Sample preparation for sequencing hits from one-bead–one-peptide combinatorial libraries by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Martínez‐Ceron, María C.; Giudicessi, Silvana L.; Marani, Mariela M.; Alberício, Fernando; Cascone, Osvaldo; Erra‐Balsells, Rosa; Camperi, Silvia A.

doi:10.1016/j.ab.2010.01.029

Cited by 22 publications

(18 citation statements)

References 12 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…This effect may complicate the correct interpretation of spectra, which was also observed in the present study. The second reference matrix, DHB, is reported to display reduced matrix cluster signals but also decreased analyte ionization and therefore reduced sentivity (see also mass spectra shown in Figure S4, supporting information). Overall, the results obtained by using monoaromatic lichen metabolites as matrices were quite close to those of DHB.…”

Section: Resultsmentioning

confidence: 99%

The inherent matrix properties of lichen metabolites in matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry

Pogam

Boustié

Richomme

et al. 2017

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

Lichen metabolites exhibit interesting MALDI matrix properties, especially for medium and low molecular weight analytes. For many of the tested molecules, matrix ion formation was very limited. This proof-of-concept study paves the way for follow-up investigations to assess the matrix properties of lichen metabolites against a wider array of analytes as well as adapting experimental settings to individually optimize the performance of successfully tested candidates.

show abstract

Section: Resultsmentioning

confidence: 99%

The inherent matrix properties of lichen metabolites in matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry

Pogam

Boustié

Richomme

et al. 2017

Rapid Comm Mass Spectrometry

View full text Add to dashboard Cite

show abstract

“…6 Graph of q* = f (c*) describing the adsorption isotherms for target protein binding to peptide immobilized on the chromatographic support, c* = free protein concentration at equilibrium, q * = the equilibrium concentration of the target protein bound to the immobilized peptide, per unit of total chromatographic matrix volume 3. Dynamic capacity is calculated as the amount of protein adsorbed by the matrix before 10 % of the initial protein concentration is detected at the column outlet (Fig.…”

Section: Breakthrough Curvesmentioning

confidence: 99%

“…The bond formed between the first amino acid and the linker HMBA is stable to all synthetic elongation reactions as well as to the conditions required for the removal of the side-chain protecting group. The solid support is amenable to easy release of the peptide for mass spectrometry (MS) analysis [5,6]. For peptides shorter than six amino acids, Ala and/or Gly residues may be introduced in the C-termini to increase peptide molecular weight in order to facilitate MS analysis.…”

Section: Introductionmentioning

confidence: 99%

Peptide Affinity Chromatography Based on Combinatorial Strategies for Protein Purification

Camperi

Martínez‐Ceron

Giudicessi

et al. 2014

Methods in Molecular Biology

Self Cite

View full text Add to dashboard Cite

We describe a method to develop affinity chromatography matrices with short peptide ligands for protein purification. The method entitles the following: (a) synthesis of a combinatorial library on the hydromethylbenzoyl (HMBA)-ChemMatrix resin by the divide-couple-recombine (DCR) method using the Fmoc chemistry, (b) library screening with the protein of interest labeled with a fluorescent dye or biotin, (c) identification of peptides contained on positive beads by tandem matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS/MS), (d) solid-phase peptide ligand synthesis and immobilization in chromatographic supports, and (e) evaluation of protein adsorption on peptide affinity matrices from the equilibrium isotherms and breakthrough curves.

show abstract

“…The analysis was performed as per Martínez-Ceron et al [26]. An amount of 1 μl aliquot of eluted peptide from a single bead was loaded onto the sample plate, air dried at room temperature, and then, 1 μl CHCA 4 mg/ml in CH 3 CN/H 2 O (1/1) with 0.1% TFA and doped with serine 20 mM [27] was added on the sample dry layer (successive dry layers deposit method).…”

Section: Maldi Ms Analysismentioning

confidence: 99%

A simple protocol for combinatorial cyclic depsipeptide libraries sequencing by matrix-assisted laser desorption/ionisation mass spectrometry

et al. 2014

Self Cite

View full text Add to dashboard Cite

Short cyclic peptides have a great interest in therapeutic, diagnostic and affinity chromatography applications. The screening of 'one-bead-one-peptide' combinatorial libraries combined with mass spectrometry (MS) is an excellent tool to find peptides with affinity for any target protein. The fragmentation patterns of cyclic peptides are quite more complex than those of their linear counterparts, and the elucidation of the resulting tandem mass spectra is rather more difficult. Here, we propose a simple protocol for combinatorial cyclic libraries synthesis and ring opening before MS analysis. In this strategy, 4-hydroxymethylbenzoic acid, which forms a benzyl ester with the first amino acid, was used as the linker. A glycolamidic ester group was incorporated after the combinatorial positions by adding glycolic acid. The library synthesis protocol consisted in the following: (i) incorporation of Fmoc-Asp[2-phenylisopropyl (OPp)]-OH to Ala-Gly-oxymethylbenzamide-ChemMatrix, (ii) synthesis of the combinatorial library, (iii) assembly of a glycolic acid, (iv) couple of an Ala residue in the N-terminal, (v) removal of OPp, (vi) peptide cyclisation through side chain Asp and N-Ala amino terminus and (vii) removal of side chain protecting groups. In order to simultaneously open the ring and release each peptide, benzyl and glycolamidic esters were cleaved with ammonia. Peptide sequences could be deduced from the tandem mass spectra of each single bead evaluated. The strategy herein proposed is suitable for the preparation of one-bead-one-cyclic depsipeptide libraries that can be easily open for its sequencing by matrix-assisted laser desorption/ionisation MS. It employs techniques and reagents frequently used in a broad range of laboratories without special expertise in organic synthesis.

show abstract

Sample preparation for sequencing hits from one-bead–one-peptide combinatorial libraries by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry

Cited by 22 publications

References 12 publications

The inherent matrix properties of lichen metabolites in matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry

The inherent matrix properties of lichen metabolites in matrix‐assisted laser desorption ionization time‐of‐flight mass spectrometry

Peptide Affinity Chromatography Based on Combinatorial Strategies for Protein Purification

A simple protocol for combinatorial cyclic depsipeptide libraries sequencing by matrix-assisted laser desorption/ionisation mass spectrometry

Contact Info

Product

Resources

About