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1.
Bioanalysis ; 9(23): 1895-1912, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-29205053

RESUMO

The 2017 11th Workshop on Recent Issues in Bioanalysis (11th WRIB) took place in Los Angeles/Universal City, California on 3-7 April 2017 with participation of close to 750 professionals from pharmaceutical/biopharmaceutical companies, biotechnology companies, contract research organizations and regulatory agencies worldwide. WRIB was once again a 5-day, weeklong event - a full immersion week of bioanalysis, biomarkers and immunogenicity. As usual, it was specifically designed to facilitate sharing, reviewing, discussing and agreeing on approaches to address the most current issues of interest including both small and large molecule analysis involving LCMS, hybrid ligand binding assay (LBA)/LCMS and LBA approaches. This 2017 White Paper encompasses recommendations emerging from the extensive discussions held during the workshop, and is aimed to provide the bioanalytical community with key information and practical solutions on topics and issues addressed, in an effort to enable advances in scientific excellence, improved quality and better regulatory compliance. Due to its length, the 2017 edition of this comprehensive White Paper has been divided into three parts for editorial reasons. This publication (Part 2) covers the recommendations for biotherapeutics, biomarkers and immunogenicity assays using hybrid LBA/LCMS and regulatory agencies' inputs. Part 1 (LCMS for small molecules, peptides and small molecule biomarkers) and Part 3 (LBA: immunogenicity, biomarkers and pharmacokinetic assays) are published in Volume 9 of Bioanalysis, issues 22 and 24 (2017), respectively.


Assuntos
Biomarcadores/análise , Imunidade Ativa , Espectrometria de Massas , Cromatografia Líquida de Alta Pressão , Conferências de Consenso como Assunto , Regulamentação Governamental , Ligantes
2.
Biopolymers ; 99(10): 686-96, 2013 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-23765412

RESUMO

Amide-linked homopolymers of sialic acid offer the advantages of stable secondary structure and increased bioavailability making them useful constructs for pharmaceutical design and drug delivery. Defining the structural characteristics that give rise to secondary structure in aqueous solution is challenging in homopolymeric material due to spectral overlap in NMR spectra. Having previously developed computational tools for heteroologomers with resolved spectra, we now report that application of these methods in combination with circular dichroism, NH/ND NMR exchange rates and nOe data has enabled the structural determination of a neutral, δ-amide-linked homopolymer of a sialic acid analogue called Neu2en. The results show that the inherent planarity of the pyranose ring in Neu2en brought about by the α,δ-conjugated amide bond serves as the primary driving force of the overall conformation of the homooligomer. This peptide surrogate has an excellent bioavailability profile, with half-life of ∼12 h in human blood serum, which offers a viable peptide scaffold that is resistant to proteolytic degradation. Furthermore, a proof-of-principle study illustrates that Neu2en oligomers are functionalizable with small molecule ligands using 1,3-dipolar cycloaddition chemistry.


Assuntos
Sequência de Aminoácidos , Estrutura Secundária de Proteína , Amidas/química , Dicroísmo Circular , Humanos , Dados de Sequência Molecular , Conformação Proteica , Soluções
3.
Anal Chem ; 83(10): 3703-8, 2011 May 15.
Artigo em Inglês | MEDLINE | ID: mdl-21473642

RESUMO

The analysis of heparan sulfate glycosaminoglycans (HSGAGs) variations in human serum at the disaccharide level has a great potential for disease diagnosis and prognosis. However, the lack of available analytical methodology for the compositional analysis of HSGAGs in human serum remains to be addressed to delineate the possible role of HSGAGs on the onset and/or progression of a disease. In this study, we have developed a method for the in-depth compositional analysis of the 12 heparin/HS-derived disaccharides from human serum using a combination of technologies--fractionation, exhaustive digestion, solid phase extraction, and LC-MS/MS. The method exhibits high recovery (72-110%) and good reproducibility (standard deviation of less than 5%) with a low limit of detection and quantification. Errors from the method validation were within 1.1%. Nondetectable non- or low-sulfated disaccharides in human serum were also detected using the optimized protocol. Further applying this method, the comprehensive analysis of HSGAGs compositions in human sera from female donors showed considerable variations in disaccharide patterns and compositions.


Assuntos
Cromatografia Líquida de Alta Pressão/métodos , Dissacarídeos/sangue , Heparina/química , Heparitina Sulfato/química , Espectrometria de Massas por Ionização por Electrospray/métodos , Adulto , Dissacarídeos/isolamento & purificação , Feminino , Heparina/sangue , Heparitina Sulfato/sangue , Humanos , Masculino , Extração em Fase Sólida/métodos
4.
J Am Chem Soc ; 133(10): 3230-3, 2011 Mar 16.
Artigo em Inglês | MEDLINE | ID: mdl-21341746

RESUMO

A fully synthetic trivalent mimotope of gp120 conjugated to pan allelic HLA DR binding epitope was prepared using solid-phase peptide synthesis and optimized copper-catalyzed azide-alkyne cycloaddition. The methodology efficiently provides chemically uniform heteromultimeric peptide constructs with enhanced binding, avidity, and specificity toward an established HIV-neutralizing human antibody, MAb b12. The versatile synthetic strategy serves as a powerful platform for the development of synthetic peptides as potential HIV-1 vaccine candidates.


Assuntos
Epitopos de Linfócito T/química , Proteína gp120 do Envelope de HIV/síntese química , Antígenos HLA-DR/química , Epitopos Imunodominantes/química , Peptídeos/síntese química , Sequência de Aminoácidos , Epitopos de Linfócito T/imunologia , Proteína gp120 do Envelope de HIV/química , Proteína gp120 do Envelope de HIV/imunologia , Antígenos HLA-DR/imunologia , Epitopos Imunodominantes/imunologia , Vacinas Antimaláricas/química , Dados de Sequência Molecular , Peptídeos/química , Peptídeos/imunologia , Linfócitos T Auxiliares-Indutores/imunologia
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