Human anti-pseudomonas-aeruginosa antibodies derived from transgenic xenomouse

Abstract

The invention described herein provides for human antibodies produced in non-human animals that specifically bind to lipopolysaccharide (LPS) from strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02). The invention further provides methods for making the antibodies in a non-human animal, expression of the antibodies in cell lines including hybridomas and recombinant host cell systems. Also provided are kits and pharmaceutical compositions comprising the antibodies and methods of treating or preventing pseudomonas infection by administering to a patient the pharmaceutical compositions described herein.

1 . An isolated human antibody or antigen-binding portion thereof that was expressed in a non-human animal and specifically binds to LPS from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02). 
     
     
         2 . The isolated human antibody or antigen-binding portion thereof according to  claim 1 , wherein said human antibody is a monoclonal antibody. 
     
     
         3 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein the antibody or portion possesses one or more properties selected from the group consisting of:
 a) is opsonic for  Pseudomonas aeruginosa  cells;   b) facilitates phagocytosis of said  Pseudomonas aeruginosa  cells;   c) enhances the immune response to  Pseudomonas aeruginosa;      d) facilitates the killing of  Pseudomonas aeruginosa  cells;   and inhibits  Pseudomonas aeruginosa  infection.   
     
     
         4 . (canceled) 
     
     
         5 . (canceled) 
     
     
         6 . (canceled) 
     
     
         7 . (canceled) 
     
     
         8 . (canceled) 
     
     
         9 . (canceled) 
     
     
         10 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody or antigen-binding portion thereof binds to  Pseudomonas aeruginosa  LPS with a K d  selected from the group consisting of
 a) 5×10 −7  M to 1×10 −7  M;   b) 1×10 −7  to 5×10 −8  M, and   c) 5×10 −8  M to 1×10 −8  M.   
     
     
         11 . (canceled) 
     
     
         12 . (canceled) 
     
     
         13 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody or antigen-binding portion thereof has a half-life in vivo of between one hour and thirty days 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody isotype is selected from the group consisting of: immunoglobulin G (IgG), IgM, IgE, IgA and IgD. 
     
     
         17 . The isolated human antibody according to  claim 16 , wherein said IgG is a subtype selected from the list consisting of IgG1, IgG2, IgG3 and IgG4. 
     
     
         18 . The isolated human antibody according to  claim 17 , wherein said IgG is the IgG2 subtype. 
     
     
         19 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody or portion is labeled. 
     
     
         20 . The isolated human antibody or antigen-binding portion thereof according to  claim 19 , wherein said label is selected from the group consisting of: a radiolabel, an enzyme label, a fluorescent label, a toxin, a magnetic agent, a second antibody, an affinity label, an epitope tag, an antibiotic, a complement protein and a cytokine. 
     
     
         21 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , comprising a light chain selected from the group consisting of:
 a) a kappa light chain; and   b) a lambda light chain.   
     
     
         22 . The isolated human antibody or antigen-binding portion thereof according to  claim 21 , wherein said kappa light chain utilizes a human gene selected from the group consisting of: Vκ2/A19/A3; Vκ1/A30; Vκ4/B3; Vκ3/A27; Vκ3/L2; Vκ1/A30; Vκ3/L2 μL16; and Vκ1/A30. 
     
     
         23 . (canceled) 
     
     
         24 . (canceled) 
     
     
         25 . (canceled) 
     
     
         26 . The isolated human antibody or antigen-binding portion thereof according to  claim 21 , wherein said kappa light chain comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; SEQ ID NO: 26; SEQ ID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; and SEQ ID NO: 30. 
     
     
         27 . (canceled) 
     
     
         28 . (canceled) 
     
     
         29 . (canceled) 
     
     
         30 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein the heavy chain utilizes a human gene selected from the group consisting of: V H 3/V4-04; V H 3/V4-59; V H 3/V3-33; V H 3/V3-15; V H 6/V6-01; and V H 5/V5-51. 
     
     
         31 . (canceled) 
     
     
         32 . (canceled) 
     
     
         33 . (canceled) 
     
     
         34 . (canceled) 
     
     
         35 . (canceled) 
     
     
         36 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , comprising a heavy chain that comprises an amino acid sequence selected from the group consisting of: SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19;
 SEQ ID NO: 20; and SEQ ID NO: 21.   
     
     
         37 . (canceled) 
     
     
         38 . The antigen-binding portion according to  claim 1  or  claim 2  selected from the group consisting of: an Fab fragment, an F(ab′)2 fragment and an Fv fragment. 
     
     
         39 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody is a single chain antibody. 
     
     
         40 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein said antibody is a chimeric antibody. 
     
     
         41 . The chimeric antibody according to  claim 40 , comprising framework regions and CDR regions from different human antibodies. 
     
     
         42 . The chimeric antibody according to  claim 40 , wherein the chimeric antibody is bispecific. 
     
     
         43 . (canceled) 
     
     
         44 . The isolated human antibody or antigen-binding portion according to  claim 1  or  claim 2 , wherein said antibody or portion is derivatized. 
     
     
         45 . The isolated human antibody or antigen-binding portion according to  claim 44 , wherein said antibody or portion is derivatized with polyethylene glycol, at least one methyl or ethyl group or at least one carbohydrate moiety. 
     
     
         46 . A pharmaceutical composition comprising the antibody or antigen-binding portion according to  claim 1  or  claim 2  and a pharmaceutically acceptable carrier. 
     
     
         47 . A kit comprising the antibody or antigen-binding portion according to  claim 1  or  claim 2 , a pharmaceutically acceptable carrier therefor, and a container. 
     
     
         48 . (canceled) 
     
     
         49 . A method for treating or preventing  Pseudomonas aeruginosa  infection, comprising the step of administering an isolated human antibody according to  claim 1  or  claim 2  to a patient at risk of being infected with, or currently infected with,  Pseudomonas aeruginosa.    
     
     
         50 . (canceled) 
     
     
         51 . (canceled) 
     
     
         52 . (canceled) 
     
     
         53 . The method according to  claim 49 , wherein said antibody is labeled with a radiolabel, a toxin, a complement protein, a cytokine or an antibiotic. 
     
     
         54 . (canceled) 
     
     
         55 . The method according to  claim 49  wherein said patient is a burn patient, a surgical patient, a prosthesis recipient, a respiratory patient, a cancer patient, a cystic fibrosis patient or an immunocompromised patient. 
     
     
         56 . The method according to  claim 49 , wherein said pharmaceutical composition further comprises toxins, complement proteins, radiolabeled proteins, cytokines, antibiotics, or any combination thereof. 
     
     
         57 . An isolated cell line that produces the antibody according to  claim 1  or  claim 2 . 
     
     
         58 . The cell line according to  claim 57  wherein said cell line is a hybridoma. 
     
     
         59 . A method of producing an isolated human antibody or antigen-binding portion thereof that specifically binds to LPS from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02), comprising:
 a) culturing a non-human cell capable of producing said antibody under conditions in which the antibody is produced;   b) isolating said antibody from said cell culture.   
     
     
         60 . (canceled) 
     
     
         61 . (canceled) 
     
     
         62 . (canceled) 
     
     
         63 . (canceled) 
     
     
         64 . A method for making an isolated human antibody that specifically binds to LPS from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), IATS016 (02/05), and 170006 (02), comprising:
 a) immunizing a non-human animal capable of producing a human antibody therein with a  Pseudomonas aeruginosa  antigenic composition;   b) allowing said non-human animal to mount a humoral response to said antigenic composition; and   c) recovering said human antibody from said non-human animal.   
     
     
         65 . A nucleic acid molecule isolated from a non-human animal encoding the heavy chain, the light chain or both, of a human antibody or an antigen-binding portion thereof that specifically binds to LPS from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02). 
     
     
         66 . (canceled) 
     
     
         67 . An isolated nucleic acid molecule, comprising a nucleotide sequence encoding an amino acid sequence selected from the group consisting of: SEQ ID NO: 13; SEQ ID NO: 14; SEQ ID NO: 15; SEQ ID NO: 16; SEQ ID NO: 17; SEQ ID NO: 18; SEQ ID NO: 19; SEQ ID NO: 20; and SEQ ID NO: 21. 
     
     
         68 . (canceled) 
     
     
         69 . (canceled) 
     
     
         70 . (canceled) 
     
     
         71 . (canceled) 
     
     
         72 . (canceled) 
     
     
         73 . An isolated nucleic acid molecule, comprising a nucleotide sequence encoding an amino acid sequence selected from the group consisting of: SEQ ID NO: 22; SEQ ID NO: 23; SEQ ID NO: 24; SEQ ID NO: 25; SEQ ID NO: 26; SEQ ID NO: 27; SEQ ID NO: 28; SEQ ID NO: 29; and SEQ ID NO: 30. 
     
     
         74 . (canceled) 
     
     
         75 . A vector comprising the nucleic acid molecule according to any one of  claims 65 ,  67  or  73 . 
     
     
         76 . The vector according to  claim 75 , wherein said vector further comprises an expression control sequence operably linked to said nucleic acid. 
     
     
         77 . An host cell comprising
 a) a nucleic acid molecule according to any one of  claims 65 ,  67  or  73 ; or   b) a vector according to  claim 75  or  76 .   
     
     
         78 . (canceled) 
     
     
         79 . (canceled) 
     
     
         80 . (canceled) 
     
     
         81 . (canceled) 
     
     
         82 . (canceled) 
     
     
         83 . A method of producing the heavy chain or the antigen-binding portion thereof, the light chain or the antigen-binding portion thereof, or both the light chain and heavy chain or antigen-binding portions thereof, of a human antibody that specifically binds to LPS from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02), comprising the step of culturing the host cell according to  claims 77  under conditions in which the nucleic acid molecules are expressed. 
     
     
         84 . (canceled) 
     
     
         85 . (canceled) 
     
     
         86 . A non-human transgenic animal comprising the nucleic acid molecule according to any one of  claims 65 ,  67  or  73 , wherein said non-human transgenic animal expresses said nucleic acid molecule. 
     
     
         87 . (canceled) 
     
     
         88 . (canceled) 
     
     
         89 . The non-human transgenic animal according to  claim 86 , wherein said animal is selected from the list consisting of a mouse, a rat, a hamster, a cow, a sheep, a primate, a horse and a pig. 
     
     
         90 . (canceled) 
     
     
         91 . (canceled) 
     
     
         92 . (canceled) 
     
     
         93 . (canceled) 
     
     
         94 . (canceled) 
     
     
         95 . (canceled) 
     
     
         96 . (canceled) 
     
     
         97 . The isolated human antibody or antigen-binding portion thereof according to  claim 1  or  claim 2 , wherein the antibody or portion thereof is produced by a process comprising the steps of:
 a) immunizing a non-human animal capable of producing a human antibody with an antigen selected from the group consisting of an isolated  Pseudomonas aeruginosa  LPS preparation, a virulent  Pseudomonas aeruginosa  cell preparation, an attenuated  Pseudomonas aeruginosa  cell preparation, and a killed  Pseudomonas aeruginosa  cell preparation;   b) allowing said non-human animal to mount an immune response to said antigen; and   c) recovering said antibody from said non-human animal.   
     
     
         98 . (canceled) 
     
     
         99 . (canceled) 
     
     
         100 . (canceled) 
     
     
         101 . A hybridoma cell line having American Type Culture Collection Deposit Designation PTA-5384, PTA-5385 or PTA-5386. 
     
     
         102 . A monoclonal antibody produced by the hybridoma cell line according to  claim 101 . 
     
     
         103 . An isolated human antibody or an antigen-binding portion thereof, that specifically binds LPS O-specific side chain from one of  Pseudomonas aeruginosa  strains Fisher Devlin (International Serogroups) It-2 (011), It-3 (02), It-4 (01), It-5 (010), It-6 (07), PA01 (05), 170003 (02), IATS016 (02/05), and 170006 (02). 
     
     
         104 . The antibody antigen-binding portion according to  claim 103 , which is monoclonal. 
     
     
         105 . The antibody or antigen-binding portion according to  claim 103  or  104 , having a relative binding avidity of about 1.0. 
     
     
         106 . The antibody or antigen-binding portion according to  claim 104 , that specifically binds the LPS O-specific side chain of one strain and does not bind to the LPS O-specific side chain of any other of the listed strains. 
     
     
         107 . A passive vaccine for preventing or inhibiting  Pseudomonas aeruginosa  infection comprising one or more human monoclonal antibodies or an antigen-binding portion thereof, selected from the group consisting of the antibody or portion according to  claim 1  or  102  or  103 . 
     
     
         108 . The passive vaccine according to  claim 107  comprising two or more human monoclonal antibodies or an antigen-binding portion thereof, wherein said human monoclonal antibodies or portions specifically bind different strains of  Pseudomonas aeruginosa.    
     
     
         109 . A method for detecting the presence of  Pseudomonas aeruginosa  in a biological sample comprising the steps of contacting said sample with an antibody or antigen-binding portion thereof according to any one of  claims 1  or  102  or  103 .