Abstract:
:A sensitive assay for the simultaneous detection of multiple serum antibodies by flow cytometry was developed. Polystyrene microspheres of 5, 7 and 9.3 micron in diameter were used as solid supports for the attachment of three different antigen preparations from Candida albicans. These antigens were a whole cell extract; a cytoplasmic protein extract and a cell wall polysaccharide. Microsphere-associated fluorescence was quantitated by flow cytometry, with the different sized microspheres analyzed separately using electronic volume gating. This procedure allowed for different antigen-coated microspheres with discrete sizes to be analyzed independently for immunofluorescence. The assay detected antibody levels in human serum at dilutions up to 10(-6) and provided complete discrimination, using all three antigen preparations, between antibody levels seen in healthy subjects and those seen in patients suspected of having a systemic Candida infection. A standard enzyme immunoassay (EIA) failed to provide complete discrimination between healthy subjects and patient samples: at least 17% of patient values fell within the healthy subject range using all three antigen preparations. The microsphere assay which allowed for the simultaneous detection of multiple antibodies, has increased dynamic range over EIA and provides for better discrimination of patients from healthy subjects in comparison to EIA. Precise quantitation of antibodies is possible and the rapid analysis of thousands of microspheres markedly enhances the statistical accuracy of the assay. We suggest this assay is likely to have many other important applications in immunologic testing.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
McHugh TM,Wang YJ,Chong HO,Blackwood LL,Stites DPdoi
10.1016/0022-1759(89)90206-8subject
Has Abstractpub_date
1989-01-17 00:00:00pages
213-9issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(89)90206-8journal_volume
116pub_type
杂志文章abstract::Dispase-treated murine hybridoma cells (SP2/0-Ag14) were transfected with the G418 resistance gene bearing plasmid pSV2-neo by electropermeabilization with a high degree of efficiency. The cells were subjected to intermittent multiple high-voltage short duration (5 microsecond) DC pulses at intervals of 1 min in a wea...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90452-8
更新日期:1988-04-22 00:00:00
abstract::Evaluation of human antibody responses to alpha-1 antitrypsin (AAT) in clinical trials and clinical practice has been limited by the lack of a validated assay. Here we describe the development and validation of an ELISA method for quantification of human and nonhuman primate antibody responses to human AAT. A referenc...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.11.008
更新日期:2013-02-28 00:00:00
abstract::IL-21 is a pleiotropic cytokine that plays a key role in modulating inflammatory responses, including the promotion of autoimmune diseases. Several groups have quantitated circulating levels of IL-21 in plasma and serum samples using various commercial ELISAs. We determined, however, that the most commonly used commer...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2018.12.005
更新日期:2019-03-01 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::A novel bioanalysis system based on immunochromatography was developed in connection with a nitrocellulose resin modified micropipette tip, such as ZipTip. The sandwich-type immunoassay was applied to our bioanalysis system. The simple handling of the micropipette enabled us to increase the sample volume and detect lo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2006.02.011
更新日期:2006-05-30 00:00:00
abstract::Pathological prion protein (PrPSc) which is a conformational isoform of a host-encoded protein designated (PrPC) serves as a specific marker protein for the immunochemical diagnosis of transmissible spongiform encephalopathies (TSE). The generation of suitable antibodies to PrPSc therefore underlies the specificity an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00121-x
更新日期:1997-08-22 00:00:00
abstract::By a rosette assay with ox red blood cells (ORBC) sensitised with rabbit IgM and IgG antibodies (EA mu and EA gamma), receptors for IgM (Fc mu-R) and IgG (Fc gamma-R) on the surface of guinea pig lymphocytes in the different lymphoid organs were demonstrated. A double rosette assay with rabbit erythrocytes (RRBC) and ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90385-4
更新日期:1982-01-01 00:00:00
abstract::A new T(H1)/T(H2) in vitro model for mechanistic studies and drug screening in human T cells was established working with ficoll-separated PBMCs or elutriated lymphocytes cultured in serum-free medium. Human T cells could be kept viable and reactive in this medium for several months. In this model, superantigens (SAs)...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(98)00142-2
更新日期:1998-10-01 00:00:00
abstract::Frozen stored human mononuclear cells were compared with non-frozen cells as responder cells in the monocyte chemotaxis assay. Frozen stored cells were found to inhibit lower background migration, resulting in greater experimental-to-control ratios. The use of frozen cells also greatly improved the speed, standardizat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)90023-5
更新日期:1977-01-01 00:00:00
abstract::Human thyroxine binding globulin (TBG) was purified to near homogeneity by chromatography on thyroxine-substituted Sepharose 4B, DEAE-Sephadex A-50, Sephadex G-150 and n-decylamine-substituted Sepharose 4B. TBG was quantitated by homogeneous fluorescence excitation transfer immunoassay; the preparation of dye-protein ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90139-8
更新日期:1980-01-01 00:00:00
abstract::Pathologic rheumatoid factor (RF) levels are hallmarks of several human diseases. Production of monoclonal RF in vitro is essential for studies of the antigenic specificities of RF, as well as for a dissection of the mechanisms of aberrant RF+ B cell activation. We have expanded upon previous methods to develop a flow...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.09.022
更新日期:2011-01-05 00:00:00
abstract::Anti-sperm cell-mediated immunity (CMI) is considered as a crucial facet of infertility in patients of both sexes. A precise and objective method is designed, based on a one-step agarose leukocyte migration inhibition factor (LMIF) assay. The migration areas are evaluated by a computer-assisted image analysis system. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(92)90186-w
更新日期:1992-10-02 00:00:00
abstract::Ten solid-phases were evaluated for their usefulness in a two-site immunoradiometric assay for serum thyroid-stimulating hormone. The criteria used to assess each reagent included the minimum detection limit attainable, the change of binding over the concentration range (signal: noise ratio), and ease of preparation o...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90491-1
更新日期:1987-11-23 00:00:00
abstract::An ultra-sensitive quantitative EPO (erythropoietin) lateral flow immunochromatographic test with a detection limit of 1.2 fM (10(-15) M), 0.035 ng EPO/L, which is 50-100 times more sensitive than a corresponding enzyme based immunoassay, is presented. In comparison with commercial lateral flow tests for other analyte...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2008.09.022
更新日期:2008-12-31 00:00:00
abstract::Antisera used in immunological assay systems for small molecular weight substances are routinely prepared by coupling the hapten to a carrier protein via a chemical linker. Often this bridge is partly recognized by the antibody, resulting in reduced sensitivity when an identically structured tracer (e.g. iodine-labell...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90296-5
更新日期:1984-11-30 00:00:00
abstract::The relative efficiency of depot (Freund's complete adjuvant (FCA) and saponin), and non-depot (Corynebacterium parvum strain P. acnes I, C. parvum strain IMI585, and levamisole) immunoadjuvants were compared in raising agglutinating and precipitating antibodies against the metacestode fluid of Taenia multiceps of she...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90230-2
更新日期:1983-07-29 00:00:00
abstract::We describe a solid-phase fluoroimmunoassay, based on competition between europium-labeled 5 MeCyd (5-methylcytidine) and sample 5MedCyd (5-methyl-2'-deoxycytidine) for polyclonal anti-5MedCyd antibodies (rabbit). Europium labeling of antigen was performed using a novel polylysine-5MeCyd conjugate. Standard and sample...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(89)90115-4
更新日期:1989-02-08 00:00:00
abstract::Recombinant antibody fragments can be produced in large quantities using bacterial expression systems and could potentially be useful for the generation of biofilters for the selective removal of viral particles from fluids. A human single chain-Fv antibody library, derived from synthetic repertoires of germ line VH-g...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00107-5
更新日期:1997-08-07 00:00:00
abstract::A two-stage culture method is described for the induction of a specific antibody response to sheep red cells (SRC) in microcultures at limiting dilutions of human peripheral blood lymphocytes (PBL). PBL from normal donors were cultured for 4 days with antigen and EBV using well defined conditions. The cells were then ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90449-8
更新日期:1988-04-22 00:00:00
abstract::We describe an intracellular cytokine staining, flow cytometry based technique for quantitative and qualitative assessment of CD8+ T lymphocyte-mediated antigen specific immune responses. This adaptation of the technique first described by Suni et al. [J. Immunol. Methods, 212 (1998) 89; Cytometry, 40 (2000) 60] utili...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(02)00025-x
更新日期:2002-04-01 00:00:00
abstract::Here we extensively describe a FACS-based protocol for isolating intact non-stained human eosinophils from peripheral blood; a stop forward from our recently published initial study. This method of purification could be accomplished in <3 h with only small volumes of whole blood necessary, even in healthy subjects gen...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2018.09.003
更新日期:2018-12-01 00:00:00
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
更新日期:1988-03-16 00:00:00
abstract::ADP-ribosylation factors (Arf) are small GTP-binding proteins involved in vesicular transport and the activation of phospholipase D (PLD). The conversion of Arf-GDP to Arf-GTP is promoted in vivo by guanine nucleotide exchange factors such as ARNO or cytohesin-1. In order to examine the expression of ARNO and cytohesi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00336-7
更新日期:2001-03-01 00:00:00
abstract::Covalently cross-linked large and intermediate-sized IgA oligomers, prepared with IgA anti-dinitrophenyl (DNP) and bis-DNP-pimelic acid ester, were used to examine the ability of different concentrations (3.5%, 5% and 7%, w/v) of polyethylene glycol (PEG) to precipitate IgA immune complexes (IgA-IC). The size of the I...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90295-x
更新日期:1987-11-05 00:00:00
abstract::Intracellular probes used for oxidative burst evaluation could be a substrate of the multidrug resistance proteins (MDR), which may cause misinterpretation of experimental data. We aimed to study the effect of the culture condition and of the MDR-interfering antioxidant quercetin on free radical measurement in serum d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2013.12.009
更新日期:2014-03-01 00:00:00
abstract::Five beta-diketone derivatives were studied for multiple labelling of proteins. The labelled proteins were characterized by absorption and fluorescence measurements. It was found that proteins labelled with chlorosulfonyl-thenoyltrifluoroacetone (CTTA) were able to form highly fluorescent complexes with Eu3+ which exh...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)00289-9
更新日期:1995-02-27 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 cell ELISA for human leukocytes has been established and standardized. The assay was based on the use of air-dried and methanol-fixed cells which were attached to microplate wells. Cellular antigens were measured through the subsequent binding of monoclonal antibody (MoAb), biotinylated sheep Ig against mouse IgG an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90219-x
更新日期:1986-11-20 00:00:00
abstract::Historically, tumour markers are substances selectively released by tumour cells in the blood stream so that they can be detected in the serum or other body fluids for clinical monitoring of various malignancies. The term is now extended to cell or tissue characteristics, such as cytogenetic markers, oncogenes or abno...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(92)90072-2
更新日期:1992-06-24 00:00:00
abstract::The techniques of flow cytometry are being increasingly used to classify lymphocytes according to their surface antigens. Accurate analyses depend on the exclusion of non-lymphocytic contaminant cells. Using a fluorescence-activated cell sorter (FACS IV), virtually complete separation of human lymphocytes from monocyt...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90389-7
更新日期:1983-11-11 00:00:00