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Updated: Oct 4, 2026

Genetic Variant Detection in the CALR gene using High Resolution Melting Analysis
Published on: August 26, 2020
Identification and characterization by high-performance liquid chromatography/electrospray ionization mass
A Motos Gallardo1, J A Hernández, J M Hernández
1Fundació per a la Recerca Biomèdica 'Germans Trias i Pujol' (HUGTIP), Badalona, Barcelona, Spain. fgallar1@pie.xtec.es
Abstract:
This work illustrates the practical use of combined microbore reversed-phase high-performance liquid chromatography (RP-HPLC) with electrospray ionization mass spectrometry (ESI-MS) in protein identification. The approach consisted of the detection of the abnormal beta-globin chain by ESI-MS analysis of mixtures of intact globins, which simultaneously provided their molecular masses. Separation of the polypeptide globin chains was carried out using microbore C4 RP-HPLC on-line with ESI-MS. Direct peptide-mapping ESI-MS without previous chromatographic separation was performed in order to identify tryptic peptides from whole blood. For the sequence confirmation of the abnormal peptide containing the mutation point, C18 RP-HPLC tryptic separation of the globin mixture on-line with collision-induced dissociation (CID) fragmentation was done. The y series ions allowed the identification of the hemoglobin (Hb) variant as [beta134(H12) Val > Ala]. This new Hb was named Hb Mataró, after the city where it was detected.
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