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Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation
Ding, Ling1,2; Guo, Zhimou1; Hu, Zhuo1; Liang, Xinmiao1
KeywordMixed-mode Stationary Phase Reversed-phase Liquid Chromaitography Positively Charged Repulsion Intact Protein Separation Click Chemistry
Source PublicationJOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS
2017-05-10
DOI10.1016/j.jpba.2017.01.004
Volume138Pages:63-69
Indexed BySCI
SubtypeArticle
WOS HeadingsScience & Technology ; Physical Sciences ; Life Sciences & Biomedicine
WOS SubjectChemistry, Analytical ; Pharmacology & Pharmacy
WOS Research AreaChemistry ; Pharmacology & Pharmacy
WOS KeywordPERFORMANCE LIQUID-CHROMATOGRAPHY ; RECOMBINANT MONOCLONAL-ANTIBODIES ; TOP-DOWN PROTEOMICS ; STATIONARY PHASES ; HORIZONTAL POLYMERIZATION ; HYDROPHOBIC INTERACTION ; THERAPEUTIC PROTEINS ; GRADIENT ELUTION ; ION-EXCHANGE ; PEPTIDES
AbstractA mixed-mode reversed phase/positively charged repulsion stationary phase C8PN composed of octyl and amino group has been developed for separation of intact protein. Before the separation of proteins, a set of probe compounds were employed to evaluate the chromatographic properties of C8PN, demonstrating typical reversed phase/positively charged repulsion interaction on this stationary phase as estimated. Then the new C8PN stationary phase was used to separate a standard protein mixture on the reversed phase mode. Compared with a commercial C4 stationary phase, it showed different selectivity for some proteins. In order to better understand the properties of C8PN, the effect of acetonitrile content was investigated based on retention equation. Higher values of the equation parameters on C8PN demonstrated that the protein retentions were more sensitive to the change of acetonitrile content. Besides, the influences of buffer salt additives on the protein retentions were also studied. The retention factors of the proteins got larger with the increase of buffer salt concentration, which confirmed the positively charged repulsion interaction on the column. Finally, the C8PN was further applied to separate oxidized and reduced- forms of Recombinant Human Growth Hormone. Our study indicated the advantages and application potential of mixed-mode reversed phase/positively charged repulsion stationary phase for intact protein separation. (C) 2017 Elsevier B.V. All rights reserved.
Language英语
WOS IDWOS:000401782400007
Citation statistics
Cited Times:2[WOS]   [WOS Record]     [Related Records in WOS]
Document Type期刊论文
Identifierhttp://cas-ir.dicp.ac.cn/handle/321008/152256
Collection中国科学院大连化学物理研究所
Affiliation1.Chinese Acad Sci, Dalian Inst Chem Phys, Key Lab Separat Sci Analyt Chem, Dalian 116023, Peoples R China
2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China
Recommended Citation
GB/T 7714
Ding, Ling,Guo, Zhimou,Hu, Zhuo,et al. Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation[J]. JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,2017,138:63-69.
APA Ding, Ling,Guo, Zhimou,Hu, Zhuo,&Liang, Xinmiao.(2017).Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation.JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS,138,63-69.
MLA Ding, Ling,et al."Mixed-mode reversed phase/positively charged repulsion chromatography for intact protein separation".JOURNAL OF PHARMACEUTICAL AND BIOMEDICAL ANALYSIS 138(2017):63-69.
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