Factors influencing the performance of peptide mapping by reversed-phase high-performance liquid chromatography

Dong, Michael W.; Tran, An D.

doi:10.1016/s0021-9673(00)96968-1

Cited by 34 publications

(10 citation statements)

References 30 publications

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“…To further purify the most promising peptides contained in the 1 kDa-UFH fraction, preparative RP-HPLC was performed, the peptides were collected every minute and the separation was monitored at 214 nm, wavelength of absorbance of peptide bonds, and 280 nm, which is the wavelength of choice to detect residues of aromatic amino acids [28]. The RP-HPLC chromatogram obtained for the 1 kDa-UFH fraction of Ulva sp.…”

Section: Resultsmentioning

confidence: 99%

In Vitro and In Silico Approaches to Generating and Identifying Angiotensin-Converting Enzyme I Inhibitory Peptides from Green Macroalga Ulva lactuca

2019

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A protein extract was generated from the macroalga Ulva lactuca, which was subsequently hydrolysed using the food-grade enzyme papain and angiotensin-converting Enzyme I and renin inhibitory peptides identified using a combination of enrichment strategies employing molecular weight cutoff filtration and mass spectrometry analysis. The generated hydrolysates with the most promising in vitro activity were further purified using preparative RP-HPLC and characterised. The 1 kDa hydrolysate (1 kDa-UFH), purified and collected by preparative RP-HPLC at minutes 41‒44 (Fr41‒44), displayed statistically higher ACE-I inhibitory activities ranging from 96.91% to 98.06%. A total of 48 novel peptides were identified from these four fractions by LC-MS/MS. A simulated gastrointestinal digestion of the identified peptide sequences was carried out using in silico enzyme cleavage simulation tools, resulting in 86 peptide sequences that were further assessed for their potential activity, toxicity and allergenicity using multiple predictive approaches. All the peptides obtained in this study were predicted to be non-toxic. However, 28 out of the 86 novel peptides released after the in silico gastrointestinal digestion were identified as potential allergens. The potential allergenicity of these peptides should be further explored to comply with the current labelling regulations in formulated food products containing U. lactuca protein hydrolysates.

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Section: Resultsmentioning

confidence: 99%

In Vitro and In Silico Approaches to Generating and Identifying Angiotensin-Converting Enzyme I Inhibitory Peptides from Green Macroalga Ulva lactuca

2019

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“…In the production of biopharmaceutical proteins peptide mapping with reversed-phase (RP) chromatography has been used for the analyses of recombinant protein biopharmaceuticals delivering comprehensive characterisation of products [[22], [23], [24], [25], [26]]. In addition, these methods are also employed for subsequent lot-to-lot identity testing (‘fingerprinting’) in support of GMP production.…”

Section: Introductionmentioning

confidence: 99%

High resolution fingerprinting of single and double-stranded RNA using ion-pair reverse-phase chromatography

Nwokeoji

Earll

Kilby

et al. 2019

Journal of Chromatography B

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The emergence of new sustainable approaches for insect management using RNA interference (RNAi) based insecticides has created the demand for high throughput analytical techniques to fully characterise and accurately quantify double stranded RNA (dsRNA) prior to downstream RNAi applications. In this study we have developed a method for the rapid characterisation of single stranded and double stranded RNA using high resolution RNase mapping in conjunction with ion-pair reverse-phase chromatography utilising a column with superficially porous particles. The high resolution oligoribonucleotide map provides an important ‘fingerprint’ for identity testing and bioprocess monitoring. Reproducible RNA mapping chromatograms were generated from replicate analyses. Moreover, this approach was used to provide a method to rapidly distinguish different RNA sequences of the same size, based on differences in the resulting chromatograms. Principal components analysis of the high resolution RNA mapping data enabled us to rapidly compare multiple HPLC chromatograms and distinguish two dsRNA sequences of different size which share 72% sequence homology. We used the high resolution RNase mapping method to rapidly fingerprint biomanufactured dsRNA across a number of different batches. The resulting chromatograms in conjunction with principal components analysis demonstrated high similarity in the dsRNA produced across the different batches highlighting the potential ability of this method to provide information for batch release in a high throughput manner.

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“…The operating conditions have a profound effect on peptide resolution because of the following equation [41,42] …”

Section: Gdiekmentioning

confidence: 99%

Electrochemical detection as an alternative to UV in RP‐HPLC peptide mapping

Chen

Krull

1994

Electroanalysis

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This article describes the possibility of electrochemical (EC) detection as an alternative to ultraviolet (C!) detection doing peptide mapping. The high sensitivity and selectivity of the electrochemical detection can be applied to determine some digested peptides. This approach demonstrated that peptides containing electroactive amino acids can be determined by EC detection. Furthermore, upon irradiation, noneiectroactive amino acids and peptides can be made electroactive, thereby increasing the sensitivity and selectivity of peptide mapping. A variety of effects are discussed to help in understanding the possibility of this useful approach.

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Factors influencing the performance of peptide mapping by reversed-phase high-performance liquid chromatography

Cited by 34 publications

References 30 publications

In Vitro and In Silico Approaches to Generating and Identifying Angiotensin-Converting Enzyme I Inhibitory Peptides from Green Macroalga Ulva lactuca

In Vitro and In Silico Approaches to Generating and Identifying Angiotensin-Converting Enzyme I Inhibitory Peptides from Green Macroalga Ulva lactuca

High resolution fingerprinting of single and double-stranded RNA using ion-pair reverse-phase chromatography

Electrochemical detection as an alternative to UV in RP‐HPLC peptide mapping

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