On-line hollow-fiber flow field-flow fractionation-electrospray ionization/time-of-flight mass spectrometry of intact proteins

Pierluigi Reschiglian, Andrea Zattoni, Barbara Roda, Leonardo Cinque, Daniela Parisi, Aldo Roda, Fabrizio Dal Piaz, Myeong Hee Moon, Byung Ryul Min

Research output: Contribution to journalArticle

58 Citations (Scopus)

Abstract

Capabilities of mass spectrometry for the analysis of intact proteins can be increased through separation methods. Flow field-flow fractionation (FlFFF) is characterized by the particularly "soft" separation mechanism, which is ideally suited to maintain the native structure of intact proteins. This work describes the original on-line coupling between hollow-fiber FlFFF (HF FlFFF), the microcolumn variant of FlFFF, and electrospray ionization/time-of- flight mass spectrometry (ESI/TOFMS) for the analysis and characterization of intact proteins. The results show that the native (or pseudonative) structure of horse heart myoglobin and horseradish peroxidase is maintained. Sample desalting is also observed for horse heart myoglobin. Correlation between the molar mass values independently measured by HF FlFFF retention and ESI/TOFMS allows us to confirm the protein aggregation features of bovine serum albumin and to indicate possible changes in the quaternary structure of human hemoglobin.

Original languageEnglish
Pages (from-to)47-56
Number of pages10
JournalAnalytical Chemistry
Volume77
Issue number1
DOIs
Publication statusPublished - 2005 Jan 1

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Electrospray ionization
Fractionation
Mass spectrometry
Flow fields
Fibers
Myoglobin
Proteins
Salt removal
Molar mass
Horseradish Peroxidase
Bovine Serum Albumin
Hemoglobins
Agglomeration

All Science Journal Classification (ASJC) codes

  • Analytical Chemistry

Cite this

Reschiglian, Pierluigi ; Zattoni, Andrea ; Roda, Barbara ; Cinque, Leonardo ; Parisi, Daniela ; Roda, Aldo ; Dal Piaz, Fabrizio ; Moon, Myeong Hee ; Min, Byung Ryul. / On-line hollow-fiber flow field-flow fractionation-electrospray ionization/time-of-flight mass spectrometry of intact proteins. In: Analytical Chemistry. 2005 ; Vol. 77, No. 1. pp. 47-56.
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On-line hollow-fiber flow field-flow fractionation-electrospray ionization/time-of-flight mass spectrometry of intact proteins. / Reschiglian, Pierluigi; Zattoni, Andrea; Roda, Barbara; Cinque, Leonardo; Parisi, Daniela; Roda, Aldo; Dal Piaz, Fabrizio; Moon, Myeong Hee; Min, Byung Ryul.

In: Analytical Chemistry, Vol. 77, No. 1, 01.01.2005, p. 47-56.

Research output: Contribution to journalArticle

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AU - Parisi, Daniela

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AU - Dal Piaz, Fabrizio

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AU - Min, Byung Ryul

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