Polynucleotides encoding a humanized anti-CD40 antibody
The present disclosure relates to anti-CD40 antibodies, such as humanized anti-CD40 antibodies, that may be used in various therapeutic, prophylactic and diagnostic methods. The antibodies generally block the ability of CD40 to bind CD154 and do so without activating the cell expressing CD40 (e.g., a B cell). The present antibodies or fragments thereof may be used to reduce complications associated with organ or tissue transplantation.
1. A polynucleotide encoding a humanized anti-CD40 antibody or antigen-binding fragment thereof comprising a heavy chain variable region comprising an amino acid sequence with at least 80% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 19, 20, 21, 24, 25, and 26 and a light chain variable region comprising an amino acid sequence with at least 80% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 22, 23, 27, 28, and 29, wherein the heavy chain variable region comprises a CDR1, CDR2, and CDR3 with the amino acid sequences set forth in SEQ ID NOs: 13, 14, and 15, respectively, and wherein the light chain variable region comprises a CDR1, CDR2, and CDR3 with the amino acid sequences set forth in SEQ ID NOs: 16, 17, and 18, respectively.
2. The polynucleotide of claim 1 , wherein the amino acid sequence of the heavy chain variable region has at least 90%, 95%, or 99% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 19, 20, 21, 24, 25, and 26, and the amino acid sequence of the light chain variable region has at least 90% sequence identity to the amino acid sequence set forth in any one of SEQ ID NOs: 22, 23, 27, 28, and 29.
3. The polynucleotide of claim 2 , wherein the amino acid sequence of the heavy chain variable region comprises the amino acid sequence set forth in any one of SEQ ID NOs: 19, 20, 21, 24, 25, and 26, and the amino acid sequence of the light chain variable region comprises the amino acid sequence set forth in any one of SEQ ID NOs: 22, 23, 27, 28 and 29, wherein the antibody or antigen-binding fragment thereof does not comprise the heavy chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 21 and the light chain variable region comprising the amino acid sequence set forth in SEQ ID NO: 23.
4. The polynucleotide of claim 1 , wherein:
a) the dissociation constant (KD) of the antibody or antigen-binding fragment thereof is less than about 1×10 −9 M;
b) the antibody or antigen-binding fragment thereof is selected from the group consisting of: (a) a whole immunoglobulin molecule; (b) an scFv; (c) a Fab fragment; (d) an F(ab′) 2 ; and (e) a disulfide linked Fv;
c) the antibody or antigen-binding fragment thereof comprises at least one constant domain selected from the group consisting of: a) an IgG constant domain; and (b) an IgA constant domain;
d) the antibody or antigen-binding fragment thereof comprises at least one human constant domain;
e) the antibody or antigen-binding fragment thereof binds to CD40 extracellular domain;
f) the CD40 is human or rhesus CD40;
g) the antibody or antigen-binding fragment thereof blocks B lymphocyte activation by CD154-expressing Jurkat cells in vitro; and/or
h) the antibody or antigen-binding fragment thereof inhibits B lymphocyte CD23, CD80, or CD86 expression.
5. A vector comprising the polynucleotide of claim 1 .
6. A cell comprising the vector of claim 5 .
7. The cell of claim 6 , wherein the cell is a eukaryotic cell or a prokaryotic cell.
8. The cell of claim 7 , wherein the eukaryotic cell is a mammalian cell.
9. A composition comprising the polynucleotide of claim 1 , a vector comprising the polynucleotide, or a cell comprising the vector, and a pharmaceutically acceptable carrier.
10. A method of producing an antibody or antigen-binding fragment thereof comprising the steps of:
(a) culturing a cell transformed with the polynucleotide of claim 1 or a vector comprising the polynucleotide in culture medium under conditions wherein the polynucleotide is expressed; and
(b) recovering the antibody or antigen-binding fragment thereof from the cell or culture medium.