Engineered Antibodies and Uses Thereof

An antibody comprising a polypeptide comprising the amino acid sequence of Formula I:

• Z-(X-Yn-Zr)p-X, wherein
• Z is a hinge;
• X is a CH2 domain;
• Y is a linker;
• n is an integer of 1 or greater;
• p is an integer of 2 or greater;
• and
• r is an integer of 0 or 1.

An antibody that immunospecifically binds to Parkin, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 4, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 5, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 6, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 13, a LCDR2 comprising the amino acid sequence RAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 15, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to OCT4, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 29, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 30, and a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 31, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 37, a LCDR2 comprising the amino acid sequence GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 39, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to IDO1, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 66, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 67, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 68, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 72, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 73, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to FLAG-tag, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 163, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 164, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 165, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 168, a LCDR2 comprising the amino acid sequence of YAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 169, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that is a Rabbit anti-Mouse IgG secondary antibody, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 127, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 128, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 129, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 132, a LCDR2 comprising the amino acid sequence of GAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 133, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to pERK1/2, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 85, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 86, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 87, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 91, a LCDR2 comprising the amino acid sequence of KVS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 92, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to PSD-95, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 111, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 112, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 113, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 116, a LCDR2 comprising the amino acid sequence of DTS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 117, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

An antibody that immunospecifically binds to TIGIT, wherein the antibody comprises a heavy chain (HC) and a light chain (LC) that pair to form a binding domain comprising a heavy chain complementarity determining region 1 (HCDR1) comprising the amino acid sequence of SEQ ID NO:

• 146, a HCDR2 comprising the amino acid sequence of SEQ ID NO:
• 147, a HCDR3 comprising the amino acid sequence of SEQ ID NO:
• 148, a light chain complementarity determining region 1 (LCDR1) comprising the amino acid sequence of SEQ ID NO:
• 151, a LCDR2 comprising the amino acid sequence of SAS, and a LCDR3 comprising the amino acid sequence of SEQ ID NO:
• 152, and wherein the heavy chain further comprises a first CH2 domain, a second CH2 domain, and a third CH2 domain, wherein the first CH2 domain and the second CH2 domain are connected by a first linker and an optional first hinge, and wherein the second CH2 domain and the third CH2 domain are connected by a second linker and an optional second hinge, and wherein the heavy chain further comprises a CH3 domain.

A method of increasing sensitivity of an antibody, the method comprising:

• introducing into a heavy chain of the antibody a polypeptide comprising amino acid sequence of Formula I:
• Z-(X-Yn-Zr)p-X, wherein
• is a hinge;
• X is a CH2 domain;
• Y is a linker;
• n is an integer of 1 or greater;
• p is an integer of 2 or greater;
• and
• r is an integer of 0 or 1,
• wherein the sensitivity of the antibody is increased by at least 1.5-fold compared to a corresponding wild type antibody.

A method of reducing background signal of an antibody, the method comprising:

• introducing into a heavy chain of the antibody a polypeptide comprising amino acid sequence of Formula I:
• Z-(X-Yn-Zr)p-X, wherein
• Z is a hinge;
• X is a CH2 domain;
• Y is a linker;
• n is an integer of 1 or greater;
• p is an integer of 2 or greater;
• and
• r is an integer of 0 or 1;
• wherein the background signal of the antibody is decreased by at least 1.5-fold compared to a corresponding wild type antibody.