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-drug conjugates followed by FITC-conjugated anti-mouse immunoglobulin antibodies in an indirect assay or with FITC-conjugated MoAbs specific for the tumor target cell line in a competition assay. The amount of fluorescent probe bound is measured by flow cytometry and the mean fluorescence intensity determined. The relative binding capacity is quantified by linear regression of the mean fluorescence versus the concentration of primary antibody or antibody conjugate. The application of these techniques to several drug and toxin conjugates of MoAb KS1/4, which defines a human adenocarcinoma-associated antigen, demonstrates that these assays can be effectively utilized to monitor the effects of covalent chemical modification on a MoAb's antigen binding reactivity.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Marder P,Apelgren LD,Bumol TFdoi
10.1016/0022-1759(87)90310-3subject
Has Abstractpub_date
1987-02-11 00:00:00pages
165-70issue
2eissn
0022-1759issn
1872-7905pii
0022-1759(87)90310-3journal_volume
96pub_type
杂志文章abstract::Antigen receptor-mediated activation of T and B lymphocytes results in activation of phospholipase C-gamma isozymes with subsequent hydrolysis of membrane inositol phospholipids. As a method of screening autoimmune or immunodeficient patients for early receptor signaling defects, we have developed a rapid technique fo...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(95)00135-w
更新日期:1995-10-12 00:00:00
abstract::This study was designed to assess the use of flow cytometry to observe the binding, under physiological conditions, of anti-lipopolysaccharide (LPS) monoclonal antibodies (mAbs) to whole bacteria, and to compare this with the more conventional whole cell ELISA and immunoblotting techniques. The bacteria consisted of t...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90363-z
更新日期:1990-10-19 00:00:00
abstract::Helper and suppressor activity of T cell lines were investigated using an assay system which is based on measurement by the enzyme-linked immunosorbent assay (ELISA) of the antibody produced in vitro. T cell lines were established from spleens of BALB/c mice immunized with ovalbumin and human serum albumin. The T cell...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(83)90063-7
更新日期:1983-02-25 00:00:00
abstract::The aim of this study was to evaluate the performance of conventional serology (Q-Preven™ and ELFAVIDAS™) and flow cytometry-based serologic tools for early serologic diagnosis of congenital toxoplasmosis. The study groups included prospectively confirmed cases of congenital toxoplasmosis (TOXO=88) and age-matching no...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.08.005
更新日期:2017-12-01 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::To reduce the cost, time and waste in screening for HIV and HBV infections a combined assay for HIV-1 and -2 antibodies and HBsAg has been developed. Monoclonal anti-HBs antibodies were co-immobilized with synthetic peptides representing immunodominant regions of HIV-1 and -2. The presence of anti-HIV antibodies in th...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90262-6
更新日期:1993-01-14 00:00:00
abstract::A method is described for the purification of monoclonal antibody from mouse ascitic fluid. The fluid is clarified and the lipid removed using silicon dioxide powder, before the immunoglobulin is precipitated using polyethylene glycol. The method provides IgM antibody in high yield and good purity. In the case of IgG ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(86)90483-7
更新日期:1986-07-24 00:00:00
abstract::Two micro enzyme immunoassays (microEIA) for circulating immune complexes (CICs) are described. The Raji cell microEIA was similar in sensitivity, reproducibility and specificity to the Raji radioimmunoassay. The F(ab')2 anti-C3 microEIA was comparable to the 2 Raji cell assays. The 2 microEIAs for CICs were used to a...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90258-3
更新日期:1985-11-07 00:00:00
abstract:BACKGROUND:Accumulating evidence intensively advises circulating endothelial progenitor cells (EPCs) and monocyte subsets as surrogate cellular biomarkers in cardiovascular and cancer disease. However, a general standard on their quantification is still elusive, thus precluding a routine monitoring and comparative inte...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2012.04.003
更新日期:2012-07-31 00:00:00
abstract::Although serum-free media have been used to expand lymphokine-activated killer cells, antigen-specific CD8 T cell cytotoxicity does not develop in vitro in the absence of serum. The immunodominant Vbeta17 response to an influenza A matrix protein epitope restricted by HLA A2.1 was used to study the serum requirement f...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00026-5
更新日期:1999-05-27 00:00:00
abstract::We have developed a large scale (10 g/batch) non-chromatographic method for the collection of IgM directly from plasma obtained from therapeutic plasmapheresis of patients. The technique uses sequential precipitations from ammonium sulfate and polyethylene glycol, and gives high yields of undenatured IgM antibodies. T...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90252-3
更新日期:1993-01-14 00:00:00
abstract::Ethanol-fixed paraffin-embedded specimens of human tissues were studied whether the surface antigens of leukocytes in these tissues can be stained and analyzed. Three-layer indirect immunoperoxidase staining was performed on the ethanol-fixed paraffin-embedded sections by the use of several monoclonal antibodies for w...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90189-3
更新日期:1990-08-07 00:00:00
abstract::Nitrocellulose was activated with divinyl sulfone, a spacer of ethylenediamine, and glutaraldehyde. The aldehyde groups on the activated nitrocellulose, Nit-CHO, were stable through one month at 4 degrees C. Peptides were attached to the membrane by reaction of the amino group with the free carbonyl, forming peptide b...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(90)90197-4
更新日期:1990-08-07 00:00:00
abstract::A 24 h, highly sensitive assay for detection of interleukin 1 (IL-1) is described. A thioguanine-resistant mutant of the murine lymphoma cell line LBRM 33 was selected (LBRM TG6). When this cell line was incubated with low concentrations of PHA and IL-1 it produced interleukin 2 (IL-2). IL-2-dependent HT2 cells were c...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(85)90389-8
更新日期:1985-05-10 00:00:00
abstract::Human interleukin-7 (IL-7) is a cytokine that appears to be critical for early T- and B-cell development and although IL-7 is currently under investigation as a therapeutic agent in a variety of hematolymphopoietic disorders, there have been few instances of the detection or investigation of this cytokine using a biol...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(99)00237-9
更新日期:2000-03-06 00:00:00
abstract::Ferromagnetic iron dextran particles were prepared by reacting a mixture of ferrous chloride and ferric chloride with dextran polymers under alkaline conditions. Particles purified by gel filtration chromatography were in the size range of 30-40 nm, had an electron dense core of about 15 nm, were stable against aggreg...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90007-2
更新日期:1982-08-13 00:00:00
abstract::Within EuroFlow, we recently developed screening tubes for hematological malignancies and immune deficiencies. Pipetting of antibodies for such 8-color 12-marker tubes however is time-consuming and prone to operational mistakes. We therefore evaluated dried formats of the lymphocytosis screening tube (LST) and of the ...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2017.03.011
更新日期:2019-12-01 00:00:00
abstract:BACKGROUND:High concentrations of propidium iodide (PI), in combination with fluorescein isothiocyanate (FITC) and R-phycoerythrin (RPE) used for multiparameter DNA flow cytometry (FCM), cause spectral cross-talk into the green fluorescence channel (FL1). We have evaluated the use of post-acquisition software compensat...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/s0022-1759(01)00550-6
更新日期:2002-02-01 00:00:00
abstract::The majority of vaccines and several treatments are administered by intramuscular injection. The aim is to engage and activate immune cells, although they are rare in normal skeletal muscle. The phenotype and function of resident as well as infiltrating immune cells in the muscle after injection are largely unknown. W...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2015.06.011
更新日期:2015-10-01 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::[75Se]selenomethionine (75SeM) has been shown to provide several advantages over Na(2)51CrO4 (51Cr) labelling of metabolizing target cells: high labelling efficiency and low spontaneous release of 75SeM-labelled target cells permit improved monitoring of cytotoxicity due to extended effector/target ratios in short- an...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(93)90297-k
更新日期:1993-05-26 00:00:00
abstract::A method for synthesising omega-stearoyldextran for the detection of plaque forming cells specific for the alpha-(1 replaces 6) determinant is described. The omega-stearoyl derivative is obtained by reacting dextran with phenylboronic acid to give a 2,4-phenylboronate, which can be subsequently esterified in a pyridin...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90206-4
更新日期:1976-01-01 00:00:00
abstract::This paper reports a new ELISA to measure the level of rabies anti-glycoprotein G antibodies after vaccination. The Platelia Rabies II kit was evaluated on different populations of dogs, cats and foxes. For each target species, sera from naive, unvaccinated and vaccinated animals were tested. Platelia Rabies II result...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2006.07.026
更新日期:2007-01-10 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::A method is described for the purification of human and guinea pig C3 from small amounts of serum. This procedure requires only two steps--polyethylene glycol (PEG) precipitation and fast protein liquid chromatography (FPLC) Mono Q HR 10/10 ion exchange chromatography. The protocol takes less than two hours to complet...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(91)90290-v
更新日期:1991-08-28 00:00:00
abstract::Macrophages are an important part of the cellular immune system and play a key role during immune responses. Thus, macrophages are interesting targets in basic and clinical research. Primary monocytes or monocyte-derived macrophages do not proliferate on a suitable scale so that their use for functional studies in vit...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2009.03.014
更新日期:2009-05-31 00:00:00
abstract::Monoclonal antibodies were used to demonstrate hybrid forms of human alkaline phosphatase (ALP) composed of subunits from both placental and intestinal loci. Four main isozymes with alkaline phosphatase activity appear on polyacrylamide gels of the HeLa cell line, Hep 2/5 after electrophoresis. The mobilities of all 4...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(82)90071-0
更新日期:1982-11-26 00:00:00
abstract::Accurately predicting peptides binding to major histocompatibility complex (MHC) I molecules is of great importance to immunologists for elucidating the underlying mechanism of immune recognition and facilitating the design of peptide-based vaccine. Various computational methods have been developed for MHC I-peptide b...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/j.jim.2010.09.007
更新日期:2011-11-30 00:00:00
abstract::A one-step immunospecific affinity chromatographic method for purification of mouse alpha-fetoprotein (AFP) directly from amniotic fluid is described. The procedure employed a rabbit anti-AFP immunoglobulin-G entrapped in a polyacrylamide gel matrix prepared as a gel slurry. The presence of impurities could not be dem...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(79)90146-7
更新日期:1979-01-01 00:00:00
abstract::A method is described for determining the percentage of rosettes formed by sheep erythrocytes and human peripheral lymphocytes (ER) using cytofluorographic analysis (CFGA). The technique utilizes acridine orange dye for differentially staining nucleus and cytoplasm of the lymphocytes to distinguish them from erythrocy...
journal_title:Journal of immunological methods
pub_type: 杂志文章
doi:10.1016/0022-1759(76)90091-0
更新日期:1976-01-01 00:00:00