Method for making heteromultimeric polypeptides
First Claim
1. An isolated 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, and wherein the amino acid sequence of an original interface has been altered so as to introduce the protuberance and/or cavity into the engineered interface such that a greater ratio of hetermultimer:
- homomultimer forms than that for a multimer having a nonengineered interface.
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Abstract
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.
302 Citations
27 Claims
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1. An isolated 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, and wherein the amino acid sequence of an original interface has been altered so as to introduce the protuberance and/or cavity into the engineered interface such that a greater ratio of hetermultimer:
- homomultimer forms than that for a multimer having a nonengineered interface.
- View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26)
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27. An isolated heteromultimer comprising a first polypeptide and a second polypeptide which meet at an engineered antibody CH 3 domain interface, wherein the interface of the first polypeptide comprises a protuberance which is positionable in a cavity in the interface of the second polyeptide, and wherein the amino acid sequence of an original antibody CH 3 domain interface has been altered to encode the protuberance and/or cavity such that a greater ratio of heteromultimer:
- homomultimer forms than that for a multimer having a nonengineered interface.
Specification