Analysis of immune complexes by capillary electrophoresis

Shihabi, Zak K.

doi:10.1002/elps.200700810

Cited by 3 publications

(1 citation statement)

References 17 publications

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“…The development time that would need to be invested to create a separate ACE method for each ChIP grade antibody would be long and costly for a research lab interested in using the purchased material immediately. Furthermore, most ChIP grade antibodies are polyclonals; unlike monoclonals, they produce broad peaks, sometimes multiple ones, on a capillary electropherogram (Shihabi 2008), making it difficult to resolve and identify pure antibodies from contaminated ones.…”

Section: Detecting Hitchhikersmentioning

confidence: 99%

Hitchhiker antigens: Inconsistent ChIP results, questionable immunohistology data, and poor antibody performance may have a common factor

Parseghian

2013

Biochem. Cell Biol.

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Questionable data and poor antibody performance may have a common factor: antigens "hitchhiking" on the very antibodies designed to target them. Here I focus on histone hitchhikers and their antibodies, given the impact of chromatin immunoprecipitation on our understanding of DNA regulation. Caused by a lack of stringency during antibody purification, hitchhikers will impede important advances in chromatin research and therapeutics derived from that research, if similar circumstances in the study of lupus decades ago are any guide. Evidence of this phenomenon is reviewed, purification modifications for antibody manufacturing are suggested, and a histone hitchhiker detection procedure is provided.

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Section: Detecting Hitchhikersmentioning

confidence: 99%

Hitchhiker antigens: Inconsistent ChIP results, questionable immunohistology data, and poor antibody performance may have a common factor

Parseghian

2013

Biochem. Cell Biol.

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Confirmation of immuno‐reactivity of the recombinant major birch pollen allergen Bet v 1a by affinity‐CIEF

et al. 2009

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Affinity-CIEF has been applied to characterize a recombinant product of the major birch pollen allergen Betula verrucosa isoform 1a (Bet v 1a) immuno-chemically. For this purpose mAbs of the IgG-type have been produced in-lab from two murine hybridoma lines, specified as clones 2 and 5.1. Both IgG clones were characterized by SDS-PAGE, MALDI-TOF-MS and CIEF. The purified IgG solutions had to be dialysed against 10 mmol/L phosphate (pH 7.4) to prevent IgG precipitation and to ensure appropriate CIEF separation. Both tested monoclonal IgGs (mIgGs) comprised four constituents covering pI ranges of 6.98-7.09 and 6.78-7.03 for clones 2 and 5.1 with major peaks at pI 7.09 and 7.03, respectively. When increasing amounts of Bet v 1a (pI 4.95) were incubated with 2.0 mumol/L mIgG, novel peaks were progressively induced in a pI range slightly more acidic than the focusing region of mIgGs. These peaks grew on the expense of original mIgG peaks. All pI values were calculated using two pI marker compounds with a repeatability of better than 0.03 units. New peaks represent complexes between Bet v 1a and mIgG either of 1:1 or of 2:1 binding stoichiometry. At a molar ratio of 2:1, saturation of both IgG paratopes with allergen (Ag) molecules was achieved as indicated by unbound Bet v 1a. The current CIEF approach addresses the proof of single epitope integrity in the course of immuno-chemical characterization of Bet v 1a. Contrary to traditional immunoassays, affinity CIEF allows for a distinction and relative quantification of mAbs, Ag-antibody complexes and Ag variants coexisting in one sample.

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A case of false positive Troponin I in a patient affected by cryoglobulinemic vasculitis

et al. 2017

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SUMMARYTroponin I (TnI) false positive results have been reported in patients affected by immune disorders. We report the case of a 74-year-old woman affected by cryoglobulinemic vasculitis, admitted to the Emergency Room because of a lipotimic episode. A marked elevation of TnI plasma concentration was confirmed in multiple determinations, despite the absence of symptoms or electrocardiogram findings suggesting myocardial infarction. TnI plasma concentration was reported normal after re-testing with a different commercial kit. A false TnI positivity should be considered in patients with immune disorders, especially if seropositive for rheumatoid factor, when the clinical context does not suggest myocardial infarction.

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Analysis of immune complexes by capillary electrophoresis

Cited by 3 publications

References 17 publications

Hitchhiker antigens: Inconsistent ChIP results, questionable immunohistology data, and poor antibody performance may have a common factor

Hitchhiker antigens: Inconsistent ChIP results, questionable immunohistology data, and poor antibody performance may have a common factor

Confirmation of immuno‐reactivity of the recombinant major birch pollen allergen Bet v 1a by affinity‐CIEF

A case of false positive Troponin I in a patient affected by cryoglobulinemic vasculitis

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