Immunoglobulins and variants directed against pathogenic microbes

Abstract

Anti-SpA murine, chimeric and humanized monoclonal antibodies, and variant antibodies having a heavy chain with at least one amino acid substitution are provided. Such antibodies may be used to prevent or treat microbial infections.

1 - 28 . (canceled) 
     
     
         29 . A humanized anti-SpA antibody comprising
 an immunoglobulin heavy chain which comprises a sequence selected from SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, or SEQ ID NO:5; and   an immunoglobulin light chain which comprises SEQ ID NO:7, SEQ ID NO:8, SEQ ID NO:9, SEQ ID NO:10, SEQ ID NO:11, or SEQ ID NO:12, SEQ ID NO:13, SEQ ID NO:14, SEQ ID NO:15, SEQ ID NO:16, SEQ ID NO:17, or SEQ ID NO:18.   
     
     
         30 . The humanized anti-SpA antibody of  claim 29 , wherein the variable light chain sequence, the variable heavy chain sequence, or both, further comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the humanized anti-SpA antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity. 
     
     
         31 . A humanized anti-SpA antibody comprising
 an immunoglobulin heavy chain which comprises a variable heavy chain sequence and a constant heavy chain sequence, wherein the immunoglobulin heavy chain comprises SEQ ID NO:77, SEQ ID NO:78, SEQ ID NO:79, SEQ ID NO:80, SEQ ID NO:86, or SEQ ID NO:87; and   an immunoglobulin light chain which comprises a variable light chain sequence and a constant light chain sequence.   
     
     
         32 . The humanized anti-SpA antibody of  claim 31 , wherein the variable light chain sequence, the variable heavy chain sequence, or both, further comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the humanized anti-SpA antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity. 
     
     
         33 . The humanized anti-SpA antibody of  claim 31 , wherein the immunoglobulin light chain comprises SEQ ID NO:68, SEQ ID NO:69, or SEQ ID NO:85. 
     
     
         34 . The humanized anti-SpA antibody of  claim 33 , wherein the variable light chain sequence, the variable heavy chain sequence, or both, further comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the humanized anti-SpA antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity. 
     
     
         35 . The humanized anti-SpA antibody of  claim 34 , wherein the immunoglobulin heavy chain comprises one or more amino acid substitutions in its constant heavy chain sequence as compared to that of a parental anti-SpA antibody, resulting in an anti-SpA variant antibody. 
     
     
         36 . The anti-SpA variant antibody of  claim 35 , wherein the anti-SpA variant antibody is an anti-SpA Fc variant antibody; and wherein the parental anti-SpA antibody is the humanized anti-SpA antibody of claim  10 , the humanized anti-SpA antibody of claim  11 , the humanized anti-SpA antibody of claim  12 , the humanized anti-SpA antibody of claim  13 , the humanized anti-SpA antibody of claim  14 , or the humanized anti-SpA antibody of claim  15 . 
     
     
         37 . The anti-SpA variant antibody of  claim 35 , wherein the anti-SpA variant antibody is an anti-SpA Fc variant antibody; and wherein the heavy chain constant domain comprises an amino acid sequence selected from SEQ ID NOs: 31-56. 
     
     
         38 . The anti-SpA variant antibody of  claim 37 , wherein the anti-SpA variant antibody is an anti-SpA Fc variant antibody; and wherein the variant immunoglobulin heavy chain comprises SEQ ID NO:81, SEQ ID NO:82, SEQ ID NO:83, SEQ ID NO:84, SEQ ID NO:88, or SEQ ID NO:89. 
     
     
         39 . The anti-SpA variant antibody of  claim 35 , wherein the variable light chain sequence, the variable heavy chain sequence, or both, further comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the anti-SpA variant antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity. 
     
     
         40 . The anti-SpA variant antibody of  claim 31 , having at least one additional amino acid substitution which results in alteration of an IgG1 allotype, and wherein the at least one additional amino acid substitution occurs at EU position 356, 358, 431, 214 or a combination thereof. 
     
     
         41 . The anti-SpA variant antibody of  claim 40 , wherein the IgG1 allotype is a G1m17 allotype, and wherein the at least one amino acid substitution comprises an Arg at position 435, a Phe at position 436, and Gln at position 274 (SEQ ID NO: 40). 
     
     
         42 . An anti-SpA variant antibody comprising an anti-SpA Fc variant antibody derived from a parental anti-SpA antibody that is a human anti-SpA antibody that has a variable light chain sequence, a variable heavy chain sequence, or both, which further comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the anti-SpA variant antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity. 
     
     
         43 . An anti- Staphylococcus aureus  ( S. aureus ) antibody comprising aparental antibody derived from an IGHV-3 family germ line V gene sequence having one or more heavy chain variable domain amino acid substitutions, resulting in attenuation of non-immune, SpA super-antigen type binding to the heavy chain variable domain of the anti- S. aureus  antibody as compared to the parental anti- S. aureus  antibody. 
     
     
         44 . The anti- S. aureus  variable heavy chain sequence variant antibody of  claim 43 , wherein the variable heavy chain sequence variant antibody is an anti-SpA antibody and wherein the heavy chain constant domain comprises an amino acid sequence selected from SEQ ID NOs: 31-56. 
     
     
         45 . An anti- S. aureus  variable heavy chain sequence variant antibody comprising an immunoglobulin light chain and a variant immunoglobulin heavy chain which comprises a variable heavy chain sequence and a constant heavy chain sequence, wherein the variant immunoglobulin heavy chain comprises one or more amino acid substitutions in its variable heavy chain sequence as compared to a parental anti- S. aureus  antibody, wherein the one or more amino acid substitutions include one or more Kabat positions selected from heavy chain positions H15, H17, H19, H57, H59, H64, H65, H66, H68, H69, H70, H80, H81 and, H82a, H82b. 
     
     
         46 . The anti- S. aureus  variable heavy chain sequence variant antibody of  claim 45 , wherein the variable heavy chain sequence variant antibody is an anti-SpA antibody. 
     
     
         47 . The anti- S. aureus  variable heavy chain sequence variant antibody of  claim 45 , wherein the variable heavy chain sequence variant antibody is an anti-SpA antibody and wherein the heavy chain constant domain comprises an amino acid sequence selected from SEQ ID NOs: 31-56. 
     
     
         48 . The anti- S. aureus  variable heavy chain sequence variant antibody of  claim 45 , wherein the parental antibody is an anti-SpA humanized antibody, an anti-SpA Fc variant antibody, an anti-SpA matured antibody, or an anti-SpA matured Fc variant antibody;
 wherein the variable light chain sequence and/or the variable heavy chain sequence of the anti-SpA matured antibody, or anti-SpA matured Fc variant antibody comprise one or more maturation mutations resulting in one or more amino acid substitutions, deletions, or insertions that improve one or more properties of the humanized anti-SpA antibody selected from affinity, avidity, stability, solubility, expression level, and/or biological activity.   
     
     
         49 . The anti- S. aureus  variable heavy chain sequence variant antibody of  claim 48  comprising:
 an immunoglobulin heavy chain which comprises a variable heavy chain sequence and a constant heavy chain sequence, wherein the variable heavy chain sequence comprises (i) a heavy chain CDR1 sequence selected from GFAFSNYD, GFTFNTNA, and GYSFTSYY; (ii) a heavy chain CDR2 sequence selected from ISSGGTYP, IRSKSNNYAT, and IDPFNGGT; and (iii) a heavy chain CDR3 sequence selected from ARGGFLITTRDYYAMDY, VTEHYDYDYYVMDY, and ARYGYDGTFYAMDY; and 
 an immunoglobulin light chain which comprises a variable light chain sequence and a constant light chain sequence, wherein the variable light chain sequence comprises (i) a light chain CDR1 sequence selected from SSVSY and ESVEYSGASL; (ii) a light chain CDR2 sequence selected from DTS, AAS, and EIS; and (iii) a light chain CDR3 sequence selected from QQWSSYPPT, QQSRKVPST, and QQWSYPFT. 
 
     
     
         50 . The anti-SpA antibody of  claim 45 , wherein the one or more amino acid substitutions are selected from: H17S to H17P; H19R to H19G. H19K or H19T; H57K to H57A, H57P, H57R, or H57S; H57I to H57A, H57P, H57R, or H57S; H57T to H57A, H57P, H57R, or H57S; H59Y to H59F, H59H, H59N, or H59S; H68T to H68A, H68I or H68S; H70S to H70F; H81Q to H81E, H81H or H81R; H82aN to H82aD, H82aH, H82aK, H82aS, H82aT; H82aG to H82aD, H82aH, H82aK, H82aS, H82aT; and H82bS to H82bG, H82bN, or H82bT.