Divalent metal cation chelators enhance chromatographic separation of structurally similar macromolecules: separation of human growth hormone isoforms.

abstract

• Human GH isoforms were separated by anion-exchange chromatography using a linear NaCl gradient in the presence and absence of EDTA and EGTA. SDS-PAGE showed that glycosylated 24-kDa hGH did not appreciably separate from other hGH variants in the absence of metal chelators. However, in the presence of metal chelators, glycosylated 24-kDa hGH separated from the bulk of the hGH isoforms. Human GH isoforms were also separated by size-exclusion chromatography in the presence and absence of metal chelators. Glycosylated 24-kDa hGH eluted with the bulk of the hGH isoforms in both separations. The inclusion of metal chelators in chromatographic buffers to alter the charge and/or size of proteins by stripping their metals may be a generally useful strategy in their fractionation.

authors

• Bustamante, Juan

published proceedings

• J Chromatogr B Biomed Sci Appl

altmetric score

• 3

author list (cited authors)

• Haro, L. S., Cubriel, A., Bustamante, J., Flores, R., & Martinez, A. O.

citation count

• 3

complete list of authors

• Haro, LS||Cubriel, A||Bustamante, J||Flores, R||Martinez, AO

publication date

• January 1998

publisher

• Elsevier  Publisher

published in

• J Chromatogr B Biomed Sci Appl  Journal

keywords

• Cations, Divalent
• Chelating Agents
• Chromatography, Gel
• Chromatography, Ion Exchange
• Electrophoresis, Polyacrylamide Gel
• Human Growth Hormone
• Humans
• Metals
• Protein Isoforms

Digital Object Identifier (DOI)

• 10.1016/s0378-4347(98)00419-8

start page

• 39

end page

• 47

volume

• 720

issue

• 1-2

URL

• http://dx.doi.org/10.1016/s0378-4347(98)00419-8