Abstract：
:Molecular interactions between the human insulin molecule and a panel of seven monoclonal antibodies (mAbs) against insulin were investigated by surface plasmon resonance. Using real-time analysis (BIAcore technology), the antigenic sites recognized on the human insulin molecule by these mAbs were mapped. Three overlapping regions could be defined on the insulin molecule coupled to the dextran matrix on the basis of its recognition by three mAbs groups: group I (mAbs 19, 10), group II (mAbs 13, 5, 25, 15) and group III (mAb P10). The antigenic determinants of the molecule in solution were mapped using a two-site assay in which insulin was presented to each mAb, bound to a rabbit anti-mouse IgG Fc gamma 1 coupled to the dextran. Four antigenic regions were defined: regions I and III were defined by the same antibodies as above whereas region II was recognized differently by mAbs 13, 5 (region IIa) and mAbs 25,15 (region IIb). The Ka value of the mAbs determined using the BIAcore were in the same range (from 3 x 10(7) M-1 to 6.6 x 10(9) M-1) whether insulin was coupled to the dextran matrix or in solution. Furthermore, mAbs 19 and P10, which recognize two distinct regions on the insulin molecule, were selected to develop a highly sensitive immunoassay for measurement of insulin in serum.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Allauzen S,Mani JC,Granier C,Pau B,Bouanani Mdoi
10.1016/0022-1759(95)00020-bsubject
Has Abstractpub_date
1995-06-14 00:00:00pages
27-32issue
1eissn
0022-1759issn
1872-7905pii
0022-1759(95)00020-Bjournal_volume
183pub_type
杂志文章abstract：:Recent data generated in our laboratory indicates that human lymphokine-activated killer (LAK) cells can be induced in several commercially available serum-free medium formulations. Interestingly, LAK cells were also generated in RPMI 1640 medium plus 1000 U/ml IL-2 without addition of human serum. However, higher lev...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(88)90239-6
更新日期：1988-05-09 00:00:00
abstract：:Dilatometry has become a useful method for the study of proteins owing to its simplicity and accuracy. However, it has seldom been used in the study of immunoglobulins. Therefore possible applications of the method for that study are being discussed. A description is also given of the most common experimental set-up a...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(75)90110-6
更新日期：1975-12-01 00:00:00
abstract：:An assay to detect circulating immune complexes is described and exemplified with serum from SLE and RA patients. The method is based on selective precipitation of immune complexes with PEG followed by a specific ELISA assay to detect the immunoglobulins present in the precipitate. The immunoglobulins of the precipita...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90186-3
更新日期：1985-02-28 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：:This study describes a four-color flow cytometric assay that detects CD4+ T cell responses to the anthrax vaccine. Whole blood from seven volunteers who previously obtained the anthrax vaccine was inoculated in vitro with varying concentrations of the anthrax antigen. TNF-alpha and IFN-gamma production from memory CD4...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.07.012
更新日期：2003-11-01 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：:A solid-phase radioimmunoassay for detection of herpes simplex virus-specific IgG and IgM antibodies in human serum specimens is presented. Virus antigen is adsorbed on polystyrene balls and antibodies which attach to the antigen are detected by 125I-labeled antihuman-gamma or antihuman-mu immunoglobulins. A total of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90008-4
更新日期：1977-01-01 00:00:00
abstract：:A method of tanned erythrocyte agglutination of wide applicability is described for the study of DNA antibodies. The practical aspects of the method have been explored in estimation of antibodies to DNA in human cases of systemic lupus erythematosus, discoid lupus erythematosus, rheumatoid arthritis, other collagenose...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90067-4
更新日期：1978-01-01 00:00:00
abstract：:CD3-specific antibodies have shown clinical efficacy in both transplantation and autoimmunity. However, targeting CD3 in this way can lead to T-cell activation and a serious cytokine release syndrome mediated by Fcγ receptor binding. An in vivo mouse model has been developed using severe combined immunodeficient (SCID...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.07.001
更新日期：2012-10-31 00:00:00
abstract：:CFSE analysis of a proliferating cell population is a popular tool for the study of cell division and divisionlinked changes in cell behavior. Recently Banks et al. (2011), Luzyanina et al. (2009), Luzyanina et al. (2007), a partial differential equation (PDE) model to describe lymphocyte dynamics in a CFSE proliferat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.08.014
更新日期：2011-10-28 00:00:00
abstract：:Human recombinant soluble 37 kDa CD23 has been expressed in insect cells and secreted into the culture medium using the IL-2 leader sequence. The 37 kDa CD23 was purified 600-fold to homogeneity by monoclonal antibody affinity chromatography and gel filtration. The pure protein is monomeric, glycosylated, depleted of ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90253-p
更新日期：1992-05-18 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：:We present a double antibody immunoassay for tumour necrosis factor alpha (TNF alpha) with a peroxidase dependent endpoint which can be detected by absorbance or chemiluminescence depending on the choice of substrate. The chemilumimetric and colorimetric assays have a detection threshold in human serum of 3.9 pg/ml an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90288-5
更新日期：1992-11-05 00:00:00
abstract：:A method for the preservation of mouse peritoneal macrophages is described. Using 5% dimethylsulphoxide with either a cooling rate of 1 degree C/min or two-step cooling with 10 min interruption at -30 degrres C allows 80% survival as judged by pinocytic activity. An important finding was that the cells must be handled...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90139-9
更新日期：1977-01-01 00:00:00
abstract：:A new method of mouse B cell isolation has been developed. It is based on the differences in buoyant density of free lymphocytes and cells bound within liposome-cellular complexes (LCC). LCC arise during the interaction of B cells bearing surface immunoglobulins (Ig) with liposomes loaded with affinity purified rabbit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90074-4
更新日期：1989-12-20 00:00:00
abstract：:The present study describes the immunomagnetic isolation of human natural killer (NK) and lymphokine activated killer (LAK) cells. Antibodies against CD56 and sheep anti-mouse IgG-coated magnetic monodisperse particles (Dynabeads M-450) were used for the positive isolation of CD56+ cells from unstimulated mononuclear ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90242-8
更新日期：1991-01-24 00:00:00
abstract：:We evaluated an adjuvant, TiterMax, as an alternative to complete Freund's adjuvant (CFA) for producing antisera in animals. TiterMax, consists of a microparticulate stabilized water-in-oil emulsion of a metabolizable oil, squalene, with the adjuvant block copolymer CRL89-41. This paper reports two evaluations of Tite...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90302-a
更新日期：1992-08-30 00:00:00
abstract：:A fluorimetric method using 4-methylumbelliferyl heptanoate (MUH) has been developed for detecting cell-mediated cytotoxicity and cell proliferation. The assay is based on the hydrolysis of the fluorochrome (MUH) by intracellular esterases of viable cells resulting in the production of highly fluorescent 4-methylumbel...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00115-q
更新日期：1995-09-25 00:00:00
abstract：:Lymphocytes preincubated with various activities of Na251CrO4 were tested for their ability to induce a local, popliteal lymph node graft-versus-host (GVH) reaction. Incubation with 100 micronCi of the isotope per ml of cell suspension impaired the GVH activity, 25 micronCi gave a reduced response in 1 out of 3 experi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90153-9
更新日期：1978-01-01 00:00:00
abstract：:The indirect immunofluorescent technique has been shown to be a more sensitive method than either immunoelectrophoresis or gel filtration for testing the efficacy of the gel filtration method in fractionating human immunoglobulins. It has been confirmed that IgM fractions of some sera from patients with multiple scler...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(75)90078-2
更新日期：1975-01-01 00:00:00
abstract：:Cell-enzyme-linked immunosorbent assay (cell-ELISA) is a technique for the rapid, convenient, and quantitative detection of molecules expressed on the cell surface. Here we present an evaluation of beta-galactosidase as an antibody-tag for cell-ELISA. In contrast to substrates for horseradish peroxidase (HRP) and alka...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00216-1
更新日期：2000-02-03 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：:The present article reports a multiparametric cytofluorimetric analysis of apoptosis in murine thymocytes and human PBMC from healthy donors or HIV-infected patients. We have evaluated four previously described cytofluorimetric methods of apoptosis quantification, each of them detecting distinct cellular alterations o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00148-2
更新日期：1996-10-30 00:00:00
abstract：:One of the major functions of mononuclear phagocytes, i.e., monocytes and macrophages, is the phagocytosis and killing of microorganisms. To obtain more insight into the pathogenesis of infectious diseases and to develop new therapies against these diseases, a better understanding of the antimicrobial mechanisms emplo...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/0022-1759(94)90021-3
更新日期：1994-09-14 00:00:00
abstract：:Monoclonal antibodies (mAbs) have emerged as a promising tool for cancer therapy. Differing approaches utilize mAbs to either deliver a drug to the tumor cells or to modulate the host's immune system to mediate tumor kill. The rate by which a therapeutic antibody is being internalized by tumor cells is a decisive feat...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.04.024
更新日期：2015-08-01 00:00:00
abstract：:Insulin was directly coupled and also indirectly coupled by a side chain to Sepharose 4B to form immunoabsorbents for affinity chromatography of insulin antibodies. The two direct derivatives were insulin lysyl and insulin phenylalanyl sepharose. The indirect derivative, insulin carboxyhexyl sepharose, was prepared in...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90130-2
更新日期：1977-01-01 00:00:00
abstract：:Recombinant fusion proteins, consisting of a monovalent anti-human RBC monoclonal antibody B6, and conserved immunodominant peptide of HIV-1 envelope glycoprotein gp41 or HIV-2 envelope glycoprotein gp36, have been designed and purified after over-expression in E. coli. These fusion proteins are Fab-based and were obt...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00435-5
更新日期：2001-10-01 00:00:00
abstract：:A cell membrane permeabilizing treatment is described which involves the use of lysolecithin at low concentration in acidic acetate buffer and paraformaldehyde fixation. It preserved well-separated scatter cytograms of small and large lymphocytes. The accuracy of the immunochemical detection of internal antigens by fl...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90162-9
更新日期：1991-04-25 00:00:00
abstract：:Recently the analytical power of the latest high throughput next generation DNA sequencing platforms has been used to analyse phage that have been selected from the panning of large combinatorial libraries displaying either peptide or antibody ligands. This process, commonly referred to as next generation phage displa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.03.005
更新日期：2015-05-01 00:00:00
abstract：:Flow cytometric methods for the evaluation of the cell surface binding properties of monoclonal antibody (MoAb)-drug/toxin conjugates defining tumor-associated antigens are presented. In these techniques, suspension cultures of solid human tumor cell lines are incubated with either varying dilutions of MoAb or MoAb-dr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90310-3
更新日期：1987-02-11 00:00:00
