a common event leading to inhalation tolerance. Distinct subsets of pulmonary dendritic cells (DC) and regulatory T cells (T(Reg)) play critical roles in mediating and maintaining such tolerance. In asthmatics, the same aeroallergens drive a maladaptive, Th2-biased immu...

    journal_title:Molecular immunology

    pub_type: 杂志文章,评审

    doi:10.1016/j.molimm.2006.06.016

    authors: Köhl J,Wills-Karp M

    更新日期:2007-01-01 00:00:00

  • Integrin CD11c contributes to monocyte adhesion with CD11b in a differential manner and requires Src family kinase activity.

    abstract::Integrin-mediated adhesion of human monocytes to fibrinogen regulated by CD11b/CD18 and the closely related integrin CD11c/CD18, play a key role in inflammation. Peripheral blood monocytes isolated from human donors despite expressing CD11c primarily utilized CD11b to mediate adhesion to fibrinogen upon stimulation wi...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2008.04.021

    authors: Georgakopoulos T,Moss ST,Kanagasundaram V

    更新日期:2008-08-01 00:00:00

  • Dmu expression causes enrichment of MZ B cells, but is non permissive for B cell maturation in Rag2-/- mice even if combined with Bcl-2.

    abstract::Rearrangements in reading frame 2 promote the expression of a truncated heavy chain, the Dmu protein. Dmu can assemble into a pre-B cell receptor like complex that appears to induce a subset of signals elicited by full length mu, but cannot promote the pro-B to pre-B cell transition of Rag-/- B cells. In order to dete...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2005.09.020

    authors: Wikström I,Bergqvist I,Holmberg D,Forssell J

    更新日期:2006-03-01 00:00:00

  • Monoclonal anti-histone H1 autoantibodies from MRL lpr/lpr mice.

    abstract::Hybridomas producing anti-histone monoclonal antibodies were generated from a 2-month old MRL lpr/lpr mouse. Five IgM antibodies showed binding to histone H3 and three of these IgM antibodies also bound histone H1. Two IgG2a antibodies (MRA3 and MRA12) were specific for histone H1 and their binding was further charact...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(89)90035-7

    authors: Monestier M,Fasy TM,Böhm L

    更新日期:1989-08-01 00:00:00

  • Amino acid sequence of the variable region of heavy chain in immunoglobulin (Mot) having unusual papain cleavage sites.

    abstract::An IgGl(lambda) Mot myeloma protein showed a unique susceptibility toward papain digestion. The Fab fragment of Mot was more digestible with papain than the Fc fragment. This phenomenon was found to occur by unusual cleavage of the Fd fragment with papain. Determination of the complete primary structure of the V regio...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/0161-5890(86)90039-8

    authors: Kojima M,Koide T,Odani S,Ono T

    更新日期:1986-02-01 00:00:00

  • Identification of the high affinity binding site in the Streptococcus intermedius toxin intermedilysin for its membrane receptor, the human complement regulator CD59.

    abstract::The unique species specificity of the bacterial cytolysin intermedilysin is explained by its requirement for the human complement regulator CD59 as the primary receptor. Binding studies using individual domains of intermedilysin mapped the CD59-binding site to domain 4 and swap mutants between human and rabbit (non-in...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2009.01.003

    authors: Hughes TR,Ross KS,Cowan GJ,Sivasankar B,Harris CL,Mitchell TJ,Morgan BP

    更新日期:2009-04-01 00:00:00

  • Conversion of trichosanthin-induced CD95 (Fas) type I into type II apoptotic signaling during Herpes simplex virus infection.

    abstract::Trichosanthin (TCS) is a type I ribosome-inactivating protein with wide spectrum of pharmacological activities. It inhibits human immunodeficiency virus type 1 (HIV-1) and Herpes simplex virus type 1 (HSV-1) replication but the mechanism is not clear. From a previous study, TCS was found to be more cytotoxic to HIV-1 ...

    journal_title:Molecular immunology

    pub_type: 杂志文章

    doi:10.1016/j.molimm.2011.06.217

    authors: He D,Yau K,He X,Shi H,Zheng Y,Tam S

    更新日期:2011-09-01 00:00:00