Abstract:
:An immunoperoxidase method has been developed which allows accurate and sensitive quantitation of the binding of monoclonal antibodies to cell surface antigens. Monolayers of fixed cells were prepared in wells of Terasaki micro-test plates and monoclonal antibodies bound to cell surface antigens were identified by the unlabeled antibody-enzyme method of Sternberger (1974). The cell-bound peroxidase could either be quantified per well or visualized on individual cells by the use of appropriate substrates for peroxidase. Experimental procedures are described in detail and results obtained with several monoclonal antibodies with specificity for different target cells are shown. Limitations and applications of the technique are discussed.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Lansdorp PM,Astaldi GC,Oosterhof F,Janssen MC,Zeijlemaker WPdoi
10.1016/0022-1759(80)90240-9subject
Has Abstractpub_date
1980-01-01 00:00:00pages
393-405issue
4eissn
0022-1759issn
1872-7905pii
0022-1759(80)90240-9journal_volume
39pub_type
杂志文章abstract::Peptide sequences that bind to a wide range of ligands such as monoclonal antibodies, receptors and carbohydrates have been successfully identified after screening phage display peptide libraries. However, these procedures tend to select mainly medium to low affinity-binding clones. A modified screening procedure has ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00318-5
更新日期:2001-01-01 00:00:00
abstract::In this study we present a new method for the elimination of mononuclear phagocytic cells from cell suspensions. By making use of liposome-encapsulated dichloromethylene diphosphonate we were able to effectively remove macrophages from spleen cell suspensions. This effect was not observed when using the free drug or c...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90376-7
更新日期:1990-12-05 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 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::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::The sensitivity of crossed immunoelectrophoresis (CIE) and rocket immunoelectrophoresis (RIE) does not usually permit the study of diluted protein solutions or the detection of trace components. Antigen-antibody precipitates obtained after immunoelectrophoresis were therefore dissociated and transferred from the agaro...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90163-9
更新日期:1988-11-10 00:00:00
abstract::Flow cytometry has been utilized to evaluate the stability of antibody production by unstable subclones of a human x human x mouse heterohybridoma. Heterogeneity of cell-associated immunoglobulin heavy chain expression was demonstrated in different subclones and an increased frequency of cells containing low levels of...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)90247-x
更新日期:1994-03-29 00:00:00
abstract::We describe a raft technique for the study of chemotaxis which possesses a number of advantages over the use of chambers. Membranes are laid on pads soaked in chemotactic agent and cells are contained in plastic cups, which are inverted on to the membrane. This method gives results which are similar to those obtained ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90033-8
更新日期:1976-01-01 00:00:00
abstract::We have developed and validated a process-specific immunoligand assay based on the Threshold system for the quantification of residual host cell proteins (HCPs) in a recombinant subunit vaccine candidate against the human respiratory syncytial virus (hRSV). The industrial process of this vaccine produced in Escherichi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00321-0
更新日期:2001-05-01 00:00:00
abstract::The influence of antibody valency in a displacement immunoassay was investigated by comparing the whole antibody molecule with the corresponding Fab-fragment. The displacement immunoassay for the herbicide 2,4-dichlorophenoxyacetic acid (2,4-D) takes advantage of the cross-reactivity of monoclonal anti-2,4-D antibodie...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(98)00211-7
更新日期:1999-03-04 00:00:00
abstract::The use of colloidal gold and colloidal gold followed by silver enhancement as marker for dot or blot immune overlays is described. Colloidal gold probes concentrating at the sites of immune reaction gradually develop a pinkish colour during incubation that can be seen with the naked eye. With a physical developer, ve...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90303-x
更新日期:1984-11-30 00:00:00
abstract:BACKGROUND:The serological screening for celiac disease (CD) is currently based on the detection of anti-transglutaminase (tTG) IgA antibodies, subsequently confirmed by positive endomysial antibodies (EMA). When an anti-tTG IgA positive/EMA IgA negative result occurs, it can be due either to the lower sensitivity of t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2020.112910
更新日期:2021-02-01 00:00:00
abstract::We describe a simplified and improved proliferation assay based on a conventional ELISA system for the in vitro measurement of cellular proliferation (ELISA proliferation assay = EPA). The assay is based on a new monoclonal antibody (Ki-S3) to the proliferation-specific Ki-67-antigen and is carried out in 96-well micr...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00175-0
更新日期:1998-02-01 00:00:00
abstract::A new antibody overlay technique for 2-dimensional immunoelectrophoresis (2-D-IEP) is described. After the first dimension electrophoresis of the antigen, the desired amount of antiserum is applied to the initial agarose layer and then evenly distributed over a defined surface of the gel with a PVC film. This modifica...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90332-6
更新日期:1984-02-10 00:00:00
abstract::A solid-phase radioimmunometric binding assay is described utilizing 125I-labeled protein A for the detection of antibody to human melanoma associated antigens. The novel aspect of this assay is the use of chemically defined spent culture medium of melanoma cells at target antigens previously depleted of fibronectin b...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90058-7
更新日期:1980-01-01 00:00:00
abstract::Nowadays many reagents are available for the biotinylation of proteins. As most of them bind to amino groups of the protein the degree of labelling differs from batch to batch and the possibility exists that the biological activity of the target protein may be affected by the labelling procedure. In the present study ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(00)00172-1
更新日期:2000-06-23 00:00:00
abstract::The cholesterol-rich lipid rafts on T cell membrane play important role in the formation of T cell receptor (TCR) signalosome upon receptor ligation. Analytical studies on the kinetics of lipid rafts formation and recruitment of protein receptors to lipid rafts are still limited by the use of a large number of cells. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(03)00014-0
更新日期:2003-04-01 00:00:00
abstract::A simple method for the simultaneous isolation of IgG2, IgG1, IgA and IgM from porcine colostral whey is described. It makes use of DEAE-cellulose chromatography followed by repeated Sepharose 6B exclusion chromatography after the removal of fat, casein and lipoproteins, Fused rocket immunoelectrophoresis is used to s...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90058-8
更新日期:1977-01-01 00:00:00
abstract::To increase the selection efficiency and productivity of stable clones expressing recombinant antibody-IL-2 fusion protein, a dicistronic expression vector containing the anti-erbB2 scFv-Fc-IL-2 fusion gene followed by a weakened neo gene, was constructed to allow for the concurrent translation of the recombinant prot...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2004.08.013
更新日期:2004-12-01 00:00:00
abstract::A method is described for measuring antibody-dependent cellular cytotoxicity (ADCC) of mononuclear cells from human peripheral blood using an established murine cell line and commercially prepared antisera. The test utilizes a standard 51Cr release technique. The ADCC activity of mononuclear cells obtained from 10 hea...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90107-3
更新日期:1985-05-23 00:00:00
abstract::The monoclonal antibody-specific immobilization of lymphocyte antigens (MAILA) assay was developed to detect antibodies present in human alloantisera against antigens of different major histocompatibility complex loci, particularly of class II specificity. The MAILA assay has been used in our laboratory to the determi...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90094-n
更新日期:1993-01-04 00:00:00
abstract::IL-2 activity is commonly estimated in cell culture supernatants by an IL-2-dependent cell proliferation assay. This method is both reproducible and sensitive. However, it often appears from the literature that statistical analysis of the titration data either is disregarded or, when performed, is based on statistical...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90463-1
更新日期:1986-02-12 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
abstract::A solid-phase enzyme immunoassay for the measurement of anti-acetylcholine receptor antibodies in the sera of patients with myasthenia gravis is reported. Sufficient amounts of acetylcholine receptor for the sensitive detection of anti-acetylcholine receptor antibody were directly fixed to Costar serocluster 96-well E...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90401-0
更新日期:1984-10-26 00:00:00
abstract::We describe the use of 7-amino-actinomycin D (7AAD) to measure phagocytosis and the opsonizing capacity of serum. Heat-inactivated Candida albicans was previously stained with 7AAD and incubated with resident peritoneal macrophages. The samples were analyzed by flow cytometry and phagocytic cells were identified by th...
journal_title:Journal of immunological methods
pub_type:
doi:10.1016/s0022-1759(01)00358-1
更新日期:2001-07-01 00:00:00
abstract::This paper describes a method to enhance the sensitivity of enzyme immunoassays by the use of solid tagged latex beads. Various bead sizes (0.21 and 0.81 micron) were tested using particles which had been covalently modified (or coated) with biotinylated rabbit anti-mouse immunoglobulin antibodies. Detection limits of...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(88)90158-5
更新日期:1988-11-10 00:00:00
abstract::Two murine IgM monoclonal antibodies (MAs) directed against the lipid A portion of bacterial lipopolysaccharide (LPS) were compared in their binding to Re LPS and lipid A and their idiotypic make-up with rabbit anti-idiotypic sera. Horseradish peroxidase (HRPO)-labelled MAs 8-2 and 26-20 bound equally well to Re LPS. ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90308-i
更新日期:1990-06-12 00:00:00
abstract::When limited amounts of antigens are available, it is often difficult to obtain large quantities of polyclonal antisera. Although antisera can be raised in small rodents, yields are usually small. To circumvent these problems, we have designed a modified method for generating polyclonal ascites fluid (AF). Using the a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90108-j
更新日期:1993-09-27 00:00:00
abstract::The performance of immunoassays for the detection of autoantibodies is of critical importance to the diagnosis and assessment of patients with systemic lupus erythematosus (SLE). Our objective was to compare 3 multiplexed assays for measurement of multiple autoantibodies and their association with global disease activ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.04.005
更新日期:2010-06-30 00:00:00
abstract::Use of fluorescently labelled multimers, particularly tetramers of peptide and MHC class I glycoprotein (pMHC-I) complexes, is essential for the analysis of CD8+ T cell immunity in basic research and clinical settings. A recently described combinatorial approach using pMHC-I multimers coupled to a unique combination o...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.06.003
更新日期:2010-08-31 00:00:00