Abstract：
:This paper describes the application of experimental design techniques to optimize a sensitive ELISA for a hapten molecule with a calibration range of 0-1000 pg/ml. Ten factors that were expected to affect the assay performance were initially screened, followed by factorial experiments to delineate the effects of the critical factors identified at the screening stage. Assay performance was evaluated using a unique rating system based on standard curve reproducibility, assay detection limits and the use of desirability functions. This rating system allowed multiple responses to be evaluated simultaneously. It was found that the substrate incubation time and enzyme label lot played an important role, while dilutions of the enzyme label and the anti-hapten antibody showed significant interaction. These observations were in good agreement with optimal assay conditions based on historical data collected over a period of two to three years. Application of experimental design techniques enabled us to confirm the significance of the factors affecting the assay within a three month period, with a minimum number of experiments. In addition, information on interaction between factors were determined.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Sittampalam GS,Smith WC,Miyakawa TW,Smith DR,McMorris Cdoi
10.1016/0022-1759(95)00262-6subject
Has Abstractpub_date
1996-04-19 00:00:00pages
151-61issue
2eissn
0022-1759issn
1872-7905pii
0022175995002626journal_volume
190pub_type
杂志文章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：:Drugs that target tumor necrosis factor-alpha (TNF) are particularly important in the treatment of severe inflammatory progression in rheumatoid arthritis, Crohn's disease and psoriasis. Despite the central role of the TNF/TNF receptor (TNFR) in various disease states, there is a paucity of information concerning TNFR...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.02.019
更新日期：2008-06-01 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：:There is an emerging trend in the pharmaceutical industry to evaluate a variety of surrogate biomarkers in Phase I/II clinical studies with the intention of determining potential activity of drugs early in clinical development. A number of cytokines expressed in pathological conditions are currently being considered a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(01)00543-9
更新日期：2002-02-01 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 new strategy has been developed for enzyme immunoassay of small molecules. The method is based on the formation and estimation of the complex Ab.H-H.Ab* where Ab and Ab* are immobilised and labeled antibodies respectively and H-H is a synthetic bis-analogue of the hapten H. The amount of the complex formed is invers...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(12)80005-6
更新日期：1992-03-04 00:00:00
abstract：:An ultra-sensitive quantitative EPO (erythropoietin) lateral flow immunochromatographic test with a detection limit of 1.2 fM (10(-15) M), 0.035 ng EPO/L, which is 50-100 times more sensitive than a corresponding enzyme based immunoassay, is presented. In comparison with commercial lateral flow tests for other analyte...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.09.022
更新日期：2008-12-31 00:00:00
abstract：:Chinese hamster lymphocyte cultures were supplemented with foetal calf serum (FCS) or hose serum (HS). Addition of 2-mercaptoethanol (2-ME) to cultures supplemented with these sera resulted in a marked increase lymphocyte proliferation. The PHA concentrations necessary for optimal lymphocyte stimulation were much lowe...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90299-x
更新日期：1982-01-01 00:00:00
abstract：:The structural diversity of the immune response may be considerably restricted by the structure of the hapten used to elicit catalytic antibodies. The ligand-binding mode and the shapes of the binding pockets of hydrolytic antibodies induced to different transition-state analogs that contain an unsubstituted arylphosp...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(02)00240-5
更新日期：2002-11-01 00:00:00
abstract：:Dendritic Cell (DC)-based vaccination approaches in man require a reproducible DC generation method that can be performed in conformity with GMP (Good Manufacturing Practice) guidelines and that circumvents the need for multiple blood drawings to generate DC. To this end we modified our previously described method to ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,评审
doi：10.1016/s0022-1759(98)00208-7
更新日期：1999-02-01 00:00:00
abstract：:The reductive methylation procedure for proteins in solution has been adapted for tritium radiolabelling of surface components of macrophages, including the Fc gamma receptor. High specific radioactivity of the receptor was obtained with no loss of IgG binding activity, tested by the solid-phase radioassay. To prepare...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90390-4
更新日期：1985-05-10 00:00:00
abstract：:The in vitro migration inhibition responses of peripheral blood leukocytes from tuberculin skin test positive and negative donors were tested to validate and determine optimal conditions for the agarose droplet method in the human. In vitro migration inhibition was observed in skin test positive donors in the presence...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90063-7
更新日期：1978-01-01 00:00:00
abstract：:A method to purify cobra venom factor (CVF) from cobra venom by sequential column chromatography is described which yields a product virtually free of phospholipase A2, a common contaminant of CVF preparations. The separation of phospholipase A2 from CVF was achieved by chromatography on Cibacron blue-agarose, a resin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90045-0
更新日期：1984-10-12 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：:We describe techniques to detect nuclear transcription factors in thymocyte subsets using flow cytometry. We have adapted a method that minimizes autofluorescence of fixed cells, thereby allowing the detection of proteins expressed at low levels. An accompanying method in which the cytoplasm is removed from stained ce...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(03)00184-4
更新日期：2003-08-01 00:00:00
abstract：:Hybridoma cells producing antibody reactive with poly(A) were isolated following fusion of spleen cells from a BALB/c mouse immunized with poly(A).poly(dT) and myeloma cells. The antibody was purified from ascitic fluid by the formation of immune complexes with poly(A). The antibody reacted with poly(A) and some homop...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90269-d
更新日期：1993-08-26 00:00:00
abstract：:Based on the observation that binding of IgM cytophilic antibodies to lymphocytes is temperature dependent, a direct plaque forming cell (PFC) assay was developed to detect IgM-receptor bearing human peripheral blood T and B lymphocytes. Lymphocytes were passively sensitized with IgM anti-SRBC molecules at 4 degrees C...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90330-2
更新日期：1979-01-01 00:00:00
abstract：:Two sensitive monoclonal antibody (MAb)-based enzyme-linked immunosorbent assays (ELISAs), one for activin A (homodimer of beta A subunits) and one for activin B (homodimer of beta B subunits) in plasma have been developed. The activin A ELISA had an effective range of 0.2-50 ng/ml while the activin B ELISA's range wa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90100-l
更新日期：1993-09-27 00:00:00
abstract：:After B lymphocyte depletion, blood lymphocytes (PBMC) from certain individuals will generate substantial quantities of immunoglobulin (Ig) when cultured with pokeweed mitogen (PWM). The present investigations were aimed at analysing the cell subset in these T cell preparations, that was responsible for Ig production ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90372-7
更新日期：1984-11-16 00:00:00
abstract：:A mild procedure for the purification of rabbit Fab' antibodies is described. A small column of normal goat IgG-Sepharose 4B (3 cm x 5mm) was saturated with F(ab')2 prepared from rabbit anti-goat IgG, and anti-goat IgG Fab was eluted from the column by splitting the disulfide bond in the hinge of the F(ab')2 molecule ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90336-1
更新日期：1980-01-01 00:00:00
abstract：:Several methods have been used to measure anti-double-stranded DNA auto-antibody (anti-dsDNA). Our aim was to determine the most efficient strategy to test anti-dsDNA in systemic lupus erythematosus (SLE). In this study, anti-dsDNA and anti-nuclear antibody (ANA) tests were requested for 644 patients. Anti-dsDNA was t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.09.003
更新日期：2015-12-01 00:00:00
abstract：:Etanercept, a recombinant human tumor necrosis factor (TNF) receptor Fc fusion protein is an effective treatment option in adults with rheumatoid arthritis, ankylosing spondylitis, psoriatic arthritis or plaque psoriasis and paediatrics with juvenile idiotypic arthritis and plaque psoriasis. Patent expiration in Europ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2017.03.007
更新日期：2017-08-01 00:00:00
abstract：:External proficiency testing programs designed to evaluate the performance of end-point laboratories involved in vaccine and therapeutic clinical trials form an important part of clinical trial quality assurance. Good clinical laboratory practice (GCLP) guidelines recommend both assay validation and proficiency testin...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2014.05.019
更新日期：2014-07-01 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：:Drug-specific monoclonal antibodies (MAbs) were produced against the very small drug hapten (162.15 Da), 5-benzimidazolecarboxylic acid, an analogue of 2-(4-Thiazolyl)benzimidazole (TBZ) but lacking the thiol group. TBZ is widely used as a broad-spectrum anthelmintic in various animal species and humans and also as a ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00340-x
更新日期：2002-12-20 00:00:00
abstract：:Three commercial second generation enzyme immunoassays (EIA) and a second generation immunoblot assay (RIBA-II) for detecting antibodies to hepatitis C virus (HCV) were evaluated in a study of confirmatory testing using sera referred by six Regional Blood Transfusion Centres (RTC). Of a total of 490 samples, 203 were ...
journal_title：Journal of immunological methods
pub_type： 杂志文章,多中心研究
doi：10.1016/0022-1759(93)90182-7
更新日期：1993-04-02 00:00:00
abstract：:A new method for measuring the uptake of Staphylococcus aureus by human polymorphonuclear neutrophils (PMN) using flow cytometry (FCM) is described. Bacteria were labeled with fluorescein isothiocyanate (FITC) and incubated with PMN in a suspension assay. At the end of the assay, phagocytosis was arrested by the addit...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90161-0
更新日期：1989-07-26 00:00:00
abstract：:In this report we show that succinic groups are far more reactive to amino compounds than the carboxylic groups derived from Asp and Glu on the protein when using coupling via 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDCI) (even by an 8 fold factor). Accordingly, a new carrier-protein was designed where both na...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2005.10.003
更新日期：2005-12-20 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：:The techniques of flow cytometry are being increasingly used to classify lymphocytes according to their surface antigens. Accurate analyses depend on the exclusion of non-lymphocytic contaminant cells. Using a fluorescence-activated cell sorter (FACS IV), virtually complete separation of human lymphocytes from monocyt...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90389-7
更新日期：1983-11-11 00:00:00
