Abstract：
:Comparing the marker enzymes horseradish peroxidase (HRP), alkaline phosphatase (AP) and beta-galactosidase (beta Gal) in IgG-coupled form with respect to their temperature-dependent kinetics over a period of 22 h the temperature of 37 degrees C warrants highest substrate turnover for all enzymes at all reaction times using fluorogens. Also applying chromogens the optimum temperature for beta Gal is 37 degrees C and depends for HRP and AP on the reaction time. The substrate turnover of HRP using ABTS as chromogen is much higher compared to the other enzymes--both related to mol enzyme (molar activity) and to gram enzyme (specific activity). The turnover decreases for all enzymes in different degrees after coupling to IgG. The turnover of fluorogenic substrates is lower for all enzymes than the turnover of chromogenic substrates but due to the more sensitive detection of fluorogenic products the detection limits for all conjugates were lowered too--especially for beta Gal-IgG by a factor of 333 compared to the colorimetric procedure. In a 2-site binding enzyme immunoassay for alpha-1-fetoprotein (AFP) the detection limit for AFP was reduced by a factor of 2 only by the fluorimetry compared to the colorimetry with all 3 marker enzymes. The HRP-IgG conjugates warranted lowest detection limits for AFP (0.5-1 microgram/1), highest analytical sensitivity (slope of standard curves) at shortest periods of substrate reaction compared to the other enzymes.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Porstmann B,Porstmann T,Nugel E,Evers Udoi
10.1016/0022-1759(85)90388-6subject
Has Abstractpub_date
1985-05-10 00:00:00pages
27-37issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(85)90388-6journal_volume
79pub_type
杂志文章abstract：:The potential value of fluorescein-conjugated human alpha 2-macroglobulin as a probe for studying macrophages in murine peritoneal exudate and spleen cell suspensions has been investigated using a fluorescence-activated cell sorter. These studies revealed that the number of alpha 2-macroglobulin-positive cells in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90360-6
更新日期：1985-10-10 00:00:00
abstract：:Bacillus anthracis spores and Escherichia coli were stained with fluorescein-conjugated antibody using direct and indirect methods, then analyzed by means of a commercial flow cytometer. To reduce the cytometer's fluorescence component resulting from unreacted conjugate, reaction mixtures were either diluted or were c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90153-0
更新日期：1987-08-03 00:00:00
abstract：:An improved sensitivity to low levels of hepatitis B surface antigen (HBsAg) was demonstrated when ELISA results were interpreted by a statistical method. The absorbance of each test well was compared with the mean absorbance of all low-colour test samples. Those wells with an absorbance of more than two standard devi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90316-x
更新日期：1988-11-25 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：:A commonly employed method to determine the function of a particular cell population and to assess its contribution to the overall system in vivo is to selectively deplete that population and observe the effects. Using monoclonal antibodies to deliver toxins to target cells can achieve this with a high degree of effic...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2020.112794
更新日期：2020-08-01 00:00:00
abstract：:Overlapping peptide scans prepared by Spot synthesis have been used to map interaction sites in several systems. Here we report our experience with this approach to identify peptides from the variable parts of anti-hapten, anti-peptide and anti-protein antibodies that retain their specific antigen-binding capacity in ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00140-0
更新日期：2002-09-01 00:00:00
abstract：:It was found that 10% horse serum (HrS) could be used to support an in vitro antibody response to SRBC. It could replace the fetal calf serum (FCS) in both the culture medium and the nutritional feed. The response could be improved by the addition of 2-mercaptoethanol or by including both FCS (5%) and HrS (5%) in the ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90042-3
更新日期：1980-01-01 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：:The present study examined the use of replication-defective adenovirus for in vitro production of an immunoadhesin. A recombinant adenovirus, rendered replication defective by deletion of the E1 gene, was constructed to contain the murine interleukin-10 gene fused in frame with the hinge, CH2, and CH3 domains of the m...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00233-5
更新日期：1997-03-10 00:00:00
abstract：:Observations that the innate arm of the immune system is upregulated in pregnancy have highlighted the need for methods of isolating pure populations of monocytes for studies into pregnancy and pre-eclampsia without activating them during the isolation process. Density gradient centrifugation using iodixanol is a usef...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2004.05.011
更新日期：2004-10-01 00:00:00
abstract：:Attempts were made to estimate peripheral T-lymphocytes in rabbits by their ability to form rosettes with sheep, human, guinea-pig, rat, mouse and chicken RBC. The reaction was studied under different conditions of pH and temperature. The test was also carried out in the presence or absence of glutaraldehyde. With she...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90165-x
更新日期：1977-01-01 00:00:00
abstract：:A technique for detecting the nitroblue tetrazolium-reducing activity of human polymorphonuclear leukocyte enzymes separated on polyacrylamide gels is reported. Slices of gel are incubated in buffer containing nitroblue tetrazolium and either NADPH or NADH. Enzymes capable of reducing nitroblue tetrazolium form a dark...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90058-8
更新日期：1982-10-29 00:00:00
abstract：:A radioimmunoassay for phenobarbital has been studied. Antiphenobarbital antisera were obtained by repeated immunization of rabbits with p-succinamidophenobarbital conjugated to bovine serum albumin. Less than 0.2 pmol of phenobarbital could be measured by this procedure. The specificity of the antibodies was directed...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90327-2
更新日期：1979-01-01 00:00:00
abstract：:A new method for quantitative antigen analysis is described which precludes necessity for long incubation periods. The new technique, Surface Immune Precipitation (SIP), entails the application of an antigen sample droplet directly onto the surface of a gel containing antibody, the gel being supported by a reflecting ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90211-8
更新日期：1976-01-01 00:00:00
abstract：:Removal and/or neutralization of preformed anti-pig antibodies in non-human primate blood have been shown to prevent the hyperacute rejection of transplanted pig organs. The purpose of this study was to establish a suitable in vitro method that would allow for screening and comparison of various agents and methods pot...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00175-6
更新日期：1999-01-01 00:00:00
abstract：:We developed a noncompetitive two-site sandwich ELISA to quantitate monoclonal antibodies in culture supernatant. This assay measures the initial enzyme activity rate during the first minute of the reaction, which ensures linear velocity relative to time and a progress curve slope proportional to analyte concentration...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.112645
更新日期：2019-11-01 00:00:00
abstract：:Efficacious typhoid vaccines for young children will significantly reduce the disease burden in developing world. The Vi polysaccharide based conjugate vaccines (Vi-rEPA) against Salmonella Typhi Vi positive strains has shown high efficacy but may be ineffective against Vi negative S. Typhi. In this study, for the fir...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.07.005
更新日期：2017-11-01 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：:A polyethylene glycol (PEG) radioimmunoprecipitation assay for human IgG is described that is sufficiently sensitive to detect 0.5 ng of IgG. This model antibody-antigen system was also used to study the stoichiometries of PEG-precipitated complexes. Our results suggest that the presence of PEG may affect the stoichio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90091-7
更新日期：1979-01-01 00:00:00
abstract：:The method of extraction of outer membrane proteins (OMPs), the conditions of electrophoretic transfer, and the conditions of antibody binding, were compared in Western blotting studies of Neisseria meningitidis outer membrane antigens. The OMP profiles obtained by SDS-PAGE of outer membrane vesicles extracted with li...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90383-7
更新日期：1990-12-05 00:00:00
abstract：:We developed two sensitive methods for identifying antimicrobial molecules in leukocytes and other tissues. One method uses a gel overlay technique and was designed to identify antimicrobial polypeptides in samples subjected to polyacrylamide gel electrophoresis. The other, a radial diffusion assay, allows multiple fr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90021-7
更新日期：1991-03-21 00:00:00
abstract：:Simian immunodeficiency virus (SIV)-infection in macaques provides an important animal model for human immunodeficiency virus-1 (HIV-1) infection. The involvement of perforin (PFN), released by cytotoxic cells to mediate killing of virus-infected cells, has been difficult to assess in this experimental model due to a ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.02.018
更新日期：2006-05-30 00:00:00
abstract：:We have developed an antigen presenting system based on the capsid protein of the Flock House virus (FHV) and used it to display, in different positions on its external surface, two neutralizing epitopes found at residues 735-752 of HIV-1 gp160. We have compared the immunoreactivity of these FHV chimeric proteins and ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00097-x
更新日期：1996-10-16 00:00:00
abstract：:The multiple antigen peptide (MAP) system has been proposed as a novel and valuable approach for eliciting antibodies to peptides and developing synthetic vaccines. The MAP system consists of a small immunogenically inert core matrix of lysine residues with alpha- and epsilon-amino groups for anchoring multiple copies...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90033-p
更新日期：1992-12-08 00:00:00
abstract：:The presence of KIR4.1 antibodies has been proposed to be a characteristic of Multiple Sclerosis (MS). This could have a significant impact on disease management. However, the validation of the initial findings has failed till date. Conflicting results have been attributed to difficulties in isolating the lower-glycos...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.03.008
更新日期：2017-06-01 00:00:00
abstract：:We have designed a cytometry-based competition assay to evaluate peptide binding to empty recombinant HLA class II molecules. The efficiency of this assay was evaluated using recombinant HLA-DP0401 molecules (HLA-DP) produced in insect cells and 13 peptides from human telomerase reverse transcriptase (hTERT). We demon...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.06.018
更新日期：2011-08-31 00:00:00
abstract：:Two different methods have been used to study immune responses in the rat to Streptococcus pneumoniae type 3 and type 4 capsular polysaccharides (PPS). First, for simultaneous detection of the specificity and isotype of anti-PPS antibody-containing cells (ACC) in cryostat sections of lymphoid tissue, a double immunocy...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90314-6
更新日期：1991-12-15 00:00:00
abstract：:Recently, a powerful approach for the detection of MHC/peptide-specific T cells has been made possible by the engineering of soluble-tetrameric MHC/peptide complexes, consisting of singly biotinylated MHC/peptide molecules bound to fluorescent-labeled streptavidin. These tetrameric molecules are thought to compensate ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00215-x
更新日期：2000-02-03 00:00:00
abstract：:Single chain antibody fragment genes are commonly created by splicing together the immunoglobulin light chain (VL) and heavy chain variable (VH) genes of a monoclonal antibody produced by a hybridoma. Selective PCR amplification of the functional immunoglobulin variable gene rearrangements can be complicated by the ex...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.04.014
更新日期：2008-07-31 00:00:00
abstract：:An ELISA method for the detection of monoclonal antibodies (MAb) to the T3-T cell antigen receptor (TCR) complex was devised. The T3-TCR complex was solubilised using digitonin and a rat anti-T3 MAb (Campath 3) was used to bind it to an ELISA plate. Normal rat serum was used to block cross-reactivity between the rat M...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90280-3
更新日期：1988-01-21 00:00:00
