Abstract：
:Typical clinical grade human IgG (intravenous immunoglobulin, IVIG), used for carbohydrate analysis, is derived from thousands of healthy donors. Quantitative high-resolution glycan profiles of IgG and its Fc-Fab fragments are presented here. Glycan profiles were established following digestions with Fc specific endoglycosidase S and generic PNGase F under denaturing and non-denaturing (native) conditions. The native PNGase F glycan profile of IgG was similar (but not identical) to that of Endo S. Endo S profiles did not contain the glycans with bisecting GlcNAc. PNGase F glycan profiles were the same for Fc fragments that were isolated from pepsin and Ide S protease digests. Both isolated Fab fragments and the previously deglycosylated IVIG (native conditions) yielded the same glycan profile. Glycan profiles were established using high resolution HPLC with 2-aminobenzoic acid (2AA) labeling. An accurate determination of sialylation levels can be made by this method. Carbohydrate content in Fc and Fab was determined using an internal standard and corrected for both protein and glycan recoveries. Fab portion contained about 14% of the total carbohydrate which translates to 2.3 sugar chains per mol in IVIG where 2 chains are located in the CH2 domain of the Fc. Fc glycans consisted of neutral (N) 84.5%; mono-sialylated (S1) 15% and di-sialylated (S2) 0.5%. In contrast, Fab contained N, 21%; S1, 43% and S2, 36%. The distribution of bisecting N-acetylglucosamine and fucose was found to be very different in various glycans (N, S1 and S2) found in Fab and Fc. Total IgG glycan profile (Fab plus Fc) contained N, 78.5%; S1, 17% and S2, 4.5%. Percent distribution of glycans G0, G1 and G2 (with 0, 1 and 2 two galactoses) was 26, 49 and 25 respectively within the 78% of the neutral glycans. Glycan profiles were nearly the same for various clinical grade IVIG preparations from various manufacturers. A fast HPLC profiling method was developed for the separation and quantitation of IgG glycans (neutral (G0, G1, and G2), mono- and di-sialylated) using simple procedures. The method should prove useful for monitoring glycan changes in clinical settings.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Anumula KRdoi
10.1016/j.jim.2012.05.022subject
Has Abstractpub_date
2012-08-31 00:00:00pages
167-76issue
1-2eissn
0022-1759issn
1872-7905pii
S0022-1759(12)00160-3journal_volume
382pub_type
杂志文章abstract：:A method for the production of human gamma interferon (HuIFN-gamma), which is both simple and efficient, has been developed. The 2 co-inducers, A-23187 and mezerein, stimulate human peripheral blood lymphocytes to produce high levels of HuIFN-gamma in both stationary cultures and spinner cultures. The compound 2,2-dim...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(83)90428-3
更新日期：1983-10-14 00:00:00
abstract：:We describe an assay and data evaluation technique for sorting a panel of murine monoclonal antibodies according to epitope specificities. The assay analyzes the simultaneous binding (pairing) of antibodies to an antigen and groups together antibodies with similar pairing profiles. Similar profiles indicate that the a...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2010.12.021
更新日期：2011-02-28 00:00:00
abstract：:A flow cytometry-based assay for analyzing cytotoxic T lymphocyte (CTL) activity is presented. This new approach is characterized by easy handling, the generation of highly reproducible data sets and is not dependent on the use of radioactivity. Before exposure to primed CTL effector cells the target cells were labele...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00047-1
更新日期：1997-05-26 00:00:00
abstract：:Monoclonal antibodies can now be generated against a wide variety of antigens. However, a potentially useful monoclonal antibody may be of the wrong class or subclass for a particular task. Antibodies of the desired class can be obtained by identifying rare subclones in which the variable region has been rearranged to...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90298-9
更新日期：1984-11-30 00:00:00
abstract：:Bacillus anthracis spores and Escherichia coli were stained with fluorescein-conjugated antibody using direct and indirect methods, then analyzed by means of a commercial flow cytometer. To reduce the cytometer's fluorescence component resulting from unreacted conjugate, reaction mixtures were either diluted or were c...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(87)90153-0
更新日期：1987-08-03 00:00:00
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：:Leukemia inhibitory factor (LIF), oncostatin-M (OSM), ciliary neurotrophic factor (CNTF) and cardiotrophin-1 (CT1) act through transmembrane receptors which share the gp190 glycoprotein chain. The understanding of its involvement in the biology of these cytokines is of importance since these systems have recently been...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(97)00074-4
更新日期：1997-07-14 00:00:00
abstract：:A rapid quantitative flow cytometric (FCM) assay for the combined kinetic measurement of phagocytosis and intracellular killing of fluorescein-isothiocyanate (FITC) labelled Candida albicans is described. The fraction of phagocytosing leukocytes and the numbers of attached or internalized Candida albicans per phagocyt...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90451-4
更新日期：1984-08-03 00:00:00
abstract：:A new assay designed to quantitate antibody reactivity to specific peptides using biospecific interaction analysis (BIAcore) has been developed. Peptides of various lengths (15-40 amino acids) and isoelectric points (pI = 4.5-13) were covalently linked (immobilized) to a biosensor and interacted with polyclonal human ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(92)90225-i
更新日期：1992-02-05 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：:As interest in the development of oral vaccines continues to rise, alternative animal models for studies of mucosal immunity are needed. The present study examines a simplified procedure for delivering antigen to rabbit Peyer's patches via an indwelling cannula. The cannula was placed 3-4 cm proximal to the Peyer's pa...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(96)00055-5
更新日期：1996-07-17 00:00:00
abstract：:Three different methods for the radioiodination of surface proteins in trypanosomatids have been comparatively analyzed. Data interpretation in the lactoperoxidase-mediated method was impaired by autoiodination of the enzyme and its tight retention by the cells. In the enzymobeads method, iodinated enzymes were releas...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(82)90051-5
更新日期：1982-07-30 00:00:00
abstract：:We describe here a specific and sensitive assay for biologically active bovine type-I interferon (IFN) in an Mx/CAT reporter gene assay. The assay is based on Madin-Darby Bovine Kidney cells transfected with a plasmid, containing a human MxA promoter driving a chloramphenicol acetyltransferase (CAT) cDNA. CAT expressi...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(00)00359-8
更新日期：2001-03-01 00:00:00
abstract：:A solid-phase chemiluminescence immunoassay for determining immunoglobulin G (IgG) in eluates of psoriatic scales is described. Polystyrene balls coated with protein A bound the IgG present in the eluates and in human sera. The bound IgG was then detected by a luminol-coupled goat anti-human IgG conjugate, characteriz...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90184-4
更新日期：1984-05-25 00:00:00
abstract：:Respiratory enteric orphan virus (reovirus) has been used to study many aspects of the biology and genetics of viruses, viral infection, pathogenesis, and the immune response to virus infection. This report describes the functional activity of virus labeled with Alexa Fluor 488, a stable fluorescent dye. Matrix assist...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2006.05.008
更新日期：2006-07-31 00:00:00
abstract：:A new radioimmunoassay has been developed for measuring IgM and IgG rheumatoid factors. Diluted sera from donors and patients were incubated with immunoprecipitates prepared from sheep serum and rabbit anti-sheep IgG antiserum. The precipitates were washed, and radioiodinated rabbit F(ab')2 fragments specific from hum...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(81)90259-3
更新日期：1981-01-01 00:00:00
abstract：:Helicobacter pylori infection is the most common human infection where approximately 50% of the world populations are infected. The diagnosis of such infection is mainly done by endoscopy where gastric biopsies are examined for the presence of H. pylori. Such invasive approach is costly, time consuming and generally r...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.08.010
更新日期：2013-10-31 00:00:00
abstract：:Plasma cells (PCs) secrete antibodies and play an essential role in protective immunity, but also in pathogenesis of antibody-mediated diseases. Physiologically, PCs mainly reside within bone marrow and spleen. In autoimmune diseases such as systemic lupus erythematosus (SLE) autoantibody-producing PCs can also be fou...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2019.06.019
更新日期：2019-11-01 00:00:00
abstract：:Macrophages can be separated from other cell types by their ability to readily attach and spread on glass or on plastic surfaces which are treated for optimal growth of cultured cells (tissue culture-treated plastic). To detach macrophages from these surfaces, techniques must be used which require prior preparation of...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(84)90142-x
更新日期：1984-03-30 00:00:00
abstract：:An enzyme-linked immunosorbent assay (ELISA) for determination of antibodies against the zona pellucida was developed and compared with the already available indirect immunofluorescence (IIF) technique. Sera from 100 women with explained and unexplained infertility were screened for the presence of autoantibodies to t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90399-0
更新日期：1985-05-10 00:00:00
abstract：:Monoclonal antibodies have been prepared against a synthetic peptide with a sequence corresponding to a repeated hydrophilic region of the protein core of the human MUC-2 gastrointestinal mucin. Peptide conjugates, prepared by glutaraldehyde cross-linking with keyhole limpet haemocyanin (KLH) and bovine serum albumin ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(93)90168-7
更新日期：1993-02-26 00:00:00
abstract：:To date, in vivo apoptosis within the thymus has been assessed using morphological criteria and/or detection of a DNA ladder indicative of oligonucleosomal fragmentation of the DNA. Here, we have used a fluorometric method to investigate activation of the caspase protease family in the thymus following in vivo inducti...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(99)00054-x
更新日期：1999-06-24 00:00:00
abstract：:Human CD1d-restricted natural killer T (NKT) cells, which are postulated to regulate the immune response in several clinical settings, can be activated by alpha-galactosylceramide (alpha-GalCer) presented by CD1d molecules on antigen presenting cells (APCs). Simple methods to quantify NKT function in fresh blood will ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(02)00497-0
更新日期：2003-01-15 00:00:00
abstract：:The performance of three basal media RPMI, DMEM/F12 (DF) and eRDF (enhanced RDF, RPMI:DMEM:F12 in 2:1:1) were evaluated in cultures with and without serum with respect to cell proliferation, metabolism and monoclonal antibody (Mab) productivity. Based on the ease of adaptation, growth rate, maximum cell density and Ma...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(94)90080-9
更新日期：1994-01-03 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：:The purpose of this study was to develop a flow cytometric method for the detection of adenosine deaminase (ADA) in a single cell suspension of mononuclear cells. Anti-human ADA antibody was purified by affinity chromatography on a column of Sepharose 4B to which calf ADA was covalently linked. This antibody was used ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(85)90017-1
更新日期：1985-06-25 00:00:00
abstract：:Foot and mouth disease is caused by a non-enveloped virus (FMDV), which disposes several antigenic sites at the surface of their capsid proteins. The most relevant and immunodominant antigenic site of FMDV (site A or AnSA) includes a key virus-cell interaction element (RGD motif) located in the Viral Protein 1 (VP1), ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2015.06.008
更新日期：2015-10-01 00:00:00
abstract：:Interferon gamma (IFNγ) is a cytokine involved in many anti-viral and immunoregulatory processes. One of the major mechanisms through which IFNγ exerts these effects is by inducing expression of indoleamine 2,3 dioxygenase-1 (IDO1), an enzyme that catalyses the first, rate-limiting step of the kynurenine pathway. In t...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/j.jim.2013.05.011
更新日期：2013-08-30 00:00:00
abstract：:We describe an ex vivo rat infection model to study protective immunity against Fasciola hepatica at the gut level. An exact number of newly excysted juveniles (NEJs) was injected into a gut segment with an intact blood supply and which was still attached to a live anaesthetized rat. NEJs that penetrated the gut wall ...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/s0022-1759(98)00026-x
更新日期：1998-04-15 00:00:00
abstract：:The reverse haemolytic plaque assay has been adapted to detect and measure the release of such cytokines as interleukin-1, -2 and -6, GM colony-stimulating factor or interferon-gamma by individual human cells derived from either peripheral blood or enzymatically dispersed breast carcinomas. Since each of these peptide...
journal_title：Journal of immunological methods
pub_type： 杂志文章
doi：10.1016/0022-1759(90)90340-2
更新日期：1990-02-20 00:00:00
