Abstract:
:A method for coating native, non-derivatized, polysaccharide (PS) onto nitrocellulose (NC) for identifying PS-specific antibodies has been developed. The new feature of this method is that PS molecules are vacuum filtered onto NC in their native state by devices that can accommodate NC of different sizes and shapes. PS-coated NC disks were used to localize antibody secreting hybridoma cells cultured on filter paper disks. These were analyzed by blotting with size-matched PS-coated NC disks and specific antibodies secreted by individual colonies were detected by enzyme-linked immunoblot. In another application of this method, immune sera were separated by isoelectric focusing and the gels were blotted with PS-coated NC sheets. The spectrotype and isotype of antibodies that bound to the NC were examined using isotype specific enzyme-linked antibody. These immunoblots showed high resolution and specificity. The advantages of this method are that the PS used for coating does not need to be derivatized in order to bind the NC, and that smaller quantities of PS may be utilized by this coating method when compared to other techniques. This provides a useful tool to ask many questions regarding the immune response to PS.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Feng SH,Rubinstein LJ,Stein KEdoi
10.1016/0022-1759(91)90032-bsubject
Has Abstractpub_date
1991-03-21 00:00:00pages
261-6issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(91)90032-Bjournal_volume
137pub_type
杂志文章abstract::A successful technique for the isolation of highly pure suspensions of viable leukocytes from the small intestine of cattle is described. Procedures ranging from mechanical mincing to enzymatic digestion of tissues were compared. The most reliable and reproducible procedure was the sequential treatment of tissues with...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(87)90410-8
更新日期:1987-12-04 00:00:00
abstract::Elucidation of local immune response at the cervix is important for understanding and evaluating STD vaccine approaches currently being proposed. However, no well-validated method exists for the collection of cervical secretions for evaluation of cervical immune response. The purpose of this study was to determine the...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00037-x
更新日期:1999-05-27 00:00:00
abstract::An approach has been devised for analysing data generated in the study of in vivo cytotoxic T cell responses. The method calculates the total number of lytic units generated in the peritoneal cavity, or in cultures of the lymph nodes draining the rear footpad, following antigenic stimulation. The assessment of group d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90387-3
更新日期:1983-11-11 00:00:00
abstract::Boronated antibodies have already been evaluated as agents in neutron capture therapy. Because the boronation procedure may alter the properties of the antibody it is important to study the immunoreactivity of the conjugated antibody before in vivo use. In our studies of two dextran-boronated monoclonal antibodies, an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90016-o
更新日期:1990-01-24 00:00:00
abstract::The Cellular Antigen Stimulation Test (CAST), a recent innovation in the laboratory detection of allergen sensitivity, is a functional assay based on the release of sulphidoleukotrienes (LTs) from allergen-activated circulating basophils. The current study was undertaken to establish optimum determinants of this proce...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2003.12.014
更新日期:2004-05-01 00:00:00
abstract::Information about the IgE receptor and IgE content of mast cells under different conditions in vivo is essential for further understanding the functions of the mast cell-IgE system. A cytofluorometric method for measuring cell-bound IgE on individual mast cells was therefore explored using peritoneal mast cells of Spr...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(94)90151-1
更新日期:1994-12-28 00:00:00
abstract::The method reported here describes a microrosetting assay which allows the early detection of antibody-synthesizing hybridomas. This method is specific, requires very few hybridoma cells and avoids artifacts resulting from the presence of antibodies secreted by spleen cells in the primary hybridoma cultures. It is eas...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90101-7
更新日期:1979-01-01 00:00:00
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::Antibody phage display technology is a well established method for selecting specific antibodies against desired targets. Although phage display is the most widely used method of generating synthetic antibodies, it is laborious to perform multiple selections with different antigens simultaneously using conventional ma...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.09.034
更新日期:2011-03-31 00:00:00
abstract::Effector-target conjugation between different cell populations of human NK cells and K562 tumor cells has been studied from binding isotherms obtained from data of effector (alpha) and target (beta) conjugate frequencies measured by flow cytometry analysis at different effector-to-target ratios. Non-linear and linear ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(97)00153-1
更新日期:1997-12-01 00:00:00
abstract::Small haptens such as methylamphetamine (MW 149) cannot, on their own, induce an immune response. It is also unlikely that they fill the binding site of any antibody that recognises them. Under such circumstances any attached label might be expected to enter the area of the binding site and exert an influence on overa...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90375-p
更新日期:1991-07-05 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::A technically simple microassay for macrophage migration inhibition studies is described. Using simple 0.08 ml of test solution per assay, meaningful statistical analysis via multiple replicates is possible. A novel feature is that test solutions may be stored frozen in the test well until the indicating macrophages a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(80)90252-5
更新日期:1980-01-01 00:00:00
abstract::The possibility that immortalisation of in vitro immunised splenocytes may result in hybridomas secreting polyreactive antibodies was investigated. A panel of nine murine hybridomas, secreting IgM(kappa) anti-goat immunoglobulin G (anti-GIgG), was produced by immortalising splenocytes that had been immunised in vitro ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00481-1
更新日期:2001-12-01 00:00:00
abstract::We report a novel competitive type immunoassay for oestradiol based on the idiotypic anti-idiotypic approach. This has been achieved by the production of an anti-idiotypic antibody (anti-Id) which is directed against the oestradiol binding site of the primary idiotypic antibody (Ab1). In this format the primary Ab1 wa...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90068-q
更新日期:1991-04-08 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 procedure to enhance incorporation of radioactive iodine into cell-surface proteins is described. The effects of chemical modification of cells with the iodinatible haptens N-succinimidyl-3-(4-hydroxyphenyl) propionate (Bolton-Hunter reagent) and methyl-p-hydroxybenzimidate HC1 (Wood reagent) are compared. The Wood ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90177-2
更新日期:1985-02-28 00:00:00
abstract::Genetic mapping of differentiation specific surface antigens has been hampered by difficulty in preparation of interspecies hybrid cells which continue to express differentiated functions. A method has been developed for production of interspecies T cell hybrids which continue to express T cell specific cell surface m...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90108-4
更新日期:1983-08-12 00:00:00
abstract::A method for cloning hybridomas is described which involves transfer of single cells with a capillary tubing connected to a suction apparatus. This method enables the efficient recovery of antibody-producing clones of differing abundances from the parent hybridoma cultures. The 4 cell lines submitted to this cloning p...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90084-5
更新日期:1985-04-22 00:00:00
abstract::The influence of various methodological variables on the CrCl3 technique for coupling antigen to the surface of erythrocytes has been investigated. Tests should be performed in protein- and phosphate-free medium. The CrCl3 stock solution should be stored at a pH of 5.0 and working dilultions prepared in acetate buffer...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90198-3
更新日期:1977-01-01 00:00:00
abstract::Knowledge of immunodominant regions in major viral antigens is important for rational design of effective vaccines and diagnostic tests. Although there have been many reports of such work done for SARS-CoV, these were mainly focused on the immune responses of humans and mice. In this study, we aim to search for and co...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2007.11.009
更新日期:2008-02-29 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::Cut point determination is an important aspect of immunogenicity assay development. The cut point can be influenced by a myriad of factors. Key among those is the analytical variability of the assay itself and biological variation due to test samples. Since a smaller cut point value may result in improved sensitivity,...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2018.10.001
更新日期:2018-12-01 00:00:00
abstract::We performed nasal lavage (NAL) combined with induced sputum to determine exudative inflammation in the upper and lower airways in patients with chronic sinusitis and in controls. To monitor plasma exudation into the respiratory lumen and loss of size-selectivity of the mucosa, we determined the sample-to-serum ratio ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00439-2
更新日期:2001-10-01 00:00:00
abstract::The effects of complement activation on the antigenic component of immune complexes have been studied, using 125I-BSA-rabbit anti-BSA-IgG complexes as models. Polyethylene glycol precipitates of 4 types of IC (those formed in native normal human serum, or NHS-containing EDTA, NHS-EDTA, and preformed soluble IC incubat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(84)90109-1
更新日期:1984-12-31 00:00:00
abstract::The simian immunodeficiency virus (SIV) model of AIDS is widely used for the development of human immunodeficiency virus (HIV) vaccine strategies, particularly for the analysis of correlates of protective immunity. As it is not always possible to establish autologous B-lymphoblastoid cell lines (B-LCL) for use as targ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00480-x
更新日期:2001-12-01 00:00:00
abstract::Radioimmunoassay of anti-tetanus antibodies produced in human serum in response to tetanus toxoid immunization was carried out by employing the non-specific precipitation effect of ammonium sulphate described by Farr (1958). The results showed that the method was sensitive and was able to differentiate between immune ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(75)90088-5
更新日期:1975-01-01 00:00:00
abstract::Complement activation is a key component of the pathogenesis of immune-mediated tissue damage in many diseases. Assessment of complement activation in current practice is largely based on the measurement of intact C3 and C4 or the determination of complement haemolytic function. These parameters reflect activation onl...
journal_title:Journal of immunological methods
pub_type: 杂志文章,评审
doi:10.1016/0022-1759(93)90063-d
更新日期:1993-01-04 00:00:00
abstract::We report a restainin method for restoring fluorescence in paraffin-embedded tissue sections previously treated with the indirect immunofluorescence technique. Antisera to gastrin and group II pepsinogens were used. Fluorescence was restored in completely faded sections retrived from storage files, as well as in secti...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(77)90121-1
更新日期:1977-01-01 00:00:00
abstract::Most current methods for measuring cell death are based on plasma membrane disintegration and the consequent release of cytoplasm. The relevant cells are usually loaded with a label (usually 51Cr or 125I), the release of which is measured. I describe here a method, based on the recent evidence that dying cells often d...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90325-a
更新日期:1991-12-15 00:00:00