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US 11,993,643 B2Granted
Method for isolation of soluble polypeptides
AssigneeNational Research Council of Canada
InventorJamshid Tanha
Life of this patent
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AssigneeNational Research Council of Canada
InventorJamshid Tanha
Estimating the term…
Polypeptides with biophysical properties such as solubility, stability, high expression, monomericity, binding specificity or non-aggregation, including monomeric human heavy and light chain variable domains (VHs and VLs), are identified using a high throughput method for screening polypeptides, comprising the steps of obtaining a phage display library, allowing infection of a bacterial lawn by the library phage, and identifying phage which form larger than average plaques on the bacterial lawn. Sequences of monomeric human VHs and VLs are identified, which may be useful for immunotherapy or as diagnostic agents. Multimer complexes of human VHs and VLs are also identified. The VHs and VLs identified may be used to create further libraries for identifying additional polypeptides. Further, the VHs and VLs may be subjected to DNA shuffling to select for improved biophysical properties.
An antigen-binding VL comprising a FR1 sequence of EIVMTQSPGTLSLSPGDRATLSC (amino acids 1-23 of SEQ ID NO:42), a FR2 sequence of WYQQKPGQAPRLLIY (amino acids 35-49 of SEQ ID NO:42), a FR3 sequence of GIPDRFSGSGSGTDFTLTISRLEPEDFAVYYC (amino acids 57-88 of SEQ ID NO:42), and a FR4 sequence of FGQGTKVTVL (amino acids 98-107 of SEQ ID NO:42).
An antigen-binding VL comprising the FR1, FR2, FR3, and FR4 portion of SEQ ID NO:42 and one or more randomized CDR sequences, wherein one or more of CDR1, CDR2 and CDR3 of SEQ ID NO:42 is replaced, respectively, with a randomized CDR1, CDR2 and CDR3.
Dependencies are inferred from the claim text.