Dataset Information

Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297.

ABSTRACT: The structural integrity and conformational stability of various IgG1-Fc proteins produced from the yeast Pichia pastoris with different glycosylation site occupancy (di-, mono-, and nonglycosylated) were determined. In addition, the physical stability profiles of three different forms of nonglycosylated Fc molecules (varying amino-acid residues at site 297 in the CH 2 domain due to the point mutations and enzymatic digestion of the Fc glycoforms) were also examined. The physical stability of these IgG1-Fc glycoproteins was examined as a function of pH and temperature by high-throughput biophysical analysis using multiple techniques combined with data visualization tools (three index empirical phase diagrams and radar charts). Across the pH range of 4.0-6.0, the di- and monoglycosylated forms of the IgG1-Fc showed the highest and lowest levels of physical stability, respectively, with the nonglycosylated forms showing intermediate stability depending on solution pH. In the aglycosylated Fc proteins, the introduction of Asp (D) residues at site 297 (QQ vs. DN vs. DD forms) resulted in more subtle changes in structural integrity and physical stability depending on solution pH. The utility of evaluating the conformational stability profile differences between the various IgG1-Fc glycoproteins is discussed in the context of analytical comparability studies.

SUBMITTER: Alsenaidy MA

PROVIDER: S-EPMC4512762 | biostudies-literature | 2014 Jun

REPOSITORIES: biostudies-literature

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Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297.

Alsenaidy Mohammad A MA Okbazghi Solomon Z SZ Kim Jae Hyun JH Joshi Sangeeta B SB Middaugh C Russell CR Tolbert Thomas J TJ Volkin David B DB

Journal of pharmaceutical sciences 20140416 6

The structural integrity and conformational stability of various IgG1-Fc proteins produced from the yeast Pichia pastoris with different glycosylation site occupancy (di-, mono-, and nonglycosylated) were determined. In addition, the physical stability profiles of three different forms of nonglycosylated Fc molecules (varying amino-acid residues at site 297 in the CH 2 domain due to the point mutations and enzymatic digestion of the Fc glycoforms) were also examined. The physical stability of th ...[more]

PMID: 24740840

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Project description:This meta-analysis was performed to evaluate the association between xeroderma pigmentosum complementary group D (XPD) Lys751Gln and Asp312Asn polymorphisms and susceptibility to hepatocellular carcinoma (HCC). PubMed, Embase, Google Scholar and the Chinese National Knowledge Infrastructure and the Chinese Biomedicine databases were systematically searched to identify relevant studies published up to June 1, 2014. Statistical analyses were performed using Stata version 12.0 software. A total of 11 case-control studies, comprising 2,852 cases and 2,936 controls, were included. The results of the meta-analysis revealed that a significant association between the risk of HCC and variant genotypes of the XPD Lys751Gln and Asp312Asn polymorphisms was evident in the homozygote comparison [Gln/Gln versus Lys/Lys: Odds ratio (OR), 1.831; 95% confidence interval (CI), 1.001-3.349], heterozygote comparison (Lys/Gln versus Lys/Lys: OR, 1.486; 95% CI, 1.044-2.114), dominant model (Gln/Gln + Lys/Gln versus Lys/Lys: OR, 1.540; 95% CI, 1.054-2.249) and allelic contrast (Gln-allele versus Lys-allele: OR, 1.453; 95% CI, 1.032-2.046) for the Lys751Gln polymorphism and the homozygote comparison for the Asp312Asn polymorphism (Asn/Asn versus Asp/Asp: OR, 1.352; 95% CI, 1.010-1.808). By contrast, no significant association was observed in the recessive model for the Lys751Gln polymorphism (Gln/Gln versus Lys/Gln + Lys/Lys: OR, 1.603; 95% CI, 0.924-2.779), or for the heterozygote comparison (Asn/Asp versus Asp/Asp: OR, 1.229; 95% CI, 0.857-1.762), dominant model (Asn/Asn + Asp/Asn versus Asp/Asp: OR, 1.249; 95% CI, 0.910-1.715), recessive model (Asn/Asn versus Asp/Asn + Asp/Asp: OR, 1.250; 95% CI, 0.940-1.663) or allelic contrast (Asn-allele versus Asp-allele: OR, 1.226; 95% CI, 0.965-1.557) for the Asp312Asn polymorphism. The present meta-analysis has indicated that the XPD Lys751Gln polymorphism could be a potential biomarker of HCC susceptibility and that the XPD Lys751Gln and Asp312Asn polymorphisms could be risk factors for HCC susceptibility in an Asian population; however, further large-scale and well-designed studies are required to reach a more precise and comprehensive conclusion.

Dataset Information

Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297.

Publications

Physical stability comparisons of IgG1-Fc variants: effects of N-glycosylation site occupancy and Asp/Gln residues at site Asn 297.

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