Isolation of monoclonal antibodies to phencyclidine from ascites fluid by preparative isoelectric focusing in the Rotofor

Egen, Ned B.; Bliss, Marla; Mayersohn, Michael; Owens, S. Michael; Arnold, Larry W.; Bier, M.

doi:10.1016/0003-2697(88)90472-1

Cited by 60 publications

(28 citation statements)

References 16 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…One possible approach described by Hochstrasser et al (16) is to obtain narrow pI cuts by using a multichamber device developed by Bier's group (17). This equipment consists of a cooling finger with a rotating cell that is divided into 20 compartments by nylon net.…”

Section: E El Le Ec Ct Tr Ro Op Ph Ho or Re Et Ti Ic C S St Te Ep Ps Smentioning

confidence: 99%

Peer Reviewed: Prefractionation Techniques in Proteome Analysis

Righetti¹,

Castagna²,

Herbert³

2001

Anal. Chem.

100

View full text Add to dashboard Cite

Section: E El Le Ec Ct Tr Ro Op Ph Ho or Re Et Ti Ic C S St Te Ep Ps Smentioning

confidence: 99%

Peer Reviewed: Prefractionation Techniques in Proteome Analysis

Righetti¹,

Castagna²,

Herbert³

2001

Anal. Chem.

100

View full text Add to dashboard Cite

“…IEF was performed by the method of Egen et al [41] with some modifications. lyophilized and then solubilized in distilled water (10% of the initial volume).…”

Section: Ige Binding Inhibition Using Bromelainmentioning

confidence: 99%

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

et al. 2012

View full text Add to dashboard Cite

The common cypress (Cupressus sempervirens) (Cups) pollen represents the first cause of respiratory allergies in the Mediterranean basin. The aim of this study was to characterize a novel 14-kDa cypress pollen allergen (BP14) allowing a clear dissociation of IgE sensitization patterns among allergic patients. The biochemical and immunochemical characterization of BP14 included determination of its isoelectric point, molecular mass, extraction kinetics, IgE binding prevalence, the presence of bromelain-type cross-reactive carbohydrate determinant and its IgE reactivity under reducing conditions. The presence of potential cross-reactive homologues in closely related cypress species, i.e. Cupressus arizonica (Cupa) and Cryptomeria japonica (Cryj), as well as in several taxonomically unrelated species was also investigated. According to our results, BP14 is easily and quickly solubilized in phosphate-buffered saline and exhibits several allergenic isoforms covering a broad range of pI (6.5-10.5). This allergen displays heat-stable conformational epitopes and does not include cross-reactive carbohydrate determinants, in contrast to high molecular weight cypress allergens. BP14 is expressed at higher levels in Cups than in Cupa and Cryj. No IgE cross-reactivity was found between the 14-kDa Cups pollen protein and proteins from some other non-Cupressaceae pollen allergenic sources such as orchard, timothy, wheat, maize, birch, ash and pine. Thus, IgE reactivity to BP14 is specific to Cupressaceae and discriminates two groups of patients allergic to cypress pollen. It might correspond to a relevant marker in relation to the sensitization process and ⁄ or the symptoms observed in some cypress-pollen-allergic patients. Furthermore, the description of BP14 should improve the diagnosis of cypress pollinosis.Abbreviations AB, ammonium bicarbonate buffer; BP14, basic protein of 14 kDa relative mass; CCD, cross-reactive carbohydrate determinant; Cryj,

show abstract

“…These non-gel-based IEF methods can accommodate large sample volumes and amounts in contrast to gelbased IEF methods. Preparative IEF as a prefractionation technique was first proposed by Bier's laboratory [60,61]. To overcome problems associated with the original device, Righetti and colleagues [62] developed a multicompartment electrolyser in which each compartment was separated For analytical imaging separations, a portion of each first-dimension FFE-IEF fraction (50 µl/total volume ∼2 ml) was injected directly from the 96 deep-well plate using the Agilent 1100 HPLC equipped with a well-plate autosampler and samples collected automatically into a multi-plate fraction collection system by a polyacrylamide gel membrane, each with a defined pH.…”

Section: Prefractionation Approaches In Proteome Analysismentioning

confidence: 99%

Proteomic analysis of colorectal cancer: prefractionation strategies using two‐dimensional free‐flow electrophoresis

Moritz

Skandarajah

et al. 2005

Comp Funct Genom

View full text Add to dashboard Cite

This review deals with the application of a new prefractionation tool, free-flow electrophoresis (FFE), for proteomic analysis of colorectal cancer (CRC). CRC is a leading cause of cancer death in the Western world. Early detection is the single most important factor influencing outcome of CRC patients. If identified while the disease is still localized, CRC is treatable. To improve outcomes for CRC patients there is a pressing need to identify biomarkers for early detection (diagnostic markers), prognosis (prognostic indicators), tumour responses (predictive markers) and disease recurrence (monitoring markers). Despite recent advances in the use of genomic analysis for risk assessment, in the area of biomarker identification genomic methods alone have yet to produce reliable candidate markers for CRC. For this reason, attention is being directed towards proteomics as a complementary analytical tool for biomarker identification. Here we describe a proteomics separation tool, which uses a combination of continuous FFE, a liquid-based isoelectric focusing technique, in the first dimension, followed by rapid reversed-phase HPLC (1–6 min/analysis) in the second dimension. We have optimized imaging software to present the FFE/RP-HPLC data in a virtual 2D gel-like format. The advantage of this liquid based fractionation system over traditional gel-based fractionation systems is the ability to fractionate large quantity protein samples. Unlike 2D gels, the method is applicable to both high-Mr proteins and small peptides, which are difficult to separate, and in the case of peptides, are not retained in standard 2D gels.

show abstract

Isolation of monoclonal antibodies to phencyclidine from ascites fluid by preparative isoelectric focusing in the Rotofor

Cited by 60 publications

References 16 publications

Peer Reviewed: Prefractionation Techniques in Proteome Analysis

Peer Reviewed: Prefractionation Techniques in Proteome Analysis

Differential IgE sensitization to cypress pollen associated to a basic allergen of 14 kDa

Proteomic analysis of colorectal cancer: prefractionation strategies using two‐dimensional free‐flow electrophoresis

Contact Info

Product

Resources

About