dc.contributor.author	Miller, Kerri
dc.contributor.author	Sarpong-Kumankomah, Sophia
dc.contributor.author	Egorov, Artem
dc.contributor.author	Gailer, Juergen
dc.date.accessioned	2021-09-13T15:49:59Z
dc.date.available	2021-09-13T15:49:59Z
dc.date.issued	2020-05-03
dc.description.abstract	The analysis of human plasma for biomarkers holds promise to revolutionize disease diagnosis, but is hampered by the inherent complexity of the plasma proteome. One way to overcome this problem is to analyze plasma for a sub-proteome, such as the metalloproteome. Previous studies employing size-exclusion chromatography (SEC) coupled on-line to an inductively coupled plasma-atomic emission spectrometer (ICP-AES) have revealed that plasma contains ~12 copper, iron and zinc metalloproteins. This included the iron metalloproteins transferrin (Tf) and a recently identified haptoglobin-hemoglobin (Hp-Hb) complex, which is formed in plasma when red blood cells rupture. Since this SEC-ICP-AES method required a sample volume of 500 µL to generate diagnostically useful results, we sought to develop an alternative SEC-based hyphenated approach using a smaller SEC column (150 x 5 mm I.D.) and a graphite furnace atomic absorption spectrometer (GFAAS) as the iron-specific detector. A designed interface enabled the integration of the SEC system with the GFAAS. Baseline separation between the Hp-Hb complex and Tf was achieved by developing a sample preparation procedure which involved the chelating agent-based mobilization of Fe from Tf to a small molecular weight Fe complex. Spiking of human plasma (1.0 mL) with red blood cell lysate (1-2 µL) increased only the intensity of the Fe peak corresponding to the Hp-Hb complex, but not that of Tf. Since the developed SEC-GFAAS method requires only 50 µL for analysis, it can now be employed for the cost-effective quantification of the clinically relevant Hb-Hp complex in human plasma in <50 min.	en_US
dc.description.grantingagency	Natural Sciences and Engineering Research Council (NSERC)	en_US
dc.description.grantingagency	Other	en_US
dc.identifier.citation	Miller, K., Sarpong-Kumankomah, S., Egorov, A., & Gailer, J. (2020). Sample preparation of blood plasma enables baseline separation of iron metalloproteins by SEC-GFAAS. Journal of Chromatography B, 1147, 122147.	en_US
dc.identifier.doi	https://doi.org/10.1016/j.jchromb.2020.122147	en_US
dc.identifier.uri	http://hdl.handle.net/1880/113856
dc.language.iso	eng	en_US
dc.publisher	Elsevier	en_US
dc.publisher.department	Chemistry	en_US
dc.publisher.faculty	Science	en_US
dc.publisher.hasversion	acceptedVersion	en_US
dc.publisher.institution	University of Calgary	en_US
dc.publisher.institution	Lumex Instruments Canada	en_US
dc.publisher.policy	https://www.elsevier.com/about/policies/sharing	en_US
dc.rights	Unless otherwise indicated, this material is protected by copyright and has been made available with authorization from the copyright owner. You may use this material in any way that is permitted by the Copyright Act or through licensing that has been assigned to the document. For uses that are not allowable under copyright legislation or licensing, you are required to seek permission.	en_US
dc.rights.uri	https://creativecommons.org/licenses/by-nc-nd/4.0	en_US
dc.title	Sample preparation of plasma enables baseline separation of iron metalloproteins by SEC-GFAAS	en_US
dc.type	journal article	en_US
ucalgary.item.requestcopy	true	en_US
ucalgary.scholar.level	Faculty	en_US

Sample preparation of plasma enables baseline separation of iron metalloproteins by SEC-GFAAS

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