Estimation of the pK a of the amine groups of benzhydrylamine resin and its lysyl derivative by measurement of swelling

Etchegaray, Augusto; Carvalho, R. S. H.; Boschcov, Paulo; Nakaie, Clóvis R.

doi:10.1007/bf02272827

Cited by 5 publications

(3 citation statements)

References 10 publications

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“…As previously mentioned, the protocol developed to estimate the pK a of resin functional groups [16] is based on the measurement of bead swelling in all pH ranges in buffered aqueous solution. By correlating the degree of swelling with the pH of a given medium, a decreasing sigmoid curve is Table 1 Extent of swelling of 4.9 mmol/g AMR with amine groups in the protonated form in several solvents obtained as the pH increases, and the mid-point of inflection of this curve corresponds to the pK a of the amino group of the resin.…”

Section: Estimation Of the Pk A Of The Amr Amino Groupmentioning

confidence: 99%

“…Since prior knowledge of the value of AMR amino group pK a is essential for its efficient anion-exchange application, this physicochemical parameter was also determined through a swelling method previously introduced for pK a determination of the BHAR amino group [16]. In this method, the degree of resin swelling is correlated with the pH of the medium.…”

Section: Introductionmentioning

confidence: 99%

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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Section: Estimation Of the Pk A Of The Amr Amino Groupmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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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“…Through a swelling versus pH of the medium approach, we have previously demonstrated that, at this and lower pH levels, the amine groups of BHAR are totally protonated, since an average pKa value of approximately 7 was determined for this basic function. 32 A pH-gradient ranging from pH 5 to 2.3 (10% HOAc) was applied. condition should be necessary for achievement of improved resolution and elution of solutes.…”

Section: Use Of Bhar As An Anion-exchange Resinmentioning

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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Highly-substituted benzhydrylamine resin: An alternative chromatographic support for ganglioside and neutral glycosphingolipid purification

et al. 2000

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Benzhydrylamine resin (BHAR) is a copolymer of (sl',/rene-1% divinylbenzene) containing phenylmethylamine groups and commonly used as a solid support for peptide synthesis in organic solvents. We have demonstrated that a larger number of NH3 + groups distribu ted throughout the BHAR matrix improves bead solvation under more polar conditions. As this characteristic might allow this aminated-polymer to be used as a stationary phase for liquid chromatogra-ph~ a highly substituted BHAR (2.4 mmol.g 1 of amine groups) was synthesized under forceful conditions. Neutral glycosphingolipids and negatively charged gangliosides and sulfatide obtained from rat brain extracts were successfully purified and fractionated in this BHAR batch by single-step, reverse-phase anion-exchange chromatography. Additionally the anion-exchange potential of BHAR was also compared to that of commercial dimethylaminoethyl (DEAE)-SephadexA25 resin.

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Estimation of the pK a of the amine groups of benzhydrylamine resin and its lysyl derivative by measurement of swelling

Cited by 5 publications

References 10 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

Highly-substituted benzhydrylamine resin: An alternative chromatographic support for ganglioside and neutral glycosphingolipid purification

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