Abstract：
:Endogenous heterophilic antibodies in blood are known to interfere with two-site enzyme-linked immunosorbent assays (ELISAs) evoking false positive signals. In the present study, we describe an assay for the assessment of components of the plasminogen activation system (uPA, tPA and PAI-1, and their complexes) in blood which is not susceptible to interference by heterophilic antibodies. In the ELISA format, two avian (duck, chicken) antibodies are employed in the pre-analyte and two mammalian (rabbit, goat) antibodies in the post-analyte stage. The assay is compared to our earlier reported ELISA for measuring uPA, tPA and PAI-1 components in tumor tissue extracts. Applying the so-called "nonsense formats", designed against non-existent components, to the NIBSC reference preparation of rheumatoid factor (RF), no response was found with the new assay, whereas a clear RF dose-dependent interfering signal was observed with the original assay designed for tumor tissue extracts. Analysis of tumor-tissue based international reference preparations (RBG EORTC 101094 and 040297), human anti-mouse antibodies (HAMA) containing sera, and sera from patients with rheumatoid arthritis (RA), also displayed no false positive signals. In conclusion, we have developed an ELISA that permits the determination of blood levels of components in the urokinase system, free from disturbance by endogenous heterophilic antibodies.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Grebenchtchikov N,Sweep CG,Geurts-Moespot A,Piffanelli A,Foekens JA,Benraad TJdoi
10.1016/s0022-1759(02)00213-2keywords:
subject
Has Abstractpub_date
2002-10-15 00:00:00pages
219-31issue
2eissn
0022-1759issn
1872-7905pii
S0022175902002132journal_volume
268pub_type
杂志文章abstract：:To date, in vivo apoptosis within the thymus has been assessed using morphological criteria and/or detection of a DNA ladder indicative of oligonucleosomal fragmentation of the DNA. Here, we have used a fluorometric method to investigate activation of the caspase protease family in the thymus following in vivo inducti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00054-x
更新日期：1999-06-24 00:00:00
abstract：:Pig peripheral blood lymphocytes, typed for LD antigens by negative reactions to homozygous cells in primary MLRs and for SD antigens by complement-mediated lymphocytotoxicity, were sensitised in vitro against X-irradiated lymphocytes. These "primed" cells were rechallenged with lymphocytes possessing a range of LD an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90151-5
更新日期：1978-01-01 00:00:00
abstract：:An assay has been developed in which the degree of in vitro spleen cell-mediated sheep red blood cell (SRBC) hemolysis is quantitated spectrophotometrically (QHS). Immune mouse spleen cells, SRBC and guinea pig complement were incubated at 37 degrees C for 1 h in buffer, centrifuged, and the supernatant solutions read...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90232-6
更新日期：1978-01-01 00:00:00
abstract：:Enzyme-linked immunoadsorbent assay (EIA) has widespread use for the measurement of antibody concentration. The affinity constant (Kaff) of the antibody has an effect upon the quantification by EIA. It is thus important to be able to measure Kaff by solid-phase EIA. Based upon the Law of Mass Action and using serial d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90187-6
更新日期：1987-06-26 00:00:00
abstract：:Human interleukin-7 (IL-7) is a cytokine that appears to be critical for early T- and B-cell development and although IL-7 is currently under investigation as a therapeutic agent in a variety of hematolymphopoietic disorders, there have been few instances of the detection or investigation of this cytokine using a biol...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00237-9
更新日期：2000-03-06 00:00:00
abstract：:Since the number of polymorphonuclear leukocytes (PMN) responding to a chemotactic stimulus in the Boyden chamber is influenced by the cell density, we studied whether this variable was important in determining chemotaxis under agarose. The chemotactic index was determined by summing the product of each cell that had ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90292-1
更新日期：1981-01-01 00:00:00
abstract：:A sensitive method for staining proteins after transfer from polyacrylamide gels to nitrocellulose paper is described. Transferred proteins are first derivatized by reaction of the nitrocellulose replica with sulfosuccinimidobiotin and are then reacted sequentially with streptavidin, rabbit anti-streptavidin, and hors...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90504-1
更新日期：1986-08-21 00:00:00
abstract：:Structural point mutations in exon 1 at codons 52, 54 and 57 and a promotor polymorphism at -221 bp of the mannose-binding lectin (MBL) gene are associated with increased susceptibility to various infectious diseases. We developed a genotyping method based on the 5' nuclease (TaqMan) assay in combination with the use ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.01.025
更新日期：2004-04-01 00:00:00
abstract：:The rise in the analytical speed of mutiparameter flow cytometers made possible by the introduction of digital instruments, has brought up the possibility to manage progressively higher number of parameters simultaneously on significantly greater numbers of individual cells. This has led to an exponential increase in ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/j.jim.2019.07.003
更新日期：2019-12-01 00:00:00
abstract：:Leukocyte scintigraphy has been used as a standard diagnostic procedure for the detection of inflammation in vivo. In this study, we developed a method of labelling purified lymphocytes with technetium99m-hexamethyl propyleneamine oxime (Tc99m-HMPAO) without significantly impairing their function. This was confirmed b...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00045-3
更新日期：1998-05-01 00:00:00
abstract：:We have previously described a monoclonal antibody (BU-1) to 5-bromo-2-deoxyuridine (BrdUrd) that is useful for measurement of cell cycle S-phase. BU-1 hybridoma supernatant reacted with incorporated BrdUrd after the cells had been ethanol fixed; without a requirement for acid or base denaturation. We have found that ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90438-2
更新日期：1986-10-23 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：:This investigation describes a new procedure for the preparation of immunogenic liposomes which circumvents the need to synthesize the N-(hapten)-substituted derivatives of phosphatidylethanolamine that were previously employed for this purpose. The method is based on the generation of liposomes containing the N-hydro...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90125-4
更新日期：1983-12-30 00:00:00
abstract：:The neutrophilic granulocyte is the most numerous leukocyte in peripheral blood. The development from a multipotent progenitor cell to a mature neutrophil takes place in the bone marrow over a period of 10-14 days. In order to understand the cellular mechanisms behind this process, it is necessary to investigate cells...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(99)00176-3
更新日期：1999-12-17 00:00:00
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
更新日期：1989-06-21 00:00:00
abstract：:Both the active ester and maleimide moieties of the cross-linking reagent, N-[(gamma-maleimidobutyryl)oxy]succinimide (GMBS), were found to react with the primary amino groups on ribonuclease (RNase). This largely inactivated RNase towards a polymeric (but not monomeric) substrate. Citraconylating the RNase first, so ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90235-n
更新日期：1990-07-20 00:00:00
abstract：:Formation of rosettes with sheep RBC by blood lymphocytes from young pigs was increased from 24.4% +/- 2.2 (mean +/- S.E.) in PBS to 54.1% +/- 1.9 in the presence of dextran. This increase was achieved without inducing appreciable rosette formation with other RBC which do not form rosettes in PBS. Lymphocytes which r...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90146-1
更新日期：1978-01-01 00:00:00
abstract：:Rabbit immunoglobulin G (RIgG) was reduced with dithioerythritol and analyzed by high performance liquid chromatography. A quantitative method for determining the percentage of reduced half-molecules in the mixture was developed. An acetic acid concentration-dependent rate of dissociation of reduced half-molecules was...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90314-0
更新日期：1987-02-11 00:00:00
abstract：:The aim of this study was to develop an in vitro test system for pyrogenic substances. Three clones derived from human monocytoid cell lines, which were selected by their high sensitivity to lipopolysaccharide (LPS), were assessed for tumor necrosis factor (TNF) production. Their response to pyrogen-containing samples...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00112-9
更新日期：1997-09-24 00:00:00
abstract：:Functional analysis of antigen-specific CD8(+) T cells is important for understanding the immune response in various immunological disorders. To analyze CD8(+) T cell responses to a variety of antigens with no readily defined peptides available, we developed a system using CD4(+) phytohemagglutinin (PHA) blasts transd...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.05.009
更新日期：2006-07-31 00:00:00
abstract：:Establishment of humoral immunity against pathogens is dependent on events that occur in the germinal center and the subsequent induction of high-affinity neutralizing antibodies. Quantitative assays that allow monitoring of affinity maturation and duration of antibody responses can provide useful information regardin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.11.026
更新日期：2014-02-01 00:00:00
abstract：:Studies in vitro of human macrophage function in health and disease have been impeded by the difficulty of obtaining such cells in sufficient number. Unlike animal species, the only readily available source of human macrophages are circulating monocytes. Herein, a method is described whereby the phagocytic rate of sma...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90172-1
更新日期：1976-03-01 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
abstract：:External proficiency testing programs designed to evaluate the performance of end-point laboratories involved in vaccine and therapeutic clinical trials form an important part of clinical trial quality assurance. Good clinical laboratory practice (GCLP) guidelines recommend both assay validation and proficiency testin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2014.05.019
更新日期：2014-07-01 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 simple, sensitive and accurate method was developed to determine the level of lymphoid chimerism in bone marrow-transplanted rodents. The method is based on flow cytometry using polyclonal alloantisera and labeled second step anti-IgG antibodies. Using mixtures of spleen cells from different mouse strains, it was de...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90044-v
更新日期：1990-07-03 00:00:00
abstract：:Human peripheral blood or tonsil lymphocytes produce T cell growth factor (TCGF), when activated with neuraminidase (NA) and galactose oxidase (GO). Partial purification of NAGO-TCGF on Sepharose G-100 columns gave a TCGF-active fraction within the same molecular weight range as the conventional lectin-induced TCGF (a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(83)80004-0
更新日期：1983-10-28 00:00:00
abstract：:Antibodies prepared by immunizing rabbits with a synthetic polypeptide representing the first 26 amino acids of the proposed NH2-terminal sequence of human erythropoietin cross-react with peptide, highly purified 125I-labeled erythropoietin and biologically active erythropoietin. We have now developed a rapid and conv...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90316-8
更新日期：1984-04-27 00:00:00
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 extra...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90206-8
更新日期：1989-01-17 00:00:00
abstract：:Early stage rheumatoid arthritis (RA) is often difficult to diagnose because initial symptoms are non-specific. To aid diagnosis, suitable serological diagnostic markers are sought. Elevated levels of soluble MHC class II (soluble human leukocyte antigen; sHLA-DR) in human serum have been associated with rheumatoid an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.06.013
更新日期：2006-08-31 00:00:00
