Use of capillary electrophoresis as a method development tool for classical gel electrophoresis

The Analyst ◽  
1998 ◽  
Vol 123 (7) ◽  
pp. 1477-1480 ◽  
Author(s):  
Richard M. C. Sutton
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Method Development ◽  
Development Tool

scholarly journals Sodium Dodecyl Sulfate Polyacrylamide Slab Gel Electrophoresis and Hydroxyethyl Cellurose Gel Capillary Electrophoresis of Luminescent Lanthanide Chelate-labeled Proteins with Time-Resolved Detection

2009 ◽  
Vol 25 (3) ◽  
pp. 327-332 ◽  
Author(s):  
Yoshinori YAMAGUCHI ◽  
Kimikazu HASHINO ◽  
Masahiro ITO ◽  
Keisuke IKAWA ◽  
Takuya NISHIOKA ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Sodium Dodecyl Sulfate ◽  
Gel Electrophoresis ◽  
Sodium Dodecyl ◽  
Time Resolved ◽  
Lanthanide Chelate ◽  
Dodecyl Sulfate

scholarly journals Quantification of Serum Proteins Using Capillary Electrophoresis

1995 ◽  
Vol 32 (5) ◽  
pp. 493-497 ◽  
Author(s):  
M A Jenkins ◽  
M D Guerin
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Serum Proteins ◽  
Clinical Laboratory ◽  
Charged Particles ◽  
Protein Electrophoresis ◽  
Agarose Gel Electrophoresis ◽  
Serum Samples ◽  
Routine Clinical Laboratory ◽  
Sample Dilution

Capillary electrophoresis is a technique that can be automated for the separation of charged particles. By investigating suitable sample dilution and injection time and adhering to a strict washing procedure we have been able to quantify paraproteins in serum samples. This has enabled us to use the technique of capillary electrophoresis for the provision of serum protein electrophoresis in a routine clinical laboratory. We present our findings of 260 serum samples, which included 76 samples with paraproteins analysed by both capillary electrophoresis (EC) and high resolution agarose gel electrophoresis (HRAGE). CE was able to detect all the monoclonal bands detected by HRAGE, and, in particular, better able to detect IgA monoclonal bands occurring in the beta region. The major advantages of CE over HRAGE relate to the automated nature of CE with the elimination of the need for a densitometer.

Immobilized metal-ion affinity systems for recovery and structure–function studies of proteins at molecular, supramolecular, and cellular levels

2010 ◽  
Vol 82 (1) ◽  
pp. 39-55 ◽  
Author(s):  
Rajasekar R. Prasanna ◽  
Mookambeswaran A. Vijayalakshmi
Keyword(s):  
Capillary Electrophoresis ◽  
Recombinant Proteins ◽  
Gel Electrophoresis ◽  
Metal Ion ◽  
Immobilized Metal Affinity Chromatography ◽  
Metal Affinity ◽  
Purification Technique ◽  
Affinity Gel Electrophoresis ◽  
Insight Into ◽  
Histidine Tag

Immobilized metal-ion affinity (IMA) adsorption is a collective term that is used to include all kinds of adsorptions where the metal ion serves as the characteristic and most essential part of adsorption center. Of all the IMA techniques, immobilized metal-affinity chromatography (IMAC) has been gaining popularity as the choice of purification technique for proteins. IMAC represents a separation technique that is primarily useful for proteins with natural surface exposed-histidine residues and for recombinant proteins with engineered histidine tag. This review also gives insight into other nonchromatographic applications of IMA adsorption such as immobilized metal-ion affinity gel electrophoresis (IMAGE), immobilized metal-ion affinity capillary electrophoresis (IMACE), and immobilized metal-ion affinity partitioning (IMAP).

scholarly journals Enhanced Gel Electrophoresis Versus Capillary Electrophoresis: Which System Meets the Needs of a High Specimen Throughput Laboratory?

2015 ◽  
Vol 144 (suppl 2) ◽  
pp. A181-A181
Author(s):  
Karie Sceiford ◽  
Sue Bradny ◽  
Mohammad Ansari
Keyword(s):  
Capillary Electrophoresis ◽  
Gel Electrophoresis

Screening for hemoglobinopathy with capillary electrophoresis in adult patients

2021 ◽  
Vol 156 (Supplement_1) ◽  
pp. S18-S18
Author(s):  
Bremansu Osa-Andrews ◽  
Nicole Desimone ◽  
Ravi Sarode ◽  
Sarita Paulino ◽  
Jing Cao
Keyword(s):  
Bone Marrow ◽  
Capillary Electrophoresis ◽  
Gel Electrophoresis ◽  
Screening Method ◽  
Adult Patients ◽  
Red Blood Cell Indices ◽  
Hemoglobin Variants ◽  
Clinically Significant ◽  
Hemoglobin A2 ◽  
Rule Out

Abstract Hemoglobinopathy screening is frequently needed in adult patients, including prenatal carrier screen, workup of unexplained anemia, and bone marrow donor and recipient screening. However, the preferred test method for screening of hemoglobinopathy is not well established due to limited guidance from professional societies. American College of Obstetricians and Gynecologists’ Committee on Genetics recommended hemoglobin electrophoresis as the screening method of hemoglobinopathy in pregnancy; nevertheless, electrophoresis employs various methodologies, including acid gel electrophoresis, alkaline gel electrophoresis, and capillary electrophoresis (CE) with alkaline buffer. For other adult patient populations who need hemoglobinopathy screening, no clear guidelines dictate the method of choice. A previous study has shown that CE captures major hemoglobinopathies with comparable performance to high-performance liquid chromatography (HPLC) in pediatric patients but no study has investigated using CE alone in adult patient screening. In this retrospective study, we evaluated the utility of CE as a screening method to rule out clinically significant hemoglobin variants. During eight months, 312 adult patients without previously identified hemoglobin variants had hemoglobinopathy screening performed using a comprehensive testing algorithm. This cascade algorithm screens for hemoglobinopathy using both CE (Capillarys, Sebia, Paris, France) and HPLC (laboratory-developed test) with reflex to more advanced variant identification such as mass spectrometry and genetic analyses. Categories of abnormal findings were reviewed to determine if hemoglobinopathy can be identified by using CE only. The patient population mainly consists of pregnant women and anemic patients with hematologic malignancies with an average age of 42. Out of the 312 screened patients, 47 had abnormal results. The most frequent condition was elevated hemoglobin F (N=25) ranging 2-5% seen in leukemia patients on chemotherapy attributed to bone marrow stress. Eight cases of beta plus thalassemia (featuring hemoglobin A2 >4%) and 3 cases of hemoglobin C trait were identified in patients with little to mild clinical manifestations (red blood cell indices suggesting anemia). Decreased hemoglobin A2 fraction was observed in 7 patients, and potential causes were alpha thalassemia or iron deficiency. Other less common hemoglobinopathies included heterozygote A2 prime (N=3, a benign delta chain variant that migrates separately from hemoglobin A2 on CE) and hemoglobin G-Philadelphia (N=1). All of the abnormal results are identifiable by CE alone, although HPLC and more advanced methods help confirm the diagnosis. Our study shows that CE as the first line of screening method would rule out major hemoglobinopathies in adults. There have been reports that rare but clinically significant hemoglobin variants like hemoglobin Malmo may not be detected by CE, and therefore, certain pre-test probability factors need to be considered when testing for hemoglobinopathies, such as race/ethnicity background, family history, red blood cell indices, and iron deficiency status.

Development and Validation of a Reversed-Phase Chiral HPLC Method to Determine the Chiral Purity of Bulk Batches of (S)-Enantiomer in Afoxolaner

2017 ◽  
Vol 100 (1) ◽  
pp. 65-73
Author(s):  
Nilusha Padivitage ◽  
Satish Kumar ◽  
Abu Rustum
Keyword(s):  
Method Development ◽  
Specific Rotation ◽  
Reversed Phase ◽  
Hplc Method ◽  
Racemic Mixture ◽  
Chiral Hplc ◽  
Development Tool ◽  
Chiral Purity ◽  
Development And Validation ◽  
First Time

Abstract Afoxolaner is a new antiparasitic molecule from the isoxazoline family that acts on insect acarine g-aminobutyric acid and glutamate receptors. Afoxolaner is a racemic mixture, which has a chiral center at the isoxazoline ring. A reversed-phase chiral HPLC method has been developed to determine the chiral purity of bulk batches of (S)-enantiomer in afoxolaner for the first time. This method can also be used to verifythat afoxolaner is a racemic mixture, which was demonstrated by specific rotation. ChromSword, an artificial intelligence method development tool, was used for initial method development. The column selected for the final method was CHIRALPAK AD-RH (150 × 4.6 mm, 5 μm particle size), maintained at 45°C, and isocratic elution using water–isopropanol–acetonitrile (40 + 50 + 10, v/v/v) as the mobile phasewith a detection wavelength of 312 nm. The run time for the method was 11 min. The resolution and selectivity factors of the two enantiomers were 2.3 and 1.24, respectively. LOQ and LOD of the method were 1.6 and 0.8 μg/mL, respectively. This method was appropriately validated according to International Conference on Harmonization guidelines for its intended use.

scholarly journals Method development for on-line species-specific sulfur isotopic analysis by means of capillary electrophoresis/multicollector ICP-mass spectrometry

2020 ◽  
Vol 412 (23) ◽  
pp. 5637-5646
Author(s):  
Sebastian Faßbender ◽  
Katerina Rodiouchkina ◽  
Frank Vanhaecke ◽  
Björn Meermann
Keyword(s):  
Mass Spectrometry ◽  
Capillary Electrophoresis ◽  
Method Development ◽  
Isotopic Analysis ◽  
On Line ◽  
Species Specific ◽  
Icp Mass Spectrometry

Enantioseparation of citalopram enantiomers by capillary electrophoresis: Method development through experimental design and computational modeling

Chirality ◽  
2020 ◽  
Vol 32 (8) ◽  
pp. 1119-1128
Author(s):  
Monica Budău ◽  
Gabriel Hancu ◽  
Daniela Lucia Muntean ◽  
Lajos Attila Papp ◽  
Anca Gabriela Cârje ◽  
...  
Keyword(s):  
Capillary Electrophoresis ◽  
Experimental Design ◽  
Computational Modeling ◽  
Method Development ◽  
Capillary Electrophoresis Method ◽  
Electrophoresis Method
Sign in / Sign up

Export Citation Format

Share Document