Protein microarrays: A new tool for profiling antibody cross-reactivity
ABSTRACT Antibody cross-reactivity can compromise interpretation of experiments and derail therapeutic antibody development. Standard techniques such as immunohistochemistry or Western analysis provide important but often inadequate approaches to assess antibody specificity. Protein microarrays are providing a new approach to rapidly characterize antibody cross-reactivity against 1,000s of proteins simultaneously. This review will focus on reported examples of antibody cross-reactivity, methods used to characterize them, and the recent development and use of protein microarrays for assessing antibody specificity.
- SourceAvailable from: Mahavir Singh
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- "However, in the present series of CSF-NIND samples, there was no antigen-specific reactivity for either of the antibodies and the PTR 3 level also was not detected. There are several reports of cross-reactive antibodies that may recognize very similar epitopes from unrelated targets and the antigen-specific antibodies detected in some CSF-OIM may be related to cross reactivity with other infectious microorganisms rather than previous mycobacterial latent infection (Michaud et al. 2003, Predki et al. 2005). In the CSF-OIM of the present series, which were infected with S. pneumoniae and Cryptococcus, a mycobacterial antigen-specific reaction was observed mainly for IgA-16 kDa and the Lionex kit. "
ABSTRACT: To evaluate commercial Lionex TB together with four antigens of Mycobacterium tuberculosis (MPT-64, MT10.3, 16 kDa and 38 kDa) for IgG and IgA cerebrospinal fluid (CSF) detection in the diagnosis of tuberculosis meningitis (TBM) with CSF negative acid-fast bacilli staining, 19 cases of TBM, 64 cases of other infectious meningoencephalitis and 73 cases of other neurological disorders were tested by enzyme linked immunosorbent assay. IgA-MPT-64 and IgG Lionex showed the highest sensitivities, specificities, positive predictive value and negative predictive value (63.2%, 47.4%; 95%, 93.7%; 40%, 98% and 28.4%, 97.1%, respectively). However, while grey zone was 12.7% and 6%, respectively, lowering sensitivity but maintains high specificity (>or= 95%). High protein concentration in CSF was associated with antibody positivity CSF/HIV+ which did not influence the sensitivity of both tests. To our knowledge, this is the first description of IgA-MPT-64 and IgG Lionex antibodies in CSF-TBM and, although there is good specificity, adjustments are needed based on antigen composition to enhance sensitivity.Memórias do Instituto Oswaldo Cruz 08/2010; 105(5):722-8. DOI:10.1590/S0074-02762010000500022 · 1.57 Impact Factor
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ABSTRACT: Current, cell‑free methodologies allow for robust and rapid protein production and screening. The use of protein arrays attached selectively or nonspecifically to various solid supports is rapidly becoming a common research tool to explore the function and potential relationships of proteins encoded within any genome. Array‑based approaches are also ideal for parallel analysis of multiple binary interactions between proteins and other molecules. In addition, engineering novel tagging techniques allows the orientation of proteins of interest and expands the capabilities and use of protein microarrays. Combining cell‑free expression with array‑based proteomics promises to be a powerful tool for protein biochemistry, molecular diagnostics and therapeutics.Cell-Free Protein Expression, 1st Edition edited by W. Antoni Kudlicki, 01/2007: chapter Chapter12: pages 145-154; Landes Biosciences., ISBN: 978-1-58706-123-3