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Optimal fusion of antibody binding domains resulted in higher affinity and wider specificity

Paper ID Volume ID Publish Year Pages File Format Full-Text
20167 43161 2015 6 PDF Available
Title
Optimal fusion of antibody binding domains resulted in higher affinity and wider specificity
Abstract

•A series of antibody binding proteins PAxPG is made from protein A and protein G.•Linkage of the domains by a flexible linker yielded higher affinity than the parents.•Optimization of the linker length resulted in higher affinity to hIgG Fab and Fc.•With its high affinity, PAxPG will be a useful probe to many Abs including mIgG1.

Antibody is a very important protein in biotechnological and biomedical fields because of its high affinity and specificity to various antigens. Due to the rise of human antibody therapeutics, its cost-effective purification is an urgent issue for bio-industry. In this study, we made novel fusion proteins PAxPG with a flexible (DDAKK)n linker between the two Ig binding domains derived from Staphylococcus protein A and Streptococcus protein G. The fusion proteins bound human and mouse IgGs and their fragments with up to 58-times higher affinity and wider specificity than the parental binding domains. Interestingly, the optimal linker for human Fab fragment was n = 4, which was close to the modeled distance between the termini of domains bound to heavy chain, implying increased avidity as a possible mechanism. For binding to Fc, the longest n = 6 linker gave the highest affinity, implying longer interchain distance between the two binding sites. The novel fusion protein with optimized interdomain linker length will be a useful tool for the purification and detection of various IgGs including mouse IgG1 that binds only weakly to natural protein A.

Keywords
Antibody binding protein; Affinity tag; Immunoassay; Avidity effect; Linker engineering
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Optimal fusion of antibody binding domains resulted in higher affinity and wider specificity
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 120, Issue 5, November 2015, Pages 504–509
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us