Engineered antibody CH2 domains binding to nucleolin: isolation, characterization and improvement of aggregation

UM > Faculty of Health Sciences > DEPARTMENT OF BIOMEDICAL SCIENCES

Status	已發表Published
	Engineered antibody CH2 domains binding to nucleolin: isolation, characterization and improvement of aggregation
	Li, D.; Gong, R.; Zheng, J.; Chen, X.; Dimitrov, D.; Zhao, Q.
	2017-04-01
Source Publication	Biochemical and Biophysical Research Communications
ISSN	0006-291X
Pages	446-453
Abstract	Smaller recombinant antibody fragments are now emerging as alternatives of conventional antibodies. Especially, immunoglobulin (Ig) constant CH2 domain and engineered CH2 with improved stability are promising as scaffolds for selection of specific binders to various antigens. We constructed a yeast display library based on an engineered human IgG1 CH2 scaffold with diversified loop regions. A group of CH2 binders were isolated from this yeast display library by panning against nucleolin, which is a tumor-associated antigen involved in cell proliferation, tumor cell growth and angiogenesis. Out of 20 mutants, we selected 3 clones exhibiting relatively high affinities to nucleolin on yeasts. However, recombinant CH2 mutants aggregated when they were expressed. To find the mechanism of the aggregation, we employed computational prediction approaches through structural homology models of CH2 binders. The analysis of potential aggregation prone regions (APRs) and solvent accessible surface areas (ASAs) indicated two hydrophobic residues, Val264 and Leu309, in the β-sheet, in which replacement of both charged residues led to significant decrease of the protein aggregation. The newly identified CH2 binders could be improved to use as candidate therapeutics or research reagents in the future.
Keyword	antibody domain CH2 nucleolin yeast display monoclonal antibody aggregation prone region
Language	英語English
The Source to Article	PB_Publication
PUB ID	29846
Document Type	Journal article
Collection	DEPARTMENT OF BIOMEDICAL SCIENCES
Corresponding Author	Chen, X.; Zhao, Q.
Recommended Citation GB/T 7714	Li, D.,Gong, R.,Zheng, J.,et al. Engineered antibody CH2 domains binding to nucleolin: isolation, characterization and improvement of aggregation[J]. Biochemical and Biophysical Research Communications, 2017, 446-453.
APA	Li, D.., Gong, R.., Zheng, J.., Chen, X.., Dimitrov, D.., & Zhao, Q. (2017). Engineered antibody CH2 domains binding to nucleolin: isolation, characterization and improvement of aggregation. Biochemical and Biophysical Research Communications, 446-453.
MLA	Li, D.,et al."Engineered antibody CH2 domains binding to nucleolin: isolation, characterization and improvement of aggregation".Biochemical and Biophysical Research Communications (2017):446-453.

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