A novel antibody engineering strategy for making monovalent bispecific heterodimeric IgG antibodies by electrostatic steering mechanism.

Authors:
Zhi Liu
Zhi Liu
State Key Laboratory of Functional Materials for Informatics
China
Esther C Leng
Esther C Leng
University of British Columbia
Canada
Kannan Gunasekaran
Kannan Gunasekaran
National Cancer Institute
United States
Min Shen
Min Shen
National Center for Advancing Translational Sciences
China
Monique Howard
Monique Howard
University of Melbourne
Australia

J Biol Chem 2015 Mar 12;290(12):7535-62. Epub 2015 Jan 12.

From the Departments of Therapeutic Discovery and Amgen Inc., Seattle, Washington 98119,

Producing pure and well behaved bispecific antibodies (bsAbs) on a large scale for preclinical and clinical testing is a challenging task. Here, we describe a new strategy for making monovalent bispecific heterodimeric IgG antibodies in mammalian cells. We applied an electrostatic steering mechanism to engineer antibody light chain-heavy chain (LC-HC) interface residues in such a way that each LC strongly favors its cognate HC when two different HCs and two different LCs are co-expressed in the same cell to assemble a functional bispecific antibody. We produced heterodimeric IgGs from transiently and stably transfected mammalian cells. The engineered heterodimeric IgG molecules maintain the overall IgG structure with correct LC-HC pairings, bind to two different antigens with comparable affinity when compared with their parental antibodies, and retain the functionality of parental antibodies in biological assays. In addition, the bispecific heterodimeric IgG derived from anti-HER2 and anti-EGF receptor (EGFR) antibody was shown to induce a higher level of receptor internalization than the combination of two parental antibodies. Mouse xenograft BxPC-3, Panc-1, and Calu-3 human tumor models showed that the heterodimeric IgGs strongly inhibited tumor growth. The described approach can be used to generate tools from two pre-existent antibodies and explore the potential of bispecific antibodies. The asymmetrically engineered Fc variants for antibody-dependent cellular cytotoxicity enhancement could be embedded in monovalent bispecific heterodimeric IgG to make best-in-class therapeutic antibodies.

Download full-text PDF

Source
http://dx.doi.org/10.1074/jbc.M114.620260DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC4367261PMC

Still can't find the full text of the article?

We can help you send a request to the authors directly.
March 2015
113 Reads
23 Citations
4.573 Impact Factor

Publication Analysis

Top Keywords

heterodimeric igg
20
bispecific heterodimeric
16
parental antibodies
12
monovalent bispecific
12
antibodies
9
heterodimeric iggs
8
mammalian cells
8
igg antibodies
8
bispecific antibodies
8
making monovalent
8
steering mechanism
8
electrostatic steering
8
strategy making
8
heterodimeric
7
bispecific
7
igg
6
compared parental
4
antibodies biological
4
affinity compared
4
functionality parental
4

References

(Supplied by CrossRef)
Coexpression of epidermal growth factor receptor and ligands in human pancreatic cancer is associated with enhanced tumor aggressiveness
Yamanaka et al.
Anticancer Res 1993

Similar Publications