CBQCA assay of primary amine losses during hemodialysis.
AuthorsKirschbaum B
JournalClin Chim Acta
PubMed ID11412827
Nitrogen loss during hemodialysis is an important issue since all steps taken to improve clearance of urea will also increase loss of amino acids and other normally recyclable sources of nitrogen. Most dialyzers exclude albumin and other proteins, which react well with dyes commonly employed to measure protein, but do ... More
Derivatization in capillary electrophoresis.
AuthorsBardelmeijer HA, Lingeman H, de Ruiter C, Underberg WJ
JournalJ Chromatogr A
PubMed ID9646486
'In recent years capillary electrophoresis (CE) has been developed into a versatile separation technique, next to gas and liquid chromatography (LC), well suited for the determination of a wide variety of e.g., pharmaceutical, biomedical and environmental samples. The main advantages of CE over chromatographic separation techniques are its simplicity and ... More
Capillary electrophoresis with laser-induced fluorescence detection: a sensitive method for monitoring extracellular concentrations of amino acids in the periaqueductal grey matter.
'The use of capillary electrophoresis with laser-induced fluorescence detection (CE-LIF) for the analysis of microdialysate samples from the periaqueductal grey matter (PAG) of freely moving rats is described. By employing 3-(4-carboxybenzoyl)-2-quinoline-carboxaldehyde (CBQCA) as a derivatization agent, we simultaneously monitored the concentrations of 8 amino acids (arginine, glutamine, valine, gamma-amino-n-butyric acid ... More
Mycothiol biosynthesis and metabolism. Cellular levels of potential intermediates in the biosynthesis and degradation of mycothiol in mycobacterium smegmatis.
AuthorsAnderberg SJ, Newton GL, Fahey RC
JournalJ Biol Chem
PubMed ID9804803
'Mycothiol (MSH; 1-D-myo-inosityl-2-(N-acetyl-L-cysteinyl)amido-2-deoxy-alpha-D- glucop yranoside (AcCys-GlcN-Ins)) is a novel thiol produced at millimolar levels by mycobacteria and other actinomycetes that do not make glutathione. We developed methods to determine the major components of MSH (AcCys, Cys-GlcN, AcCys-GlcN, Cys-GlcN-Ins, GlcN-Ins) in cell extracts. Mycobacterium smegmatis was shown to produce measurable levels ... More
Identification of a recombinant synaptobrevin-thioredoxin fusion protein by capillary zone electrophoresis using laser-induced fluorescence detection.
AuthorsAsermely KE, Broomfield CA, Nowakowski J, Courtney BC, Adler M
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID9271130
'Capillary zone electrophoresis (CZE) was utilized to identify a synaptobrevin-thioredoxin fusion protein (TSB-51). TSB-51 is a substrate for cleavage by botulinum toxin B at the Q(76)-F(77) site. TSB-51 was derivatized with a fluorophore, CBQCA [3-(4-carboxy-benzoyl)-2-quinoline-carboxaldehyde], for 4 h at room temperature. Optimal conditions for CZE separation of the TSB-51-CBQCA complex ... More
Facile synthesis of multivalent nitrilotriacetic acid (NTA) and NTA conjugates for analytical and drug delivery applications.
AuthorsHuang Z, Park JI, Watson DS, Hwang P, Szoka FC
JournalBioconjug Chem
PubMed ID17105240
'High-affinity nitrilotriacetic acids (NTA) have great potential in the molecular manipulation of His-tagged proteins. We have developed a facile method to synthesize multivalent NTA and its conjugates. Starting with appropriately protected lysine, we synthesized the mono-NTA synthons functionalized with either an amino group or a carboxylic group. We then obtained ... More
Capillary electrophoresis laser-induced fluorescence for screening combinatorial peptide libraries in assays of botulinum neurotoxin A.
AuthorsLaing TD, Marenco AJ, Moore DM, Moore GJ, Mah DC, Lee WE,
JournalJ Chromatogr B Analyt Technol Biomed Life Sci
PubMed ID16828348
'Botulinum neurotoxin serotype A (BoNT/A) is a proteolytic enzyme that induces muscle paralysis. It is a cause of food poisoning, a potential bioterrorist threat and, in low doses an emerging pharmaceutical product. No effective treatment is currently available for BoNT intoxication. Previously we developed a BoNT/A light chain enzyme assay ... More
Analysis of human cerebrospinal fluid by capillary electrophoresis with laser-induced fluorescence detection.
AuthorsBergquist J, Gilman SD, Ewing AG, Ekman R
JournalAnal Chem
PubMed ID7978320
'Capillary electrophoresis with laser-induced fluorescence detection is used to analyze 10 microL samples of human cerebrospinal fluid. Primary amine-containing compounds in untreated cerebrospinal fluid are labeled with 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde prior to analysis, producing fluorescent isoindoles. Electropherograms containing approximately 50 peaks are obtained in less than 35 min from cerebrospinal fluid samples. ... More
Evaluation of silica resins for direct and efficient extraction of DNA from complex biological matrices in a miniaturized format.
AuthorsTian H, Hühmer AF, Landers JP
JournalAnal Biochem
PubMed ID10906238
'For DNA purification to be functionally integrated into the microchip for high-throughput DNA analysis, a miniaturized purification process must be developed that can be easily adapted to the microchip format. In this study, we evaluate the effectiveness of a variety of silica resins for miniaturized DNA purification and gauge the ... More
Evaluation of amino sugar, low molecular peptide and amino acid impurities of biotechnologically produced amino acids by means of CE.
AuthorsNovatchev N, Holzgrabe U
JournalJ Pharm Biomed Anal
PubMed ID12008126
'As the second part of our studies on the impurity profiles of amino acids produced by biotechnological processes, a micellar electrokinetic chromatography (MEKC) was applied to determine amino sugars, low molecular peptides and amino acids as potential expected impurities at a level 0.1% w/w. 3-(4-Carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) was found to be ... More
Ultrasensitive fluorometric detection of carbohydrates as derivatives in mixtures separated by capillary electrophoresis.
AuthorsLiu JP, Shirota O, Wiesler D, Novotny M
JournalProc Natl Acad Sci U S A
PubMed ID1706520
'Reducing monosaccharides and oligosaccharides, after reductive amination, were separated and detected as their 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde (CBQCA) derivatives by capillary electrophoresis/laser-induced fluorescence. Under optimized conditions, the minimum detectable quantities for monosaccharide solutes were assessed at low attomole levels (0.5 amol for the CBQCA derivative of galactose). The system has shown considerable promise ... More
High-sensitivity peptide mapping with CE using laser-induced fluorescence detection.
Authorsvan de Goor AA, Wanders BJ, Henzel WJ
JournalAm Biotechnol Lab
PubMed ID1368313
Sensitive, laser-assisted determination of complex oligosaccharide mixtures separated by capillary gel electrophoresis at high resolution.
AuthorsLiu J, Shirota O, Novotny MV
JournalAnal Chem
PubMed ID1622005
Recent advances in capillary electrophoresis of carbohydrates.
AuthorsEl Rassi Z, Mechref Y
JournalElectrophoresis
PubMed ID8900934
Derivatization of posttranslationally modified amino acids.
AuthorsTeerlink T
JournalJ Chromatogr B Biomed Appl
PubMed ID7820276
After a brief overview of posttranslational modifications of protein amino acids, the use of various derivatizing reagents for amino acid analysis is discussed. Derivatization and chromatographic separation of hydroxyproline, methylhistidine, and phosphorylated amino acids are discussed in detail to illustrate some of the strategies that can be applied to the ... More
Design of 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde as a reagent for ultrasensitive determination of primary amines by capillary electrophoresis using laser fluorescence detection.
AuthorsLiu JP, Hsieh YZ, Wiesler D, Novotny M
JournalAnal Chem
PubMed ID2064006
Amino acids and peptides, from both standard solutions and biological samples, were successfully reacted with 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde, at low concentration, to form highly fluorescent isoindole derivatives. The formed mixtures are effectively separated by high-performance capillary electrophoresis and their constituents detected by their laser-induced fluorescence signals. The minimum detectable quantities in the ... More
Recent advances in capillary electrophoresis of proteins, peptides and amino acids.
AuthorsNovotny MV, Cobb KA, Liu JP
JournalElectrophoresis
PubMed ID2257844
The current status of high-performance capillary electrophoresis as an analytical separation method for proteins, peptides and amino acids is assessed. Recent advances in suppressing the effects of electroosmotic flow and irreversible adsorption of proteins at the capillary wall are reviewed, together with procedures for optimal separations of peptides and amino ... More
Separation of amino acid homopolymers by capillary gel electrophoresis.
AuthorsDolník V, Novotny MV
JournalAnal Chem
PubMed ID8452245
Gel-filled capillaries utilizing highly concentrated and moderately cross-linked acrylamide-type gels in capillary electrophoresis were successfully applied to the separation of the individual oligomers of various poly(amino acids). Mixtures of both anionic and cationic nature were adequately resolved. While UV detection at 220 nm was mostly utilized, the polyanions with N-terminal ... More
High-performance capillary electrophoresis of glycoconjugates.
AuthorsNovotny MV, Sudor J
JournalElectrophoresis
PubMed ID8354224
Capillary electrophoresis (CE) has recently emerged as a highly promising technique for the analysis and structural study of glycoconjugates. This article reviews the current status of CE and its potential in glycobiology. The separation media suitable for the analysis of monosaccharides, oligosaccharides and very large carbohydrate-based biomolecules are discussed. The ... More
Capillary electrophoresis of catecholamines with laser-induced fluorescence intensified charge-coupled device detection.
AuthorsXiong S, Han H, Zhao R, Chen Y, Liu G
JournalBiomed Chromatogr
PubMed ID11268047
A capillary electrophoresis-laser-induced fluorescence-intensified charge-coupled device system was used for the separation and determination of catecholamines. Optimization of derivatization and separation conditions was investigated in order to reach a high separation efficiency and sensitivity. All fluorecein isothiocyanate derivatives of catecholamines were satisfactorily separated within 12 min. The detection limits were ... More
Capillary electrophoresis of amino sugars with laser-induced fluorescence detection.
AuthorsLiu JP, Shirota O, Novotny M
JournalAnal Chem
PubMed ID2064007
3-(4-Carboxybenzoyl)-2-quinolinecarboxaldehyde has been utilized as a precolumn derivatization agent for various amino sugars. Constituents of various biological mixtures can be converted to highly fluorescent isoindole derivatives, separated by high-performance capillary electrophoresis and determined at attomole (10(-18) mol) levels by a laser-induced fluorescence detector. This method has been applied to the ... More
Separation of fluorescent oligosaccharide derivatives by microcolumn techniques based on electrophoresis and liquid chromatography.
AuthorsLiu JP, Shirota O, Novotny M
JournalJ Chromatogr
PubMed ID1761624
Various aldose oligosaccharides can be quantitatively derivatized into primary amines for subsequent reaction with fluorogenic reagents, such as 3-(4-carboxybenzoyl)-2-quinolinecarboxaldehyde or 3-benzoyl-2-naphthaldehyde. Capillary electrophoresis (CE) and microcolumn liquid chromatography (LC), coupled with laser-induced fluorescence detection, were evaluated as a means of separating complex oligosaccharide mixtures. Whereas microcolumn LC and open-tubular CE ... More
Modification of the electrophoretic mobility of neutral and charged polysaccharides.
AuthorsStefansson M, Novotny M
JournalAnal Chem
PubMed ID7978318
Suitable buffer additives can either induce or reduce the electrophoretic mobility of certain polysaccharides during capillary electrophoresis. With water-soluble cellulose derivatives, adsorption of detergents on the analyte molecules governs the migration rates. Different types of adsorption sites, including multilayer formation, were found to be present on the cellulose-derived polymers. For ... More
Detection of carbohydrates in capillary electrophoresis.
AuthorsPaulus A, Klockow A
JournalJ Chromatogr A
PubMed ID8601201
This review focuses on recent developments in sensitive detection modes for carbohydrates after separation by capillary electrophoretic methods. To bring detection sensitivity for carbohydrates analysis in line with current methods in protein sequencing, concentration detection limits of 10(-6) molar or better are required. A discussion of mass detection limits and ... More
Capillary electrophoretic separations of peptides using micelle-forming compounds and cyclodextrins as additives.
AuthorsLiu J, Cobb KA, Novotny M
JournalJ Chromatogr
PubMed ID2077044
The value of electrokinetic capillary chromatography for separating structurally similar model peptides and tryptic digests is demonstrated. The behavior of model peptides in buffer systems containing dodecyltrimethylammonium bromide, hexadecyltrimethylammonium bromide, sodium dodecyl sulfate and two cyclodextrins as additives is described. These additives, under different analytical circumstances, exhibit certain beneficial effects ... More
Electromigration behavior of polysaccharides in capillary electrophoresis under pulsed-field conditions.
AuthorsSudor J, Novotny M
JournalProc Natl Acad Sci U S A
PubMed ID8415721
Various polysaccharides can successfully migrate through entangled polymer solutions during high-voltage capillary electrophoresis. For neutral polysaccharides, complexation with borate provides the electric charge needed for electromigration, while a fluorescent tag is needed to detect the solute bands with adequate sensitivity. At constant potentials between 50 and 300 V/cm, the charged ... More
Maltooligosaccharides as chiral selectors for the separation of pharmaceuticals by capillary electrophoresis.
AuthorsSoini H, Stefansson M, Riekkola ML, Novotny MV
JournalAnal Chem
PubMed ID7978319
Complexation between the linear maltodextrin oligosaccharides and certain enantiomeric compounds of pharmaceutical interest in buffered solutions can lead to an analytically desirable chiral recognition. Different maltodextrins were assessed in their capacity to cause enantiomeric separations under various conditions of capillary electrophoresis. The mechanism of chiral recognition has been probed through ... More
Separation of free amino acids in human plasma by capillary electrophoresis with laser induced fluorescence: potential for emergency diagnosis of inborn errors of metabolism.
AuthorsBoulat O, McLaren DG, Arriaga EA, Chen DD
JournalJ Chromatogr B Biomed Sci Appl
PubMed ID11318418
Free amino acids (AAs) in human plasma are derivatized with 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA) and analyzed by capillary electrophoresis (CE) with laser induced fluorescence (LIF) detection. The labeling procedure is significantly improved over results reported previously. Derivatization can be completed in 40 min, with concentrations as low as 4 x 10(-8) M ... More
3-(4-Carboxybenzoyl)quinoline-2-carboxaldehyde, a reagent with broad dynamic range for the assay of proteins and lipoproteins in solution.
AuthorsYou WW, Haugland RP, Ryan DK, Haugland RP
JournalAnal Biochem
PubMed ID9025944
This paper describes the application of 3-(4-carboxybenzoyl)quinoline-2-carboxaldehyde (CBQCA), a sensitive fluorogenic reagent for detection of amines, to the assay of proteins in solution. The sensitivity and dynamic range of CBQCA in the determination of protein concentration is modulated by the number of accessible amines present in the protein assayed. Thus, ... More
Electromigration behavior of poly-(L-glutamate) conformers in concentrated polyacrylamide gels.
AuthorsDolník V, Novotny M, Chmelík J
JournalBiopolymers
PubMed ID7689865
During electrophoretic separation of anionic polyamino acids, resolution according to the number of peptide units can be achieved in capillaries filled with hydrophilic gels. While polyaspartate preparations yield single peaks for the individual oligomers at pH above 8.0, polyglutamates exhibit an anomalous behavior of peak splitting, which is attributed here ... More
Experimental evaluation of the separation efficiency in capillary electrophoresis using open tubular and gel-filled columns.
AuthorsLiu J, Dolnik V, Hsieh YZ, Novotny M
JournalAnal Chem
PubMed ID1503214
The band dispersion phenomena in capillary zone electrophoresis (CZE) using untreated and surface-treated open tubular and gel-filled capillaries were experimentally evaluated, with emphasis on small capillary diameters (10-100 microns). Laser-induced fluorescence detection was used for high-sensitivity detection of the isoindoles originated from model amino acids. The plots of plate height ... More
Direct fluorescent staining and analysis of proteins on microspheres using CBQCA.
AuthorsGraves SW, Woods TA, Kim H, Nolan JP
JournalCytometry A
PubMed ID15812850
BACKGROUND: General methods for accurate determination of microsphere surface protein loading are needed for applications from protein arrays to molecular assembly studies. Current methods include bulk absorption measurements of stained microspheres or use of known fluorescently tagged binding partners, which limit sensitivity and general applicability, respectively. METHODS: Microspheres bearing covalently ... More
Capillary electrophoresis of carbohydrates.
AuthorsOefner PJ, Chiesa C
JournalGlycobiology
PubMed ID7827401
Capillary electrophoresis has emerged as a highly promising technique for the analysis of mono- and oligosaccharides. The approaches developed for overcoming the lack of chromophoric and fluorophoric functions in most carbohydrates involve the use of indirect photometric detection, amperometry, mass spectrometry, and precolumn derivatization with various tags. The merits and ... More