Abstract：
:Multivalent antibody fragments (scFv dimers, trimers and tetramers) provide high avidity and ideal pharmacokinetics for tumour targeting applications. This protocol describes our optimised protocol for high-level bacterial synthesis of soluble antibody scFv fragments, as either monomers or multimers, using the heat-inducible bacterial expression vector pPOW3. Our protocol is rapid, which minimizes protein degradation, and utilises inexpensive reagents for cost-effective product synthesis. The strong, temperature-regulated promoters in pPOW3 provide efficient production of either monomeric or multimeric single-chain antibody fragments as dictated by the gene construct design.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Power BE,Hudson PJdoi
10.1016/s0022-1759(00)00201-5keywords:
subject
Has Abstractpub_date
2000-08-28 00:00:00pages
193-204issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(00)00201-5journal_volume
242pub_type
杂志文章abstract：:Transplantation of ascitic P-815 mastocytoma cells intradermally in the ears of syngeneic DBA/2J mice allows a simple and precise quantitation of tumor growth by ear swelling. Tumor growth was retarded and then arrested in allogeneic DBA/1J mice, whereas syngeneic hosts died as a result of tumor dissemination in drain...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90086-2
更新日期：1982-11-26 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) has been developed for the quantitation of part-per-million levels of the most probable E. coli polypeptide (ECP) contaminants of E. coli produced biosynthetic human growth hormone (hGH). The antibody preparation, used for both coat and conjugate in this ELISA, was demonstr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90481-3
更新日期：1986-07-24 00:00:00
abstract：:In the guinea pig system, there are no T lymphocyte tumors available. In addition, in this species the production of T cell lines in vitro for analysis of T cell markers or function proved to be difficult. As an alternative approach, guinea pig X mouse and guinea pig X rat T cell hybridomas were obtained by fusion of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90003-3
更新日期：1989-03-31 00:00:00
abstract：:As interest in the development of oral vaccines continues to rise, alternative animal models for studies of mucosal immunity are needed. The present study examines a simplified procedure for delivering antigen to rabbit Peyer's patches via an indwelling cannula. The cannula was placed 3-4 cm proximal to the Peyer's pa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00055-5
更新日期：1996-07-17 00:00:00
abstract：:The unique features of protein recognition of membrane-anchored glycolipids were investigated by surface plasmon resonance (SPR) monitoring of antibody interactions with glycolipids contained in liposomes. Several positive hybridomas belonging to the IgM and IgG classes were identified when tested for binding to the g...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00012-x
更新日期：1998-03-01 00:00:00
abstract：:Efficient monitoring of HIV-1-specific T-cells is crucial for the development of HIV-1 vaccines and immunotherapies. Currently, mainly peptides and vaccinia vectors are used for detection of HIV-1-specific cytotoxic T-lymphocytes (CTL), however, as HIV-1 is a variable virus, it is unknown to what extent the T-cell res...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.03.005
更新日期：2012-06-29 00:00:00
abstract：:Investigation of MHC class I endogenous antigen processing and presentation is important for understanding non-self immune recognition, vaccine design, immune evasion by pathogens and tumours, and elucidation of the mechanisms of certain auto-immune diseases. To study the MHC class I antigen-processing pathway, the pr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.02.017
更新日期：2008-06-01 00:00:00
abstract：:This article describes a novel method for detecting anti-endothelial cell antibodies (AECAs). Sera from patients with systemic vasculitis or inflammatory conditions have been reported to contain antibodies (Abs) that bind to endothelial cells (EC), i.e., AECAs. AECAs are known to play immunogenic effects by triggering...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.05.002
更新日期：2012-08-31 00:00:00
abstract：:A simple method to obtain rat hybridomas producing specific IgA antibodies is reported. By fusing the IR983F rat myeloma cell line with mesenteric lymph node cells from LOU/C rats immunized via the Peyer's patches with DNP-Salmonella typhimurium, twenty hybrids secreting monoclonal IgA antibodies specific for DNP were...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90034-7
更新日期：1986-05-01 00:00:00
abstract：:We report a simple procedure for the maintenance and retrieval of rodent peritoneal macrophages on dialysis membrane supports. Culture units are easily constructed from plastic petri dishes and dialysis membranes. Macrophages adhere readily but are detached easily from the dialysis membrane by mild mechanical agitatio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90322-2
更新日期：1982-11-12 00:00:00
abstract：:The colorimetric MTT assay based on the selective ability of living cells to reduce 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide into formazan was adapted for measuring antibody independent monocyte mediated cytotoxicity. In view of the suggested application of adoptive immunotherapy we studied the an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90205-t
更新日期：1991-07-26 00:00:00
abstract：:We describe the setting up and validation of a reporter gene assay for type I IFN based on monkey Vero cells transfected with pMx-Luc, a plasmid carrying the luciferase gene under the control of the type I IFN inducible mouse Mx1 promoter. Vero cells were stably transfected with pMx-Luc and clone 3-143/5 was selected ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00168-8
更新日期：1996-11-29 00:00:00
abstract：:In the context of allogeneic bone marrow transplantation, an accurate estimate of the risk of developing graft-versus-host disease (GVHD) is of major interest. The pre-transplant frequency of donor's helper T-lymphocyte precursors (HTLp) directed against host's antigens may be helpful in predicting this risk. This tec...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00065-9
更新日期：1998-06-01 00:00:00
abstract：:A new and simple method for quantitation of IgM of low molecular weight in sera by radial immunodiffusion in 7% agar gel is described. The test is easy to perform and permits exact determination of low molecular weight IgM concentration down to very small quantities (1 mg%). Results obtained with the method described ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90094-7
更新日期：1978-01-01 00:00:00
abstract：:A radioimmunoassay (RIA) for antibodies to the Ro/SSA particle is described using Iodogen to radiolabel the antigenic protein moiety of Ro/SSA with 125I. RIA methods utilizing either 33% saturated ammonium sulfate (NH4)2SO4 or 3.5% polyethylene glycol to separate bound from free antigen are comparable. Either method i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90240-v
更新日期：1990-07-20 00:00:00
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
更新日期：2007-07-31 00:00:00
abstract：:Eosinophils can be isolated from a mixed suspension of granulocytes by different procedures. We compared functional responses of human eosinophils purified according to two different principles: (1) an fMLP-induced difference in specific gravity between eosinophils and neutrophils and (2) selective removal of neutroph...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)00255-u
更新日期：1995-01-27 00:00:00
abstract：:This paper examines the parameters necessary for the efficient measurement of anti-Theiler's murine encephalomyelitis virus (TMEV) antibodies in an affinity-dependent manner using a variation of a solid-phase particle concentration fluorescence immunoassay (PCFIA). By allowing antibody to react with fluorochrome-label...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90384-0
更新日期：1989-04-21 00:00:00
abstract：:The single-chain Fv fragment (scFv) is the most frequently used form of recombinant antibody. It is possible to establish clones specific to a certain target by displaying the scFv library on phages followed by biopanning against the target. For the construction of superior scFv libraries, the light-chain variable reg...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.07.003
更新日期：2013-10-31 00:00:00
abstract：:A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modifica...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90332-6
更新日期：1984-02-10 00:00:00
abstract：:A method for synthesising omega-stearoyldextran for the detection of plaque forming cells specific for the alpha-(1 replaces 6) determinant is described. The omega-stearoyl derivative is obtained by reacting dextran with phenylboronic acid to give a 2,4-phenylboronate, which can be subsequently esterified in a pyridin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90206-4
更新日期：1976-01-01 00:00:00
abstract：:Among the many methods currently available for quantifying mRNA transcript abundance, reverse transcription-polymerase chain reaction (RT-PCR) has proved to be the most sensitive. Recently, several protocols for real-time relative RT-PCR using the reporter dye SYBR Green I have appeared in the literature. In these met...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00103-0
更新日期：2003-12-01 00:00:00
abstract：:The method reported here describes a microrosetting assay which allows the early detection of antibody-synthesizing hybridomas. This method is specific, requires very few hybridoma cells and avoids artifacts resulting from the presence of antibodies secreted by spleen cells in the primary hybridoma cultures. It is eas...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90101-7
更新日期：1979-01-01 00:00:00
abstract：:A method is presented for measuring the hemolytic activity of undiluted guinea pig serum. Undiluted serum is placed in contact with a column of sensitized sheep red blood cells in an agar matrix in a capillary tube. After incubation, the length of the zone of lysis is measured. Comparison with a standard curve prepare...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90212-6
更新日期：1979-01-01 00:00:00
abstract：:Natural CD4(+)CD25(+)Foxp3(+) regulatory T cells (Tregs) control the activation of the immune system and therefore have become a major area of research in immunology. The generation of monoclonal antibodies against human Tregs offers the possibility to discover novel Treg-specific or Treg-associated surface markers an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.01.002
更新日期：2010-02-28 00:00:00
abstract：:A method of data analysis for flow cytometry is presented which enables up to eight-dimensional data to be handled by a microcomputer with 28K addressable plus a further 32K non-addressable memory. The multi-parameter coordinates are coded into single numbers using a minimal modification of the array vector mapping eq...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90333-x
更新日期：1988-10-26 00:00:00
abstract：:A microscale ELISA system, designated as the 'C1q-micro ELISA,' has been developed for the measurement of circulating immune complexes. The sensitivity of the method ranged between 0.1 and 3 microgram equivalents of aggregated human gamma-globulins. The procedure requires small quantities of serum and reagents, and it...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90308-8
更新日期：1982-01-01 00:00:00
abstract：:Over the past 10 years an immunoperoxidase method using dinitrophenyl (DNP) hapten-labelled primary or secondary probes has been devised. Its widely successful application in research and diagnostic work has depended upon the development of certain key reagents. These include a novel non-deleterious DNP labelling comp...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(92)90078-8
更新日期：1992-06-24 00:00:00
abstract：:Hapten (trinitrophenyl) was coupled to antigen (ovalbumin). The haptenated antigen was bound by anti-ovalbumin antibody and binding was quantitated with 125I-labelled anti-hapten antibodies. Thus, with a single radioactive reagent antibodies against a variety of antigens can be detected, whilst the problems inherent i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90011-x
更新日期：1978-01-01 00:00:00
abstract：:A new method for measuring the uptake of Staphylococcus aureus by human polymorphonuclear neutrophils (PMN) using flow cytometry (FCM) is described. Bacteria were labeled with fluorescein isothiocyanate (FITC) and incubated with PMN in a suspension assay. At the end of the assay, phagocytosis was arrested by the addit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90161-0
更新日期：1989-07-26 00:00:00
