Abstract:
:Two highly sensitive immunoassays for measuring soluble DNA are described. These methods employ a purified mouse monoclonal anti-DNA antibody in enzyme-linked immunoassays. One assay is an antigen capture method utilizing immobilized antibody. Bound DNA is subsequently detected with biotinylated monoclonal anti-DNA antibody, avidin-coupled horseradish peroxidase and a chromogenic substrate. The limit of detection of the assay was 2 ng/ml of DNA. A competition assay relying on immobilized heat-denatured DNA was also designed. In this assay the solution to be analyzed is mixed in equal proportions with monoclonal anti-DNA antibody and the mixture is incubated in the DNA-containing microtiter plates. The inhibition of antibody binding as detected with peroxidase-conjugated anti-mouse IgG was proportional to the concentration of DNA in the sample. The competition assay was tested for its ability to detect DNA in human plasma. The detection limit of DNA in plasma was approximately 150 ng/ml. The assays were compared in their ability to detect various size fragments of DNA. The competition assay was capable of detecting DNA fragments as small as 30 base pairs. In contrast, the capture assay failed to detect low molecular weight fragments up to 150 base pairs although its sensitivity for undigested DNA was comparable to the competition assay. The assay may be of use in the rapid quantitation of low levels of DNA, especially low molecular weight DNA and may also be useful in measuring DNA in human plasma.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Du Clos TW,Rubin RL,Tan EMdoi
10.1016/0022-1759(86)90005-0subject
Has Abstractpub_date
1986-04-17 00:00:00pages
185-92issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(86)90005-0journal_volume
88pub_type
杂志文章abstract::We describe here a spot test to detect antibodies to non-erythroid cell surface antigens, using ethidium bromide staining to visualize areas of target cell lysis. This assay allows rapid screening of large numbers of samples of culture supernatants, antisera, or ascitic fluids. Antibody titers determined by this spot ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90367-2
更新日期:1982-01-01 00:00:00
abstract:OBJECTIVE:Interleukin-13 (IL-13) has been difficult to quantify within human serum due to low abundance. Available assays have not been sensitive enough to detect IL-13 at the femtogram level. Thus, there are inconsistencies within the published literature as to IL-13 concentrations in normal or disease populations. To...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.08.006
更新日期:2015-11-01 00:00:00
abstract::A flow cytometric method for enumeration of intracytoplasmic antibody-containing cells was developed. Flow cytometry was found to yield results comparable to traditional fluorescence microscopy in cells stimulated to produce intracytoplasmic antibody by either pokeweed mitogen or Epstein-Barr virus. This technique off...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90083-8
更新日期:1984-02-24 00:00:00
abstract::Concanavalin A-activated murine suppressor T cells act via the mediator, soluble immune response suppressor (SIRS) which non-specifically suppresses IgM and IgG antibody responses to a variety of antigens, cytotoxic T lymphocyte responses and proliferative responses to alloantigens and mitogens in vitro. SIRS is a pro...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90235-6
更新日期:1982-08-27 00:00:00
abstract::Feasibility, amount of sample aliquots, processing time and cost are critical considerations for optimizing and conducting assays for large-population based studies. Well designed statistical approaches that quickly identify optimal conditions for a given assay could assist efficient completion of the laboratory assay...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.11.001
更新日期:2009-02-28 00:00:00
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 dilu...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90301-b
更新日期:1990-06-12 00:00:00
abstract::The development of monoclonal antibodies to human differentiation antigens and cytofluorographs have added a new dimension to immune monitoring. However, the technology is such that regionalization will most likely occur. We have demonstrated that human lymphocyte antigens OKT3, OKT4, OKT8 and OKT11 are stable in bloo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90179-5
更新日期:1983-09-16 00:00:00
abstract::The concentrations of major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO) have been associated with eosinophilic disease severity. Whereas a variety of techniques have been used to measure individual eosinophil granule protein concentration,...
journal_title:Journal of immunological methods
pub_type: 临床试验,杂志文章
doi:10.1016/j.jim.2014.05.020
更新日期:2014-09-01 00:00:00
abstract::A simple method to study 19S anti-SRBC antibody production is described. This technique, performed by incubation of immune spleen cells, SRBC and C in the fluid phase, utilises photomeric estimation of antibody mediated haemoglobin release. The test is quicker and more accurate than the two-step haemolytic plaque assa...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90143-0
更新日期:1977-01-01 00:00:00
abstract::In analyses of antigen-specific immune responses, it is essential to estimate the frequency of individual T cell clonotypes. This frequency has been estimated, however, only indirectly by the frequency of T cell receptor (TCR) mRNA. We have developed a method to determine T cell frequency directly by cell count using ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00022-8
更新日期:1999-04-22 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
abstract::Neuraminidase-induced removal of sialic acid from natural substrates (desialylation) unmasks saccharides that are specifically recognized by the lectin peanut agglutinin (PNA). We demonstrate that, when a neuraminidase substrate is coated on to the wells of a microplate, it is possible to quantitate the binding of PNA...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90255-t
更新日期:1990-12-31 00:00:00
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
更新日期:1980-01-01 00:00:00
abstract::Specific rabbit and goat antibodies to tobacco mosaic virus, bovine serum albumin and human gamma globulin have been isolated by acid dissociation from the antigen in the absence of salt. Maximum recovery of 96-100% of active antibody occurred at pH 2.8-2.9. The amount of antibody recovered within any given pH range w...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90092-8
更新日期:1977-01-01 00:00:00
abstract::Liposomes have been widely used as carriers of protein or peptide antigens. Antigenic materials can be attached to the outer surface, encapsulated within the internal aqueous spaces, or reconstituted within the lipid bilayers of the liposomes. The natural tendency of liposomes to interact with macrophages has served a...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(91)90120-5
更新日期:1991-06-24 00:00:00
abstract::Leucocyte procoagulant activity (PCA) has previously been advocated as a useful measurement of cell-mediated immune reactivity. The assay is, however, susceptible to inter- and intra-experimental variation. This investigation identifies several factors which influence the stability and reproducibility of the test. Maj...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90151-6
更新日期:1985-12-27 00:00:00
abstract::A method that permits high energy beta-irradiation to be applied to individual lymphocyte microplate cultures is described. The principle involves the introduction of single energy sources (in this case 32P) contained within tubes which rest inside wells containing the lymphocyte cultures. Cell viability and mitogen-i...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90205-9
更新日期:1984-06-08 00:00:00
abstract::An enzyme immunoassay for the detection of IgG and IgM anti-polymerized albumin autoantibodies (AAA) is described. It was found that polyalbumin receptors on HBsAg particles interfere in the detection of IgG AAA when polymerized human albumin (pHSA), but not polymerized bovine albumin (pBSA), is used as coating antige...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90484-3
更新日期:1985-01-21 00:00:00
abstract::Nonspecific effects of reagents used to selectively isolate lymphocyte subpopulations are most often analyzed according to their influence on cell viability and proliferation. While characterizing monoclonal anti-human B cell antibodies, we detected consistent changes in B cell function between individual donors, no m...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90352-1
更新日期:1984-02-10 00:00:00
abstract::A method for coating native, non-derivatized, polysaccharide (PS) onto nitrocellulose (NC) for identifying PS-specific antibodies has been developed. The new feature of this method is that PS molecules are vacuum filtered onto NC in their native state by devices that can accommodate NC of different sizes and shapes. P...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90032-b
更新日期:1991-03-21 00:00:00
abstract::The reverse haemolytic plaque assay has been adapted to detect and measure the release of such cytokines as interleukin-1, -2 and -6, GM colony-stimulating factor or interferon-gamma by individual human cells derived from either peripheral blood or enzymatically dispersed breast carcinomas. Since each of these peptide...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90340-2
更新日期:1990-02-20 00:00:00
abstract::Although serum-free media have been used to expand lymphokine-activated killer cells, antigen-specific CD8 T cell cytotoxicity does not develop in vitro in the absence of serum. The immunodominant Vbeta17 response to an influenza A matrix protein epitope restricted by HLA A2.1 was used to study the serum requirement f...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00026-5
更新日期:1999-05-27 00:00:00
abstract::Rapid development in design and production of recombinant antibodies and antibody fragments specific for cell surface markers opens new opportunities for targeted delivery of therapeutic or imaging agents. However, the progress in this field is slowed by inactivation of many antibodies by chemical conjugation of paylo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.03.011
更新日期:2004-06-01 00:00:00
abstract:BACKGROUND:High concentrations of propidium iodide (PI), in combination with fluorescein isothiocyanate (FITC) and R-phycoerythrin (RPE) used for multiparameter DNA flow cytometry (FCM), cause spectral cross-talk into the green fluorescence channel (FL1). We have evaluated the use of post-acquisition software compensat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00550-6
更新日期:2002-02-01 00:00:00
abstract::We have developed a standardized procedure for the isolation of monocytes from peripheral blood by negative selection using magnetic polymer particles coated with monoclonal antibodies against T and B lymphocytes. The average purity of the monocyte suspension was 85%, and monocyte recovery was 72% from Ficoll-Hypaque ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90397-x
更新日期:1991-03-01 00:00:00
abstract::We describe a stable chelating linkage for the reversible immobilization of oligohistidine tagged proteins in the flow cell of the 'BIAcore' surface plasmon resonance (SPR) biosensor. The carboxymethylated dextran surface of the flow cell was covalently derivatized with N-(5-amino-1-carboxypentyl)iminodiacetic acid (N...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00032-6
更新日期:1995-06-14 00:00:00
abstract::An enzyme-linked direct antiglobulin test (DAGT) for assessing erythrocyte-bound IgG, IgM and IgA is described. The test is carried out in microtitre plates using heavy chain-specific, alkaline phosphatase-linked, goat anti-human globulin reagents with p-nitrophenyl phosphate as substrate. Results are expressed in opt...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90268-2
更新日期:1988-01-21 00:00:00
abstract::Bovine serum albumin (BSA) was used as a non-amphipathic model protein to be included into iscoms. Pretreatment at an acidic pH (2.5) was used to reveal hydrophobic regions, after which BSA could be integrated. In immunization experiments in mice the BSA iscoms induced long lasting and considerably higher serum antibo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90208-d
更新日期:1990-04-17 00:00:00
abstract::A double-sandwich ELISA, developed for detection of IgM antibodies to the major surface protein of Toxoplasma gondii (P30), is proposed for the diagnosis of acute acquired toxoplasmosis. The method is based on the capture of serum IgM antibodies, which are revealed indirectly by the sequential addition of a Toxoplasma...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90068-7
更新日期:1985-10-24 00:00:00
abstract::A radioimmunoassay is described that allows rapid determination of a urinary oligosaccharide -- Glc alpha 1-6Glc alpha 1-4Glc alpha 1-4Glc [(Glc)4] -- at concentrations greater than 2 pmol/microliter. Antibodies produced in rabbits immunized with the phenethylamine derivative of (Glc)4 coupled to keyhole limpet hemocy...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90215-0
更新日期:1982-01-01 00:00:00