ArticlePloS one2011
Impact of immunization technology and assay application on antibody performance--a systematic comparative evaluation.
Article in PloS one, 2011. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 25 papers.
What it found
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The abstract states no effect estimate the extractor could read, or names no intervention and outcome on the map, so this paper lights no cell and moves no belief. It is still indexed, cited and linked below.
The trial behind it
Trials whose registry record cites this paper, or whose number appears in the abstract. A trial that started after this paper was published is citing it as background, not reporting it.
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Who cites it
25 citing papers in PubMed, 45 citations in OpenAlex.
- Evolutionary Dynamics-AwareACS omega · 2026Article
- Immobilization-free SELEX for aptamer discovery targeting colorectal cancer-derived small extracellular vesicles.Journal of nanobiotechnology · 2025Article
- Data-driven evaluation of suitable immunogens for improved antibody selection.Protein science : a publication of the Protein Society · 2025Article
- Antibodies and cryptographic hash functions: quantifying the specificity paradox.Frontiers in immunology · 2025Article
- Computational prediction of multiple antigen epitopes.Bioinformatics (Oxford, England) · 2024Article
- Computational Methods to Predict Conformational B-Cell Epitopes.Biomolecules · 2024Review
- The v8-10 variant isoform of CD44 is selectively expressed in the normal human colonic stem cell niche and frequently is overexpressed in colon carcinomas during tumor development.Cancer biology & therapy · 2023Article
- B-Cell Epitope Predictions Using Computational Methods.Methods in molecular biology (Clifton, N.J.) · 2023Article
- Bioinformatics tools and data resources for assay development of fluid protein biomarkers.Biomarker research · 2022Review
- Molecular Cross-Talk between Gravity- and Light-Sensing Mechanisms inInternational journal of molecular sciences · 2022Article
- Retbindin mediates light-damage in mouse retina while its absence leads to premature retinal aging.Experimental eye research · 2021Article
- Generation of a Polyclonal Antibody against the Mouse Metal Transporter ZIP8.Antibodies (Basel, Switzerland) · 2021Article
- Immunological detection of AcAMP antimicrobial peptide secreted byIranian journal of microbiology · 2021Article
- Development of hypoallergenic variants of the major horse allergen Equ c 1 for immunotherapy by rational structure based engineering.Scientific reports · 2019Article
- Isolation of state-dependent monoclonal antibodies against the 12-transmembrane domain glucose transporter 4 using virus-like particles.Proceedings of the National Academy of Sciences of the United States of America · 2018Article
- Identification of a flagellar protein implicated in the gravitaxis in the flagellate Euglena gracilis.Scientific reports · 2018Article
- Large-scale 3-dimensional quantitative imaging of tissues: state-of-the-art and translational implications.Translational research : the journal of laboratory and clinical medicine · 2017Review
- An Introduction to B-Cell Epitope Mapping and In Silico Epitope Prediction.Journal of immunology research · 2016Review
- Cloning and Stable Expression of cDNA Coding For Platelet Endothelial Cell Adhesion Molecule -1 (PECAM-1, CD31) in NIH-3T3 Cell Line.Advanced pharmaceutical bulletin · 2015Article
- Preparation of Proper Immunogen by Cloning and Stable Expression of cDNA coding for Human Hematopoietic Stem Cell Marker CD34 in NIH-3T3 Mouse Fibroblast Cell Line.Advanced pharmaceutical bulletin · 2015Article
Corrections and comments
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Authors and funding
12 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Antibodies are quintessential affinity reagents for the investigation and determination of a protein's expression patterns, localization, quantitation, modifications, purification, and functional understanding. Antibodies are typically used in techniques such as Western blot, immunohistochemistry (IHC), and enzyme-linked immunosorbent assays (ELISA), among others. The methods employed to generate antibodies can have a profound impact on their success in any of these applications. We raised antibodies against 10 serum proteins using 3 immunization methods: peptide antigens (3 per protein), DNA prime/protein fragment-boost ("DNA immunization"; 3 per protein), and full length protein. Antibodies thus generated were systematically evaluated using several different assay technologies (ELISA, IHC, and Western blot). Antibodies raised against peptides worked predominantly in applications where the target protein was denatured (57% success in Western blot, 66% success in immunohistochemistry), although 37% of the antibodies thus generated did not work in any of these applications. In contrast, antibodies produced by DNA immunization performed well against both denatured and native targets with a high level of success: 93% success in Western blots, 100% success in immunohistochemistry, and 79% success in ELISA. Importantly, success in one assay method was not predictive of success in another. Immunization with full length protein consistently yielded the best results; however, this method is not typically available for new targets, due to the difficulty of generating full length protein. We conclude that DNA immunization strategies which are not encumbered by the limitations of efficacy (peptides) or requirements for full length proteins can be quite successful, particularly when multiple constructs for each protein are used.
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Registered trials
Read under generation 80e0d062 · epoch 390. Bibliography from PubMed, PubMed Central and OpenAlex; grants from NIH RePORTER; trial links from ClinicalTrials.gov; estimates, votes and beliefs from the OpenQuestion graph.