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Improving the solubility of anti-LINGO-1 monoclonal antibody Li33 by isotype switching and targeted mutagenesis.
- Source :
-
Protein science : a publication of the Protein Society [Protein Sci] 2010 May; Vol. 19 (5), pp. 954-66. - Publication Year :
- 2010
-
Abstract
- Monoclonal antibodies (Mabs) are a favorite drug platform of the biopharmaceutical industry. Currently, over 20 Mabs have been approved and several hundred others are in clinical trials. The anti-LINGO-1 Mab Li33 was selected from a large panel of antibodies by Fab phage display technology based on its extraordinary biological activity in promoting oligodendrocyte differentiation and myelination in vitro and in animal models of remyelination. However, the Li33 Fab had poor solubility when converted into a full antibody in an immunoglobulin G1 framework. A detailed analysis of the biochemical and structural features of the antibody revealed several possible reasons for its propensity to aggregate. Here, we successfully applied three molecular approaches (isotype switching, targeted mutagenesis of complementarity determining region residues, and glycosylation site insertion mutagenesis) to address the solubility problem. Through these efforts we were able to improve the solubility of the Li33 Mab from 0.3 mg/mL to >50 mg/mL and reduce aggregation to an acceptable level. These strategies can be readily applied to other proteins with solubility issues.
- Subjects :
- Amino Acid Sequence
Antibodies, Monoclonal genetics
Antibodies, Monoclonal immunology
Antibodies, Monoclonal metabolism
Area Under Curve
Crystallography, X-Ray
Electrophoresis, Polyacrylamide Gel
Immunoglobulin Class Switching
Immunoglobulin Fab Fragments chemistry
Immunoglobulin Fab Fragments genetics
Immunoglobulin Fab Fragments metabolism
Immunoglobulin G genetics
Immunoglobulin G metabolism
Models, Molecular
Molecular Sequence Data
Protein Conformation
Protein Stability
Solubility
Temperature
Antibodies, Monoclonal chemistry
Immunoglobulin G chemistry
Membrane Proteins immunology
Mutagenesis, Site-Directed methods
Nerve Tissue Proteins immunology
Protein Engineering methods
Subjects
Details
- Language :
- English
- ISSN :
- 1469-896X
- Volume :
- 19
- Issue :
- 5
- Database :
- MEDLINE
- Journal :
- Protein science : a publication of the Protein Society
- Publication Type :
- Academic Journal
- Accession number :
- 20198683
- Full Text :
- https://doi.org/10.1002/pro.372