Abstract:
:A new method based on flow cytometry has been developed to determine IgG alloantibodies in the serum of immunized rodents. The method utilizes target lymphoid cells, diluted serum and labeled anti-mouse or anti-rat IgG antibodies. In serum from highly immunized animals alloantibodies could be demonstrated up to a dilution of (3 x 10(4))-1 which makes the test approximately 300 times more sensitive than a simultaneously performed complement-dependent cytotoxicity assay (CDCA). Moreover a linear relationship between the amount of alloantibody and the log value of the mean fluorescence of the target cells was found. This linearity permits the comparison of alloantibody production between individual samples by comparing the mean fluorescence values. The reproducibility of the assay was excellent since the coefficients of variation for all dilutions were less than 15%. By using two-color fluorescence the method can discriminate alloantibodies directed against class I and class II MHC antigens. Finally, in addition to its high sensitivity and good reproducibility, the method was found to be at least twice as fast as CDCA.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Leenaerts PL,De Ruysscher D,Vandeputte M,Waer Mdoi
10.1016/0022-1759(90)90301-bsubject
Has Abstractpub_date
1990-06-12 00:00:00pages
73-9issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(90)90301-Bjournal_volume
130pub_type
杂志文章abstract::The effects of complement activation on the antigenic component of immune complexes have been studied, using 125I-BSA-rabbit anti-BSA-IgG complexes as models. Polyethylene glycol precipitates of 4 types of IC (those formed in native normal human serum, or NHS-containing EDTA, NHS-EDTA, and preformed soluble IC incubat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90109-1
更新日期:1984-12-31 00:00:00
abstract::The ubiquitin-proteasome pathway plays major roles in all aspects of biology and contributes to various disease processes. Due to the lack of assays that permit proteasome quantification in crude cell extracts, its concentrations in health and disease states as well as the relationship between free 20S core particles ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.02.004
更新日期:2008-05-20 00:00:00
abstract::The aim of the present study was to analyze the relations between the serum anti-erythropoietin antibody (AEAb) levels and the antibodies' neutralizing activity in 20 patients with renal anemia and rhEPO-induced antibodies. AEAb levels were determined by the enzyme-linked immunosorbent assay (ELISA, double antigen-bri...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.04.009
更新日期:2008-07-31 00:00:00
abstract::The technology of the mixed antiglobulin rosetting (MARR) and the direct antiglobulin rosetting reaction (DARR) is reviewed. These tests have been found to be more sensitive than the standard direct immunofluorescence (DIF) method for delineation of Ig-bearing B lymphocytes and to have practical advantages over DIF. F...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(81)90268-4
更新日期:1981-01-01 00:00:00
abstract::Human tumor infiltrating lymphocytes (TIL) from metastatic melanoma of six patients were grown using a new hollow fiber bioreactor system. After inoculating 0.35-10 X 10(8) TIL into the extra-fiber space (EFS), each Cellmax bioreactor was perfused with AIM-V medium, supplemented with rIL-2. The cells subsequently expa...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90337-u
更新日期:1990-02-20 00:00:00
abstract::Tetramolecular monoclonal antibody complexes were used to selectively cross-link a subset of human peripheral blood T cells (CD8 positive) to glass beads coated with hapten (fluorescein) modified bovine serum albumin. Tetramolecular antibody complexes were prepared with anti-CD8 (Leu 2a), anti-FITC mouse IgG1 monoclon...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90361-4
更新日期:1988-09-13 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 fluorimetric method using 4-methylumbelliferyl heptanoate (MUH) has been developed for detecting cell-mediated cytotoxicity and cell proliferation. The assay is based on the hydrolysis of the fluorochrome (MUH) by intracellular esterases of viable cells resulting in the production of highly fluorescent 4-methylumbel...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00115-q
更新日期:1995-09-25 00:00:00
abstract::An assay for detection of monoclonal hybridoma antibodies against cell surface antigens is described. Samples of spent medium from the hybridoma cultures are incubated in microtest wells with cells, either as adherent monolayers or in suspension. Antibodies bound to surface antigens are detected by successive incubati...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90132-7
更新日期:1979-01-01 00:00:00
abstract::A standardized, reproducible two-step method for separation of human peripheral blood monocytes on continuous Percoll gradients has been developed. The first step involves separation of mononuclear cell on Percoll of density 1.075 g/ml and the second step separation of monocytes from lymphocytes on a continuous Percol...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90194-6
更新日期:1982-01-01 00:00:00
abstract::A simple and reproducible technique for murine platelet isolation is described. Such preparations then become useful in determining in vitro platelet secretory responses and platelet aggregation. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90226-4
更新日期:1980-01-01 00:00:00
abstract::Antibodies directed against elliptinium acetate, a quaternary ammonium compound with antineoplastic activity in man, were obtained in rabbits, after conjugation of the drug with haemocyanin. These antibodies are specific for the quaternary ammonium structure. However, the recognition of the drug can be markedly decrea...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90124-4
更新日期:1987-05-20 00:00:00
abstract:BACKGROUND:The role of CD4(+) regulatory T cells (Tregs) and their subsets during HIV infection is controversial. Cryopreserved peripheral blood mononuclear cells (PBMC) are an important source for assessing number and function of Tregs. However, it is unknown if PBMC isolation and cryopreservation affect the expressio...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2016.02.004
更新日期:2016-04-01 00:00:00
abstract::The Immune Epitope Database and Analysis Resource (IEDB, http://www.iedb.org) hosts a continuously growing set of immune epitope data curated from the literature, as well as data submitted directly by experimental scientists. In addition, the IEDB hosts a collection of prediction tools for both MHC class I and II rest...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/j.jim.2010.10.010
更新日期:2011-11-30 00:00:00
abstract::Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90130-2
更新日期:1977-01-01 00:00:00
abstract::Monoclonal antibodies have been prepared against a synthetic peptide with a sequence corresponding to a repeated hydrophilic region of the protein core of the human MUC-2 gastrointestinal mucin. Peptide conjugates, prepared by glutaraldehyde cross-linking with keyhole limpet haemocyanin (KLH) and bovine serum albumin ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90168-7
更新日期:1993-02-26 00:00:00
abstract::The major isoform of hGH is a polypeptide of 191 amino acids. Human GH1-43 is an amino terminal segment of hGH1-191 which comprises the first 43 amino acids. This peptide is a potent regulator of glucose homeostasis. To facilitate our understanding of the physiological regulation of hGH1-43 an assay to measure its lev...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(98)00086-6
更新日期:1998-06-01 00:00:00
abstract::The purpose of this study was to compare and contrast two enzyme immunoassay systems: the enzyme-linked immunosorbent assay (ELISA), which utilizes polystyrene microtiter plates as the adsorptive surface and the enzyme-linked immunoflow assay (ELIFA), which utilizes nitrocellulose membranes. The principal parameter un...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90152-6
更新日期:1991-08-09 00:00:00
abstract::Detection of anti-drug antibodies (ADA) can be difficult, if not impossible, in the presence of drug in the sample. This is a particular concern with therapeutic monoclonal antibodies (mAbs), which have typically longer half-lives than other proteins. For detection of ADA in presence of high drug concentrations, assay...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.01.010
更新日期:2008-05-20 00:00:00
abstract::A procedure to enhance incorporation of radioactive iodine into cell-surface proteins is described. The effects of chemical modification of cells with the iodinatible haptens N-succinimidyl-3-(4-hydroxyphenyl) propionate (Bolton-Hunter reagent) and methyl-p-hydroxybenzimidate HC1 (Wood reagent) are compared. The Wood ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90177-2
更新日期:1985-02-28 00:00:00
abstract::The circumsporozoite (CS) protein is the major surface protein of Plasmodium sporozoites. Antibodies to the immunodominant repeat domain of CS immobilize sporozoites and prevent infection of hepatocytes. Plasmodium falciparum vaccines containing CS repeats are undergoing human trials in endemic areas, and proof of eff...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.06.017
更新日期:2004-09-01 00:00:00
abstract::Protocols for purification of mouse monoclonal antibodies (MAbs) from nude mice ascites were investigated in order to assess the yield and to compare the purified products in two-dimensional gel electrophoresis (2DGE). Three MAbs (one IgG2 and two IgG1), selected for their differing behaviours towards protein A, were ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90379-0
更新日期:1986-06-10 00:00:00
abstract::A competition radioimmunoassay for human superoxide dismutase (SOD-1) is described. Radioiodinated SOD-1 is incubated with unlabeled competitor and limiting antibody, then immune complexes are isolated using Staphylococcus aureus. The assay is sensitive (detecting 5 ng enzyme) and reproducible. Average values for SOD-...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90313-2
更新日期:1979-01-01 00:00:00
abstract::The identification of naturally processed peptides presented by molecules of the major histocompatibility complex (MHC) has progressed significantly over the past decade. The elution of peptides from immunoaffinity purified complexes of MHC class I or class II molecules has provided highly specific biochemical informa...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/s0022-1759(00)00361-6
更新日期:2001-03-01 00:00:00
abstract::Using the advantages of phenol red, a signal enhancer, and bovine serum albumin (BSA), a stabilizer of horseradish peroxidase (HRP), added in HRP enzyme reaction of Amplex Red and H2O2, highly sensitive 1,1'-oxalyldiimidazole chemiluminescence enzyme immunoassay (ODI-CLEIA) was developed to rapidly quantify trace leve...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2019.05.003
更新日期:2019-08-01 00:00:00
abstract::Six monoclonal and two polyclonal antibodies to fluorescein (FLU) were affinity purified and immobilized on Immulon 2 polystyrene as capture antibodies (CAbs): (a) by passive adsorption at pH 9.6, (b) via a streptavidin bridge to a biotinylated carrier molecule, and (c) via an antiglobulin which had been previously ad...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(92)90066-3
更新日期:1992-06-24 00:00:00
abstract::Various methods have been developed recently for predicting antigenic determinants in a protein by analyzing its hydropathicity, flexibility or secondary structure. The predicted results by these methods were compared directly to the observed structure of the proteins to determine how accurate they were. Regression an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90423-s
更新日期:1990-05-08 00:00:00
abstract::Whilst several centres have reported lymphocytotoxic antibody detection using single and dual fluorescent stains with analysis of the fluorescence emitted from the cell population present in a well of a multiwell plate, problems are encountered with cell concentration and light emission overlap. A method we have devel...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90042-1
更新日期:1987-05-04 00:00:00
abstract::Intracellular DNA triggers interferon release during the innate immune response. Cyclic GMP-AMP synthase (cGAS) senses intracellular double-stranded DNA not only in response to viral infection but also under autoimmune conditions. Measuring the levels of cyclic GMP-AMP (cGAMP) as a second messenger of cGAS activation ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2020.112751
更新日期:2020-05-01 00:00:00
abstract::The aim of this study was to establish a cytokine-free, serum-free system which would enable the long-term survival and proliferation of human peripheral blood monocytes. Monocytes were isolated from peripheral blood mononuclear cells (PBMC) by adherence to untreated plastic petri dishes and maintained up to 6 weeks i...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(92)90323-l
更新日期:1992-08-30 00:00:00