First purification of heparan sulfate disaccharides with an amine resin used as solid support for peptide synthesis

Carvalho, R. S. H.; Tersariol, Ivarne L.S.; Nader, Helena B.; Nakaie, Clóvis R.

doi:10.1016/s0003-2670(99)00631-5

Cited by 6 publications

(5 citation statements)

References 12 publications

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“…This has been demonstrated in the case of the anion-exchange capacity of BHAR [5][6][7] and in the unexpected peptide synthesis and affinity capacities of some commercial anion-exchange resins [10]. Additional physicochemical and chromatographic experiments are currently being undertaken in order to further evaluate the potential of the AMR support.…”

Section: Affinity Assays With Peptide-amrmentioning

confidence: 96%

“…These results indicate that the AMR support, to date used only for peptide synthesis, can be applied as a cationic resin for chromatography, as was previously reported for 2.4 mmol/g BHAR, another polystyrene-type resin that attaches phenylaminomethyl groups instead. This latter solid support presented similar efficiency in the fractionation of negatively charged gangliosides [5] or carbohydrates [6,7] if compared with commercial anion exchanger resins such as the dextran-based DEAESephadex A25 (Amersham Biosciences) or the quaternary ammonium-bearing polystirene-2% divinylbenzene-based AG1-X1 (Bio Rad Co.). Despite its comparatively low exclusion limit if compared, for instance, to those containing highly hydrophilic dextranor agarose (Sepharose)-type anion exchanger matrices, the presence of more reactive primary amino groups in the AMR structure can be advantageous since this basic function could be easily derived in order to obtain novel resin derivatives.…”

Section: Anion-exchange Chromatography With the 49 Mmol/g Amrmentioning

confidence: 97%

“…Negatively charged gangliosides [5] and carbohydrates [6,7] have been successfully fractionated with highly substituted BHAR batches. This resin is indeed, the first developed exclusively for use in peptide synthesis but applied as solid support for anion-exchange chromatography.…”

Section: Introductionmentioning

confidence: 99%

See 2 more Smart Citations

Polystyrene-type resin used for peptide synthesis: application for anion-exchange and affinity chromatography

Carvalho

Ianzer

Malavolta

et al. 2005

Journal of Chromatography B

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Section: Affinity Assays With Peptide-amrmentioning

confidence: 96%

Section: Anion-exchange Chromatography With the 49 Mmol/g Amrmentioning

confidence: 97%

See 1 more Smart Citation

Polystyrene-type resin used for peptide synthesis: application for anion-exchange and affinity chromatography

Carvalho

Ianzer

Malavolta

et al. 2005

Journal of Chromatography B

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“…We have previously demonstrated that very highly ammonium substituted BHAR batches (>1.5-2.0 mmol g -1 ) actually display improved swelling in polar solvents, including water. 16 These findings prompted us to initiate the evaluation of its use as ion exchange supports for purification of negatively charged disaccharides 17 and gangliosides 18 in aqueous solution. Some physicochemical characteristics of this type of cationic resin have also been described.…”

Section: Introductionmentioning

confidence: 99%

Use of the same polymer for synthesis and purification of peptides

Silva¹,

Etchegaray²,

Carvalho³

et al. 2005

J. Braz. Chem. Soc.

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Este trabalho revela uma estratégia biotecnológica incomum mas valiosa e relacionada com o uso de um mesmo polímero (benzidrilamino-resina, BHAR) para fins de síntese e purificação por troca aniônica de peptídeos. Inicialmente, o octapeptídeo DRVYIHPF-NH 2 foi sintetizado em BHAR com 1% e 3% de intercruzamento e contendo 2,5 mmol g -1 de grupos amônio. Devido certamente a uma menor rigidez da sua estrutura polimérica, um maior rendimento na síntese (cerca de 80%) foi obtido com a primeira resina. A seguir, os peptídeos DEVYEHPF-NH 2 e DEVYEDPF-NH 2 carregados negativamente (-1 e -3 em pH neutro, respectivamente), ambos sintetizados em 1% BHAR, foram submetidos a um teste de separação cromatográfica no mesmo tipo de resina (1% e 3%). Concordante com resultados da síntese de peptídeos e de valores de inchamento dos grãos das resinas, obtidos por microscopia, uma melhor separação entre ambos os peptídeos ocorreu com o lote de 1% BHAR. Estes achados demonstraram que a BHAR, aplicada até o momento somente para a síntese peptídica, se contiver elevado teor de grupos amônio positivos, pode ser utilizada alternativamente como suporte sólido para purificação cromatográfica deste tipo de molécula biológica.This work reveals an uncommon but valuable biotechnological approach regarding the use of a same polymer (benzhydrylamine-resin, BHAR) for synthesis and anion exchange purification of peptides. Initially, the octapeptide DRVYIHPF-NH 2 was synthesized in 1% and 3% cross-linked BHAR, attaching 2.5 mmol g -1 ammonium groups. Due certainly to its less rigid polymeric backbone, higher synthesis yield (about 80%) was achieved with the former resin. Next, the negatively charged peptides DEVYEHPF-NH 2 and DEVYEDPF-NH 2 (-1 and -3 in neutral pH, respectively), both synthesized in 1% BHAR were submitted to chromatographic separation test in this same type of resin (1% and 3%). Following comparative results of peptide synthesis and swelling data of resin beads obtained by microscopy, an improved separation of both peptides occurred with 1% BHAR batch. These findings demonstrated that BHAR applied so far for peptide synthesis, when containing high amount of positively charged ammonium groups, can be also used alternatively as a solid support for chromatographic purification of this type of biological molecule.Keywords: peptide synthesis, resin, anion exchange chromatography, polymer, solvent polarity IntroductionThe inception of solid phase peptide synthesis (SPPS) 1 almost four decades ago 2 represented a landmark in the use of cross-linked beaded polymers, which were, until that time, restricted to stationary phases in column chromatography. Since then, various studies have been conducted in order to develop more varied applications for such polymeric materials. In addition to the peptide synthesis method itself, the concept of performing chemical reactions on an insoluble polymeric structure has, for instance, been successfully extended to oligonucleotides 3 and polysaccharides. 4 In recent years, a process denoted solid phase o...

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“…This effort in improving the polymer solvation knowledge has also recently led us to demonstrate that benzhydrylamine-resin (BHAR) 12 , a phenylmethylamine group-containing copolymer of styrene-1%-divinilbenzene, so far used as the solid support for peptide synthesis, can be alternatively employed as a novel anion exchange resin for liquid chormatography 13,14 .…”

Section: Introductionmentioning

confidence: 99%

Importance of the solvation degree of peptide-resin beads for amine groups determination by the picric acid method

Cilli¹,

Jubilut²,

Ribeiro³

et al. 2000

J. Braz. Chem. Soc.

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O clássico e importante método do ácido pícrico, usado nas áreas de bioquímica e química de polímeros para a quantificação de grupos aminos, foi escolhido neste trabalho como modelo para investigar a importância do grau de solvatação de grãos poliméricos durante o protocolo analítico. Verificou-se que este método, proposto há cerca de três décadas atrás, falha na quantificação de grupos amino de peptidil-resinas contendo seqüência agregante e polar. Isto ocorre devido à solvatação insuficiente dos grãos quando apenas o CH 2 Cl 2 é utilizado na etapa de ligação do ânion picrato e na subsequente etapa de lavagem. Demonstrou-se que a utilização nestas etapas, de soluções de CH 2 Cl 2 /DMF (dimetilformamida) e de CH 2 Cl 2 /EtOH permite uma determinação correta dos grupos amínicos de peptídil-resinas. Além da importância em si para o método da síntese de peptídeos em fase sólida, estes resultados representam também a primeira comprovação experimental da correlação quantitativa existente entre o grau de solvatação e a eficiência de um método analítico efetuado em grãos de resinas.The classic and important picric acid method used in polymers biochemical and chemical fields of polymers for amine group quantification was chosen in this work as a model for evaluating the influence of the resin bead solvation during an analytical procedure. It was observed that this method, proposed almost three decades ago, failed to quantify amine groups of peptidyl-resin containing aggregating and polar sequence. This was due to inefficient solvation of resin beads when only CH 2 Cl 2 was used for picrate anion binding and subsequent washing steps. It was demonstrated that the use of CH 2 Cl 2 /DMF (dimethylformamide) and CH 2 Cl 2 /EtOH solutions during these steps allows correct determination of peptidyl-resin amine groups. Besides the importance for the solid phase peptide synthesis methodology itself, these findings also represent the first quantitative demonstration of the relationship between solvation degree and the efficiency of a polymer-supported analytical method.

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First purification of heparan sulfate disaccharides with an amine resin used as solid support for peptide synthesis

Cited by 6 publications

References 12 publications

Polystyrene-type resin used for peptide synthesis: application for anion-exchange and affinity chromatography

Polystyrene-type resin used for peptide synthesis: application for anion-exchange and affinity chromatography

Use of the same polymer for synthesis and purification of peptides

Importance of the solvation degree of peptide-resin beads for amine groups determination by the picric acid method

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