Aldehyde-tagged immunoglobulin polypeptides and methods of use thereof
AssigneeRedwood Bioscience, Inc.
InventorsRobyn M. Barfield, Mark Alan Breidenbach, Gregory W. deHart, David Rabuka
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Abstract
The present disclosure provides aldehyde-tagged immunoglobulin (Ig) polypeptides that can be converted by a formylglycine-generating enzyme to produce a 2-formylglycine (FGly)-modified Ig polypeptide. An FGly-modified Ig polypeptide can be covalently and site-specifically bound to a moiety of interest to provide an Ig conjugate. The disclosure also encompasses methods of production of such aldehyde-tagged Ig polypeptides, FGly-modified Ig polypeptides, and Ig conjugates, as well as methods of use of same.
Claims44
An immunoglobulin (Ig) polypeptide comprising a sulfatase motif comprising an amino acid sequence LCTPSR (SEQ ID NO:17), the Ig polypeptide comprising:
- a) an IgG1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- ASTKGLCTPSRV (SEQ ID NO:
- 231),
- LCTPSRSKSTSGGT (SEQ ID NO:
- 234),
- NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO:235),
- EPKSCDKTHTCPPCPLCTPSRELLGG (SEQ ID NO:
- 236),
- NLCTPSRAP (SEQ ID NO:
- 206),
- SKAKGLCTPSRE (SEQ ID NO:
- 237), and
- DVSHEDLCTPSREV (SEQ ID NO:
- 250);
- or
- b) an IgG4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- NSGALCTPSRGVHTFPAVLQSSGL (SEQ ID NO:235), and
- SKAKGLCTPSRE (SEQ ID NO:237);
- or
- c) an Ig kappa light chain constant region comprising the amino acid sequence:
- DNALCTPSRQSGN (SEQ ID NO:
- 220).
An immunoglobulin (Ig) polypeptide comprising a 2-formylglycine (FGly) moiety, comprising:
- a) an IgG1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- ASTKGL(FGly)TPSRV (SEQ ID NO:
- 232),
- L(FGly)TPSRSKSTSGGT (SEQ ID NO:
- 238),
- NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO:
- 239),
- EPKSCDKTHTCPPCPL(FGly)TPSRELLGG (SEQ ID NO:
- 240),
- NL(FGly)TPSRAP (SEQ ID NO:
- 241),
- SKAKGL(FGly)TPSRE (SEQ ID NO:
- 242), and
- DVSHEDL(FGly)TPSREV (SEQ ID NO:251);
- or
- b) an IgG4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- NSGAL(FGly)TPSRGVHTFPAVLQSSGL (SEQ ID NO:239), and
- SKAKGL(CGly)TPSRE (SEQ ID NO:242);
- or
- c) an Ig light chain constant region comprising the amino acid sequence:
- DNAL(FGly)TPSRQSGN (SEQ ID NO:
- 248).
An immunoglobulin (Ig) conjugate comprising:
- an Ig polypeptide and a covalently bound moiety,
- wherein the Ig polypeptide comprises:
- a) an IgG1 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- ASTKGL(FGly′)TPSRV (SEQ ID NO:
- 233),
- L(FGly′)TPSRSKSTSGGT (SEQ ID NO:
- 243),
- NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO:
- 244),
- EPKSCDKTHTCPPCPL(FGly′)TPSRELLGG (SEQ ID NO:
- 245),
- NL(FGly′)TPSRAP (SEQ ID NO:
- 246),
- SKAKGL(FGly′)TPSRE (SEQ ID NO:
- 247), and
- DVSHEDL(FGly′)TPSREV (SEQ ID NO:252);
- or
- b) an IgG4 heavy chain constant region comprising an amino acid sequence selected from the group consisting of:
- NSGAL(FGly′)TPSRGVHTFPAVLQSSGL (SEQ ID NO:244), and
- SKAKGL(FGly′)TPSRE (SEQ ID NO:247);
- or
- c) an Ig light chain constant region comprising DNAL(FGly′)TPSRQSGN (SEQ ID NO:
- 249);
- and wherein FGly′ is of the formula:
- wherein J1 is the covalently bound moiety;
- each L1 is independently selected from alkylene, substituted alkylene, alkenylene, substituted alkenylene, alkynylene, arylene, substituted arylene, cycloalkylene, substituted cycloalkylene, heteroarylene, substituted heteroarylene, heterocyclene, substituted heterocyclene, acyl, amido, acyloxy, urethanylene, thioester, sulfonyl, sulfonamide, sulfonyl ester, O, S, NH, and substituted amine;
- and
- n is a number selected from zero to 40.
Dependencies are inferred from the claim text.