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US 6,673,580 B2Granted
Identification and modification of immunodominant epitopes in polypeptides
AssigneeGenentech, Inc.
InventorsEugen Koren, John Hok Nin Lowe
Life of this patent
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AssigneeGenentech, Inc.
InventorsEugen Koren, John Hok Nin Lowe
Estimating the term…
The invention provides for methods of modifying immunodominant epitopes on polypeptides, preferably polypeptides intended for therapeutic use. Knowledge of immunodominant epitopes prior to clinical use of polypeptides would be useful to design and engineer less immunogenic molecules. The invention provides for methods of identifying immunodominant epitopes and modifying an immunodominant epitope to reduce the immune response to the polypeptide while still retaining a substantial therapeutic activity of the polypeptide. The modified polypeptides are useful therapeutically.
A method of modifying a polypeptide, comprising:
A method of modifying a therapeutic polypeptide, comprising:
A method of modifying a nucleic acid encoding a modified polypeptide comprising:
A host cell comprising a modified isolated nucleic acid obtained by a method comprising:
A method of modifying a polypeptide, comprising:
A method of modifying a therapeutic polypeptide, comprising:
A method of modifying a nucleic acid encoding a modified polypeptide comprising:
A host cell comprising a modified isolated nucleic acid obtained by a method comprising:
Dependencies are inferred from the claim text.
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Against material from animals or humans: against cytokines, lymphokines or interferons
Drugs for disorders of the blood or the extracellular fluid
Cytokines; Lymphokines; Interferons: thrombopoietin, i.e. C-MPL ligand
Medicinal preparations containing peptides
Term ended 1 Jun 2021 (estimated)
A number of polypeptides from a variety of sources are used to treat disease in humans. If the polypeptide is derived from a heterologous or non-self source, patients readily develop an immune response to the polypeptide. In fact, in many cases, an immune response to the polypeptide is a desired therapeutic outcome. However, there are situations where an immune response to a polypeptide intended for therapeutic use is undesirable and could be very detrimental. For example, polypeptides intended for therapeutic use also include polypeptides that have endogenous or “self” counterparts. It is generally thought that an immune response is not generated to self or endogenous polypeptides, except in the case of autoimmune disease. If an immune response were generated upon administration of such polypeptides, adverse consequences could result including development of an autoimmune response.
Many factors contribute to immune responses to polypeptides. The presence of immunodominant epitopes in a polypeptide is one of the key factors in antibody responses. An immunodominant epitope is an epitope that more frequently elicits and binds to antibodies in a human or animal or population thereof when compared with other epitopes. Immunodominant epitopes in polypeptides are typically identified after administration of the polypeptide to the human or animal.
Immunodominant epitopes have been mapped on proteins using antibodies from treated animals. For example, staphylokinase is a bacterial polypeptide derived from Staphylococcus aureus and is used to treat myocardial infarction. Because staphylokinase is a heterologous or non-self polypeptide when administered to humans, treated patients readily develop an immune response to this polypeptide. This immune response can result in adverse reactions to further therapeutic treatment with the polypeptide. As described in U.S. Pat. No. 5,951,980, the immunodominant epitopes identified using antibodies from treated patients can be modified to reduce immunogenicity of the polypeptide.
FIG. 1 shows the structure of full-length recombinant human TPO (TPO FL) and truncated TPO (TPO TR);glycosylation sites are shown on C terminal.
FIG. 2 shows the effect of administration of rhesus thrombopoietin on platelet levels in normal rhesus monkeys. Antibody responses to rh TPO were also measured over time.
FIG. 3 shows the effect on platelet counts of passive transfer of anti-TPO antibodies to CD-1 mice on platelet counts.
An “epitope” means a portion or a site on an antigen, such as a polypeptide, with the ability or potential to elicit and combine with an antibody. Polypeptides often include more than one epitope. For the purpose of this disclosure, a polypeptide epitope will usually include at least 3 amino acids, preferably 8 to 50 amino acids, and more preferably between about 10-20 amino acids in the peptide. There is no critical upper limit to the length of the peptide, which could comprise nearly the full length of the polypeptide sequence. Epitopes can be either linear or conformational epitopes. A linear epitope is comprised of a single segment of a primary sequence of a polypeptide chain. Linear epitopes can be contiguous or overlapping. Conformational epitopes are comprised of amino acids brought together by folding of the polypeptide to form a tertiary structure and the amino acids are not necessarily adjacent to one another in the linear sequence.
“Immunodominant epitope” means an epitope of a polypeptide that more frequently elicits or binds to antibodies from an antibody positive human or animal subject or population of thereof compared with other epitopes in the polypeptide. A polypeptide can have one or more immunodominant epitopes.
“Epitope specific or specificity” means the unique association between an antibody and its epitope. Epitope specificity is typically determined using competitive binding assays so that the enhanced binding between an antibody and its specific epitope is detected as compared to lower binding of the antibody to other epitopes.