Analysis of non-covalent aggregation of synthetic hPTH (1-34) by size-exclusion chromatography and the importance of suppression of non-specific interactions for a precise quantitation.
There are few methods available for the rapid and precise quantitation of non-covalent aggregation. Size-exclusion chromatography (SEC), a traditional approach, used to measure the non-covalent aggregation can easily disrupt the weak forces holding an aggregate together. Under the conditions described in this paper the disaggregation of non-covalent aggregate of the synthetic human parathyroid hormone hPTH (1-34) due to hydrophobic/electrostatic interactions with the size-exclusion chromatography column packing was completely suppressed. This report details the effectiveness of adding salts and organic solvents in the mobile phase to overcome non-specific interactions that disrupt the aggregate during the SEC process and may aid in the understanding precise quantitation of non-covalent aggregation.
Kamberi
Marika
M
Department of Analytical Sciences, ALZA Corporation, Mountain View 94039-7210, 1501 California Avenue, Palo Alto, CA 94304, USA. mkamberi@alzus.jnj.com
Chung
Paul
P
DeVas
Richard
R
Li
Lily
L
Li
Zengji
Z
Ma
Xiaoyan Sharon
XS
Fields
Steven
S
Riley
Christopher M
CM
D000595:Amino Acid Sequence; D002850:Chromatography, Gel; D006801:Humans; D007202:Indicators and Reagents; D008027:Light; D008969:Molecular Sequence Data; D012542:Scattering, Radiation; D012965:Sodium Chloride; D012997:Solvents; D013056:Spectrophotometry, Ultraviolet; D019379:Teriparatide / Q000032:analysis*
D007202:Indicators and Reagents; D012997:Solvents; D019379:Teriparatide; D012965:Sodium Chloride