Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Blood proteins or globulins – e.g. – proteoglycans – platelet...
Reexamination Certificate
2000-11-02
2004-10-05
Kunz, Gary (Department: 1642)
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Blood proteins or globulins, e.g., proteoglycans, platelet...
C530S387100, C530S387700, C530S388150, C530S388220, C530S388800, C530S391100, C530S389700, C530S391300, C530S391500, C530S391700, C424S130100, C424S133100, C424S134100, C424S135100, C424S136100, C424S138100, C424S141100, C424S142100, C424S143100, C424S152100, C424S155100, C424S172100, C424S174100, C424S177100
Reexamination Certificate
active
06800738
ABSTRACT:
FIELD OF THE INVENTION
This invention relates to methods for the preparation and use of variant antibodies and finds application particularly in the fields of immunology and cancer diagnosis and therapy.
BACKGROUND OF THE INVENTION
Naturally occurring antibodies (immunoglobulins) comprise two heavy chains linked together by disulfide bonds and two light chains, one light chain being linked to each of the heavy chains by disulfide bonds. Each heavy chain has at one end a variable domain (V
H
) followed by a number of constant domains. Each light chain has a variable domain (V
L
) at one end and a constant domain at its other end; the constant domain of the light chain is aligned with the first constant domain of the heavy chain, and the light chain variable domain is aligned with the variable domain of the heavy chain. Particular amino acid residues are believed to form an interface between the light and heavy chain variable domains, see e.g. Chothia et al.,
J. Mol. Biol
. 186:651-663 (1985); Novotny and Haber,
Proc. Natl. Acad. Sci. USA
82:4592-4596 (1985).
The constant domains are not involved directly in binding the antibody to an antigen, but are involved in various effector functions, such as participation of the antibody in antibody-dependent cellular cytotoxicity. The variable domains of each pair of light and heavy chains are involved directly in binding the antibody to the antigen. The domains of natural light and heavy chains have the same general structure, and each domain comprises four framework (FR) regions, whose sequences are somewhat conserved, connected by three hyper-variable or complementarity determining regions (CDRs) (see Kabat, E. A. et al.,
Sequences of Proteins of Immunological Interest
, National Institutes of Health, Bethesda, Md., (1987)). The four framework regions largely adopt a &bgr;-sheet conformation and the CDRs form loops connecting, and in some cases forming part of, the &bgr;-sheet structure. The CDRs in each chain are held in close proximity by the framework regions and, with the CDRs from the other chain, contribute to the formation of the antigen binding site.
Widespread use has been made of monoclonal antibodies, particularly those derived from rodents including mice, however they are frequently antigenic in human clinical use. For example, a major limitation in the clinical use of rodent monoclonal antibodies is an anti-globulin response during therapy (Miller, R. A. et al.,
Blood
62:988-995 (1983); Schroff, R. W. et al.,
Cancer Res
. 45:879-885 (1985)).
The art has attempted to overcome this problem by constructing “chimeric” antibodies in which an animal antigen-binding variable domain is coupled to a human constant domain (Cabilly et al., U.S. Pat. No. 4,816,567; Morrison, S. L. et al.,
Proc. Natl. Acad. Sci. USA
81:6851-6855 (1984); Boulianne, G. L. et al.,
Nature
312:643-646 (1984); Neuberger, M. S. et al.,
Nature
314:268-270 (1985)). The term “chimeric” antibody is used herein to describe a polypeptide comprising at least the antigen binding portion of an antibody molecule linked to at least part of another protein (typically an immunoglobulin constant domain).
The isotype of the human constant domain may be selected to tailor the chimeric antibody for participation in antibody-dependent cellular cytotoxicity (ADCC) and complement-dependent cytotoxicity (see e.g. Bruggemann, M. et al.,
J. Exp. Med
. 166:1351-1361 (1987); Riechmann, L. et al.,
Nature
332:323-327 (1988); Love et al.,
Methods in Enzymology
178:515-527 (1989); Bindon, et al.,
J. Exp. Med
. 168:127-142 (1988).
In the typical embodiment, such chimeric antibodies contain about one third rodent (or other non-human species) sequence and thus are capable of eliciting a significant anti-globulin response in humans. For example, in the case of the murine anti-CD3 antibody, OKT3, much of the resulting anti-globulin response is directed against the variable region rather than the constant region (Jaffers, G. J. et al.,
Transplantation
41:572-578 (1986)).
In a further effort to resolve the antigen binding functions of antibodies and to minimize the use of heterologous sequences in human antibodies, Winter and colleagues (Jones, P. T. et al.,
Nature
321:522-525 (1986); Riechmann, L. et al.,
Nature
332:323-327 (1988); Verhoeyen, M. et al.,
Science
239:1534-1536 (1988)) have substituted rodent CDRs or CDR sequences for the corresponding segments of a human antibody. As used herein, the term “humanized” antibody is an embodiment of chimeric antibodies wherein substantially less than an intact human variable domain has been substituted by the corresponding sequence from a non-human species. In practice, humanized antibodies are typically human antibodies in which some CDR residues and possibly some FR residues are substituted by residues from analogous sites in rodent antibodies.
The therapeutic promise of this approach is supported by the clinical efficacy of a humanized antibody specific for the CAMPATH-1 antigen with two non-Hodgkin lymphoma patients, one of whom had previously developed an anti-globulin response to the parental rat antibody (Riechmann, L. et al.,
Nature
332:323-327 (1988); Hale, G. et al.,
Lancet
i:1394-1399 (1988)). A murine antibody to the interleukin 2 receptor has also recently been humanized (Queen, C. et al.,
Proc. Natl. Acad. Sci. USA
86:10029-10033 (1989)) as a potential immunosuppressive reagent. Additional references related to humanization of antibodies include Co et al.,
Proc. Natl. Acad. Sci. USA
88:2869-2873 (1991); Gorman et al.,
Proc. Natl. Acad. Sci. USA
88:4181-4185 (1991); Daugherty et al.,
Nucleic Acids Research
19(9):2471-2476 (1991); Brown et al.,
Proc. Natl. Acad. Sci. USA
88:2663-2667 (1991); Junghans et al.,
Cancer Research
50:1495-1502 (1990).
In some cases, substituting CDRs from rodent antibodies for the human CDRs in human frameworks is sufficient to transfer high antigen binding affinity (Jones, P. T. et al.,
Nature
321:522-525 (1986); Verhoeyen, M. et al.,
Science
239:1534-1536 (1988)), whereas in other cases it has been necessary to additionally replace one (Riechmann, L. et al.,
Nature
332:323-327 (1988)) or several (Queen, C. et al.,
Proc. Natl. Acad. Sci. USA
86:10029-10033 (1989)) framework region (FR) residues. See also Co et al., supra.
For a given antibody a small number of FR residues are anticipated to be important for antigen binding. Firstly for example, certain antibodies have been shown to contain a few FR residues which directly contact antigen in crystal structures of antibody-antigen complexes (e.g., reviewed in Davies, D. R. et al.,
Ann. Rev. Biochem
. 59:439-473 (1990)). Secondly, a number of FR residues have been proposed by Chothia, Lesk and colleagues (Chothia, C. & Lesk, A. M.,
J. Mol. Biol
. 196:901-917 (1987); Chothia, C. et al.,
Nature
342:877-883 (1989); Tramontano, A. et al.,
J. Mol. Biol
. 215:175-182 (1990)) as critically affecting the conformation of particular CDRs and thus their contribution to antigen binding. See also Margolies et al.,
Proc. Natl. Acad. Sci. USA
72:2180-2184 (1975).
It is also known that, in a few instances, an antibody variable domain (either V
H
or V
L
) may contain glycosylation sites, and that this glycosylation may improve or abolish antigen binding, Pluckthun,
Biotechnology
9:545-51 (1991); Spiegelberg et al.,
Biochemistry
9:4217-4223 (1970); Wallic et al,
J. Exp. Med
. 168:1099-1109 (1988); Sox et al.,
Proc. Natl. Acad. Sci. USA
66:975-982 (1970); Margni et al.,
Ann. Rev. Immunol
. 6:535-554 (1988). Ordinarily, however, glycosylation has no influence on the antigen-binding properties of an antibody, Pluckthun, supra, (1991).
The three-dimensional structure of immunoglobulin chains has been studied, and crystal structures for intact immunoglobulins, for a variety of immunoglobulin fragments, and for antibody-antigen complexes have been published (see e.g., Saul et al.,
Journal of Biological Chemistry
25:585-97 (1978); Sheriff et al.,
Proc. Natl. Acad. Sci. USA
84:8075-79 (1987); Segal et al.,
Proc. Nat
Carter Paul J.
Presta Leonard G.
Genentech Inc.
Holleran Anne L.
Kunz Gary
Lee Wendy M.
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