Profiling preparations of recombinant birch pollen allergen Bet v 1a with capillary zone electrophoresis in pentamine modified fused-silica capillaries

Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences
M PunzetHanno Stutz

Abstract

Three preparation batches of the recombinant birch pollen allergen Bet v 1a have been analyzed by capillary zone electrophoresis (CZE) using a separation electrolyte consisting of 100 mmol L(-1) phosphate at pH 6.50 with 2.0 mmol L(-1) tetraethylenepentamine (TEPA) added. TEPA improved the resolution by wall shielding and selective attachment to allergens, but reduced migration repeatability at concentrations >2.0 mmol L(-1). Heterogeneity of preparations determined by CZE and electrospray ionization-quadrupole-time-of flight-MS were in accordance and revealed chemically modified (carbamylated) allergens in one of the preparations. The method was validated according to the ICH-guidelines. Repeatability of effective electrophoretic mobility (mu(eff)) was <0.55% R.S.D. (n = 5). Migration time corrected peak areas were used for quantification. Limit of quantification (LOQ) was 25 microg mL(-1) for the major isoform Bet v 1a, based on a signal-to-noise ratio of 10, and detector response was linear between LOQ and 0.90 mg mL(-1). Purity of the different rBet v 1a preparations was determined to be between 40 and 92% depending on the manufacturing protocol.

References

May 31, 1996·Journal of Chromatography. B, Biomedical Applications·V PacákováK Stulík
Mar 1, 1997·The Journal of Allergy and Clinical Immunology·Z ChenX Baur
Feb 1, 1997·Protein Expression and Purification·K Hoffmann-SommergruberH Breiteneder
May 1, 1997·Electrophoresis·A PetersenW M Becker
Oct 29, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·K A Denton, S A Tate
Dec 10, 1997·Journal of Chromatography. B, Biomedical Sciences and Applications·V PacákováM Tichá
Dec 22, 1998·Annals of Allergy, Asthma & Immunology : Official Publication of the American College of Allergy, Asthma, & Immunology·J BousquetH Yssel
Jul 3, 1999·Indoor Air·L Holmquist, O Vesterberg
Dec 22, 1999·Electrophoresis·A L Lagu
Sep 13, 2000·The Journal of Allergy and Clinical Immunology·M D ChapmanA Pomés
Apr 12, 2001·Electrophoresis·P G RighettiL Castelletti
May 1, 2001·The Science of the Total Environment·M VeriniF Chiarelli
Sep 6, 2001·Proceedings of the National Academy of Sciences of the United States of America·C CalonderP R Van Tassel
May 16, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Petra SázelováGabriel Peltre
May 16, 2002·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Jetse C ReijengMarcella Chiari
Aug 9, 2002·Auris, Nasus, Larynx·Mikikazu YamagiwaTsuyoshi Nishizumi
Sep 21, 2002·International Journal of Biometeorology·J EmberlinA Rantio-Lehtimäki
Dec 12, 2002·FASEB Journal : Official Publication of the Federation of American Societies for Experimental Biology·Martin HimlyFátima Ferreira
Mar 26, 2003·Electrophoresis·Erin A S DohertyAnnelise E Barron
Jul 5, 2003·Food and Chemical Toxicology : an International Journal Published for the British Industrial Biological Research Association·N WolffS Yannai
Aug 13, 2003·Clinical and Experimental Allergy : Journal of the British Society for Allergy and Clinical Immunology·S TreschP Schmid-Grendelmeier
Feb 14, 2004·Methods : a Companion to Methods in Enzymology·Oliver CromwellHelmut Fiebig
Apr 6, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Miao-Fang LinPhilip A Ropp
Jul 9, 2004·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Petra SázelováGabriel Peltre

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Citations

Nov 2, 2014·Journal of Separation Science·Theresa Kristl, Hanno Stutz
Jul 27, 2007·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Milan BlanusaTanja Cirkovic Velickovic

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