Format

Send to

Choose Destination
Protein Eng Des Sel. 2013 Feb;26(2):151-64. doi: 10.1093/protein/gzs090. Epub 2012 Nov 19.

An integrated approach to extreme thermostabilization and affinity maturation of an antibody.

Author information

1
AnaptysBio, Inc., 10421 Pacific Center Court, Suite 200, San Diego, CA 92121, USA. amcconnell@anaptysbio.com

Abstract

Antibodies are important tools for a broad range of applications due to their high specificity and ability to recognize virtually any target molecule. However, in order to be practically useful, antibodies must be highly stable and bind their target antigens with high affinity. We present a combinatorial approach to generate high-affinity, highly stable antibodies through the design of stable frameworks, specificity grafting and maturation via somatic hypermutation in vitro. By collectively employing these methods, we have engineered a highly stable, high-affinity, full-length antibody with a T(m) over 90°C that retains significant activity after heating to 90°C for 1 h, and has ~95-fold improved antigen-binding affinity. The stabilized IgG framework is compatible with affinity maturation, and should provide a broadly useful scaffold for grafting a variety of complementarity-determining region loops for the development of stable antibodies with desired specificities.

PMID:
23173178
DOI:
10.1093/protein/gzs090
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Silverchair Information Systems
Loading ...
Support Center