Determination of penicillamine and other thiols by combined high-performance liquid chromatography and post-column reaction with Ellman's reagent: application to human urine

Beales, Denise; Finch, Rosemary E.; McLean, A. E. M.; Smith, M. J. H.; Wilson, Ian D.

doi:10.1016/s0378-4347(00)86088-0

Cited by 47 publications

(10 citation statements)

References 13 publications

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“…The dimerizations by disulfide bonds of the monomer MAP were carried out, at basic pH, directly in the extraction buffer. The reaction was followed by Ellman's test [17]. The homodimer MAPs were then dialyzed in 2 M urea and 100 mM Tris -HCl buffer, pH 8.0 for 24 h. Finally, the MAPs were dialyzed in 1 M HOAc, to remove all the urea, and were then lyophilized.…”

Section: Synthesis Of Mapmentioning

confidence: 99%

Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeficiency virus type 1 with TAB9 protein in mice

et al. 2000

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The multiple antigenic peptide system (MAP) has been proposed as a novel and valuable approach for eliciting antibodies for peptides and developing synthetic vaccines. Multi-epitope polypeptides (MEP) have also been developed as an alternative to the recombinant approach for vaccines. The V3 loop from the HIV type 1 (HIV-1) external glycoprotein (gp120) contains the principal neutralization domain (PND). Antibodies against this region neutralize HIV-1 in vitro and in vivo. In this work, a novel presentation of di-epitope MAP was synthesized. A monomeric MAP carrying two identical JY1 V3 sequences as B-cell epitopes and the 830-843 region of tetanus toxoid as a T-helper cell epitope was synthesized. This basic structure was covalently linked to produce a four-JY1-branched homodimer (JY1-MAP4). Additionally, six different monomeric MAPs, bearing four copies of V3 from isolates LR150, JY1, RF, MN, BRVA and IIIB, were synthesized. These monomers were conveniently linked among themselves to produce homodimeric and heterodimeric MAPs of eight V3 branches (V3-MAP8). JY1-MAP8 elicited higher antibody titers in Balb/c mice than JY1-MAP4. The immunogenicity of two different, hexavalent V3-MAP8 mixtures and the MEP TAB9, which tandems the same six V3 sequences in a single molecule, were compared. The antibody response against the mixtures of the heterodimeric MAP showed a wider recognition pattern of the V3 region, while the homodimeric cocktail showed an intermediate pattern. Antibodies elicited by TAB9 recognized only the JY1, LR150 peptides. These results emphasize the influence of V3 epitope presentation upon the characteristics of the antibody response generated.

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Section: Synthesis Of Mapmentioning

confidence: 99%

Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeficiency virus type 1 with TAB9 protein in mice

et al. 2000

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“…Cysteine and glutamic acid were detected by post-column labeling with 5,5′-dithiobis(2-nitrobenzoate) (Beales et al 1981) and with o-phthalaldehyde (Fujiwara et al 1987), respectively.…”

Section: Enzyme Assaymentioning

confidence: 99%

Purification and characterization of 5-oxo- l -prolinase from Paecilomyces varioti F-1, an ATP-dependent hydrolase active with l -2-oxothiazolidine-4-carboxylic acid

Mochizuki

1999

Archives of Microbiology

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An enzyme cleaving L-2-oxothiazolidine-4-carboxylic acid to L-cysteine was purified 75-fold with 8% recovery to near homogeneity from crude extracts of Paecilomyces varioti F-1, which had been isolated as a fungus able to assimilate L-2-oxothiazolidine-4-carboxylic acid. The molecular mass was estimated to be 260 kDa by gel filtration. The purified preparation migrated as a single band of molecular mass 140 kDa upon SDS-PAGE. The maximum activity was observed at a range of pH 7.0-8.0 and at 50 degrees C. The enzyme activity was completely inhibited by SH-blocking reagents such as AgNO(3), p-chloromercuribenzoic acid, N-ethylmaleimide, and N-bromosuccinimide. The enzyme required ATP, Mg(2+), and KCl for the cleavage of L-2-oxothiazolidine-4-carboxylic acid. The enzyme also cleaved 5-oxo-L-proline to L-glutamic acid and is considered to be 5-oxo-L-prolinase.

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“…It is known [18,19] that selected disulfides such as 5,5'-dithiobis-(2-nitrobenzoic acid), better known as Ellman's reagent, are particularly suited for spectrophotometric determination of thiols. When the thiol reacts with excess reagent a mixed disulfide is formed (the rate depending on the particular thiol):…”

Section: Resultsmentioning

confidence: 99%

Comparison of analytical methods for quality control of pharmaceutical formulations containing glutathione

et al. 1997

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SummaryThe aim of this investigation was the study and development of analytical procedures suitable for the assay of glutathione (GSH) in pharmaceutical formulations. Two are based on isocratic HPLC with a 250 mmx 4.6 mm i.d., 5 ~m C18 column and UV detection. In the first procedure sample solutions were injected without pretreatment whereas in the second the samples were injected after derivatization with Ellman's reagent which forms an easily detectable adduct with GSH. Good linearity was obtained over the range 0.12-6.00 x 10 --4 M for the direct procedure and 0.25--3.00 x 10 -4 M for the derivatization procedure. The precision and rapidity of analysis were also good for both methods. The third method is based on capillary zone electrophoresis (CZE) in a 27 cm x 75 pm i.d untreated fused silica capillary containing pH 7 phosphate buffer. All results are in good agreement with a spectrophotometric procedure used as reference method.

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Determination of penicillamine and other thiols by combined high-performance liquid chromatography and post-column reaction with Ellman's reagent: application to human urine

Cited by 47 publications

References 13 publications

Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeficiency virus type 1 with TAB9 protein in mice

Immunogenicity comparison of a multi-antigenic peptide bearing V3 sequences of the human immunodeficiency virus type 1 with TAB9 protein in mice

Purification and characterization of 5-oxo- l -prolinase from Paecilomyces varioti F-1, an ATP-dependent hydrolase active with l -2-oxothiazolidine-4-carboxylic acid

Comparison of analytical methods for quality control of pharmaceutical formulations containing glutathione

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