Anti-VASA antibodies, and methods of production and use thereof
Anti-VASA antibodies (mAbs), particularly humanized mAbs that specifically bind to VASA with high affinity, are disclosed. The amino acid sequences of the CDRs of the light chains and the heavy chains, as well as consensus sequences for these CDRs, of these anti-VASA mAbs are provided. The disclosure also provides nucleic acid molecules encoding the anti-VASA mAbs, expression vectors, host cells, methods for making the anti-VASA mAbs, and methods for expressing the anti-VASA mAbs. Finally, methods of using the anti-VASA mAbs to isolate and/or purify cells expressing VASA are disclosed.
1. An antibody that specifically binds to a human VASA protein comprising an immunoglobulin heavy chain and an immunoglobulin light chain,
a) wherein the variable region of said light chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-88;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-95; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96-104; and
b) wherein the variable region of said heavy chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105-112;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 113-121; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 122-131.
2. An antibody preparation comprising: an antibody that specifically binds to a human VASA protein comprising an immunoglobulin heavy chain and an immunoglobulin light chain,
a) wherein the variable region of said light chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-88;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-95; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96-104; and
b) wherein the variable region of said heavy chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105-112;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 113-121; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 122-131.
3. The antibody preparation of claim 2 wherein said preparation is a monoclonal antibody preparation.
4. The antibody preparation of claim 2 wherein said preparation is a mixture of at least two monoclonal antibody preparations.
5. An isolated nucleic acid molecule encoding a heavy chain or light chain of an antibody that specifically binds to a human VASA protein comprising: a) an immunoglobulin heavy chain and an immunoglobulin light chain,
a) wherein the variable region of said light chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-88;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-95; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96-104; and
b) wherein the variable region of said heavy chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105-112;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 113-121; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 122-131.
6. The isolated nucleic acid of claim 5 wherein said nucleic acid is selected from the group consisting of a cloning vector, an expression vector, a heterologous recombination vector and a viral integration vector.
7. A method of isolating a cell expressing a VASA protein comprising:
(A) obtaining a population of cells;
(B) contacting the population of cells with a multiplicity of antibodies comprising an antibody that specifically binds to a human VASA protein comprising an immunoglobulin heavy chain and an immunoglobulin light chain,
a) wherein the variable region of said light chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 83-88;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 89-95; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 96-104; and
b) wherein the variable region of said heavy chain comprises:
(i) a CDR1 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 105-112;
(ii) a CDR2 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 113-121; and
(iii) a CDR3 region comprising an amino acid sequence selected from the group consisting of SEQ ID NOs: 122-131; and
(C) separating cells in the population that specifically bind the antibodies from cells in the population that do not specifically bind the antibodies.
8. The method of claim 7 wherein the cells are separated by fluorescence activated cell sorting.
9. The method of claim 7 wherein the cells are separated using an immobilized secondary antibody by fluorescence activated cell sorting.