Boosting antibody developability through rational sequence optimization.


Abstract

The application of monoclonal antibodies as commercial therapeutics poses substantial demands on stability and properties of an antibody. Therapeutic molecules that exhibit favorable properties increase the success rate in development. However, it is not yet fully understood how the protein sequences of an antibody translates into favorable in vitro molecule properties. In this work, computational design strategies based on heuristic sequence analysis were used to systematically modify an antibody that exhibited a tendency to precipitation in vitro. The resulting series of closely related antibodies showed improved stability as assessed by biophysical methods and long-term stability experiments. As a notable observation, expression levels also improved in comparison with the wild-type candidate. The methods employed to optimize the protein sequences, as well as the biophysical data used to determine the effect on stability under conditions commonly used in the formulation of therapeutic proteins, are described. Together, the experimental and computational data led to consistent conclusions regarding the effect of the introduced mutations. Our approach exemplifies how computational methods can be used to guide antibody optimization for increased stability.

Submission Details

ID: oRPNZUHY

Submitter: Marie Ary

Submission Date: Nov. 29, 2018, 5:01 p.m.

Version: 1

Publication Details
Seeliger D;Schulz P;Litzenburger T;Spitz J;Hoerer S;Blech M;Enenkel B;Studts JM;Garidel P;Karow AR,MAbs (2015) Boosting antibody developability through rational sequence optimization. PMID:25759214
Additional Information

Structure view and single mutant data analysis

Study data

No weblogo for data of varying length.
Colors: D E R H K S T N Q A V I L M F Y W C G P
 

Data Distribution

Studies with similar sequences (approximate matches)

Correlation with other assays (exact sequence matches)


Relevant PDB Entries

Structure ID Release Date Resolution Structure Title
4HAF 2012-09-26T00:00:00+0000 2.04 Crystal structure of fc-fragment of human IgG2 antibody (primitive crystal form)
4HAG 2012-09-26T00:00:00+0000 3.4 Crystal structure of fc-fragment of human IgG2 antibody (centered crystal form)
4L4J 2013-06-07T00:00:00+0000 1.92 Crystal structure of fc-fragment of human IgG2-Sigma antibody
7LUS 2021-02-23T00:00:00+0000 2.45 IgG2 Fc Charge Pair Mutation version 1 (CPMv1)
7LUS 2021-02-23T00:00:00+0000 2.45 IgG2 Fc Charge Pair Mutation version 1 (CPMv1)
1YPO 2005-01-31T00:00:00+0000 3.0 Human Oxidized Low Density Lipoprotein Receptor LOX-1 P3 1 21 Space Group
1YPQ 2005-01-31T00:00:00+0000 1.4 Human Oxidized Low Density Lipoprotein Receptor LOX-1 Dioxane Complex
1YPU 2005-01-31T00:00:00+0000 2.05 Human Oxidized Low Density Lipoprotein Receptor LOX-1 C2 Space Group
1YXJ 2005-02-22T00:00:00+0000 1.78 Crystal structure of human lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) at low pH
1YXK 2005-02-22T00:00:00+0000 2.4 Crystal structure of human lectin-like oxidized low-density lipoprotein receptor 1 (LOX-1) disulfide-linked dimer

Relevant UniProtKB Entries

Percent Identity Matching Chains Protein Accession Entry Name
99.1 Oxidized low-density lipoprotein receptor 1 (LOX-1) P78380 OLR1_HUMAN
99.1 CL Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P0DOX7 IGK_HUMAN
99.1 CL Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P01834 IGKC_HUMAN
91.5 CH Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P01861 IGHG4_HUMAN
91.5 CH Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P01859 IGHG2_HUMAN
99.4 CH Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P01857 IGHG1_HUMAN
99.4 CH Anti-LOX-1 (lectin-type optimized LDL receptor 1) mAb1 P0DOX5 IGG1_HUMAN