Human IgE monoclonal antibodies to antibodies to alpha-gal (galactose-α-1,3-galactose) and uses therefor
The present disclosure is directed to human IgE antibodies and fragments thereof binding to galactose-α-1, 3-galactose, a known red meat allergen, and methods for use thereof.
1 . A method of detecting galactose-α-1,3-galactose (alpha-gal) in a sample comprising:
(a) contacting a sample with an antibody or antibody fragment comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively; and
(b) detecting alpha-gal in said sample by binding of said antibody or antibody fragment to alpha-gal in said sample.
2 . A method of detecting IgE anti-galactose-α-1,3-galactose (alpha-gal) antibodies in a sample comprising:
(a) contacting a sample suspected IgE anti-alpha gal antibodies with alpha-gal in a competitive assay with an antibody or antibody fragment comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively; and
(b) detecting competition for binding of alpha-gal by a suspected IgE anti-alpha gal in said sample by an antibody or antibody fragment comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively.
3 . A method of preventing or treating an alpha-gal-related allergic reaction in a subject comprising delivering to said subject an IgG antibody or antibody fragment comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively.
4 . An IgG monoclonal antibody or antibody fragment thereof, wherein the antibody or antibody fragment comprises (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10, and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively.
5 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody or antibody fragment is encoded by heavy and light chain variable regions comprising SEQ ID NOS: 1 and 2, respectively, or SEQ ID NOS: 3 and 4, respectively.
6 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody or antibody fragment is encoded by heavy and light chain variable regions having at least 80%, 90%, or 95% identity to SEQ ID NOS: 1 and 2, respectively, or SEQ ID NOS: 3 and 4, respectively.
7 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody or antibody fragment comprises heavy and light chain variable sequences comprising SEQ ID NOS: 5 and 6, respectively, or SEQ ID NOS: 7 and 8, respectively.
8 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody or antibody fragment comprises heavy and light chain variable regions having 95% identity to SEQ ID NOS: 5 and 6, respectively, or SEQ ID NOS: 7 and 8, respectively.
9 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein the antibody fragment is a recombinant scFv (single chain fragment variable) antibody, Fab fragment, F(ab′) 2 fragment, or Fv fragment.
10 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody is a chimeric antibody or bispecific antibody.
11 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody comprises an Fc portion mutated to alter FcR interactions, to increase half-life and/or increase therapeutic efficacy, or glycan modified to alter FcR interactions.
12 . The monoclonal antibody or antibody fragment of claim 11 , wherein the Fc portion mutated to alter FcR interactions, to increase half-life and/or increase therapeutic efficacy is selected from the group consisting of a LALA mutation, a LALA-PG mutation, an N297 mutation, a GASD/ALIE mutation, a DHS mutation, a YTE mutation or a LS mutation, and wherein the glycan modification to alter FcR interactions is enzymatic or chemical addition or removal of glycans or expression in a cell line engineered with a defined glycosylating pattern.
13 . The IgG monoclonal antibody or antibody fragment of claim 4 , wherein said antibody or antibody fragment further comprises a cell penetrating peptide and/or is an intrabody.
14 . An engineered cell encoding an antibody or antibody fragment wherein the antibody or antibody fragment comprises (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively.
15 . The engineered cell of claim 14 , wherein said antibody or antibody fragment is encoded by heavy and light chain variable regions comprising SEQ ID NOS: 1 and 2, respectively, or SEQ ID NOS: 3 and 4, respectively.
16 . The engineered cell of claim 14 , wherein said antibody or antibody fragment is encoded by heavy and light chain variable regions having at least 80%, 90%, or 95% identity to SEQ ID NOS: 1 and 2, respectively, or SEQ ID NOS: 3 and 4, respectively.
17 . The engineered cell of claim 14 , wherein said antibody or antibody fragment comprises heavy and light chain variable regions comprising SEQ ID NOS: 5 and 6, respectively, or SEQ ID NOS: 7 and 8, respectively.
18 . The engineered cell of claim 14 , wherein said antibody or antibody fragment comprises heavy and light chain variable regions having at least 95% identity to variable regions comprising SEQ ID NOS: 5 and 6, respectively, or SEQ ID NOS: 7 and 8, respectively.
19 . The hybridoma or engineered cell of claim 14 , wherein the antibody fragment is a recombinant scFv (single chain fragment variable) antibody, Fab fragment, F(ab′) 2 fragment, or Fv fragment.
20 . The hybridoma or engineered cell of claim 14 , wherein said antibody is a chimeric or bispecific antibody.
21 . A method of de-sensitizing a subject to galactose-α-1,3-galactose (alpha-gal) comprising:
(a) administering to said subject galactose-α-1,3-galactose (alpha-gal); and
(b) administering to said subject an IgG antibody that has a binding specificity to galactose-α-1,3-galactose (alpha-gal) fragment and comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10 and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively.
22 . A monoclonal antibody or antibody fragment thereof comprising (i) heavy chain CDR1-3 comprising SEQ ID NOS: 9, 10, and 11, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 15, 16 and 17, respectively, or (ii) heavy chain CDR1-3 comprising SEQ ID NOS: 12, 13 and 14, respectively, and light chain CDR1-3 comprising SEQ ID NOS: 18, 19 and 20, respectively, wherein the antibody or antibody fragment comprises an Fc portion mutated to alter FcR interactions, to increase half-life and/or increase therapeutic efficacy or is glycan modified to alter FcR interactions.
23 . The monoclonal antibody or antibody fragment of claim 22 , wherein the Fc portion mutated to alter FcR interactions, to increase half-life and/or increase therapeutic efficacy is selected from the group consisting of a LALA mutation, a LALA-PG mutation, an N297 mutation, a GASD/ALIE mutation, a DHS mutation, a YTE mutation or a LS mutation, and wherein the glycan modification to alter FcR interactions is enzymatic or chemical addition or removal of glycans or expression in a cell line engineered with a defined glycosylating pattern.