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Antigenic Specificity

Antigenic Specificity refers to the unique immune recognition patterns exhibited by antibodies or T cells towards specific antigenic targets.
This highly selective process is fundamental to the adaptive immune response, allowing the body to mount targeted defenses against invading pathogens or altered self-cells.
Understanding the nuances of antigenic specificity is crucial for developing effective vaccines, diagnostic tools, and personalized immunotherapies.
Reasearchers leveraging PubCompare.ai's AI-powered platform can streamline their workflow, locate the best protocols, and maximize the accuracy of their antigenic specificity investigations.

Most cited protocols related to «Antigenic Specificity»

1
Immunodetection of IgLON5 Protein Specificity

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Publication 2014
Antibodies Antigenic Specificity Brain Cells Clone Cells Epithelial Cells Fluorescent Antibody Technique Green Fluorescent Proteins HEK293 Cells Homo sapiens Immunohistochemistry Immunoprecipitation Kidney Neurons Plasmids Serum
2
Evaluation of TIL Phenotype and Antigen Specificity
TIL cultures for patient treatment were generated as previously described.3 (link),13 (link) The phenotype of TIL was determined by fluorescence-activated cell sorting (FACS) analysis and the antigen specificity was assessed by cytokine-release assays using standard methods.3 (link),13 (link) The mean length of telomere repeats at chromosome ends in individual T-cell cultures was measured by quantitative flow-fluorescent in situ hybridization as previously described.14 (link)
Serum was collected from patients and frozen before initiation of therapy and after lymphodepletion but before cell infusion. Sera were thawed and assayed at the same time in a blinded fashion using commercially available enzyme-linked immunosorbent assay kits.
Dudley M.E., Yang J.C., Sherry R., Hughes M.S., Royal R., Kammula U., Robbins P.F., Huang J., Citrin D.E., Leitman S.F., Wunderlich J., Restifo N.P., Thomasian A., Downey S.G., Smith F.O., Klapper J., Morton K., Laurencot C., White D.E, & Rosenberg S.A. (2008). Adoptive Cell Therapy for Patients With Metastatic Melanoma: Evaluation of Intensive Myeloablative Chemoradiation Preparative Regimens. Journal of Clinical Oncology, 26(32), 5233-5239.
Publication 2008
Antigenic Specificity Cells Chromosomes Cytokine Enzyme-Linked Immunosorbent Assay Fluorescent in Situ Hybridization Freezing Patients Phenotype Serum T-Lymphocyte Telomere Therapeutics
3
Expansion and Characterization of Tumor-Infiltrating Lymphocytes
Lymphocytes were grown from resected metastatic melanoma lesions in high-dose IL-2 as previously described16 (link)–18 (link). In brief, individual cultures in 24 well culture plates were established from either single cell suspensions or 1–2 mm3 fragments. The wells were individually grown and expanded and tested for the presence of antigen specificity utilizing overnight cytokine release coculture assays against either autologous tumor or HLA matched melanoma cell lines. Cultures with evidence of specific reactivity compared to allogeneic non-MHC matched controls that exceeded 200 pg/ml of interferon gamma and were at least twice control values were selected for rapid expansion as previously described16 (link), 17 (link).
The average length of telomere repeats in chromosomes from the infused cells was measured by quantitative flow-FISH as previously described19 (link). The number and percent of CD8+ CD27+ cells in the infused TIL was measured after withdrawal of IL-2 for two days20 (link). The persistence of the infused cells in the circulation was measured by amplifying TCR BV region sequences using the SMART RACE cDNA Amplification kit (Clontech Laboratories, Inc., Mountain View, CA) and comparing clonotypes in the infused TIL with those in the circulation at one month using techniques previously described21 , 22 (link).
Rosenberg S.A., Yang J.C., Sherry R.M., Kammula U.S., Hughes M.S., Phan G.Q., Citrin D.E., Restifo N.P., Robbins P.F., Wunderlich J.R., Morton K.E., Laurencot C.M., Steinberg S.M., White D.E, & Dudley M.E. (2011). Durable Complete Responses in Heavily Pretreated Patients with Metastatic Melanoma Using T Cell Transfer Immunotherapy. Clinical cancer research : an official journal of the American Association for Cancer Research, 17(13), 4550-4557.
Publication 2011
Antigenic Specificity Biological Assay CD8-Positive T-Lymphocytes Cell Lines Chromosomes Coculture Techniques Cytokine DNA, Complementary Fishes Interferon Type II Lymphocyte Melanoma Neoplasms Telomere
4
Robust Clustering of Immune Repertoire Sequences
Related reads from each sample were clustered using parameters that best clustered together reads arising from the same cell. To be considered part of the same cluster, reads were required to have the same V and J gene annotation, the same length CDR3, and a similar CDR3 AA sequence, with one AA mismatch per 12 AAs allowed (length < 12 AA = 1 mismatch, 12 AA <= length < 24 AA = 2 mismatches, etc). The same D gene annotation was not required for inclusion in the same cluster, as somatic hypermutation in this region makes accurate assignment of a gene segment difficult. We focused on the CDR3 region of the sequence, as it is highly variable, and has the dominant role in determining antigenic specificity of the sequence.18 (link) Clusters were iteratively defined using an approach to identify cluster centers that gave the largest possible clusters. Briefly, clustering started with a set of unique sequences of the same length U(L) and an empty set of clusters C(L). The first cluster center is defined as the sequence x∈U(L) that had the most neighbor’s |N(x)|; the set N(x) is then added to C(L), with cluster center x. N(x) is then removed from U(L), and the process repeated until U(L) is empty and C(L) is full. Clusters were then collapsed into single reads, with each cluster represented by its cluster center sequence, V D and J gene usage, isotype subclass, and average V gene mutation. Where a single cluster contained multiple reads with different isotype subclass or D gene usage, the most frequent usage within the cluster was used for cluster annotation. These collapsed clusters were used in all subsequent analysis, and defined as “sequences”.
Galson J.D., Clutterbuck E.A., Trück J., Ramasamy M.N., Münz M., Fowler A., Cerundolo V., Pollard A.J., Lunter G, & Kelly D.F. (2015). BCR repertoire sequencing: different patterns of B cell activation after two Meningococcal vaccines. Immunology and cell biology, 93(10), 885-895.
Publication 2015
Antigenic Specificity Cells Diploid Cell Gene Annotation Genes Immunoglobulin Isotypes Mutation
5
Epidemiology of ANCA-Associated Vasculitis
The French cohort (n = 533) included patients with ANCA vasculitis diagnosed between 1970 and February 2006 at more than 100 university and general hospitals throughout France. Patients were identified for the study when they were enrolled in one of the FVSG single-center or multicenter trials or when referred to the Internal Medicine Department at Bobigny, France until 2003, and thereafter to the French National Referral Center for Necrotizing Vasculitides, Department of Internal Medicine, Hôpital Cochin. Data were compiled in a standardized form in a centralized registry at the French National Referral Center for Systemic Vasculitides. Patients were invited to participate via their treating physician, and written informed consent was obtained from all patients. Followup was conducted through phone calls to patients and/or their physicians, starting in 1980. The patient registry and database were approved for research purposes by the Commission Nationale de l'Informatique et des Libertés (Paris, France).
Patients with ANCA vasculitis in the US cohort (n = 350) were diagnosed between 1985 and 2003 and followed up by physicians in the GDCN, which included 63 private practice offices and 5 academic medical centers (7 (link)). Methods of identifying and enrolling patients have been described previously (7 (link)). Briefly, patients were primarily identified for the study through the University of North Carolina (UNC) Nephropathology Laboratory. Therefore, most of the patients (n = 307) had biopsy-proven renal involvement in ANCA vasculitis (pauci-immune necrotizing and crescentic glomerulonephritis). Forty-three patients who had not undergone renal biopsy were recruited through the multidisciplinary UNC Vasculitis Clinic and by other GDCN nephrologists who collaborate with various medical specialists who care for patients with ANCA vasculitis. The GDCN study was approved by The UNC Committee on the Protection of Human Subjects.
To be included in the study, patients in both cohorts had to be positive for ANCA, as determined by immunofluorescence microscopy and/or antigen-specific enzyme-linked immunosorbent assay (14 (link)). Patients were categorized as having cytoplasmic and/or PR3 ANCA or perinuclear and/or myeloperoxidase ANCA (MPO ANCA). Patients who had perinuclear ANCA alone had to be negative for antinuclear antibodies in order to be included in the study. Patients with target antigen specificities to both MPO and PR3 were excluded, because it was not possible to assess the effect of PR3 ANCA on the risk of relapse.
Pagnoux C., Hogan S.L., Chin H., Jennette J.C., Falk R.J., Guillevin L, & Nachman P.H. (2008). Predictors of Treatment Resistance and Relapse in Antineutrophil Cytoplasmic Antibody–Associated Small-Vessel Vasculitis: Comparison of Two Independent Cohorts. Arthritis and rheumatism, 58(9), 2908-2918.
Publication 2008
Antibodies, Antinuclear Antigenic Specificity Antigens Antineutrophil Cytoplasmic Antibodies Biopsy Cytoplasm Enzyme-Linked Immunosorbent Assay Glomerulonephritis Immunofluorescence Microscopy Kidney Necrotizing Arteritis Nephrologists Patients Peroxidase Physicians Relapse Specialists Vasculitis

Most recents protocols related to «Antigenic Specificity»

1
Antibody Identification and RBC Typing
Antibody identification and RBC typing were performed by indirect antiglobulin gel‐test (ID‐Card LISS/Coombs; DiaMed, Bio‐Rad). The serum of proband 1 containing anti‐RIF was used to screen compatible subjects from cryopreserved RBCs lacking a high‐prevalence antigen of unknown specificity.
Koehl B., Vrignaud C., Mikdar M., Nair T.S., Yang L., Landry S., Laiguillon G., Giroux‐Lathuile C., Anselme‐Martin S., El Kenz H., Hermine O., Mohandas N., Cartron J.P., Colin Y., Detante O., Marlu R., Le Van Kim C., Carey T.E., Azouzi S, & Peyrard T. (2023). Lack of the human choline transporter‐like protein SLC44A2 causes hearing impairment and a rare red blood phenotype. EMBO Molecular Medicine, 15(3), e16320.
Publication 2023
Antigenic Specificity Coombs Test Erythrocytes Immunoglobulins Indirect Coombs Test Serum
2
Cryopreserved Blood Group Samples
Samples from subjects lacking a high‐prevalence antigen of unknown specificity were cryopreserved at the National Reference Center for Blood Groups (CNRGS, biocollection #DC‐2016‐2872). Informed consent for research studies was obtained for all subjects in accordance with the principles set out in the WMA Declaration of Helsinki and the Department of Health and Human Services Belmont Report. The study was approved by local institutional review boards.
Koehl B., Vrignaud C., Mikdar M., Nair T.S., Yang L., Landry S., Laiguillon G., Giroux‐Lathuile C., Anselme‐Martin S., El Kenz H., Hermine O., Mohandas N., Cartron J.P., Colin Y., Detante O., Marlu R., Le Van Kim C., Carey T.E., Azouzi S, & Peyrard T. (2023). Lack of the human choline transporter‐like protein SLC44A2 causes hearing impairment and a rare red blood phenotype. EMBO Molecular Medicine, 15(3), e16320.
Publication 2023
Antigenic Specificity Ethics Committees, Research
3
Spatial Mapping of Immune Cell Signatures
Sequencing libraries for gene expression and TCR/BCR were jointly processed using cellranger multi (v6.0.0) and the GRCh38 genome annotation, and analyzed with Seurat v4.0.11. We next used Seurat’s reference mapping workflow to jointly transfer celltype labels at different granularity (“levels”) and embedding coordinates from a PBMC reference (24 (link)), after filtering out cells with more than 10% mitochondrial gene content, less than 250 or more than 5000 genes and those with a level 1 cell type prediction score of less than 0.75. We used level 2 annotation for B and T-cells, and level 1 annotation otherwise. Next, we used scRepertoire v1.1.22 to process cellranger VDJ output. Persisting clonotypes (both chains) were defined as those appearing in at least one recipient and one donor sample each. Clonal diversity was assessed using the inverse Simpson score, and clonal overlap with the Morisita index. Antigen specificity was assessed using vdjmatch (v1.3.1) (25 (link)). Functional enrichment analysis was done with tmod v0.46.24 with gene sets from the Hallmark, Reactome, Kegg and Gene Ontology BP databases. We investigated differential cell-cell signaling between donors and recipients using scDiffCom v0.1.05. The cytotoxicity score was computed following Zhang et al. (26 (link)), i.e., by projecting our pseudobulk data onto the PCA space defined by their reference dataset (GSE124731). The effectorness score was computed analogously to the approach of Cano-Gamez et al. (27 (link)), i.e., by computing a pseudotime ordering of all CD8 T-cells with monocle3 v0.2.3.08 on the “integrated” assay obtained using Seurat’s Integrate Data workflow to remove batch variation between different samples (24 (link)). Computational enrichment of persisting cells from donor CD4+/CD8+ T cells was done with logistic regression and random forest classifiers (randomForest package v4.6-14). We first used 10fold cross-validation with a 75:25 train:test split across all cells to evaluate the classifiers and feature sets, and then another round of cross-validation, training on 3 donors and testing on the fourth.
Obermayer B., Keilholz L., Conrad T., Frentsch M., Blau I.W., Vuong L., Lesch S., Movasshagi K., Tietze-Stolley C., Loyal L., Henze L., Penack O., Stervbo U., Babel N., Haas S., Beule D., Bullinger L., Wittenbecher F, & Na I.K. (2023). Single-cell clonal tracking of persistent T-cells in allogeneic hematopoietic stem cell transplantation. Frontiers in Immunology, 14, 1114368.
Publication 2023
Antigenic Specificity Biological Assay CD8-Positive T-Lymphocytes Cells Clone Cells Cytoplasmic Granules Cytotoxin Donors Gene Expression Genes Genes, Mitochondrial Genome T-Lymphocyte Tissue Donors
4
Detecting LSDV Antibodies via ELISA
Serum samples were tested for the presence of antibodies to LSDV using ID Screen Capripox Double Antigen Multi-species ELISA (IDVet, Grabels, France, https://www.innovative-diagnostics.com/produit/id-screen-capripox-double-antigen-multi-species/; accessed on 23 January 2023) [28 ] recommended by the OIE Guidelines for Diagnostic Tests and Vaccines for Terrestrial Animals and performed according to the manufacturer instructions. This ELISA is designed to detect antibodies against Capripoxviruses including LSDV, sheep pox virus, and goat pox virus. Samples are read at 450nm and considered positive when the S/P% of the sample ≥30%. The S/P% is calculated according to: (OD of the sample/OD of the positive control) × 100.
Hakobyan V., Sargsyan K., Kharatyan S., Elbakyan H., Sargsyan V., Markosyan T., Vardanyan T., Badalyan M, & Achenbach J.E. (2023). The Serological Response in Cattle following Administration of a Heterologous Sheep Pox Virus Strain Vaccine for Protection from Lumpy Skin Disease; Current Situation in Armenia. Veterinary Sciences, 10(2), 102.
Publication 2023
Animals Antibodies Antigenic Specificity Antigens Capripoxvirus Diagnosis Enzyme-Linked Immunosorbent Assay Goatpox virus Serum Sheeppox virus Tests, Diagnostic Vaccines
5
Generation of SARS-CoV-2 Spike-Specific Monoclonal Antibodies
The spleens of selected mice were used to isolate S-specific mAbs as previously described [18 (link)]. Briefly, spleen cells were incubated with 5 mL lysis buffer on ice for 5 min, and after washing, were mixed with the mouse myeloma cell line SP2 in a 2:1 = splenocytes:myeloma cells ratio. They were then centrifuged, and the cell pellet was slowly resuspended with 1 mL of polyethilenglycol (PEG, Sigma-Aldrich, St. Louis, MI, USA). Afterwards, 7 mL of complete RPMI medium supplemented with 25 mM of Hepes was slowly added, and the centrifuged pellet was resuspended in complete RPMI medium containing selective hypoxanthine, aminopterin, and thymidine (HAT, Sigma-Aldrich) and cultured in a flat-bottomed 96-well plate for 14 days at 37 °C with 5% CO2. The fused growing cell lines were selected by microscope examination, transferred to new 96-well plates and expanded in complete RPMI medium containing selective HAT. Hybridomas were screened for antigen specificity by ELISA using HEK293T S protein-coated plates. The selected polyclonal Ab-producing hybridomas were single-cell-cloned by limiting dilution in the presence of 5 × 104 cells/well feeder splenocytes in a flat-bottom 96-well plate in complete RPMI medium containing selective HAT for 12 days. The growing clones were tested for antigen specificity by ELISA. The clones producing S-specific mAbs were expanded in static T75 flasks in the presence of 50 mL of DCCM2 (Biological Industries, Cromwell, CT, USA) supplemented with kanamycin, and the mAbs were purified and concentrated using chromatography cartridges’ protein G columns (Thermo Fisher). The concentration of purified mAbs was evaluated by a NanoPhotometer (Implen, Münich, Germany) spectrophotometer at 280 nm.
Mariotti S., Chiantore M.V., Teloni R., Iacobino A., Capocefalo A., Michelini Z., Borghi M., Baggieri M., Marchi A., Bucci P., Gioacchini S., D’Amelio R., Brouwer P.J., Sandini S., Acchioni C., Sgarbanti M., Di Virgilio A., Grasso F., Cara A., Negri D., Magurano F., Di Bonito P, & Nisini R. (2023). New Monoclonal Antibodies Specific for Different Epitopes of the Spike Protein of SARS-CoV-2 and Its Major Variants: Additional Tools for a More Specific COVID-19 Diagnosis. Biomedicines, 11(2), 610.
Publication 2023
Aminopterin Antigenic Specificity Biopharmaceuticals Buffers Cell Lines Cells Chromatography Clone Cells Enzyme-Linked Immunosorbent Assay Feeder Cells G-substrate HEPES Hybridomas Hypoxanthine Kanamycin Microscopy Monoclonal Antibodies Multiple Myeloma Mus spike protein, SARS-CoV-2 Spleen Technique, Dilution Thymidine

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More about "Antigenic Specificity"

Immune Recognition, Epitope Specificity, Antigen-Antibody Interactions, Lymphocyte Activation, Humoral Immunity, Cell-Mediated Immunity, Tolerance, FACS, ELISA, Lymphoproliferation Assays, IL-7, IL-4, FBS, GlutaMAX, Lipopolysaccharide, W6/32, ID Screen® Capripox Double Antigen Multi-species ELISA, FACSAria Fusion, Click's medium