COMPOSITIONS AND METHODS FOR THE THERAPY AND DIAGNOSIS OF CYTOMEGALOVIRUS INFECTIONS
The present invention provides novel anti-cytomegalovirus antibodies and related compositions and methods. These antibodies may be used in the diagnosis, prevention, and treatment of cytomegalovirus infection.
1 . An isolated anti-CMV antibody or fragment thereof, wherein said antibody comprises:
I:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SNHGIH (SEQ ID NO: 36);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDGDDDRYADSVKG (SEQ ID NO: 37);
(iii) a V H CDR3 region comprising the amino acid sequence of DGRCGEPKCYSGLPDY (SEQ ID NO: 38); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of DASNRAT (SEQ ID NO: 44);
(iii) a V L CDR3 region comprising the amino acid sequence of LQRNTWPPLT (SEQ ID NO: 45);
II:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SNYGMH (SEQ ID NO: 48);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDGSNEHYADSVKG (SEQ ID NO: 49);
(iii) a V H CDR3 region comprising the amino acid sequence of DGRCPDVNCYSGLIDY (SEQ ID NO: 50); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYLA (SEQ ID NO: 53);
(ii) a V L CDR2 region comprising the amino acid sequence of DASNRAT (SEQ ID NO: 44);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSNWPPLT (SEQ ID NO: 54);
III:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SSNGIH (SEQ ID NO: 57);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDANDKQYADSVKG (SEQ ID NO: 58);
(iii) a V H CDR3 region comprising the amino acid sequence of DGTCSGGNCYSGLIDY (SEQ ID NO: 59); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of ASIRAT (SEQ ID NO: 64);
(iii) a V L CDR3 region comprising the amino acid sequence of HQRSNWPPLT (SEQ ID NO: 65);
IV:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SNHGIH (SEQ ID NO: 36);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNAHYADSVRG (SEQ ID NO: 68);
(iii) a V H CDR3 region comprising the amino acid sequence of EGRCIEENCYSGQIDY (SEQ ID NO: 69); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYMA (SEQ ID NO: 74);
(ii) a V L CDR2 region comprising the amino acid sequence of DASIRAT (SEQ ID NO: 75);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSSWPPLT (SEQ ID NO: 76);
V:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SNHGIH (SEQ ID NO: 36);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNAHYADSVRGR (SEQ ID NO: 79);
(iii) a V H CDR3 region comprising the amino acid sequence of EGRCIEEKCYSGQIDY (SEQ ID NO: 80); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYMA (SEQ ID NO: 74);
(ii) a V L CDR2 region comprising the amino acid sequence of DASIRAT (SEQ ID NO: 75);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSSWPPLT (SEQ ID NO: 76);
VI:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SDYGMH (SEQ ID NO: 85);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNTHYADSVRG (SEQ ID NO: 86);
(iii) a V H CDR3 region comprising the amino acid sequence of DGKCPDLKCYSGLIDY (SEQ ID NO: 87);
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of DASKRAT (SEQ ID NO: 92);
(iii) a V L CDR3 region comprising the amino acid sequence of HQRSSWPPLT (SEQ ID NO: 93);
VII:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of SXXGXH (SEQ ID NO: 95), SXXGIH (SEQ ID NO: 98), or SXYGMH (SEQ ID NO: 101);
(ii) a V H CDR2 region comprising the amino acid sequence of VISXD YADSVRG (SEQ ID NO: 96) or VISXDGXNXHYADSVXG (SEQ ID NO: 99);
(iii) a V H CDR3 region comprising the amino acid sequence of DGXCSXXXCYSGLXDY (SEQ ID NO: 100), EGRCIEEXCYSGQIDY (SEQ ID NO: 102), or DGXCPDXXCYSGLIDY (SEQ ID NO: 103); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGXYXA (SEQ ID NO: 111) or RASQSVGXYLA (SEQ ID NO: 114);
(ii) a V L CDR2 region comprising the amino acid sequence of XASXRAT (SEQ ID NO: 112) or DASXRAT (SEQ ID NO: 115);
(iii) a V L CDR3 region comprising the amino acid sequence of XQRXXWPPLT (SEQ ID NO: 113), HQRSXWPPLT (SEQ ID NO: 116), or QQRSXWPPLT (SEQ ID NO: 117).
2 . An isolated anti-CMV antibody or fragment thereof, wherein said antibody comprises:
I:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of GFTFSN (SEQ ID NO: 39);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDGDDDR (SEQ ID NO: 40);
(iii) a V H CDR3 region comprising the amino acid sequence of DGRCGEPKCYSGLPDY (SEQ ID NO: 38); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of DASNRAT (SEQ ID NO: 44);
(iii) a V L CDR3 region comprising the amino acid sequence of LQRNTWPPLT (SEQ ID NO: 45);
II:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of GLTFSN (SEQ ID NO: 118);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDGSNEH (SEQ ID NO: 51);
(iii) a V H CDR3 region comprising the amino acid sequence of DGRCPDVNCYSGLIDY (SEQ ID NO: 50); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYLA (SEQ ID NO: 53);
(ii) a V L CDR2 region comprising the amino acid sequence of DASNRAT (SEQ ID NO: 44);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSNWPPLT (SEQ ID NO: 54);
III:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of GFTFSS (SEQ ID NO: 60);
(ii) a V H CDR2 region comprising the amino acid sequence of VISSDANDKQ (SEQ ID NO: 61);
(iii) a V H CDR3 region comprising the amino acid sequence of DGTCSGGNCYSGLIDY (SEQ ID NO: 59); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of ASIRAT (SEQ ID NO: 64);
(iii) a V L CDR3 region comprising the amino acid sequence of HQRSNWPPLT (SEQ ID NO: 65);
IV:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of KFIFSN (SEQ ID NO: 70);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNAH (SEQ ID NO: 71);
(iii) a V H CDR3 region comprising the amino acid sequence of EGRCIEENCYSGQIDY (SEQ ID NO: 69); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYMA (SEQ ID NO: 74);
(ii) a V L CDR2 region comprising the amino acid sequence of DASIRAT (SEQ ID NO: 75);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSSWPPLT (SEQ ID NO: 76);
V:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of KFIFSN (SEQ ID NO: 70);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNAH (SEQ ID NO: 71);
(iii) a V H CDR3 region comprising the amino acid sequence of EGRCIEEKCYSGQIDY (SEQ ID NO: 80); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGRYMA (SEQ ID NO: 74);
(ii) a V L CDR2 region comprising the amino acid sequence of DASIRAT (SEQ ID NO: 75);
(iii) a V L CDR3 region comprising the amino acid sequence of QQRSSWPPLT (SEQ ID NO: 76);
VI:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of GLTFSD (SEQ ID NO: 88);
(ii) a V H CDR2 region comprising the amino acid sequence of VISKDGTNTH (SEQ ID NO: 89);
(iii) a V H CDR3 region comprising the amino acid sequence of DGKCPDLKCYSGLIDY (SEQ ID NO: 87); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGGYLA (SEQ ID NO: 43);
(ii) a V L CDR2 region comprising the amino acid sequence of DASKRAT (SEQ ID NO: 92);
(iii) a V L CDR3 region comprising the amino acid sequence of HQRSSWPPLT (SEQ ID NO: 93);
VII:
(a) a V H region comprising
(i) a V H CDR1 region comprising the amino acid sequence of XXXFSX (SEQ ID NO: 104) or GXTFSX (SEQ ID NO: 107);
(ii) a V H CDR2 region comprising the amino acid sequence of VISXDXXXXX (SEQ ID NO: 105) or VISKDGTNXH (SEQ NO: 108);
(iii) a V H CDR3 region comprising the amino acid sequence of XGXCXXXXCYSGXXDY (SEQ ID NO: 106), DGXCXXXXCYSGLXDY (SEQ ID NO: 109), or EGRCIEEXCYSGQIDY (SEQ ID NO: 110); and
(b) a V L region comprising
(i) a V L CDR1 region comprising the amino acid sequence of RASQSVGXYXA (SEQ ID NO: 111) or RASQSVGXYLA (SEQ ID NO: 114);
(ii) a V L CDR2 region comprising the amino acid sequence of XASXRAT (SEQ ID NO: 112) or DASXRAT (SEQ ID NO: 115);
(iii) a V L CDR3 region comprising the amino acid sequence of XQRXXWPPLT (SEQ ID NO: 113), HQRSXWPPLT (SEQ ID NO: 116), or QQRSXWPPLT (SEQ ID NO: 117).
3 . An isolated anti-CMV antibody comprising:
a) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 35 and a light chain sequence comprising amino acid sequence SEQ ID NO: 42 or;
b) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 47 and a light chain sequence comprising amino acid sequence SEQ ID NO: 52 or;
c) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 56 and a light chain sequence comprising amino acid sequence SEQ ID NO: 63 or;
d) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 67 and a light chain sequence comprising amino acid sequence SEQ ID NO: 73 or;
e) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 78 and a light chain sequence comprising amino acid sequence SEQ ID NO: 82 or;
f) a heavy chain sequence comprising the amino acid sequence of SEQ ID NO: 84 and a light chain sequence comprising amino acid sequence SEQ ID NO: 91.
4 . The anti-CMV antibody of claim 1 , wherein said antibody binds an epitope of the glycoprotein b (gB) envelope protein of the CMV virus.
5 . The anti-CMV antibody of claim 4 , wherein said epitope comprises the antigenic determinant 2 site 1 of gB, gp 116.
6 . A composition comprising the antibody of claim 1 .
7 . The composition of claim 6 , further comprising an anti-viral treatment.
8 . The composition of claim 7 , wherein said anti-viral treatment is ganciclovir, foscarnet, cidofovir, valganciclovir, and intravenous immunoglobulin (IVIG).
9 . The composition of claim 6 , further comprising a second anti-CMV antibody.
10 . The antibody of claim 1 , wherein said antibody is operably-linked to a therapeutic agent or a detectable label.
11 . A method for the treatment or prevention of a CMV infection in a subject, comprising administering to the subject the composition of claim 6 .
12 . The method of claim 11 , wherein the method further comprises administering an anti-viral treatment.
13 . The method of claim 11 , wherein said anti-viral treatment is ganciclovir, foscarnet, cidofovir, valganciclovir, and intravenous immunoglobulin (IVIG).
14 . The method of claim 11 , wherein said antibody is administered prior to or after exposure to CMV.
15 . The method of claim 11 , wherein said antibody is administered at a dose sufficient to neutralize CMV infection.
16 . A method for determining the presence of a CMV infection in a patient, comprising the steps of:
(a) contacting a biological sample obtained from the patient with the antibody according to claim 1 ;
(b) detecting an amount of the antibody that binds to the biological sample; and
(c) comparing the amount of antibody that binds to the biological sample to a control value, and therefrom determining the presence of the influenza virus in the patient.
17 . A diagnostic kit comprising the antibody according to claim 1 .