Abstract：
:A serum-free in vitro immunization method for the generation of hybridomas producing specific antibodies to an antigen is described. The method was tested with human thyroglobulin as antigen. The serum-free medium used (Yssel et al., 1984) consisted of Iscove's modification of Dulbecco's modified Eagle's medium, supplemented with albumin, transferrin, insulin, ethanolamine and linoleic, oleic and palmitic acids. An optimal response was obtained when splenocytes from BALB/c mice were cultured for 3 days in the presence of 1.5 nM thyroglobulin and thymocyte-conditioned medium prior to fusion with SP2/0 myeloma cells and seeding of the fused cells in microtitre plates. The frequency of positive wells, defined as the number of wells secreting anti-(thyroglobulin) antibodies/number of viable cells used for the fusion, was 1.6 X 10(-6) +/- 0.25 X 10(-6) (mean +/- SD; n = 4). Eight stable clones producing anti-(thyroglobulin) antibodies were isolated. One clone (3D12) produced antibodies reacting only with human thyroglobulin. The antibodies produced by the other clones reacted with human, murine and porcine thyroglobulins. Seven of the clones produced antibodies of the IgM class and one clone produced IgG. The specificity of 3D12 (IgM) for human thyroglobulin and the absence of any reactivity with murine thyroglobulin provides evidence for a primary response of splenocytes in culture to the presence of an antigen.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Ossendorp FA,De Boer M,Al BJ,Hilgers J,Bruning PF,Tager JMdoi
10.1016/0022-1759(86)90487-4subject
Has Abstractpub_date
1986-07-24 00:00:00pages
257-64issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(86)90487-4journal_volume
91pub_type
杂志文章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：: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：:Natural killer (NK) cells are important mediators of virus- and tumor-specific immune responses. The transfection of genes into NK cells has been proven difficult and so far requires infection with virus-based vectors. Here, the application of a novel nonviral, electroporation-based gene transfer method is described f...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00431-3
更新日期：2003-03-01 00:00:00
abstract：:There are several double immunolabelling methods but each has its drawbacks. More often than not, antibodies with the required specificities are available in only one species and their use normally produces false labels due to cross-reactivity. We describe a new and reliable technique for staining with primary antibod...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(95)00281-2
更新日期：1996-04-19 00:00:00
abstract：:Helicobacter pylori is a bacterial pathogen, estimated to infect half the world's population. The bacterium is the aetiological cause of gastritis, the common precursor for peptic ulcer disease and gastric cancer. Immunisation of at-risk individuals is the most cost-effective means of dealing with such a widespread pa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00141-5
更新日期：1997-11-10 00:00:00
abstract：:A new micro-glass-tube leukocyte adherence inhibition (LAI) assay which is appropriate for detecting delayed type hypersensitivity in vitro has been developed for human leukocytes. Enumeration of adherent cells is replaced by a cellular radioimmunoassay determining antibody binding of the monoclonal reagents, OKT4, OK...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90482-5
更新日期：1986-07-24 00:00:00
abstract：:We describe the preparation of a new rat monoclonal antibody (CRC1) to the N-terminal sequence of the 43 kDa subunit of human ovarian inhibin, and its use together with other anti-peptide monoclonal antibodies, in two-site immunoassays for the detection of inhibin-related material in biological fluids. The Fab fractio...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90342-5
更新日期：1993-10-15 00:00:00
abstract：:The relative efficiency of depot (Freund's complete adjuvant (FCA) and saponin), and non-depot (Corynebacterium parvum strain P. acnes I, C. parvum strain IMI585, and levamisole) immunoadjuvants were compared in raising agglutinating and precipitating antibodies against the metacestode fluid of Taenia multiceps of she...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90230-2
更新日期：1983-07-29 00:00:00
abstract：:In the present work we demonstrate the efficient display of functional scFv antibodies on the bacteriophage lambda capsid. A single-chain (scFv) anti-CEA antibody gene was cloned in two different vectors to obtain fusion of the scFv antibody to the N- or C-terminus of the bacteriophage lambda capsid protein D (gpD). L...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.01.003
更新日期：2006-03-20 00:00:00
abstract：:We have developed a sensitive and flexible method for the qualitative evaluation of IgG-Fc receptor interactions in cell suspensions. The assay is based on the flow cytometric quantitation of antibody-coated erythrocyte (EA) rosetting using fluorescein-labelled indicator erythrocytes (E). The number of IgG molecules o...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90070-c
更新日期：1990-03-09 00:00:00
abstract：:The concentrations of major basic protein (MBP), eosinophil cationic protein (ECP), eosinophil derived neurotoxin (EDN) and eosinophil peroxidase (EPO) have been associated with eosinophilic disease severity. Whereas a variety of techniques have been used to measure individual eosinophil granule protein concentration,...
journal_title：Journal of immunological methods
pub_type： 临床试验,杂志文章
doi：10.1016/j.jim.2014.05.020
更新日期：2014-09-01 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：:The measurement of the Epstein-Barr viral load in peripheral blood has been recognised as an important way of monitoring the response to treatment in patients with Epstein-Barr virus (EBV)-related malignancies. In particular, EBV load in transplant recipients can be used as a predictive parameter for Post-transplant L...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00333-2
更新日期：2002-12-15 00:00:00
abstract：:A separation procedure based on Percoll gradient centrifugation is described here that allows enrichment for two main populations of granules, with average densities of 1.05 and 1.1 g/ml. The heavy-density granules, which comprise 5-20% of the total amount of membrane-lytic activity, are morphologically and biochemica...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90179-8
更新日期：1989-11-13 00:00:00
abstract：:The cell mediated immune response to herpes simplex virus was assessed by a modification of the leukocyte migration inhibition test, developed to increase its sensitivity and correlated with the in vitro stimulation of lymphocytes. The introduction of a preincubation step allowed us to demonstrate significant inhibiti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(76)90202-7
更新日期：1976-01-01 00:00:00
abstract：:In standard displacement flow immunoassays the analyte in the sample creates an active dissociation of labelled antigens (or antigen homologues) from an antigen binding site of an immobilized antibody, after which the labelled substance is measured downstream. Such systems have been described for molecules up to 1 kDa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00106-9
更新日期：1998-08-01 00:00:00
abstract：:A simple method is described for the preparation of vimentin-type intermediate filaments for solid-phase immunoassays. Bovine aortic endothelial cells were grown to confluency in multiwells, and extracted with Triton X-100 and a high-salt buffer. Residual protein in the wells consisted of 53% vimentin as judged by SDS...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90149-8
更新日期：1985-12-27 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：: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：:The open reading frame of the α-subunit (amino acids 1-211) of human muscle nicotinic acetylcholine receptor (hAChR(α211)) was inserted into eukaryotic expression vector of pPIC9K to form a recombinant plasmid. After transformation and expression, the target protein rhAChR(α211) (recombinant hAChR(α211)) was secretive...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2011.04.015
更新日期：2011-09-30 00:00:00
abstract：:Hapten (trinitrophenyl) was coupled to antigen (ovalbumin). The haptenated antigen was bound by anti-ovalbumin antibody and binding was quantitated with 125I-labelled anti-hapten antibodies. Thus, with a single radioactive reagent antibodies against a variety of antigens can be detected, whilst the problems inherent i...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90011-x
更新日期：1978-01-01 00:00:00
abstract：:A sensitive time-resolved fluoroimmunoassay of nerve growth factor (NGF) has been developed. The method is based on the unique property of the lanthanides for delayed fluorescence, which reduces substantially the endogenous fluorescence of biological substances, because the excitation of the sample and detection of th...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90334-7
更新日期：1989-08-15 00:00:00
abstract：:The colorimetric MTT assay based on the selective ability of living cells to reduce 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide into formazan was adapted for measuring antibody independent monocyte mediated cytotoxicity. In view of the suggested application of adoptive immunotherapy we studied the an...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90205-t
更新日期：1991-07-26 00:00:00
abstract：:A sensitive procedure based on the use of iodinated antibody allows the detection of antigenic material by double-diffusion analysis. An autoradiographed immunodiffusion pattern reveals distinct radio-labeled precipitation lines for an extract of yeast with wild type HIS4 protein and for extracts of mutants containin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90048-0
更新日期：1978-01-01 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：:Neisseria meningitidis is a leading cause of morbidity and mortality worldwide. The presently available capsular polysaccharide vaccine is poorly immunogenic in children under the age of 2 due to its T-independent (TI) nature. Efforts to overcome the TI response elicited by the polysaccharide vaccine have led to the d...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2003.08.005
更新日期：2004-02-01 00:00:00
abstract：:We report the development of an expression system for the production of soluble, calmodulin (CaM)-tagged proteins in Drosophila Schneider S2 cells and the subsequent use of these proteins for the selection of phage displayed antibodies. The CaM-tag permitted the purification of recombinant protein to >90% purity in a ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.08.003
更新日期：2006-10-20 00:00:00
abstract：:Monoclonal antibodies (Mab) are the fastest growing group of biopharmaceuticals in development. For production in mammalian cells, the four polypeptide chains of the immunoglobulin diheterotetramer must be assembled prior to exit from the endoplasmic reticulum. Various recombinant Mab expression vectors have been deve...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.10.010
更新日期：2007-01-10 00:00:00
abstract：:γδ T cells function as sentinels in early host response to infections and malignancies. Although γδ T cells are regarded as innate immune cells and recognize antigens in a non-MHC restricted manner, they possess a huge diversity of complementary determinant region 3 (CDR3) of T cell receptor (TCR) generated by the rea...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.01.009
更新日期：2017-04-01 00:00:00
abstract：:A microscale ELISA system, designated as the 'C1q-micro ELISA,' has been developed for the measurement of circulating immune complexes. The sensitivity of the method ranged between 0.1 and 3 microgram equivalents of aggregated human gamma-globulins. The procedure requires small quantities of serum and reagents, and it...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90308-8
更新日期：1982-01-01 00:00:00
