Abstract：
:Testing of diphtheria vaccines for routine lot release relies heavily on the use of in vivo potency assays. However, consistency of production is also recognised as an important feature of vaccine manufacture, and in vitro assays are superior to in vivo assays for providing this information. In adsorbed vaccines, antigen and adjuvant are the major components contributing to immunogenicity and are therefore critical factors to be evaluated as part of consistency testing. Here we describe a simple and sensitive Enzyme Linked Immunosorbent Assay (ELISA) which has been developed to quantify diphtheria toxoid antigen in combined vaccine products and can also be used to monitor the degree of adsorption. This assay can be applied to a variety of multi-component vaccines and is robust, specific and highly sensitive, with a limit of quantification of approximately 0.005 Lf/ml. The antigen assay is an excellent test to characterise vaccines and monitor trends in production. For well established vaccines, the antigen assay could be used alongside other in vitro methods to provide a more informative product profile, with the ultimate aim of reducing the requirement for in vivo potency assays and therefore the number of animals required for routine batch release testing.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Coombes L,Stickings P,Tierney R,Rigsby P,Sesardic Ddoi
10.1016/j.jim.2009.09.002subject
Has Abstractpub_date
2009-10-31 00:00:00pages
142-9issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(09)00282-8journal_volume
350pub_type
杂志文章abstract：:Antibody avidity, defined as the strength of binding between antibody and antigen, represents a functional measure of affinity maturation of antibodies. Determination of the antibody avidity is usually performed separating high and low avidity antibodies by dissociating agents, but measurement of the antibody avidity ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2012.09.008
更新日期：2013-01-31 00:00:00
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：:A microtechnique to study lymphocyte stimulation in solid cultures by autoradiography is described. Using this technique at a concentration of 2 x 10(6) lymphocytes/chamber, it was found that the percentage of proliferating cells was 8.4% with anti-lymphocyte serum, 3.9% with calcium ionophore A23187, 2.5% with phytoh...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(79)90251-5
更新日期：1979-01-01 00:00:00
abstract：:We have previously described a monoclonal antibody (BU-1) to 5-bromo-2-deoxyuridine (BrdUrd) that is useful for measurement of cell cycle S-phase. BU-1 hybridoma supernatant reacted with incorporated BrdUrd after the cells had been ethanol fixed; without a requirement for acid or base denaturation. We have found that ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90438-2
更新日期：1986-10-23 00:00:00
abstract：:Staphylococcus aureus causes a variety of infections. Knowledge about the physiological role of most S. aureus antigens in colonization and infection is only limited. This can be studied by measuring antigen-specific antibody responses. In this study, we optimized the multiplex microsphere bead-based flow cytometry te...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.12.013
更新日期：2011-02-28 00:00:00
abstract：:A procedure is described for sensitive titration of antibodies, macromolecular antigens and haptens by enzyme immunoassay. It involves using first antigen or antibody labelled with bovine serum albumin (BSA) and then an anti-BSA antibody conjugated with an enzyme. The performance characteristics of this assay are indi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90270-3
更新日期：1983-03-11 00:00:00
abstract：:Immunochemical quantitative determination of a lipid antigen, ganglioside GM2, has been developed, based on the inhibition of the immune lysis of liposomes containing the antigen in their lipid bilayer. It has been shown that the full expression of the antigenicity of the competing lipid requires its dispersion in acc...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(77)90071-0
更新日期：1977-01-01 00:00:00
abstract：:In vitro differentiation of human monocytes (Mo) provides large amounts of mature and functionally competent macrophages (M phi) which may be used as potentially powerful anticancer agents for adoptive immunotherapy. Granulocyte macrophage-colony stimulating factor (GM-CSF) was evaluated for its ability to influence l...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(91)90046-i
更新日期：1991-10-25 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：:A solid-phase radioimmunometric binding assay is described utilizing 125I-labeled protein A for the detection of antibody to human melanoma associated antigens. The novel aspect of this assay is the use of chemically defined spent culture medium of melanoma cells at target antigens previously depleted of fibronectin b...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(80)90058-7
更新日期：1980-01-01 00:00:00
abstract：:An antibody binding-inhibition test is described, which allows the detection of P. falciparum in red blood cells (RBC) infected in vitro, using a crossreacting, monoclonal anti-P. berghei antibody and P. berghei coated microtiter plates. Experiments carried out to determine the coating efficiency of various P. berghei...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90231-5
更新日期：1985-09-03 00:00:00
abstract：:An immunoblot (Western) assay was developed employing a species-specific monoclonal antibody to a 43 kDa Mycoplasma pneumoniae membrane polypeptide and a species-specific monoclonal antibody to 29 kDa Legionella pneumophila outer membrane protein. This assay could simultaneously detect these two different antigens dir...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90291-2
更新日期：1987-11-05 00:00:00
abstract：:A radioimmunoassay (RIA) was developed for the quantitative measurement of a glycoprotein which was purified from human milk-fat-globule membrane (MFGM) and termed MFGM-pg 70. The assay with a sensitivity of detecting 30 ng of MFGM-pg 70/ml was employed to quantitate the levels of MFGM-gp 70 shed in supernatants from ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90231-0
更新日期：1986-11-20 00:00:00
abstract：:A novel enzyme-linked bio-immunoassay (bio-ELISA) has been developed to detect interferon-gamma (IFN-gamma) induced HLA-DR antigen on the surface of human tumor cells. Cells are cultured at 37 degrees C in 96-well microtiter plates in the presence of IFN-gamma for 2 days. After fixation with reagent alcohol the HLA-DR...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90083-5
更新日期：1989-12-20 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：:The existing methods for phagocytosis evaluation are inadequate for assessing all the real events occurring during phagocytosis, or for continuously following the kinetics of the process. Our purpose is to establish the use of luminous bacteria as an object for phagocytosis. The bioluminescence test offers an easy and...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90443-x
更新日期：1983-11-25 00:00:00
abstract：:A series of monoclonal antibodies has been prepared against a granulocyte antigen previously shown to be present in the serum of individuals homozygous or heterozygous for the cystic fibrosis (CF) gene. The specificity of the monoclonals was established by their ability to remove the antigen (CF antigen) from serum. T...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(86)90109-2
更新日期：1986-07-11 00:00:00
abstract：:The aim of the present study was to analyze the relations between the serum anti-erythropoietin antibody (AEAb) levels and the antibodies' neutralizing activity in 20 patients with renal anemia and rhEPO-induced antibodies. AEAb levels were determined by the enzyme-linked immunosorbent assay (ELISA, double antigen-bri...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2008.04.009
更新日期：2008-07-31 00:00:00
abstract：:The amino acid glutamine is an essential nutrient for cells in culture. In aqueous solutions such as liquid culture media, glutamine spontaneously decomposes into ammonia. In this study, we examined the toxicity of ammonia for two different cell lines. In mouse hybridoma cell cultures, viable cell counts were reduced ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90331-z
更新日期：1993-11-05 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
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(89)90238-x
更新日期：1989-06-21 00:00:00
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journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90119-e
更新日期：1992-07-06 00:00:00
abstract：:Measurement of the absorbance due to light scattering at 40 nm proved to be a simple and reliable way of assessing the extent of aggregation in heat-treated IgG solutions. Using this technique the rate of aggregation was demonstrated to be markedly temperature dependent with a sharp inflexion in the curve close to 63 ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90274-x
更新日期：1981-01-01 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：:One major obstacle in antibody production is the lack of highly purified immunogen. In this study, we describe an alternative strategy to circumvent this problem. A nucleotide sequence encoding a full-length of human survivin was cloned into pAK400cb. After transforming into an E. coli Origami B strain, survivin-bioti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.07.024
更新日期：2006-12-20 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 new and simple method for quantitation of IgM of low molecular weight in sera by radial immunodiffusion in 7% agar gel is described. The test is easy to perform and permits exact determination of low molecular weight IgM concentration down to very small quantities (1 mg%). Results obtained with the method described ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(78)90094-7
更新日期：1978-01-01 00:00:00
abstract：:Serologically and biochemically pure HLA antigens and other beta 2-microglobulin-associated proteins were isolated using a rapid immunoprecipitation technique with a monoclonal anti-beta 2-microglobulin reagent A88. Elution of the immunoprecipitate with beta 2-microglobulin isolated from kidney patients proved to be e...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90237-4
更新日期：1981-01-01 00:00:00
abstract：:The aim of the study was to develop a novel system permitting automated analysis of multicolor immunofluorescence-staining of cells in solid tissues which would be comparable to the analytical capacity of flow cytometry. In the user friendly automated data acquisition and image processing system which was established,...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00240-9
更新日期：2000-04-03 00:00:00
abstract：:The diagnostic and therapeutic applications of antibody single-chain Fv (sFv) fragments often require large amounts of protein that can be problematic and expensive to obtain. Here we report the secretion of two sFv fragments by the yeast Pichia pastoris at levels up to 250 mg/l. Soluble sFv fragments were purified fr...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(96)00213-x
更新日期：1997-02-14 00:00:00
