# US 7,608,260 B2 · Stabilized immunoglobulins > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 7,608,260 B2 - Document ID: US7608260B2 - Status: Granted - Kind: B2 - Application number: 10/751,744 - Filed: 2004-01-05 - Published: 2004-09-30 - Granted: 2009-10-27 - Earliest claimed: 2003-01-06 - Assignees: MedImmune, LLC - Inventors: Mark A. Schenerman; Jose Casas-Finet; Jinhua Feng; Guillermo Tous - Web page: https://patentgrep.com/patent/US7608260B2 ## CPC - C07K 16/2806: {against CD2} - C07K 16/00: Immunoglobulins [IG], e.g. monoclonal or polyclonal antibodies - C07K 2317/53: Hinge - C07K: PEPTIDES (peptides in foodstuffs A23; obtaining protein compositions for foodstuffs, working-up proteins for foodstuffs A23J; preparations for medicinal purposes A61K; peptides containing beta-lactam rings C07D; cyclic dipeptides not having in their molecule any other peptide link than those which form their ring, e.g. piperazine-2,5-diones, C07D; ergot alkaloids of the cyclic peptide type C07D519/02; macromolecular compounds having statistically distributed amino acid units in their molecules, i.e. when the preparation does not provide for a specific; but for a random sequence of the amino acid units, homopolyamides and block copolyamides derived from amino acids C08G69/00; macromolecular products derived from proteins C08H1/00; preparation of glue or gelatine C09H; single cell proteins, enzymes C12N; genetic engineering processes for obtaining peptides C12N15/00; compositions for measuring or testing processes involving enzymes C12Q; investigation or analysis of biological material G01N33/00) ## Abstract The present invention provides stabilized immunoglobulin molecules that have increased storage stability and/or in vivo half-lives due to the mutation of one or more amino acids that would otherwise render the immunoglobulin molecules susceptible to degradation. In a preferred embodiment, the stabilized immunoglobulins of the invention have mutations at the heavy chain constant domain hinge region. Such stabilized immunoglobulin molecules, i.e., immunoglobulin molecules with increased storage stability have one or more of the following advantages they are more readily transported and/storable for longer periods and/or less stringent conditions than non-stabilized counterparts; that smaller amounts and or less frequent dosing is required in the therapeutic, prophylactic or diagnostic use of such stabilized molecules. ## Claims ### Claim 1 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position cysteine 233 with either alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 2 (depends on 1) The modified IgG1 of claim 1, wherein said amino acid substitutions further comprise an amino acid substitution at position cysteine 239 with either alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine, as numbered according to Kabat. ### Claim 3 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position cysteine 239 with either alanine, arginine, asparagine, aspartic acid, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 4 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position aspartic acid 234 with either alanine, arginine, asparagine, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 5 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position lysine 235 with either alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, serine. threonine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 6 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position threonine 236 with either alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, methionine, phenylalanine, proline, serine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 7 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position histidine 237 with either alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, glycine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitutions. ### Claim 8 (depends on 7) The modified IgG1 of claim 7, wherein said amino acid substitution at position histidine 237 is a valine or isoleucine. ### Claim 9 A modified human or humanized IgG1 comprising a modified hinge region containing an amino acid substitution at position glycine 249 with either alanine, arginine, asparagine, aspartic acid, cysteine, glutamic acid, glutamine, histidine, isoleucine, leucine, lysine, methionine, phenylalanine, proline, serine, threonine, tryptophan, tyrosine, or valine, as numbered according to Kabat, said modified IgG1 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG1 not comprising said amino acid substitution. ### Claim 10 (depends on 9) The modified IgG1 of claim 9, wherein said amino acid substitution at position glycine 249 is a valine or isoleucine. ### Claim 11 A modified human or humanized IgG4 comprising a modified hinge region containing an amino acid substitution at position serine 236 with proline, as numbered according to Kabat, said modified IgG4 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG4 not comprising said amino acid substitution. ### Claim 12 A modified human or humanized IgG4 comprising a modified hinge region containing an amino acid substitution at position serine 241 with proline, as numbered according to Kabat, said modified IgG4 exhibiting reduced degradation upon heating to 55° C. for one week, as determined by mass spectrometry, as compared to a corresponding wild-type IgG4 not comprising said amino acid substitution.