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US 5,731,168 AGranted
Method for making heteromultimeric polypeptides
AssigneeGenentech, Inc.
InventorsPaul J. Carter, Leonard G. Presta, John B. Ridgway
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AssigneeGenentech, Inc.
InventorsPaul J. Carter, Leonard G. Presta, John B. Ridgway
Estimating the term…
The invention relates to a method of preparing heteromultimeric polypeptides such as bispecific antibodies, bispecific immunoadhesins and antibody-immunoadhesin chimeras. The invention also relates to the heteromultimers prepared using the method. Generally, the method involves introducing a protuberance at the interface of a first polypeptide and a corresponding cavity in the interface of a second polypeptide, such that the protuberance can be positioned in the cavity so as to promote heteromultimer formation and hinder homomultimer formation. "Protuberances" are constructed by replacing small amino acid side chains from the interface of the first polypeptide with larger side chains (e.g. tyrosine or tryptophan). Compensatory "cavities" of identical or similar size to the protuberances are created in the interface of the second polypeptide by replacing large amino acid side chains with smaller ones (e.g. alanine or threonine). The protuberance and cavity can be made by synthetic means such as altering the nucleic acid encoding the polypeptides or by peptide synthesis.
A method of preparing a heteromultimer comprising a first polypeptide and a second polypeptide which meet at an interface which has been engineered to promote heteromultimer formation, wherein the interface of the first polypeptide comprises a protuberance which is positionable in a cavity in the interface of the second polypeptide the method comprising the steps of:
A method of preparing a heteromultimer which comprises a first polypeptide and a second polypeptide which meet at an interface which has been engineered to promote heteromultimer formation the method comprising the steps of:
Isolated nucleic acid encoding a heteromultimer comprising a first polypeptide and a second polypeptide which meet at an interface, wherein the interface of the first polypeptide comprises a protuberance which is positionable in a cavity in the interface of the second polypeptide, and wherein nucleic acid encoding the interface of the first polypeptide has been altered from nucleic acid encoding the original interface of the first polypeptide to encode the protuberance or the nucleic acid encoding the interface of the second polypeptide has been altered from nucleic acid encoding the original interface of the second polypeptide to encode the cavity, or both, such that a greater ratio of hetermultimer:homomultimer forms than that for a multimer having a nonengineered interface.
A composition comprising two or more recombinant nucleic acids encoding a heteromultimer comprising a first polypeptide and a second polypeptide which meet at an interface, wherein the interface of the first polypeptide comprises a protuberance which is positionable in a cavity in the interface of the second polypeptide, and wherein nucleic acid encoding the interface of the first polypeptide has been altered from nucleic acid encoding the original interface of the first polypeptide to encode the protuberance or the nucleic add encoding the interface of the second polypeptide has been altered from nucleic acid encoding the original interface of the second polypeptide to encode the cavity, or both, such that a greater ratio of hetermultimer:homomultimer forms than that for a multimer having a nonengineered interface.
Dependencies are inferred from the claim text.