Synthetic Peptide Purification via Solid-Phase Extraction with Gradient Elution: A Simple, Economical, Fast, and Efficient Methodology

Cepeda, Diego Sebastián Insuasty; Pineda-Castañeda, Héctor Manuel; Mayor, Andrea Verónica Rodríguez; Villamil, Mauricio Maldonado; Medina, Ricardo; Rivera‐Monroy, Zuly Jenny

doi:10.3390/molecules24071215

Cited by 28 publications

(29 citation statements)

References 23 publications

(34 reference statements)

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“…The collected fractions were analyzed via RP‐HPLC and MALDI‐TOF MS. The fractions containing the pure peptide were mixed and then lyophilized [33] …”

Section: Methodsmentioning

confidence: 99%

Designing Chimeric Peptides: A Powerful Tool for Enhancing Antibacterial Activity

Pineda-Castañeda

Huertas-Ortiz

Leal-Castro

et al. 2021

Chemistry & Biodiversity

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Chimeric peptides containing short sequences derived from bovine Lactoferricin (LfcinB) and Buforin II (BFII) were synthetized using solid‐phase peptide synthesis (SPPS) and characterized via reversed‐phase liquid chromatography and mass spectrometry. The chimeras were obtained with high purity, demonstrating their synthetic viability. The chimeras’ antibacterial activity against Gram‐positive and Gram‐negative strains was evaluated. Our results showed that all the chimeras exhibited greater antibacterial activity against the evaluated strains than the individual sequences, suggesting that chemical binding of short sequences derived from AMPs significantly increased the antibacterial activity. For each strain, the chimera with the best antibacterial activity exerted a bacteriostatic and/or bactericidal effect, which was dependent on the concentration. It was found that: (i) the antibacterial activity of a chimera is mainly influenced by the linked sequences, the palindromic motif RLLRRLLR being the most relevant one; (ii) the inclusion of a spacer between the short sequences did not significantly affect the chimera's synthesis process; however, it enhanced its antibacterial activity against Gram‐negative and Gram‐positive strains; on the other hand, (iii) the replacement of Arg with Lys in the LfcinB or BFII sequences improved the chimeras’ synthesis process without significantly affecting their antibacterial activity. These results illustrate the great importance of the synthesis of chimeric peptides for the generation of promising antibacterial peptides.

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“…The collected fractions were analyzed via RP‐HPLC and MALDI‐TOF MS. The fractions containing the pure peptide were mixed and then lyophilized [33] …”

Section: Methodsmentioning

confidence: 99%

Designing Chimeric Peptides: A Powerful Tool for Enhancing Antibacterial Activity

Pineda-Castañeda

Huertas-Ortiz

Leal-Castro

et al. 2021

Chemistry & Biodiversity

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“…Peptides dissolved in solvent A were filtered through a 0.44 μm membrane and seeded in the column. Elution involved using a step gradient with increasing concentrations of solvent B; the fractions were collected and analysed by RP-HPLC, combining the fractions having the highest chromatographic purity [ 34 ].…”

Section: Methodsmentioning

confidence: 99%

Shorter Antibacterial Peptide Having High Selectivity for E. coli Membranes and Low Potential for Inducing Resistance

et al. 2020

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Antimicrobial peptides (AMPs) have been recognised as a significant therapeutic option for mitigating resistant microbial infections. It has been found recently that Plasmodium falciparum-derived, 20 residue long, peptide 35409 had antibacterial and haemolytic activity, making it an AMP having reduced selectivity, and suggesting that it should be studied more extensively for obtaining new AMPs having activity solely targeting the bacterial membrane. Peptide 35409 was thus used as template for producing short synthetic peptides (<20 residues long) and evaluating their biological activity and relevant physicochemical characteristics for therapeutic use. Four of the sixteen short peptides evaluated here had activity against E. coli without any associated haemolytic effects. The 35409-1 derivative (17 residues long) had the best therapeutic characteristics as it had high selectivity for bacterial cells, stability in the presence of human sera, activity against E. coli multiresistant clinical isolates and was shorter than the original sequence. It had a powerful membranolytic effect and low potential for inducing resistance in bacteria. This peptide’s characteristics highlighted its potential as an alternative for combating infection caused by E. coli multiresistant bacteria and/or for designing new AMPs.

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“…For the purification of the peptides, 5 g RP-SPE columns (particle size: 40-60 µm) were used [50]. The peptide was dissolved in solvent A, and the sample was seeded and then eluted with solutions containing different percentages of solvent B.…”

Section: Peptide Purification By Solid Phase Extraction (Spe)mentioning

confidence: 99%

Peptides Derived from (RRWQWRMKKLG)2-K-Ahx Induce Selective Cellular Death in Breast Cancer Cell Lines through Apoptotic Pathway

Insuasty‐Cepeda

Barragán‐Cárdenas

Ochoa‐Zarzosa

et al. 2020

IJMS

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The effect on the cytotoxicity against breast cancer cell lines of the substitution of 26Met residue in the sequence of the Bovine Lactoferricin-derived dimeric peptide LfcinB (20-30)2: (20RRWQWRMKKLG30)2-K-Ahx with amino acids of different polarity was evaluated. The process of the synthesis of the LfcinB (20-30)2 analog peptides was similar to the original peptide. The cytotoxic assays showed that some analog peptides exhibited a significant cytotoxic effect against breast cancer cell lines HTB-132 and MCF-7, suggesting that the substitution of the Met with amino acids of a hydrophobic nature drastically enhances its cytotoxicity against HTB-132 and MCF-7 cells, reaching IC50 values up to 6 µM. In addition, these peptides have a selective effect, since they exhibit a lower cytotoxic effect on the non-tumorigenic cell line MCF-12. Interestingly, the cytotoxic effect is fast (90 min) and is maintained for up to 48 h. Additionally, through flow cytometry, it was found that the obtained dimeric peptides generate cell death through the apoptosis pathway and do not compromise the integrity of the cytoplasmic membrane, and there are intrinsic apoptotic events involved. These results show that the obtained peptides are extremely promising molecules for the future development of drugs for use against breast cancer.

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Synthetic Peptide Purification via Solid-Phase Extraction with Gradient Elution: A Simple, Economical, Fast, and Efficient Methodology

Cited by 28 publications

References 23 publications

Designing Chimeric Peptides: A Powerful Tool for Enhancing Antibacterial Activity

Designing Chimeric Peptides: A Powerful Tool for Enhancing Antibacterial Activity

Shorter Antibacterial Peptide Having High Selectivity for E. coli Membranes and Low Potential for Inducing Resistance

Peptides Derived from (RRWQWRMKKLG)2-K-Ahx Induce Selective Cellular Death in Breast Cancer Cell Lines through Apoptotic Pathway

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