Solubility optimization of immunobinders
AssigneeNovartis AG
InventorsLeonardo Borras, David Urech
Life of this patent
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Abstract
The invention provides methods of using sequence based analysis and rational strategies to improve the solubility of immunobinders, and in particular of single chain antibodies (scFvs). The invention provides methods of engineering immunobinders, and in particular scFvs, by performing one or more substitutions with hydrophilic residues identified by analysis of a database of selected, stable scFv sequences. The invention also provides immunobinders with optimized solubility prepared according to the engineering methods of the invention.
Claims15
An immunobinder comprising one of the following solubility enhancing motifs in the heavy chain amino acid positions 12, 103 and 144 (AHo numbering):
- (a) Serine (S) at heavy chain amino acid position 12;
- Serine (S) at heavy chain amino acid position 103;
- and Threonine (T) at heavy chain amino acid position 144;
- or
- (b) Serine (S) at heavy chain amino acid position 12;
- Threonine (T) at heavy chain amino acid position 103;
- and Serine (S) at heavy chain amino acid position 144;
- or
- (c) Serine (S) at heavy chain amino acid position 12;
- Threonine (T) at heavy chain amino acid position 103;
- and Threonine (T) at heavy chain amino acid position 144.
A method of enhancing the solubility of an immunobinder in native state, the immunobinder comprising a heavy chain variable (VH) region, or fragment thereof, the method comprising:
- (a) selecting at least two amino acid positions within the VH region for mutation;
- and
- (b) mutating the at least two amino acid positions selected for mutation,
- wherein the at least two amino acid positions are selected from the group of heavy chain amino acid positions consisting of 12, 103 and 144 (according to AHo numbering convention) and the mutating comprises the substitution of the amino acid at the selected amino acid position against a hydrophilic amino acid.
Dependencies are inferred from the claim text.