Expression of full-length immunoglobulins in Escherichia coli: rapid and efficient production of aglycosylated antibodies.

Abstract:

:Many research and clinical applications require large quantities of full-length antibodies with long circulating half-lives, and production of these complex multi-subunit proteins has in the past been restricted to eukaryotic hosts. In this report, we demonstrate that efficient secretion of heavy and light chains in a favorable ratio leads to the high-level expression and assembly of full-length IgGs in the Escherichia coli periplasm. The technology described offers a rapid, generally applicable and potentially inexpensive method for the production of full-length therapeutic antibodies, as verified by the expression of several humanized IgGs. One E. coli-derived antibody in particular, anti-tissue factor IgG1, has been thoroughly evaluated and has all of the expected properties of an aglycosylated antibody, including tight binding to antigen and the neonatal receptor. As predicted, the protein lacks binding to C1q and the FcgammaRI receptor, making it an ideal candidate for research purposes and therapeutic indications where effector functions are either not required or are actually detrimental. In addition, a limited chimpanzee study suggests that the E. coli-derived IgG1 retains the long circulating half-life of mammalian cell-derived antibodies.

journal_name

J Immunol Methods

authors

Simmons LC,Reilly D,Klimowski L,Raju TS,Meng G,Sims P,Hong K,Shields RL,Damico LA,Rancatore P,Yansura DG

doi

10.1016/s0022-1759(02)00036-4

keywords:

subject

Has Abstract

pub_date

2002-05-01 00:00:00

pages

133-47

issue

1-2

eissn

0022-1759

issn

1872-7905

pii

S0022175902000364

journal_volume

263

pub_type

杂志文章
  • Production of monoclonal antibodies that recognize the extracellular domain of mouse langerin/CD207.

    abstract::Langerin CD207 is a type II transmembrane protein. It is responsible for the formation of Birbeck granules, which are intracellular organelles within Langerhans cells, the dendritic cells of stratified squamous epithelia like the epidermis. Because current anti-CD207 antibodies have limitations, we prepared new monocl...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2007.05.001

    authors: Cheong C,Idoyaga J,Do Y,Pack M,Park SH,Lee H,Kang YS,Choi JH,Kim JY,Bonito A,Inaba K,Yamazaki S,Steinman RM,Park CG

    更新日期:2007-07-31 00:00:00

  • Immunoaffinity purification of hepatitis B surface antigen using anti-idiotype as a specific eluent.

    abstract::An immunoaffinity method for the purification of hepatitis B surface antigen (HBsAg) using monoclonal anti-idiotype (anti-Id) as a specific eluent is described. Ammonium sulfate fractions of HBsAg-containing plasma were absorbed on monoclonal idiotype (Id) immunoadsorbents. The bound material was eluted from the colum...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(90)90019-r

    authors: Kim WB,Kim BM,Choi EC,Kim BK

    更新日期:1990-01-24 00:00:00

  • Characterization of FcγRIIIA effector cells used in in vitro ADCC bioassay: Comparison of primary NK cells with engineered NK-92 and Jurkat T cells.

    abstract::Antibody-dependent cell-mediated cytotoxicity (ADCC) is an important mechanism of action (MOA) of several therapeutic antibody drugs and evaluation in ADCC bioassays is important in antibody drug development and maintenance. Three types of effector cells now routinely used in bioassay evaluation of ADCC are natural ki...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2016.12.002

    authors: Hsieh YT,Aggarwal P,Cirelli D,Gu L,Surowy T,Mozier NM

    更新日期:2017-02-01 00:00:00

  • The use of denaturing conditions for gel diffusion of insoluble epidermal proteins.

    abstract::Modifications of the classical technique for double diffusion in agar plates are presented. These procedures allow insoluble proteins to be dissolved in a variety of denaturing solvents and antibody production specificity to be monitored by the appearance of precipitin lines in agarose gels as in the conventional Ouch...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(77)90192-2

    authors: Lee LD,Baden HP

    更新日期:1977-01-01 00:00:00

  • Enumeration of t rosette-forming cell population in rabbit peripheral blood using human, sheep, guinea-pig, rat, mouse and chicken red blood cells under different experimental conditions.

    abstract::Attempts were made to estimate peripheral T-lymphocytes in rabbits by their ability to form rosettes with sheep, human, guinea-pig, rat, mouse and chicken RBC. The reaction was studied under different conditions of pH and temperature. The test was also carried out in the presence or absence of glutaraldehyde. With she...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(77)90165-x

    authors: Vakharia DD,Vakharia MR,Rao SS

    更新日期:1977-01-01 00:00:00

  • Enzyme immunoassays for the detection of bacterial antigens utilizing biotin-labeled antibody and peroxidase biotin--avidin complex.

    abstract::This report describes the use of biotin-labeled antibodies in sensitive enzyme immunoassay systems for the measurement of bacterial antigens. Biotinylated immunoglobulins could be reproducibly formulated using N-hydroxysuccinimide biotin ester to link biotin to the immunoglobulin. Binding of the biotinylated antibody ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(83)80021-0

    authors: Yolken RH,Leister FJ,Whitcomb LS,Santosham M

    更新日期:1983-02-11 00:00:00

  • Cell-specific heterogeneity in sensitivity of phosphatidylinositol-anchored membrane antigens to release by phospholipase C.

    abstract::Cell surface antigens thought to be linked to the membrane via phosphatidylinositol (PI) are incompletely, and variably, released by treatment with phosphatidylinositol-specific phospholipase C (PI-PLC). The basis for this was investigated with cloned tumor cell lines and PI-PLCs isolated from two species of bacteria....

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(88)90386-9

    authors: Low MG,Stiernberg J,Waneck GL,Flavell RA,Kincade PW

    更新日期:1988-10-04 00:00:00

  • The lymphocytosis promoting action of pertussis toxin can be mimicked in vitro. Holotoxin but not the B subunit inhibits invasion of human T lymphoma cells through fibroblast monolayers.

    abstract::Pertussis toxin is known to elicit lymphocytosis in whooping cough patients and experimental animals, by blocking the extravasation of lymphocytes and stimulating their release from lymphoid organs such as the thymus. The mechanisms responsible for these unique effects of PT are not fully understood. The effect of per...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90090-3

    authors: Verschueren H,Dewit J,Van der Wegen A,De Braekeleer J,Van Heule G,Dekegel D,De Baetselier P

    更新日期:1991-11-22 00:00:00

  • Influence of molecular size of IgA on its immunoassay by various techniques. III. Immunonephelometry.

    abstract::The influence of size heterogeneity of IgA on its immunonephelometric (IN) assay was studied using highly purified preparations of monoclonal (MC) or polyclonal (PC) monomeric (M), dimeric (D), trimeric (T) and tetrameric (Q) IgA, as well as of secretory IgA (sIgA). Concentrations measured by optical density (OD) were...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(82)90381-7

    authors: Delacroix DL,Vaerman JP

    更新日期:1982-01-01 00:00:00

  • Analysis of the interaction between monoclonal antibodies and human hemoglobin (native and cross-linked) using a surface plasmon resonance (SPR) biosensor.

    abstract::To develop a stable immuno-assay system for quantification of human hemoglobin (Hb), the interaction between various antibodies and Hb was studied using a surface plasmon resonance (SPR) biosensor in the BIAcore equipment (Amersham Pharmacia Biotech) with an immobilized anti-Hb antibody sensor chip. When polyclonal an...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(00)00144-7

    authors: Sonezaki S,Yagi S,Ogawa E,Kondo A

    更新日期:2000-04-21 00:00:00

  • A novel approach using streptavidin magnetic bead-sorted in vivo biotinylated survivin for monoclonal antibody production.

    abstract::One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-bioti...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2006.07.024

    authors: Tayapiwatana C,Chotpadiwetkul R,Kasinrerk W

    更新日期:2006-12-20 00:00:00

  • Detection of antibodies to mycobacterium tuberculosis by solid phase radioimmunoassay.

    abstract::The sandwich solid phase radioimmunoassay procedure has been adapted to the serological diagnosis of tuberculosis by the use of disposable plastic haemagglutination trays in which the wells were sensitized with soluble mycobacterial antigen and the subsequent uptake of globulin from test sera detected with radioiodina...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(75)90069-1

    authors: Nassau E,Parsons ER,Johnson GD

    更新日期:1975-01-01 00:00:00

  • An enzyme immunoassay design using labelled antibodies for the determination of haptens. Application to methotrexate assay.

    abstract::A competitive enzyme immunoassay with labelled antibodies has been developed for methotrexate (MTX). Methotrexate in the sample and a constant quantity of this hapten physically absorbed to polystyrene spheres through a methylated bovine albumin carrier were allowed to compete for a limiting amount of peroxidase label...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(83)90353-8

    authors: Ferrua B,Milano G,Ly B,Guennec JY,Masseyeff R

    更新日期:1983-05-27 00:00:00

  • Ultrasensitive assays for endogenous antimicrobial polypeptides.

    abstract::We developed two sensitive methods for identifying antimicrobial molecules in leukocytes and other tissues. One method uses a gel overlay technique and was designed to identify antimicrobial polypeptides in samples subjected to polyacrylamide gel electrophoresis. The other, a radial diffusion assay, allows multiple fr...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90021-7

    authors: Lehrer RI,Rosenman M,Harwig SS,Jackson R,Eisenhauer P

    更新日期:1991-03-21 00:00:00

  • Long-term direct visualization of passively transferred fluorophore-conjugated antibodies.

    abstract::The use of therapeutic antibodies, delivered by intravenous (IV) instillation, is a rapidly expanding area of biomedical treatment for a variety of conditions. However, little is known about how the antibodies are anatomically distributed following infusion and the underlying mechanism mediating therapeutic antibody d...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2017.07.009

    authors: Schneider JR,Carias AM,Bastian AR,Cianci GC,Kiser PF,Veazey RS,Hope TJ

    更新日期:2017-11-01 00:00:00

  • Centrifugation of normal and rheumatoid arthritis blood on Ficoll-Hypaque and Ficoll-Nycodenz solutions.

    abstract::Attempts to use the rapid single-step Ficoll-Hypaque centrifugation procedure for the purification of mononuclear and polymorphonuclear leucocytes from the blood of normal individuals and rheumatoid arthritis patients have sometimes been unsuccessful, largely because the erythrocytes would not sediment through the cen...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90416-2

    authors: McCarthy DA,Perry JD,Holburn CM,Kirk AP,James DW,Moore SR,Holborow EJ

    更新日期:1984-10-26 00:00:00

  • An enzyme-linked immunosorbent assay (ELISA) for quantification of human collectin 11 (CL-11, CL-K1).

    abstract::Collectin 11 (CL-11), also referred to as collectin kidney 1 (CL-K1), is a pattern recognition molecule that belongs to the collectin group of proteins involved in innate immunity. It interacts with glycoconjugates on pathogen surfaces and has been found in complex with mannose-binding lectin-associated serine proteas...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2011.10.010

    authors: Selman L,Henriksen ML,Brandt J,Palarasah Y,Waters A,Beales PL,Holmskov U,Jørgensen TJ,Nielsen C,Skjodt K,Hansen S

    更新日期:2012-01-31 00:00:00

  • An impedance-based cytotoxicity assay for real-time and label-free assessment of T-cell-mediated killing of adherent cells.

    abstract::The in vitro assessment of T-cell-mediated cytotoxicity plays an important and increasingly relevant role both in preclinical target evaluation and during immunomonitoring to accompany clinical trials employing targeted immunotherapies. For a long time, the gold standard for this purpose has been the chromium release ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2014.01.012

    authors: Peper JK,Schuster H,Löffler MW,Schmid-Horch B,Rammensee HG,Stevanović S

    更新日期:2014-03-01 00:00:00

  • Efficient eukaryotic expression of fluorescent scFv fusion proteins directed against CD antigens for FACS applications.

    abstract::Two sets of expression vectors were constructed that permitted the efficient expression of single-chain Fv fragments (scFvs) fused N-terminally to an enhanced mutant of the green fluorescent protein GFP+ or the red fluorescent protein DsRed in insect and mammalian cells. The vectors allowed rapid cloning of scFv fragm...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2003.12.001

    authors: Peipp M,Saul D,Barbin K,Bruenke J,Zunino SJ,Niederweis M,Fey GH

    更新日期:2004-02-15 00:00:00

  • A reliable method for reconstituting thymectomized, lethally irradiated guinea pigs with bone marrow cells.

    abstract::We developed a reliable method for reconstituting thymectomized, lethally irradiated guinea pigs. Injection of 2.5-10 X 10(7) syngeneic bone marrow cells into adult thymectomized, lethally irradiated guinea pigs produced survival of 46-100% of treated animals. Gentamycin sulfate (5 mg/kg of body weight) for 10 days wa...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(84)90187-x

    authors: Terata N,Tanio Y,Zbar B

    更新日期:1984-05-25 00:00:00

  • Differential binding properties of protein A and protein G for dog immunoglobulins.

    abstract::We studied the binding of dog immunoglobulins G, A, M and E to protein A and protein G. Passive cutaneous anaphylaxis (PCA) testing was used for the measurement of dog IgE and enzyme-linked immunosorbent assays (ELISA) were used for the measurements of dog IgG, IgA and IgM. Protein A from lyophilized cells of Staphylo...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(91)90335-d

    authors: Peng ZK,Simons FE,Becker AB

    更新日期:1991-12-15 00:00:00

  • Retinoblastoma protein monoclonal antibodies with novel characteristics.

    abstract::We have developed a family of monoclonal antibodies directed against the retinoblastoma gene product (p110RB). One of these monoclonal antibodies, 3C8, binds p110RB near the C-terminal end of the protein (aa886-aa905). It was characterized by immunoblotting, ELISA, fluorescence-activated flow cytometry and immunohisto...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(94)90267-4

    authors: Wen SF,Nodelman M,Nared-Hood K,Duncan J,Geradts J,Shepard HM

    更新日期:1994-03-10 00:00:00

  • Development of a custom pentaplex sandwich immunoassay using Protein-G coupled beads for the Luminex® xMAP® platform.

    abstract::Multiplex bead-based assays have many advantages over ELISA, particularly for the analyses of large quantities of samples and/or precious samples of limited volume. Although many commercial arrays covering multitudes of biologically significant analytes are available, occasionally the development of custom arrays is n...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/j.jim.2016.02.018

    authors: McDonald JU,Ekeruche-Makinde J,Ho MM,Tregoning JS,Ashiru O

    更新日期:2016-06-01 00:00:00

  • A raft technique for chemotaxis: a versatile method suitable for clinical studies.

    abstract::We describe a raft technique for the study of chemotaxis which possesses a number of advantages over the use of chambers. Membranes are laid on pads soaked in chemotactic agent and cells are contained in plastic cups, which are inverted on to the membrane. This method gives results which are similar to those obtained ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(76)90033-8

    authors: Addison IE,Babbage JW

    更新日期:1976-01-01 00:00:00

  • A method for the production of cryopreserved aliquots of antigen-preloaded, mature dendritic cells ready for clinical use.

    abstract::Dendritic cells (DC) are increasingly used as a vaccine. Unfortunately, a satisfactory cryopreservation of DC in the absence of FCS is not yet available, so that laborious repeated generation of DC from fresh blood or frozen peripheral blood mononuclear cells for each vaccination has been required to date. We now aime...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/s0022-1759(00)00269-6

    authors: Feuerstein B,Berger TG,Maczek C,Röder C,Schreiner D,Hirsch U,Haendle I,Leisgang W,Glaser A,Kuss O,Diepgen TL,Schuler G,Schuler-Thurner B

    更新日期:2000-11-01 00:00:00

  • The immunochemistry of sandwich-ELISAs. IV. The antigen capture capacity of antibody covalently attached to bromoacetyl surface-functionalized polystyrene.

    abstract::The antigen capture capacity of antibodies covalently immobilized on injection-molded polystyrene beads was evaluated. Bromoacetyl groups on the bead surfaces rendered them reactive to protein nucleophilic groups. The bromoacetyl surface exhibited up to a ten-fold greater capacity for protein compared to unmodified po...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(88)90136-6

    authors: Peterman JH,Tarcha PJ,Chu VP,Butler JE

    更新日期:1988-07-22 00:00:00

  • A novel method for the purification of sheep red cell rosetting lymphocytes.

    abstract::A rapid, direct method for the purification of sheep red cell rosetting lymphocytes (ERFC) was developed. The whole procedure, including rosette formation, density separation and hemolysis could be completed within 10 min. A mixture of human peripheral blood mononuclear cells (PBMC) and 2-aminoethylisothiouronium brom...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(88)90217-7

    authors: Mikamo S

    更新日期:1988-03-16 00:00:00

  • Quantitation of IgG antibody to Streptococcus pneumoniae vaccine by ELISA and FAST-ELISA using tyraminated antigen.

    abstract::We examined two ELISA methods for measuring antibodies to Streptococcus pneumoniae using tyraminated S. pneumoniae polysaccharide types 3, 7N, 9F and 14 as antigens. The ELISA has the usual format with a relatively long incubation time whereas the FAST-ELISA has a short incubation time and employs a different solid-ph...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(89)90238-x

    authors: Boctor FN,Barka NE,Agopian MS

    更新日期:1989-06-21 00:00:00

  • An anti-human t-lymphocyte antiserum. Further characterization of the specificity for T-cell subpopulations and a comparison of methods for identification of T-lymphocytes in tissue sections.

    abstract::An anti-human T-lymphocyte antiserum has been further studied for specificity for T-cell subpopulations and application in tissue sections. Using a complement-dependent microcytotoxicity assay about 60% of normal peripheral blood T-cells were found to be sensitive to lysis, while 40% were resistant. When the T-cell su...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(80)90149-0

    authors: Braathen LR

    更新日期:1980-01-01 00:00:00

  • Use of the periodate oxidation coupling method for the detection of antibody and antibody-producing cells specific for staphylococcal lipoteichoic acid.

    abstract::The periodate oxidation and chromium chloride coupling methods were compared for their ability to sensitize indicator erythrocytes with staphylococcal lipoteichoic acid (LTA) for the detection of specific antibody. Erythrocytes, sensitized with periodiate-activated lipoteichoic acid, were found to be superior for use ...

    journal_title:Journal of immunological methods

    pub_type: 杂志文章

    doi:10.1016/0022-1759(80)90070-8

    authors: Beining PR,Flannery GM,Caldes G,Prescott B,Baker PJ

    更新日期:1980-01-01 00:00:00