Constructs specifically recognizing glypican-3 and uses thereof

Abstract

The present application provides constructs comprising an antibody portion that specifically recognizes glypican 3 (GPC3), such as GPC3 bound to the cell surface. Also provided are methods of preparing and using these constructs.

Description

[0001] Cross - reference to related applications

[0002] This application claims priority to U.S. Provisional Application No. 62 / 490,586, filed Apr. 26, 2017, the contents of which are hereby incorporated by reference in their entirety.

[0003] Sequence listing submitted as an ASCII text file

[0004] The following content submitted as an ASCII text file is hereby incorporated by reference in its entirety: a sequence listing in computer-readable form (CRF) (file name: 750042001240SEQLIST.txt, date of record: Apr. 24, 2018, size: 395 KB). Technical Field

[0005] The present invention relates to antibody constructs that specifically recognize glypican 3 (GPC3), methods for preparing the same, and uses thereof, including treating and diagnosing diseases. Background Art

[0006] Glypican 3 (GPC3, also known as SGB, DGSX, MXR7, SDYS, SGBS, OCI-5, SGBS1, GTR2-2) is a cell surface protein that is overexpressed in various types of cancers, including various solid tumors, such as hepatocellular carcinoma (HCC), melanoma (Nakatsura T et al., Clin Cancer Res. 2004), squamous cell lung cancer (Yu X et al., Genet Mol Res. 2015), ovarian cancer (Stadlmann S et al., Int J Gynecol Pathol. 2007), yolk sac tumor, choriocarcinoma (Zynger DL et al., Am J Surg Pathol. 2006), Wilms' tumor, and liposarcoma (Baumhoer D et al., Am J Clin Pathol. 2008). There is still a medical need to develop therapies for various cancers, including cancers that overexpress GPC3.

[0007] The GPC3 gene is located on the X chromosome and encodes a 70 kDa precursor protein of 580 amino acids (aa). This precursor protein can be cleaved by furin at Arg 358 and Cys 359It splits between them and generates a 40 kDa N-terminal subunit and a 30 kDa C-terminal subunit linked by a disulfide bond. Mature GPC3 is attached to the cell surface through a glycosylphosphatidylinositol (GPI) anchor with two heparan sulfate (HS) chains in the C-terminal region close to the cell membrane. As a member of the heparan sulfate proteoglycan family, GPC3 is an oncofetal protein that is widely expressed in human embryos and regulates morphogenesis or growth through interactions with signaling factors (such as Wnt, Hedgehog signaling, etc.). GPC3 is not expressed in normal adult liver; however, during hepatocarcinogenesis, GPC3 reactivation has been reported in HCC patients. Yamauchi N et al. examined the expression of GPC3 in normal, non-neoplastic, and neoplastic liver tissues and found that GPC3 was present in 84% (47 / 56) of HCC patients but not in normal adult liver, cirrhosis, or hepatitis (Yamauchi N et al., Modern Pathology 2005). Results reported by other research groups were similar (Nakatsura T et al., Biochem Biophys Res Commun. 2003; Baumhoer D et al., Am J Clin Pathol 2008; Shirakawa H et al., Cancer Sci. 2009; Wang L et al., Hepatobiliary Pancreat Dis Int. 2015). Studies also showed that GPC3 can be released from the cell surface into the extracellular environment in different forms in HCC patients but not in healthy donors. Therefore, GPC3 is currently used as a serum diagnostic marker for HCC (Hippo Y et al., Cancer Res. 2004; Capurro M et al., Gastroenterology 2003; Haruyama Y and Kataoka H, World J Gastroenterol. 2016).

[0008] Hepatocellular carcinoma (HCC) is the most common type of liver cancer, accounting for approximately 75% of all liver cancers. Liver cancer is the fifth most common cancer in the world and the second most common cause of cancer death. The incidence of liver cancer has increased by more than threefold since 1980, and the mortality rate of liver cancer has increased by almost 3% per year since 2000 (https: / / www.cancer.org / cancer / liver-cancer / about / what-is-key-statistics.html). The prognosis of liver cancer is very poor, and the overall ratio of mortality to incidence is 0.95. Currently, the treatment of liver cancer is mainly limited to chemotherapy or surgery. However, the effect of chemotherapy is usually limited because hepatocytes express ATP-binding cassette (ABC) transporters, which can export a wide range of commonly used chemotherapeutic agents. Another option - surgery can only be used for early-stage cancer, where the 5-year survival rate is 31%. If the cancer is diagnosed at an advanced stage (5-year survival rate: 3 - 11%), then the only approved chemotherapy is the tyrosine kinase inhibitor sorafenib. This therapy only increases the survival period by 2 - 3 months (Fleming BD and Ho M, Toxins 2016). Therefore, there is a need to develop a novel method for treating liver cancer.

[0009] The entire disclosures of all publications, patents, patent applications, and published patent applications mentioned herein are hereby incorporated by reference in their entirety. Summary of the Invention

[0010] In one aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) that specifically recognizes GPC3 bound to the cell surface (referred to herein as "native form GPC3" or "native GPC3 (nGPC3)"). In some embodiments, the construct (referred to herein as "anti-GPC3 construct") includes an antibody portion (referred to herein as "anti-GPC3 antibody portion") that specifically recognizes native form GPC3.

[0011] Therefore, in some embodiments, there is provided an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) that includes an antibody portion that specifically recognizes GPC3 bound to the cell surface. In some embodiments, the antibody portion specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity. In some embodiments, the anti-GPC3 construct is directed against the K of GPC3 bound to the cell surface dis from about 0.1 nM to about 2 nM (such as from about 0.1 nM to about 1 nM, or from about 0.1 nM to about 1.5 nM). In some embodiments, the IC50 of soluble GPC3 competing for binding between the anti-GPC3 construct and GPC3 binding to the cell surface is from about 1 μg / ml to about 100 μg / ml (such as from about 1 μg / ml to about 10 μg / ml, or from about 2 μg / ml to about 5 μg / ml). In some embodiments, GPC3 binding to the cell surface comprises the amino acid sequence of SEQ ID NO: 460 or SEQ ID NO: 462. In some embodiments, GPC3 binding to the cell surface is GPC3 expressed on HepG2 cells. In some embodiments, soluble GPC3 comprises the amino acid sequence of SEQ ID NO: 461 or SEQ ID NO: 463.

[0012] In some embodiments according to any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within human GPC3 comprising the amino acid sequence SEQ ID NO: 460.

[0013] In some embodiments of any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within the amino acid sequence SEQ ID NO: 461.

[0014] In some embodiments according to any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within the amino acid sequence SEQ ID NO: 462.

[0015] In some embodiments according to any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within the amino acid sequence SEQ ID NO: 463.

[0016] In some embodiments according to any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within the N-terminal fragment of GPC3. In some embodiments, the antibody portion specifically recognizes an epitope within the amino acid sequence SEQ ID NO: 464.

[0017] In some embodiments according to any of the above anti-GPC3 constructs (such as the isolated anti-GPC3 construct), the antibody portion specifically recognizes an epitope within the C-terminal fragment of GPC3. In some embodiments, the antibody portion specifically recognizes an epitope within the C-terminal fragment of GPC3 lacking heparan sulfate side chains.

[0018] In some embodiments of any of the above anti-GPC3 constructs (e.g., isolated anti-GPC3 constructs), the antibody portion does not competitively and specifically bind to the same or substantially the same GPC3 epitope as GC33. In some embodiments, the antibody portion does not bind to an epitope within the amino acid sequence SEQ ID NO:536. In some embodiments, the antibody portion does not bind to a fragment comprising the amino acid sequence SEQ ID NO:536.

[0019] In some embodiments of any of the above anti-GPC3 constructs (e.g., isolated anti-GPC3 constructs), GPC3 is expressed on the surface of cells selected from the group consisting of HepG2, Hep3B, Huh7, JHH-7, and 293.

[0020] In some embodiments of any of the above anti-GPC3 constructs (e.g., isolated anti-GPC3 constructs), GPC3 is expressed on the surface of cancer cells. In some embodiments, the cancer cells are liver cancer cells, such as hepatocellular carcinoma (HCC).

[0021] In another aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) comprising an antibody portion that specifically recognizes GPC3 bound to the cell surface, wherein the antibody portion comprises: i) a heavy chain variable domain (V H ), which comprises a heavy chain complementarity determining region (HC-CDR) 1 having an amino acid sequence comprising any one of SEQ ID NOs: 1-31 or a variant thereof comprising up to about 5 amino acid substitutions, an HC-CDR2 having an amino acid sequence comprising any one of SEQ ID NOs: 52-82 or a variant thereof comprising up to about 5 amino acid substitutions, and an HC-CDR3 having an amino acid sequence comprising any one of SEQ ID NOs: 103-133 or a variant thereof comprising up to about 5 amino acid substitutions; and ii) a light chain variable domain (V L ), which comprises a light chain complementarity determining region (LC-CDR) 1 having an amino acid sequence comprising any one of SEQ ID NOs: 154-184 or a variant thereof comprising up to about 5 amino acid substitutions, an LC-CDR2 having an amino acid sequence comprising any one of SEQ ID NOs: 205-235 or a variant thereof comprising up to about 3 amino acid substitutions, and an LC-CDR3 having an amino acid sequence comprising any one of SEQ ID NOs: 256-286 or a variant thereof comprising up to about 5 amino acid substitutions. In some embodiments, the antibody portion comprises: i) V H, which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286.

[0022] In some embodiments of any of the above anti-GPC3 constructs (e.g., isolated anti-GPC3 constructs), the antibody portion comprises: i) a V comprising an amino acid sequence of any one of SEQ ID NOs: 307-337 H , or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NOs: 307-337; and ii) a V comprising an amino acid sequence of any one of SEQ ID NOs: 358-388 L , or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NOs: 358-388. In some embodiments, the antibody portion comprises: i) a V comprising an amino acid sequence of any one of SEQ ID NOs: 307-337 H ; and ii) a V comprising an amino acid sequence of any one of SEQ ID NOs: 358-388 L .

[0023] In another aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) that comprises an antibody portion that specifically recognizes GPC3 bound to the cell surface, wherein the antibody portion comprises the HC-CDR of V H and the LC-CDR of V L in the isolated anti-GPC3 construct, the V H comprises an amino acid sequence of any one of SEQ ID NOs: 307-337 and the V L comprises an amino acid sequence of any one of SEQ ID NOs: 358-388.

[0024] In another aspect, the present application provides an isolated anti-GPC3 construct that comprises an antibody portion that specifically recognizes GPC3, wherein the antibody portion comprises: i) V H, which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 32-51 or a variant thereof comprising up to about 5 amino acid substitutions, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 83-102 or a variant thereof comprising up to about 5 amino acid substitutions, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153 or a variant thereof comprising up to about 5 amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 185-204 or a variant thereof comprising up to about 5 amino acid substitutions, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 236-255 or a variant thereof comprising up to about 3 amino acid substitutions, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306 or a variant thereof comprising up to about 5 amino acid substitutions. In some embodiments, the antibody portion comprises: i) V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 32-51, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 83-102, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 185-204, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 236-255, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306.

[0025] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the antibody portion comprises: i) V comprising an amino acid sequence of any one of SEQ ID NO: 338-357 H , or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NO: 338-357; and ii) V comprising an amino acid sequence of any one of SEQ ID NO: 389-408 L , or a variant thereof having at least about 95% sequence identity to any one of SEQ ID NO: 389-408. In some embodiments, the antibody portion comprises: i) V comprising an amino acid sequence of any one of SEQ ID NO: 338-357 H ; and ii) V comprising an amino acid sequence of any one of SEQ ID NO: 389-408L 。

[0026] In another aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) comprising an antibody portion that specifically recognizes GPC3, wherein the antibody portion comprises the HC-CDR of V H in the isolated anti-GPC3 construct and the LC-CDR of V L in the isolated anti-GPC3 construct, the V H comprises an amino acid sequence of any one of SEQ ID NO: 338-357 and the V L comprises an amino acid sequence of any one of SEQ ID NO: 389-408.

[0027] In another aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) comprising an antibody portion that competitively and specifically binds to GPC3 with the isolated anti-GPC3 construct of any one of the above anti-GPC3 constructs.

[0028] In another aspect, the present application provides an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) comprising an antibody portion that competitively and specifically binds to the same or substantially the same GPC3 epitope with the isolated anti-GPC3 construct of any one of the above anti-GPC3 constructs.

[0029] In some embodiments according to any one of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the antibody portion that specifically recognizes GPC3 is chimeric, human, partially humanized, fully humanized, or semi-synthetic.

[0030] In some embodiments according to any one of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the antibody portion that specifically recognizes GPC3 is a full-length antibody, Fab, Fab', F(ab')2, Fv, or single-chain Fv (scFv). In some embodiments, the antibody portion that specifically recognizes GPC3 is scFv. In some embodiments, the antibody portion that specifically recognizes GPC3 is Fab or Fab'.

[0031] In some embodiments according to any one of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the antibody portion that specifically recognizes GPC3 is fused to an Fc fragment, optionally via a linker. In some embodiments, the Fc fragment is an IgG1 Fc fragment.

[0032] In some embodiments according to any one of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is a full-length antibody.

[0033] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is monospecific.

[0034] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is multispecific, e.g., bispecific. In some embodiments, the isolated anti-GPC3 construct is a tandem scFv, a bifunctional antibody (Db), a single-chain bifunctional antibody (scDb), a dual-affinity retargeting (DART) antibody, an F(ab')2, a dual-variable domain (DVD) antibody, a kappa-in-hinge (KiH) antibody, a docking and locking (DNL) antibody, a chemically crosslinked antibody, a hetero-polymeric antibody, or a heteroconjugate antibody. In some embodiments, the isolated anti-GPC3 construct is a tandem scFv, which comprises two scFvs linked by a peptide linker. In some embodiments, the peptide linker comprises the amino acid sequence TSGGGGS (SEQ ID NO: 474). In some embodiments, the isolated anti-GPC3 construct further comprises a second antibody portion that specifically recognizes a second antigen. In some embodiments, the second antigen is an antigen on the surface of a T cell (e.g., a cytotoxic T cell, a helper T cell, or a natural killer T cell). In some embodiments, the second antigen is an antigen on the surface of a B cell, a natural killer cell, a dendritic cell, a macrophage, a monocyte, or a neutrophil. In some embodiments, the second antigen is selected from the group consisting of CD3γ, CD3δ, CD3ε, CD3ζ, CD28, OX40, GITR, CD137, CD27, CD40L, and HVEM. In some embodiments, the second antigen is CD3ε. In some embodiments, the isolated anti-GPC3 construct is a tandem scFv, which comprises an N-terminal scFv that specifically recognizes GPC3 and a C-terminal scFv that specifically recognizes CD3ε. In some embodiments, the expression of the anti-GPC3 construct is induced by activating engineered T cells. In some embodiments, the engineered T cells are T cells comprising a chimeric antigen receptor (CAR). In some embodiments, the engineered T cells are T cells comprising a chimeric antibody-T cell receptor (TCR) construct (caTCR).

[0035] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is a chimeric antigen receptor (CAR), which comprises: (a) an extracellular domain comprising an antibody portion; (b) a transmembrane domain; and (c) an intracellular signaling domain. In some embodiments, the intracellular signaling domain comprises a CD3ζ intracellular signaling sequence and a CD28 intracellular signaling sequence.

[0036] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is a chimeric antibody-T cell receptor (TCR) construct (caTCR) comprising: (a) an extracellular domain comprising an antibody portion; and (b) a T cell receptor module (TCRM) comprising a first TCR domain (TCRD) containing a first TCR transmembrane domain (TCR-TM) and a second TCRD containing a second TCR-TM, wherein the TCRM promotes the recruitment of at least one TCR-associated signaling molecule. In some embodiments, the first TCR-TM is derived from one of the transmembrane domains of a naturally occurring first TCR and the second TCR-TM is derived from another transmembrane domain of the naturally occurring first TCR. In some embodiments, at least one of the TCR-TMs is non-naturally occurring. In some embodiments, the TCRM can enhance the recruitment of at least one TCR-associated signaling molecule as compared to a TCRM comprising a naturally occurring first T cell receptor transmembrane domain. In some embodiments, the naturally occurring first TCR is a γ / δ TCR. In some embodiments, the naturally occurring first TCR is an α / β TCR. In some embodiments, the signaling molecules that bind to the TCR are selected from the group consisting of CD3δε, CD3γε, and ζζ (also known as CD3ζ or CD3ζζ).

[0037] In some embodiments of any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct), the isolated anti-GPC3 construct is an immunoconjugate comprising an antibody portion and an effector molecule. In some embodiments, the effector molecule is a therapeutic agent selected from the group consisting of a drug, a toxin, a radioisotope, a protein, a peptide, and a nucleic acid. In some embodiments, the therapeutic agent is a drug or a toxin. In some embodiments, the effector molecule is a label.

[0038] In another aspect, the present application provides an isolated nucleic acid encoding a polypeptide component of any of the above isolated anti-GPC3 constructs.

[0039] In another aspect, the present application provides a vector comprising an isolated nucleic acid encoding a polypeptide component of any of the above isolated anti-GPC3 constructs.

[0040] In another aspect, the present application provides an isolated host cell comprising any of the above anti-GPC3 constructs.

[0041] In another aspect, the present application provides an isolated host cell comprising an isolated nucleic acid encoding a polypeptide component of any of the above isolated anti-GPC3 constructs.

[0042] In another aspect, the present application provides an isolated host cell comprising a vector containing an isolated nucleic acid encoding a polypeptide component in any of the above-described isolated anti-GPC3 constructs.

[0043] In another aspect, the present application provides an effector cell expressing any of the above-described isolated anti-GPC3 constructs. In some embodiments, the effector cell is a T cell.

[0044] In another aspect, the present application provides a pharmaceutical composition comprising any of the above anti-GPC3 constructs (e.g., an isolated anti-GPC3 construct) and a pharmaceutically acceptable carrier.

[0045] Also provided is a kit comprising any of the above-described isolated anti-GPC3 constructs, an isolated nucleic acid encoding a polypeptide component in any of the above-described isolated anti-GPC3 constructs, a vector comprising a nucleic acid encoding a polypeptide component in any of the above-described isolated anti-GPC3 constructs, any of the above-described isolated cells, or any of the above-described effector cells.

[0046] In another aspect, the present application provides a method for detecting GPC3 in a sample, comprising: contacting the sample with any of the above-described isolated anti-GPC3 immunoconjugates comprising an antibody portion and a label, and detecting the presence of the label. In some embodiments, the sample comprises cells having GPC3 bound to the cell surface. In some embodiments, the sample comprises soluble GPC3.

[0047] In another aspect, the present application provides a method for treating an individual suffering from a GPC3-positive disease, comprising administering to the individual: a) an effective amount of any of the above pharmaceutical compositions; or b) an effective amount of any of the above effector cells. In some embodiments, administration is via an intravenous or intratumoral route. In some embodiments, the injection site is distal to the first disease site. In some embodiments, the method for treating an individual suffering from a GPC3-positive disease further comprises administering to the individual another therapy. In some embodiments, the GPC3-positive disease is cancer, such as HCC, melanoma, squamous cell lung cancer, ovarian cancer, yolk sac tumor, choriocarcinoma, neuroblastoma, hepatoblastoma, Wilms' tumor, testicular non-seminomatous germ cell tumor, gastric cancer, or liposarcoma. In some embodiments, the cancer is HCC, such as metastatic HCC.

[0048] In another aspect, the present application provides a method for diagnosing an individual with a GPC3-positive disease, comprising: a) administering an effective amount of any of the isolated anti-GPC3 immunoconjugates comprising an antibody portion and a label; and b) determining the amount of label in the individual, wherein an amount of label above a threshold indicates that the individual has a GPC3-positive disease. In some embodiments, the GPC3-positive disease is cancer, such as HCC, melanoma, squamous cell lung cancer, ovarian cancer, yolk sac tumor, choriocarcinoma, neuroblastoma, hepatoblastoma, Wilms' tumor, testicular non-seminomatous germ cell tumor, gastric cancer or liposarcoma. In some embodiments, the cancer is HCC, such as metastatic HCC.

[0049] In another aspect, the present application provides a method for diagnosing an individual with a GPC3-positive disease, comprising: a) contacting a sample derived from the individual with any of the isolated anti-GPC3 comprising an antibody portion and a label; and b) determining the number of cells in the sample that bind to the isolated anti-GPC3 construct, wherein a value of the number of cells that bind to the isolated anti-GPC3 construct above a threshold indicates that the individual has a GPC3-positive disease. In some embodiments, the GPC3-positive disease is cancer, such as HCC, melanoma, squamous cell lung cancer, ovarian cancer, yolk sac tumor, choriocarcinoma, neuroblastoma, hepatoblastoma, Wilms' tumor, testicular non-seminomatous germ cell tumor, gastric cancer or liposarcoma. In some embodiments, the cancer is HCC, such as metastatic HCC.

[0050] Also provided are methods of making any of the constructs described herein, articles of manufacture, and kits suitable for the methods described herein. BRIEF DESCRIPTION OF THE DRAWINGS

[0051] Figures 1A to 1G FACS analysis showing the binding of 14 exemplary GPC3A phage clones to GPC3 + HepG2 cells. The binding of helper phage to GPC3 + HepG2 cells was used as a control.

[0052] Figures 2A to 2E FACS analysis showing the binding of 5 exemplary GPC3A phage clones to GPC3 + HepG2 cell line and HepG2 GPC3 gene knockout cell lines (HepG2-GPC3-KO-2 and HepG2-GPC3-KO-3). Helper phage was used as a negative control.

[0053] Figures 3A to 3D FACS analysis showing the binding of 4 exemplary GPC3B phage clones to GPC3 +FACS analysis of the binding of the HepG2 cell line and the HepG2-GPC3-KO-2 cell line. Helper phage was used as a negative control.

[0054] Figures 4A to 4B FACS analysis showing the binding of an exemplary GPC3A L2K bispecific antibody (anti-GPC3×CD3 bis scFv derived from GPC3A screening) to SK-Hep1-GPC3 cells and GPC3-negative SK-Hep1 cells. The y-axis shows the median fluorescence intensity (MFI).

[0055] Figures 5A to 5B FACS analysis showing the binding of an exemplary GPC3A L2K bispecific antibody (anti-GPC3×CD3 bis scFv derived from GPC3A screening) to GPC3+HepG2 cells. The binding of a control L2K bispecific antibody to GPC3+HepG2 cells was used as a negative control.

[0056] Figure 6 FACS analysis of the binding of a GPC3A L2K bispecific antibody to GPC3+HepG2 cells in a competition assay with or without soluble GPC3 antigen. A negative control L2K bispecific antibody and a commercially available anti-GPC3 (1G12) antibody were used for comparison.

[0057] Figure 7A Demonstration that T cells mediate the killing of target cells by a GPC3A L2K bispecific antibody, tested against GPC3+HepG2 cells, GPC3-negative SK-Hep1 cells, SK-Hep1-GPC3 cells, HepG2-GPC3-KO cells (#2 and #11). Figure 7B Demonstration that T cells mediate the killing of target cells by a GPC3B L2K bispecific antibody, tested against GPC3+HepG2 cells, GPC3-negative SK-Hep1 cells, SK-Hep1-GPC3 cells, and HepG2-GPC3-KO-2 cells.

[0058] Figure 8 FACS analysis of HepG2 GPC3 gene knockout cell lines using a commercially available mouse anti-human GPC3 antibody (1G12), demonstrating the successful generation of the HepG2-GPC3-KO cell line.

[0059] Figure 9A FACS analysis of the binding of a human anti-GPC3 (hGPC3) monospecific IgG antibody with a murine constant domain / Fc region (mIgG1) to GPC3-positive HepG2 cells. Figure 9BFACS analysis showing the binding of anti-hGPC3 mIgG1 to GPC3-negative SK-Hep1 cells.

[0060] Figure 10 FACS fluorescence intensity curves are shown, which indicate that GPC3 L2K antibodies (GPC3B-87L2K or GC33L2K) competitively bind to HepG2 through soluble GPC3.

[0061] Figure 11 FACS fluorescence intensity curves are shown, which indicate the binding affinity of GPC3 LK2 clones to HepG2 cells.

[0062] Figure 12 ELISA analysis showing the reactivity of GPC3 mIgG1 clones to rat GPC3 and human GPC3 (hGPC3).

[0063] Figure 13 GPC3 epitope binning analysis of GC33-mIgG, GPC3A-37mIgG, and GPC3A-55mIgG using the linear form of epitope binning.

[0064] Figure 14 GPC3 epitope binning analysis of GC33-mIgG, GPC3A-37mIgG, and GPC3A-55mIgG using the sandwich form of epitope binning.

[0065] Figure 15A and 15B Data plots of single-cycle kinetics analysis showing the binding between GPC3A L2K clones and GPC3 antigen.

[0066] Figure 16 LDH cytotoxicity analysis of GPC3-positive HepG2 cells, GPC3-negative SK-Hep1 cells, SK-Hep1-GPC3 cells, and HepG2-GPC3-KO cells treated with GPC3A or GPC3B CAR-T cells and GC33 CAR-T cells.

[0067] Figure 17A LDH cytotoxicity analysis of GPC3+ HepG2 cells, GPC3-negative SK-Hep1 cells, SK-Hep1-GPC3 cells, and HepG2-GPC3-KO cells treated with GPC3A or GPC3B CAR-T cells and GC33 CAR-T cells. Figure 17BShow the LDH cytotoxicity analysis of GPC3-positive JHH5 cells, GPC3-negative A-498 cells, and GPC3-negative PANC-1 cells treated with GPC3A or GPC3B CAR-T cells and GC33 CAR-T cells.

[0068] Figure 18 Show T cell-mediated killing of the cancer cell line HepG2 (AFP + / GPC3 + ) and HepG2-GPC3-KO (AFP + / GPC3 - ) by percent specific lysis, where the T cells are transduced with the indicated percentages (5% to 40%) of caTCR-positive single anti-AFP158 / HLA-A*2:01 caTCR or anti-AFP158 / HLA-A*2:01 caTCR + anti-CD3 / anti-GPC3 BsAb.

[0069] Figure 19 Show the concentrations of cytokines (IL-2, IFN-γ, and TNF-α) found in the supernatant after T cell-mediated in vitro killing of the cancer cell lines HepG2 and HepG2-GPC3-KO, where the T cells are transduced with the indicated percentages (5% to 40%) of caTCR-positive single anti-AFP158 / HLA-A*2:01 caTCR or anti-AFP158 / HLA-A*2:01 caTCR + anti-CD3 / anti-GPC3 BsAb.

[0070] Figure 20 Show enhanced target-specific cancer cell line killing mediated by anti-CD3 / anti-GPC3 BsAb released from activated anti-AFP158 / HLA-A*2:01 caTCR + anti-CD3 / anti-GPC3 BsAb T cells. Incubate the indicated transduced T cells with target cells and separate from the indicated target cells and mock T cells by a membrane permeable to BsAb but not to T cells.

[0071] Figure 21 Show T cell-mediated killing of the cancer cell line HepG2 (AFP + / GPC3 + ) and HepG2-GPC3.ko (AFP + / GPC3 - ) by percent specific lysis, where the T cells are transduced with single anti-AFP158 / HLA-A*2:01 caTCR-1-0, single anti-GPC3 CSR, or anti-AFP158 / HLA-A*2:01 caTCR (1-0 and 1-TM5) + anti-GPC3 CSR.

[0072] Figure 22 Shows the concentrations of cytokines (IL-2, GM-CSF, IFN-γ and TNF-α) found in the supernatant after T cell-mediated in vitro killing of the cancer cell lines HepG2 and HepG2-GPC3-KO, where the T cells were transduced with the anti-AFP158 / HLA-A*2:01caTCR-1-0 alone, the anti-GPC3 CSR alone, or the anti-AFP158 / HLA-A*2:01caTCR (1-0 and 1-TM5) + anti-GPC3 CSR.

[0073] Figure 23 Shows the degranulation activity (as determined by CD107a expression) in T cells transduced with the anti-AFP158 / HLA-A*2:01caTCR-1-0 alone, the anti-GPC3 CSR alone, or the anti-AFP158 / HLA-A*2:01caTCR (1-0 and 1-TM5) + anti-GPC3 CSR after stimulation with the cancer cell line HepG2.

[0074] Figure 24 Shows the proliferation of T cells transduced with the anti-AFP158 / HLA-A*2:01caTCR-1-0 alone, the anti-GPC3 CSR alone, or the anti-AFP158 / HLA-A*2:01caTCR (1-0 and 1-TM5) + anti-GPC3 CSR after stimulation with the cancer cell line HepG2 (as determined by CFSE dye dilution).

[0075] Figure 25 Shows tumor growth of HepG2 in a subcutaneous mouse model treated with a mock or a single intratumoral injection of T cells transduced with anti-AFPCAR, or a combination of anti-AFPCAR and anti-GPC3 CSR. Detailed Description

[0076] In one aspect, the present application provides constructs (referred to herein as "anti-GPC3 constructs") (e.g., isolated anti-GPC3 constructs) that include an antibody portion (referred to herein as the "anti-GPC3 antibody portion") that specifically recognizes GPC3 bound to the cell surface (referred to herein as "native form GPC3" or "native GPC3 (nGPC3)"). These anti-GPC3 constructs specifically recognize GPC3 bound to the cell surface relative to non-cell surface-bound GPC3 (referred to herein as "soluble GPC3 (sGPC3)" or "non-native GPC3"), such as circulating GPC3 protein or free GPC3 peptides in serum.

[0077] When provided in the form of an anti-CD3 bispecific antibody or present in a chimeric antigen receptor (CAR) expressed by T cells, the anti-GPC3 antibody moiety specifically redirects human T cells to kill target cells expressing GPC3 (e.g., GPC3-expressing cancer cells). The technical advantages of this strategy are significantly superior to using antibodies against any form of GPC3 (especially soluble GPC3) because T cell targeting of tumor sites can become much more effective and precise, thereby effectively occurring T cell-mediated cancer cell killing without harming normal tissues. Additionally, when the anti-GPC3 antibody moiety is fused with a detectable moiety, it can diagnose and prognose GPC3-positive diseases or disorders that are highly sensitive to changes in the number and distribution (a potential measure of disease progression, with a greater correlation than circulating GPC3 content) of cells expressing GPC3 on the cell surface (e.g., GPC3-positive tumor cells).

[0078] This application also provides anti-GPC3 constructs (e.g., isolated anti-GPC3 constructs) comprising an antibody moiety that specifically recognizes GPC3 (e.g., nGPC3 and / or sGPC3). These constructs can also be provided as anti-CD3 bispecific antibodies or present in a CAR expressed by T cells. In cancer patients (e.g., HCC patients), GPC3 can be expressed on the cell surface or released from the cell surface in different forms into the extracellular environment. Therefore, these anti-GPC3 constructs can also be fused with a detectable moiety and used for diagnostic and prognostic purposes.

[0079] Using phage display technology, a variety of monoclonal antibodies with specificity and high affinity for human GPC3 bound to the cell surface were generated. Flow cytometry and T cell-mediated cytotoxicity assays demonstrated that these antibodies recognize GPC3-expressing cancer cell lines in a manner restricted to the native form of GPC3. When provided in the form of an anti-CD3 bispecific antibody or CAR-T cells, the antibodies redirect human T cells to kill GPC3-positive target cancer cells. The data presented herein demonstrate that anti-GPC3 constructs comprising an antibody moiety that specifically recognizes GPC3 bound to the cell surface described herein can be effective therapeutic agents for cancer indications (e.g., solid tumor indications such as HCC).

[0080] Using phage display technology, a variety of monoclonal antibodies with specificity and high affinity for human GPC3 (e.g., nGPC3 and / or sGPC3) were also generated. Flow cytometry and T cell-mediated cytotoxicity assays demonstrated that these antibodies specifically recognize GPC3.

[0081] The present application thus provides constructs (e.g., isolated constructs) comprising an antibody portion that specifically recognizes GPC3 (e.g., GPC3 bound to the cell surface). The constructs can be, for example, anti-GPC3 scFv, anti-GPC3 Fc fusion proteins, full-length anti-GPC3 antibodies, multispecific (e.g., bispecific) anti-GPC3 molecules (e.g., tandem bis-scFv bispecific T cell engagers), anti-GPC3 chimeric antigen receptors (CARs), anti-GPC3 chimeric antibody-T cell receptors (caTCRs), and anti-GPC3 immunoconjugates.

[0082] In another aspect, provided are nucleic acids encoding an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) or a portion of an anti-GPC3 antibody portion of the construct (e.g., those portions that specifically recognize GPC3 bound to the cell surface).

[0083] In another aspect, provided are compositions (e.g., pharmaceutical compositions) comprising an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct), the construct comprising an antibody portion that specifically recognizes GPC3 (e.g., GPC3 bound to the cell surface). The compositions can be pharmaceutical compositions comprising an anti-GPC3 construct (e.g., GPC3 bound to the cell surface) or effector cells expressing an anti-GPC3 construct or binding to the anti-GPC3 construct (e.g., T cells expressing an anti-GPC3 CAR) (CARs that specifically recognize GPC3 bound to the cell surface).

[0084] Also provided are methods of making and using an anti-GPC3 construct (e.g., an isolated anti-GPC3 construct, or cells expressing an anti-GPC3 construct or binding to the anti-GPC3 construct) for therapeutic or diagnostic purposes, as well as kits and articles of manufacture suitable for such methods.

[0085] Definitions

[0086] As used herein, "treatment / treating" is a method for obtaining a beneficial or desired result, including a clinical result. For the purposes of the present invention, beneficial or desired clinical results include, but are not limited to, one or more of the following: alleviating one or more symptoms caused by a disease, reducing the severity of the disease, stabilizing the disease (e.g., preventing or delaying disease progression), preventing or delaying the spread of the disease (e.g., metastasis), preventing or delaying disease recurrence, delaying or slowing disease progression, improving the disease state, achieving remission (partial or complete) of the disease, reducing the dose of one or more other drugs required to treat the disease, delaying disease progression, improving or enhancing quality of life, increasing weight gain, and / or extending survival. "Treatment" also encompasses a reduction in a pathological outcome (e.g., tumor volume) of cancer. The methods of the present invention encompass any or all of these aspects of treatment.

[0087] As used herein, "activation" of T cells refers to a state of T cells that have been sufficiently stimulated to induce detectable cell proliferation. Activation can also be associated with induced cytokine production and detectable effector functions.

[0088] The term "antibody portion" encompasses full-length antibodies and their antigen-binding fragments. A full-length antibody includes two heavy chains and two light chains. The variable regions of the light and heavy chains are responsible for antigen binding. The variable regions in both chains generally contain three highly variable loops, called complementarity-determining regions (CDRs) (light chain (LC) CDRs include LC-CDR1, LC-CDR2, and LC-CDR3, and heavy chain (HC) CDRs include HC-CDR1, HC-CDR2, and HC-CDR3). The CDR boundaries of the antibodies and antigen-binding fragments disclosed herein can be defined or identified according to the Kabat, Chothia, or Al-Lazikani conventions (Al-Lazikani 1997; Chothia 1985; Chothia 1987; Chothia 1989; Kabat 1987; Kabat 1991). The three CDRs of the heavy or light chain are inserted between flanking segments called framework regions (FRs), which are more conserved than the CDRs and form a scaffold that supports the hypervariable loops. The constant regions of the heavy and light chains do not participate in antigen binding but exhibit various effector functions. Antibodies are classified based on the amino acid sequence of their heavy chain constant regions. The five major classes or isotypes of antibodies are IgA, IgD, IgE, IgG, and IgM, which are characterized by the presence of α, δ, ε, γ, and μ heavy chains, respectively. Some of the major antibody classes are divided into subclasses, such as IgG1 (γ1 heavy chain), IgG2 (γ2 heavy chain), IgG3 (γ3 heavy chain), IgG4 (γ4 heavy chain), IgA1 (α1 heavy chain), or IgA2 (α2 heavy chain).

[0089] As used herein, the term "antigen-binding fragment" refers to an antibody fragment that includes, for example, bispecific antibodies, Fab, Fab', F(ab')2, Fv fragments, disulfide-stabilized Fv fragments (dsFv), (dsFv)2, bispecific dsFv (dsFv-dsFv'), disulfide-stabilized bispecific antibodies (ds bispecific antibodies), single-chain Fv (scFv), scFv dimers (bivalent bispecific antibodies), multispecific antibodies formed from a portion of an antibody that includes one or more CDRs, camelized single-domain antibodies, nanobodies, domain antibodies, bivalent domain antibodies, or any other antibody fragment that binds to an antigen but does not include the complete antibody structure. An antigen-binding fragment is capable of binding to the same antigen as the parental antibody or parental antibody fragment (e.g., parental scFv). In some embodiments, an antigen-binding fragment can include one or more CDRs from a particular human antibody transplanted into a framework region from one or more different human antibodies.

[0090] As used herein, the term "epitope" refers to a specific atom or amino acid group on an antigen to which an antibody or antibody portion binds. If two antibodies or antibody portions exhibit competitive binding to an antigen, then they can bind to the same epitope within the antigen.

[0091] As used herein, in the presence of an equimolar concentration of a first antibody portion, the first antibody portion "competes" with a second antibody portion for binding to a target GPC3 (e.g., nGPC3 and / or sGPC3), or vice versa, when the first antibody portion inhibits the binding of the second antibody portion to the target GPC3 by at least about 50% (e.g., any one of at least about 55%, 60%, 65%, 70%, 75%, 80%, 85%, 90%, 95%, 98%, or 99%). High-throughput methods for "grouping" antibodies based on cross-competition are described in PCT Publication No. WO 03 / 48731.

[0092] As used herein, the terms "specifically bind," "specifically recognize," or "being specific" refer to a measurable and reproducible interaction for determining the presence of a target in the presence of a heterogeneous population of molecules, including biomolecules, such as the binding between a target and an antibody or antibody portion. For example, an antibody or antibody portion that specifically recognizes a target (which can be an epitope) binds to the target with greater affinity, avidity, facility, and / or duration than it binds to other targets. In some embodiments, the binding affinity of an antibody or antibody portion that specifically recognizes an antigen to one or more antigenic determinants of the antigen (e.g., native form GPC3) is at least about 10-fold that for other targets (e.g., soluble GPC3).

[0093] As used herein, an "isolated" anti-GPC3 construct refers to an anti-GPC3 construct that (1) is not bound to proteins found in nature, (2) does not contain other proteins from the same source, (3) is expressed by cells from different species, or (4) does not exist in nature.

[0094] As used herein, the term "isolated nucleic acid" means a nucleic acid of genomic, cDNA, or synthetic origin, or some combination thereof, by virtue of its origin, the "isolated nucleic acid" (1) is not bound to all or a portion of the polynucleotide in which the "isolated nucleic acid" exists in nature, (2) is operably linked to a polynucleotide to which it is not linked in nature, or (3) does not exist as part of a larger sequence in nature.

[0095] As used herein, the term "CDR" or "complementary determining region" means the non - contiguous antigen - combining sites found within the variable regions of heavy and light chain polypeptides. These specific regions have been described in Kabat et al., J. Biol. Chem. 252:6609 - 6616 (1977); Kabat et al., U.S. Dept. of Health and Human Services, "Sequences of proteins of immunological interest" (1991); Chothia et al., J. Mol. Biol. 196:901 - 917 (1987); Al - Lazikani B. et al., J. Mol. Biol. 273:927 - 948 (1997); MacCallum et al., J. Mol. Biol. 262:732 - 745 (1996); Abhinandan and Martin, Mol. Immunol., 45:3832 - 3839 (2008); Lefranc M.P. et al., Dev. Comp. Immunol., 27:55 - 77 (2003); and Honegger and Plückthun, J. Mol. Biol. 309:657 - 670 (2001), where, when compared to each other, the definitions include overlapping or subsets of amino acid residues. However, the application of any of the definitions to refer to the CDRs or variants thereof of an antibody or a grafted antibody is intended to fall within the scope of the term as defined and used herein. The amino acid residues encompassing the CDRs as defined in each of the above - mentioned references are set forth in Table 1 below for comparison. CDR prediction algorithms and interfaces are known in the art, including, for example, Abhinandan and Martin, Mol. Immunol., 45:3832 - 3839 (2008); Ehrenmann F. et al., Nucleic Acids Res., 38:D301 - D307 (2010); and Adolf - Bryfogle J. et al., Nucleic Acids Res., 43:D432 - D438 (2015). The content of the references cited in this paragraph is hereby incorporated by reference in its entirety for use in this invention and may be included in one or more claims herein.

[0096] Table 1: CDR Definitions

[0097]

[0098] 1 Residue numbering follows the nomenclature of the aforementioned Kabat et al.

[0099] 2 The residue numbering follows the nomenclature of Chothia et al. as described above.

[0100] 3 The residue numbering follows the nomenclature of MacCallum et al. as described above.

[0101] 4 The residue numbering follows the nomenclature of Lefranc et al. as described above.

[0102] 5 The residue numbering follows the nomenclature of Honegger and Plückthun as described above.

[0103] The term "chimeric antibody" refers to an antibody in which part of the heavy and / or light chain is identical or homologous to the corresponding sequence in an antibody derived from a particular species or belonging to a particular antibody class or subclass, while the remaining part of the chain is identical or homologous to the corresponding sequence in an antibody derived from another species or belonging to another antibody class or subclass, and fragments of such antibodies, provided that they exhibit the biological activity of the present invention (see U.S. Patent No. 4,816,567; and Morrison et al., Proc. Natl. Acad. Sci. USA, 81:6851-6855 (1984)).

[0104] The term "semi-synthetic" with respect to an antibody or antibody portion means that the antibody or antibody portion has one or more naturally occurring sequences and one or more non-naturally occurring (i.e., synthetic) sequences.

[0105] "Fv" is the smallest antibody fragment that contains the complete antigen recognition and binding site. This fragment consists of a dimer of a heavy chain variable region and a light chain variable region that are non-covalently and tightly associated. The two domains fold to give six hypervariable loops (3 loops each from the heavy and light chains), which contribute the amino acid residues for antigen binding and confer antigen-binding specificity to the antibody. However, even a single variable domain (or half of the Fv that includes only three CDRs specific for the antigen) can recognize and bind the antigen, but with lower affinity than the entire binding site.

[0106] "Single-chain Fv" (also abbreviated as "sFv" or "scFv") is an antibody fragment that comprises a V H and a V L antibody domain joined into a single polypeptide chain. In some embodiments, the scFv polypeptide further comprises a linker between the V H and the V LA polypeptide linker between domains that allows the scFv to form the required structure for antigen binding. For a review of scFv, see Plückthun in The Pharmacology of Monoclonal Antibodies, Volume 113, Rosenberg and Moore eds., Springer-Verlag, New York, pp. 269-315 (1994).

[0107] The term "bispecific antibody" refers to small antibody fragments prepared as follows: In V H and V L Typically, a short linker (e.g., about 5 to about 10 residues) is used to construct the scFv fragment between domains (see the preceding paragraph) such that the V domains achieve interchain pairing rather than intrachain pairing, resulting in a bivalent fragment, i.e., a fragment with two antigen-binding sites. A bispecific bispecific antibody is a heterodimer of two "swapped" scFv fragments, where the V H and V L domains are on different polypeptide chains. Bispecific antibodies are more fully described, for example, in EP 404,097; WO 93 / 11161; and Hollinger et al., Proceedings of the National Academy of Sciences USA 90:6444-6448 (1993).

[0108] "Humanized" forms of non-human (e.g., rodent) antibodies are chimeric antibodies that contain minimal sequences derived from non-human antibodies. To a large extent, humanized antibodies are human immunoglobulins (recipient antibodies) in which the residues of the hypervariable regions (HVRs) from the recipient are replaced with residues of the hypervariable regions from a non-human species (donor antibody), such as a mouse, rat, rabbit, or non-human primate having the desired antibody specificity, affinity, and capacity. In some instances, the framework region (FR) residues of the human immunoglobulin are replaced with the corresponding non-human residues. Additionally, humanized antibodies can include residues not found in the recipient antibody or the donor antibody. These modifications are made to further improve antibody performance. Generally, a humanized antibody will include substantially all of at least one and usually two variable domains, wherein all or substantially all of the hypervariable loops correspond to those of the non-human immunoglobulin and all or substantially all of the FRs are FRs having the sequences of human immunoglobulins. A humanized antibody optionally will also include at least a portion of the immunoglobulin constant region (Fc), usually at least a portion of the constant region of a human immunoglobulin. For additional details, see Jones et al., Nature 321:522-525 (1986); Riechmann et al., Nature 332:323-329 (1988); and Presta, Curr. Op. Struct. Biol. 2:593-596 (1992).

[0109] The "percent amino acid sequence identity (%)" or "homology" with respect to the polypeptide and antibody sequences identified herein is defined as the percentage of amino acid residues in a candidate sequence that are identical to the amino acid residues in the polypeptide being compared, after alignment, with any conservative substitutions being considered part of the sequence identity. The alignment for purposes of determining the percent amino acid sequence identity can be achieved in various ways within the skill in the art, e.g., using publicly available computer software such as BLAST, BLAST-2, ALIGN, Megalign (DNASTAR), or MUSCLE software. Those skilled in the art can determine the parameters suitable for measuring the alignment, including any algorithms required to achieve the maximum alignment over the full length of the sequences being compared. However, for purposes herein, the percent amino acid sequence identity values are generated using the sequence comparison computer program MUSCLE (Edgar, R.C., Nucleic Acids Research 32(5):1792-1797, 2004; Edgar, R.C., BMC Bioinformatics 5(1):113, 2004).

[0110] The term "Fc receptor" or "FcR" is used to describe a receptor that binds to the Fc region of an antibody. In some embodiments, the FcR of the invention is an FcR that binds to an IgG antibody (gamma receptor) and includes Fc gamma RI, Fc gamma RII, and Fc gamma RIII subclasses of receptors (including allelic variants and alternative splicing forms of these receptors). The Fc gamma RII receptor includes Fc gamma RIIA ("activating receptor") and Fc gamma RIIB ("inhibitory receptor"), which have similar amino acid sequences that differ primarily in their cytoplasmic domains. The activating receptor Fc gamma RIIA contains an immunoreceptor tyrosine-based activation motif (ITAM) in its cytoplasmic domain. The inhibitory receptor Fc gamma RIIB contains an immunoreceptor tyrosine-based inhibitory motif (ITIM) in its cytoplasmic domain (see review by M. Yu Annu.Rev.Immunol. 15:203-234 (1997)). The term includes isotypes, such as the Fc gamma RIIIA isotypes: Fc gamma RIIIA-Phe158, Fc gamma RIIIA-Val158, Fc gamma RIIA-R131, and / or Fc gamma RIIA-H131. FcRs are reviewed in Ravetch and Kinet, Annu.Rev.Immunol 9:457-92 (1991); Capel et al., Immunomethods 4:25-34 (1994); and de Haas et al., J.Lab.Clin.Med. 126:330-41 (1995). The term "FcR" as used herein encompasses other FcRs, including FcRs to be identified in the future. The term also includes the neonatal receptor FcRn that is responsible for the transfer of maternal IgG to the fetus (Guyer et al., J.Immunol. 117:587 (1976) and Kim et al., J.Immunol. 24:249 (1994)).

[0111] The term "FcRn" refers to the neonatal Fc receptor (FcRn). FcRn is structurally similar to the major histocompatibility complex (MHC) and is composed of an alpha chain non-covalently bound to beta2-microglobulin. The various functions of the neonatal Fc receptor FcRn are reviewed in Ghetie and Ward (2000) Annu.Rev.Immunol. 18, 739-766. FcRn plays a role in the passive delivery of immunoglobulin IgG from mother to infant and in the regulation of serum IgG levels. FcRn can act as a salvage receptor that binds intracellularly to transcytosed and endocytosed intact IgG and rescues it from the default degradation pathway.

[0112] The "CH1 domain" of the human IgG Fc region (also referred to as "C1" of the "H1" domain) generally extends from approximately amino acid 118 to approximately amino acid 215 (EU numbering system).

[0113] The "hinge region" is typically defined as extending from Glu216 to Pro230 of human IgG1 (Burton, Molec. Immunol. 22:161-206 (1985)). The hinge regions of other IgG isotypes can be aligned with the IgG1 sequence by placing the first and last cysteine residues that form the inter-heavy chain S-S bond in the same positions.

[0114] The "CH2 domain" of the human IgG Fc region (also referred to as "C2" of the "H2" domain) typically extends from approximately amino acid 231 to approximately amino acid 340. The unique feature of the CH2 domain is that it does not pair tightly with another domain. The fact is that two N-linked branched carbohydrate chains are inserted between the two CH2 domains of the intact native IgG molecule. It has been speculated that the carbohydrate may provide a substitute for domain-domain pairing and contribute to the stabilization of the CH2 domain. Burton, Molec Immunol. 22:161-206 (1985).

[0115] The "CH3 domain" (also referred to as the "C2" or "H3" domain) comprises the C-terminal residues to the fragment of the CH2 domain in the Fc region (i.e., from approximately amino acid residue 341 of the antibody sequence to the C-terminus, which is typically at amino acid residue 446 or 447 of IgG).

[0116] A "functional Fc fragment" has the "effector functions" of the native sequence Fc region. Exemplary "effector functions" include C1q binding; complement-dependent cytotoxicity (CDC); Fc receptor binding; antibody-dependent cell-mediated cytotoxicity (ADCC); phagocytosis; downregulation of cell surface receptors (e.g., B cell receptor; BCR), etc. Such effector functions generally require the combination of the Fc region with a binding domain (e.g., an antibody variable domain) and can be evaluated using various assays known in the art.

[0117] An antibody with a variant IgG Fc having an "altered" FcR binding affinity or ADCC activity is an antibody that has enhanced or diminished FcR binding activity (e.g., FcγR or FcRn) and / or ADCC activity as compared to a parental polypeptide or a polypeptide comprising the native sequence Fc region. A variant Fc that "exhibits" "enhanced binding" to an FcR binds at least one FcR with a higher affinity (e.g., a lower apparent K d or IC 50value) the parental polypeptide or the native sequence IgG Fc. According to some embodiments, the binding is improved by about 3-fold over the parental polypeptide, such as any one of about 5, 10, 25, 50, 60, 100, 150, 200 or up to 500-fold, or a binding improvement of about 25% to 1000%. A polypeptide variant that "exhibits reduced binding to FcR" binds to at least one FcR with an affinity lower than (e.g., a higher apparent K d or a higher IC 50 value) the parental polypeptide. The reduction in binding compared to the parental polypeptide can be a reduction in binding of about 40% or more than 40%.

[0118] "Antibody-dependent cell-mediated cytotoxicity" or "ADCC" refers to a form of cytotoxicity in which secreted Ig that binds to Fc receptors (FcRs) present on certain cytotoxic cells (e.g., natural killer (NK) cells, neutrophils, and macrophages) allows these cytotoxic effector cells to specifically bind to antigen-bearing target cells and subsequently kill the target cells via cytotoxins. The antibody "arms" the cytotoxic cells and is absolutely required for this killing. Primary NK cells, which mediate ADCC, express only FcγRIII, while monocytes express FcγRI, FcγRII, and FcγRIII. The expression of FcRs on hematopoietic cells is summarized in Table 3 on page 464 of Ravetch and Kinet, Annu. Rev. Immunol. 9:457-92 (1991). To assess the ADCC activity of a molecule of interest, an in vitro ADCC assay can be performed, such as those described in U.S. Patent No. 5,500,362 or 5,821,337. Effector cells suitable for such assays include peripheral blood mononuclear cells (PBMCs) and natural killer (NK) cells. Alternatively or additionally, the ADCC activity of the molecule of interest can be evaluated in vivo (e.g., in an animal model, such as the animal model disclosed in Clynes et al., PNAS (USA) 95:652-656 (1998)).

[0119] When the amounts of a polypeptide with a variant Fc region and a polypeptide with a wild-type Fc region (or parental polypeptide) are substantially the same in an assay, a polypeptide comprising a variant Fc region that "exhibits enhanced ADCC" or mediates ADCC more effectively than a polypeptide with a wild-type IgG Fc or parental polypeptide in the presence of human effector cells is a polypeptide that mediates ADCC more effectively in vitro or in vivo. Generally, such variants will be identified using any in vitro ADCC assay known in the art, such as assays or methods for determining ADCC activity, for example, in an animal model. In some embodiments, the effectiveness of the variant in mediating ADCC is about 5-fold to about 100-fold that of the wild-type Fc (or parental polypeptide), such as about 25 to about 50-fold.

[0120] "Complement-dependent cytotoxicity" or "CDC" refers to the lysis of target cells in the presence of complement. Activation of the classical complement pathway begins with the binding of the first component of the complement system (C1q) to an antibody (of the appropriate subclass) that is bound to its cognate antigen. To assess complement activation, a CDC assay can be performed, such as that described by Gazzano-Santoro et al., J. Immunol. Methods 202:163 (1996). Polypeptide variants having an altered amino acid sequence in the Fc region and an increased or decreased ability to bind C1q are described in U.S. Patent No. 6,194,551 B1 and WO 99 / 51642. The contents of those patent disclosures are hereby specifically incorporated by reference. See also Idusogie et al., J. Immunol. 164:4178-4184 (2000).

[0121] Unless otherwise specified, a "nucleotide sequence encoding an amino acid sequence" includes all nucleotide sequences that are degenerate to each other and encode the same amino acid sequence. Insofar as a nucleotide sequence encoding a protein can contain introns in some forms, the phrase nucleotide sequence encoding the protein or RNA may also include introns.

[0122] The term "operably linked" refers to a functional linkage between a regulatory sequence and a heterologous nucleic acid sequence such that the latter is expressed. For example, a first nucleic acid sequence is operably linked to a second nucleic acid sequence when the two are placed in a functional relationship. For example, a promoter is operably linked to a coding sequence if it affects the transcription or expression of the coding sequence. Generally, operably linked DNA sequences are contiguous and, where necessary, join two protein-coding regions in the same reading frame.

[0123] "Homologous" refers to sequence similarity or sequence identity between two polypeptides or between two nucleic acid molecules. When the positions in two compared sequences are occupied by the same base or amino acid monomer subunit, e.g., if the position in each of two DNA molecules is occupied by adenine, then the molecules are homologous at that position. The % homology between two sequences is a function of the number of matching or homologous positions shared by the two sequences divided by the number of positions compared times 100. For example, if 6 / 10 positions in two sequences match or are homologous, then the two sequences are 60% homologous. For example, the DNA sequences ATTGCC and TATGGC share 50% homology. Generally, two sequences are compared when aligned to give maximum homology.

[0124] An "effective amount" of an anti-GPC3 construct or composition as disclosed herein is an amount sufficient to effect a particular stated purpose. An "effective amount" can be determined empirically and by known methods related to that purpose.

[0125] The term "therapeutically effective amount" refers to the amount of an anti-GPC3 construct or composition as disclosed herein that is effective to "treat" an individual for a disease or disorder. In the case of cancer, a therapeutically effective amount of an anti-GPC3 construct or composition as disclosed herein can reduce the number of cancer cells; reduce the size or weight of a tumor; inhibit (i.e., slow down and preferably prevent to some extent) the infiltration of cancer cells into surrounding organs; inhibit (i.e., slow down and preferably prevent to some extent) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with cancer to some extent. To the extent that an anti-GPC3 construct or composition as disclosed herein can prevent growth and / or kill existing cancer cells, it can be cytostatic and / or cytotoxic. In some embodiments, the therapeutically effective amount is a growth inhibitory amount. In some embodiments, the therapeutically effective amount is an amount that prolongs the survival of a patient. In some embodiments, the therapeutically effective amount is an amount that improves the progression-free survival of a patient.

[0126] As used herein, "pharmaceutically acceptable" or "pharmacologically compatible" means a material that is biologically or otherwise without adverse effects, i.e., the material can be incorporated into a pharmaceutical composition administered to a patient without causing any significant adverse biological effects or interacting in a harmful manner with any other component in the composition containing it. Pharmaceutically acceptable carriers or excipients preferably meet the required standards of toxicological and manufacturing tests and / or are included in the Inactive Ingredient Guide established by the U.S. Food and Drug Administration.

[0127] The term "label" as used herein refers to a detectable compound or composition that can bind directly or indirectly to an anti-GPC3 antibody moiety. The label itself can be detectable (e.g., a radioisotope label or a fluorescent label), or in the case of an enzyme label, can catalyze a detectable chemical change in a substrate compound or composition.

[0128] It should be understood that the embodiments of the invention described herein include "consisting of the embodiments" and / or "consisting essentially of the embodiments".

[0129] References herein to "about" a value or parameter include (and describe) variations that are about the value or parameter itself. For example, a description of "about X" includes a description of "X".

[0130] As used herein, a reference to "not" a particular value or parameter generally means and describes "except for" that particular value or parameter. For example, a method is not used to treat cancer of type X means that the method is used to treat cancer types other than type X.

[0131] Unless the context clearly indicates otherwise, as used herein and in the appended claims, the singular forms "a", "an", and "the" include plural referents.

[0132] Anti-GPC3 construct

[0133] In one aspect, the present invention provides a GPC3-specific construct (e.g., an isolated anti-GPC3 construct) comprising an antibody portion that specifically binds to GPC3. The specificity of the anti-GPC3 construct is derived from the anti-GPC3 antibody portion that specifically binds to GPC3, such as a full-length antibody or an antigen-binding fragment thereof. In some embodiments, reference to a portion that specifically binds to GPC3 (e.g., an antibody portion) means that the portion binds to GPC3 with an affinity that is at least about 10-fold (including, for example, at least about 10, 10 2 、10 3 、10 4 、10 5 、10 6 or 10 7 times any of them) greater than its binding affinity for a non-target. In some embodiments, the non-target is an antigen that is not GPC3. In some embodiments, the non-target of an anti-nGPC3 antibody portion is soluble GPC3. In some embodiments, the non-target of an anti-sGPC3 antibody portion is cell-surface-bound GPC3. Binding affinity can be determined by methods known in the art, such as ELISA, fluorescence-activated cell sorting (FACS) analysis, or radioimmunoprecipitation assay (RIA). K d can be determined by methods known in the art, such as surface plasmon resonance (SPR) analysis using, for example, a BIACORE TM instrument, or kinetic exclusion analysis (KinExA) using, for example, a Sapidyne instrument.

[0134] Exemplary anti-GPC3 constructs include, for example, anti-GPC3 scFv, anti-GPC3 Fc fusion proteins, full-length anti-GPC3 antibodies, multispecific (e.g., bispecific) anti-GPC3 molecules (e.g., tandem bis-scFv bispecific T cell engagers), anti-GPC3 chimeric antigen receptors (CARs), anti-GPC3 chimeric antibody-T cell receptors (caTCRs), and anti-GPC3 immunoconjugates.

[0135] The different aspects are discussed in more detail in the following sections.

[0136] Although embodiments using anti-GPC3 constructs are discussed extensively herein, the anti-GPC3 constructs including an anti-GPC3 antibody portion containing human sequences (i.e., human heavy and light chain variable region sequences including human CDR sequences), the present invention also provides non-human anti-GPC3 constructs. In some embodiments, the non-human antibody agent includes human CDR sequences from the antibody agents as described herein and non-human framework sequences. In some embodiments, the non-human framework sequences include any sequences that can be used to generate synthetic heavy and / or light chain variable regions by using one or more human CDR sequences as described herein, including, for example, mammals such as mice, rats, rabbits, pigs, bovines (e.g., cows, bulls, water buffalo), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (e.g., marmosets, rhesus monkeys), etc. In some embodiments, the antibody agent provided includes an antibody agent generated by grafting one or more human CDR sequences as described herein onto a non-human framework sequence (e.g., a mouse or chicken framework sequence). In some embodiments, the antibody agent provided is a human antibody agent (e.g., a human monoclonal antibody or fragment thereof, a human antigen-binding protein or polypeptide, a human multispecific binding agent [e.g., a human bispecific antibody], a human polypeptide having one or more structural components of a human immunoglobulin polypeptide).

[0137] anti-GPC3 antibody portion

[0138] An anti-GPC3 construct (e.g., an isolated anti-GPC3 construct) includes an anti-GPC3 antibody portion that specifically recognizes GPC3. In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface (referred to herein as the "anti-nGPC3 antibody portion"). In some embodiments, the anti-nGPC3 antibody portion has a higher binding affinity for nGPC3 than for sGPC3. In some embodiments, the anti-nGPC3 antibody portion specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity. In some embodiments, the cells expressing GPC3 on their surface are HepG2, Hep3B, Huh7, JHH-7, or 293. In some embodiments, the cells present an abnormally high amount of GPC3 on their surface. In some embodiments, the cells are cancer cells. In some embodiments, the cancer cells are present in a solid tumor (e.g., liver cancer, e.g., HCC). In some embodiments, the cancer cells are metastatic cancer cells (e.g., metastatic HCC). In some embodiments, the anti-GPC3 antibody portion specifically recognizes soluble GPC3 (referred to herein as the "anti-sGPC3 antibody portion"). In some embodiments, the anti-GPC3 antibody portion specifically recognizes nGPC3 and sGPC3.

[0139] Glypican-3 (GPC3)

[0140] Glypican 3 (GPC3) is a carcinoembryonic antigen belonging to the heparan sulfate proteoglycan family that is present on the cell surface and is involved in cell signaling occurring at the cell-extracellular matrix interface. GPC3 is expressed in the fetal liver and placenta during development and is downregulated or silent in normal adult tissues. This development stage-specific and tissue-specific expression pattern suggests that GPC3 may be involved in morphogenesis.

[0141] The GPC3 gene encodes a 70 kDa precursor protein of 580 amino acids. After translocation into the endoplasmic reticulum, the N-terminal signal peptide (SS; residues 1-24) and the C-terminal glycosylphosphatidylinositol (GPI) anchor addition signal (predicted cleavage site: S 560 ) are removed and the latter is replaced by a GPI anchor. The GPC3 precursor protein can be cleaved by furin between Arg 358 and Ser 359 to generate a disulfide-linked 40 kDa N-terminal subunit (Q 25 -R 358 ) and a 30 kDa C-terminal subunit (starting at S 359 ). The C-terminus of GPC3 is linked to the membrane via a GPI anchor and is post-translationally modified to have two O-linked heparan sulfate (HS) side chains that are close to the cell surface. Based on the initial amino acid sequence, the N-terminal subunit, the C-terminal subunit, and the two HS glycan chains form the three functional domains of GPC3. The N-terminal and C-terminal subunits form the core protein of GPC3.

[0142] The negatively charged HS chains on GPC3 are functionally important glycosaminoglycans. They can bind positively charged growth factors such as HGF, fibroblast growth factor (FGF), Wnts, hedgehog, and bone morphogenetic protein. Thus, the HS chains may act as "docking sites" for these growth factors, depending on the cell context.

[0143] The N-terminal subunit of GPC3 is a ~40 kDa N-terminal peptide of GPC3 that is found in a soluble form of the GPC3 core protein. In some embodiments, the N-terminal subunit is a peptide comprising the amino acid sequence from Met 1 to Arg 358 . In some embodiments, the N-terminal subunit is a peptide comprising the amino acid sequence from Gln 25 to Arg 358A peptide of the amino acid sequence. According to the present invention, fragments of such N-terminal peptides can also be used, which are referred to herein as GPC3 N-terminal fragments. In some embodiments, the anti-GPC3 antibody portion specifically recognizes an epitope present on the N-terminal subunit of the GPC3 protein. In some embodiments, the antibody portion that specifically recognizes the N-terminal subunit (or a fragment thereof) of the GPC3 protein described herein specifically binds to the soluble form of GPC3. In some embodiments, the antibody portion that specifically recognizes the N-terminal subunit (or a fragment thereof) of the GPC3 protein specifically binds to the native form of GPC3. In some embodiments, the antibody portion that specifically recognizes the N-terminal subunit (or a fragment thereof) of the GPC3 protein can bind to nGPC3 and sGPC3.

[0144] The C-terminal subunit of GPC3 is a C-terminal peptide of approximately 30 kDa of GPC3. Based on the above cleavage site, in some embodiments, the C-terminal subunit is a peptide comprising the amino acid sequence from Ser 359 to His 580 In some embodiments, the C-terminal subunit is a peptide comprising the amino acid sequence from Ser 359 to Ser 560 According to the present invention, fragments of such C-terminal peptides can also be used, which are referred to herein as GPC3 C-terminal fragments. In some embodiments, the anti-GPC3 antibody portion specifically recognizes an epitope present on the C-terminal subunit of the GPC3 protein. In some embodiments, the antibody portion that specifically recognizes the C-terminal subunit (or a fragment thereof) of the GPC3 protein described herein specifically binds to the soluble form of GPC3. In some embodiments, the antibody portion that specifically recognizes the C-terminal subunit (or a fragment thereof) of the GPC3 protein specifically binds to the native form of GPC3. In some embodiments, the antibody portion that specifically recognizes the C-terminal subunit (or a fragment thereof) of the GPC3 protein can bind to nGPC3 and sGPC3.

[0145] The anti-GPC3 constructs described herein do not bind to the same epitopes on GPC3 as the antibodies known in the art, such as GC33 (Ishiguro et al., Cancer Res 2008; 68:9832-9838) (e.g., SEQ ID NO:510). In some embodiments, the anti-GPC3 antibody portion does not competitively and specifically bind to the same or substantially the same GPC3 epitope as GC33. In some embodiments, the anti-GPC3 antibody portion does not bind to an epitope within the amino acid sequence of SEQ ID NO:536. In some embodiments, the anti-GPC3 antibody portion does not bind to a fragment comprising the amino acid sequence of SEQ ID NO:536.

[0146] GPC3 has been reported to be expressed in various cancers and specifically, in HCC, melanoma, squamous cell lung cancer, ovarian cancer, yolk sac tumor, choriocarcinoma, neuroblastoma, Wilms' tumor, and liposarcoma. Thus, GPC3 can potentially serve as a therapeutic target for antibody-based immunotherapy and cell-based immunotherapy. Specifically, given that GPC3 is highly expressed in HCC and is expressed in more than 70% of HCC tumors but not in normal liver tissue, GPC3 can be expected to be a candidate target for liver cancer therapy. For some patients with GPC3-positive cancers, a soluble form of GPC3 can be detected in the blood. Thus, both the soluble and native forms of GPC3 can serve as suitable biomarkers for cancer diagnosis (e.g., HCC diagnosis). The 5-year survival rate of GPC3-positive HCC patients has been found to be significantly lower than that of GPC3-negative HCC patients. Thus, GPC3 expression is associated with a poor prognosis in HCC.

[0147] GPC3 cell surface localization is crucial for cell growth and Wnt activation in HCC. GPC3 binds to Wnt via its core protein. It is hypothesized that GPC3 stimulates Wnt signaling by promoting and / or stabilizing the interaction of Wnt with its signaling receptor Frizzled (Fz). Interestingly, it has been noted that GPC3 is secreted in the serum of approximately 50% of HCC patients. However, it is unclear which form of GPC3 is present in the circulating blood of cancer patients. The extracellular lipase Notum can be responsible for the cleavage of GPC3 from tumor cells into the extracellular environment.

[0148] In some embodiments, the anti-GPC3 antibody moieties described herein (against nGPC3 and / or sGPC3) specifically recognize epitopes within human GPC3. The complete amino acid sequence of exemplary human GPC3 has UniProt accession number P51654 (SEQ ID NO: 460). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within human GPC3 that include the amino acid sequence of SEQ ID NO: 460. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 1-560 (SEQ ID NO: 461) of SEQ ID NO: 460. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 25-580 of SEQ ID NO: 460 (SEQ ID NO: 462). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 25-560 (SEQ ID NO: 463) of SEQ ID NO: 460. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within the N-terminal fragment of GPC3. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 1-358 (SEQ ID NO: 468) of SEQ ID NO: 460. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 25-358 of SEQ ID NO: 460 (SEQ ID NO: 464). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within the C-terminal fragment of GPC3. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 359-560 of SEQ ID NO: 460 (SEQ ID NO: 465). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 359-580 of SEQ ID NO: 460 (SEQ ID NO: 466). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within GPC3 lacking heparan sulfate side chains. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within GPC3 bearing heparan sulfate side chains. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within the C-terminal fragment of GPC3 lacking heparan sulfate side chains. In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes within amino acids 510-560 of SEQ ID NO: 460 (SEQ ID NO: 467). In some embodiments, the anti-GPC3 antibody moieties specifically recognize epitopes spanning amino acid R located at SEQ ID NO: 460 358 / S 359The epitope of the furin cleavage site. In some embodiments, the anti-GPC3 antibody portion can specifically bind to human mature full-length GPC3 (e.g., amino acids 25-560 or 25-580 of SEQ ID NO: 460), but not to the N-terminal fragment of human GPC3 (e.g., amino acids 25-358 of SEQ ID NO: 460) or the C-terminal fragment of human GPC3 (e.g., amino acids 359-560 or 359-580 of SEQ ID NO: 460). In some embodiments, the anti-GPC3 antibody portion specifically recognizes the epitope within recombinant human GPC3. In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity. In some embodiments, GPC3 bound to the cell surface comprises (and in some embodiments, consists of or consists predominantly of) the amino acid sequence of SEQ ID NO: 460 or SEQ ID NO: 462. In some embodiments, soluble GPC3 is the circulating GPC3 protein in serum or free GPC3 peptide. In some embodiments, soluble GPC3 is the GPC3 protein or peptide present in solution. In some embodiments, soluble GPC3 comprises (and in some embodiments, consists of or consists predominantly of) the amino acid sequence of SEQ ID NO: 461 or SEQ ID NO: 463. In some embodiments, the anti-GPC3 antibody portion can cross-react with GPC3 from species other than human. In some embodiments, the anti-GPC3 antibody portion can be fully specific for one or more human GPC3 proteins and may not exhibit species or other types of non-human cross-reactivity.

[0149] In some embodiments, the anti-GPC3 antibody portion cross-reacts with at least one allelic variant of the GPC3 protein (or a fragment thereof). In some embodiments, the allelic variant has up to about 30 (e.g., about 1, 2, 3, 4, 5, 6, 7, 8, 9, 10, 15, 20, 25, or 30) amino acid substitutions (e.g., conservative substitutions) compared to the naturally occurring GPC3 (or a fragment thereof). In some embodiments, the anti-GPC3 antibody portion does not cross-react with any allelic variant of the GPC3 protein (or a fragment thereof).

[0150] In some embodiments, the anti-GPC3 antibody portion cross-reacts with at least one interspecies variant of the GPC3 protein. In some embodiments, for example, the GPC3 protein (or a fragment thereof) is human GPC3 and the interspecies variant of the GPC3 protein (or a fragment thereof) is its mouse or rat variant. In some embodiments, the anti-GPC3 antibody portion does not cross-react with any interspecies variant of the GPC3 protein.

[0151] In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 expressed on the cell surface of HepG2, Hep3B, Huh7, JHH-7, or 293 cells. In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 expressed on the cell surface of cancer cells (e.g., solid tumors). In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 expressed on the cell surface of liver cancer (e.g., HCC), melanoma, squamous cell lung cancer, ovarian cancer, yolk sac tumor, choriocarcinoma, neuroblastoma, Wilms' tumor, hepatoblastoma, testicular non-seminomatous germ cell tumor, gastric cancer, or liposarcoma.

[0152] Binding affinity

[0153] Binding affinity can be K d 、K 解离 、K 结合 or K a indicated. As used herein, the term "K 解离 " means the dissociation rate constant at which the antibody portion dissociates from the antibody portion / antigen complex, as determined using a kinetic selection configuration. As used herein, the term "K 结合 " means the association rate constant at which the antibody portion binds to the antigen to form an antibody portion / antigen complex. As used herein, the term equilibrium dissociation constant "K d " refers to the dissociation constant for a particular antibody portion-antigen interaction and describes the antigen concentration required to occupy half of all antibody binding domains present in a solution of antibody molecules at equilibrium, and is equal to K 解离 / K 结合 . The measurement of K d assumes that all binders are in solution. In the case where the antibody portion is tethered to the cell wall, such as in a yeast expression system, the corresponding equilibrium rate constant is represented by EC 50 , thereby obtaining a good approximation of K d . The affinity constant K a is the reciprocal of the dissociation constant K d .

[0154] The dissociation constant (K d ) is used as an indicator of the affinity of the antibody portion for the antigen. For example, by the Scatchard method, using antibodies labeled with various labels, and by using BIACORE TM (manufactured by Amersham Biosciences), analyzing biomolecular interactions by surface plasmon resonance, according to the user manual and the accompanying kit, the analysis can be made simple. The K dValues are expressed in units of M (Mols). The antibody portion that specifically binds to the target may have, for example, ≤ 10 -7 M, ≤ 10 -8 M, ≤ 10 -9 M, ≤ 10 -10 M, ≤ 10 -11 M, ≤ 10 -12 M or ≤ 10 -13 M of K d .

[0155] The binding specificity of the antibody portion can be determined experimentally by methods known in the art. Such methods include (but are not limited to) Western blotting, ELISA, RIA, ECL, IRMA, EIA, BIACORE TM testing and peptide scanning. In some embodiments, the binding affinity of the anti-GPC3 antibody portion is measured by testing the binding affinity of the anti-GPC3 antibody portion to cells expressing GPC3 on the surface (such as HepG2 cells).

[0156] In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface (for example, the binding affinity of the antibody portion for GPC3 bound to the cell surface is higher than that for soluble GPC3). In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity. In some embodiments, in a competitive binding assay for soluble GPC3 antigen using, for example, flow cytometry, the anti-nGPC3 antibody portion specifically binds to GPC3 bound to the cell surface. In some embodiments, when the anti-nGPC3 antibody portion has been pre-incubated with soluble GPC3 antigen in a competitive binding assay, the anti-nGPC3 antibody portion specifically binds to GPC3 bound to the cell surface. For example, in some embodiments, an anti-GPC3 construct comprising the anti-nGPC3 antibody portion can be pre-incubated with various concentrations of soluble GPC3 antigen (such as recombinant GPC3 fragments), and then GPC3 +Cells (e.g., HepG2 cells) are added to the antibody / antigen mixture and incubated, and then the anti-GPC3 construct bound to the cell surface can be detected: The binding of the anti-GPC3 construct pre-incubated with soluble GPC3 to GPC3+ cells may not show a significant difference compared to the binding of the anti-GPC3 construct to GPC3+ cells in the absence of pre-incubation with soluble GPC3, demonstrating that the anti-GPC3 antibody portion can specifically recognize GPC3 bound to the cell surface, or that the anti-nGPC3 antibody portion has a higher binding affinity for GPC3 bound to the cell surface than for soluble GPC3, and that the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface with a high binding affinity and binds to soluble GPC3 with a low binding affinity.

[0157] In some embodiments, the anti-GPC3 antibody portion binds specifically to the target GPC3 (e.g., nGPC3 and / or sGPC3) with a K -7 of about 10 -13 M to about 10 -7 M (e.g., about 10 -13 M to about 10 -9 M, about 10 -13 M to about 10 -10 M, or about 10 -12 M to about 10 d Therefore, in some embodiments, the K d of the binding between the anti-nGPC3 antibody portion and nGPC3, the K d of the binding between the anti-sGPC3 antibody portion and sGPC3, or the K d of the binding between the anti-GPC3 antibody portion and GPC3 (in any form) is about 10 -7 M to about 10 -13 M, about 1×10 -7 M to about 5×10 -13 M, about 10 -7 M to about 10 -12 M, about 10 -7 M to about 10 -11 M, about 10 -7 M to about 10 -10 M, about 10 -7 M to about 10 -9 M, about 10 -8 M to about 10 -13 M, about 1×10 -8 M to about 5×10 -13 M, about 10 -8 M to about 10 -12 M, about 10 -8 M to about 10 -11 M, about 10 -8M to about 10 -10 M, about 10 -8 M to about 10 -9 M, about 5×10 -9 M to about 1×10 -13 M, about 5×10 -9 M to about 1×10 -12 M, about 5×10 -9 M to about 1×10 -11 M, about 5×10 -9 M to about 1×10 -10 M, about 10 -9 M to about 10 -13 M, about 10 -9 M to about 10 -12 M, about 10 -9 M to about 10 -11 M, about 10 -9 M to about 10 -10 M, about 5×10 -10 M to about 1×10 -13 M, about 5×10 -10 M to about 1×10 -12 M, about 5×10 -10 M to about 1×10 -11 M, about 10 -10 M to about 10 -13 M, about 1×10 -10 M to about 5×10 -13 M, about 1×10 -10 M to about 1×10 -12 M, about 1×10 -10 M to about 5×10 -12 M, about 1×10 -10 M to about 1×10 -11 M, about 10 -11 M to about 10 -13 M, about 1×10 -11 M to about 5×10 -13 M, about 10 -11 M to about 10 -12 M, or about 10 -12 M to about 10 -13 M. In some embodiments, the K of the binding between the anti-nGPC3 antibody portion and nGPC3 d is about 10 -7 M to about 10 -13 M.

[0158] In some embodiments, the K of the binding between the anti-GPC3 antibody portion and a non-target d is greater than the K of the binding between the anti-GPC3 antibody portion and the targetd and, in this document, in some embodiments, the binding affinity of the anti-GPC3 antibody portion for a target (e.g., GPC3 bound to the cell surface) is higher than the binding affinity for a non-target (e.g., soluble GPC3). In some embodiments, the non-target is a non-GPC3 antigen. In some embodiments, for the anti-nGPC3 antibody portion, the non-target is soluble GPC3. In some embodiments, for the anti-sGPC3 antibody portion, the non-target is GPC3 bound to the cell surface. For example, the K of the binding between the anti-nGPC3 antibody portion and soluble GPC3 d can be at least about 10 times the K of the binding between the anti-nGPC3 antibody portion and GPC3 bound to the cell surface d , such as about 10 - 100 times, about 100 - 1000 times, about 10 3 -10 4 times, about 10 4 -10 5 times, about 10 5 -10 6 times, about 10 6 -10 7 times, about 10 7 -10 8 times, about 10 8 -10 9 times, about 10 9 -10 10 times, about 10 10 -10 11 times or about 10 11 -10 12 times. Similarly, for example, the K of the binding between the anti-sGPC3 antibody portion and GPC3 bound to the cell surface d can be at least about 10 times the K of the binding between the anti-sGPC3 antibody portion and soluble GPC3 d , such as about 10 - 100 times, about 100 - 1000 times, about 10 3 -10 4 times, about 10 4 -10 5 times, about 10 5 -10 6 times, about 10 6 -10 7 times, about 10 7 -10 8 times, about 10 8 -10 9 times, about 10 9 -10 10 times, about 10 10 -10 11 times or about 1011 -10 12 。In some embodiments, the K of the binding between the anti-GPC3 antibody portion (against nGPC3 and / or sGPC3) and a non-GPC3 target d can be at least about 10 times the K of the binding between the anti-GPC3 antibody portion and the target GPC3 d , such as about 10-100 times, about 100-1000 times, about 10 3 -10 4 times, about 10 4 -10 5 times, about 10 5 -10 6 times, about 10 6 -10 7 times, about 10 7 -10 8 times, about 10 8 -10 9 times, about 10 9 -10 10 times, about 10 10 -10 11 times or about 10 11 -10 12 times. In some embodiments, the K of the binding between the anti-nGPC3 antibody portion and soluble GPC3 d is at least about 10 times the K of the binding between the anti-nGPC3 antibody portion and GPC3 bound to the cell surface d .

[0159] In some embodiments, the K of the binding between the anti-GPC3 antibody portion and a non-target d is about 10 -1 M to about 10 -6 M (such as about 10 -1 M to about 10 -6 M, about 10 -1 M to about 10 -5 M, or about 10 -2 M to about 10 -4 M). In some embodiments, the non-target is a non-GPC3 antigen. In some embodiments, for the anti-nGPC3 antibody portion, the non-target is soluble GPC3. In some embodiments, for the anti-sGPC3 antibody portion, the non-target is GPC3 bound to the cell surface. Thus in some embodiments, the K of the binding between the anti-GPC3 antibody portion and a non-GPC3 target d , the K of the binding between the anti-nGPC3 antibody portion and soluble GPC3 d or the K of the binding between the anti-sGPC3 antibody portion and GPC3 bound to the cell surface d is about 10-1 M to about 10 -6 M, about 1×10 -1 M to about 5×10 -6 M, about 10 -1 M to about 10 -5 M, about 1×10 -1 M to about 5×10 -5 M, about 10 -1 M to about 10 -4 M, about 1×10 -1 M to about 5×10 -4 M, about 10 -1 M to about 10 -3 M, about 1×10 -1 M to about 5×10 -3 M, about 10 -1 M to about 10 -2 M, about 10 -2 M to about 10 - 6 M, about 1×10 -2 M to about 5×10 -6 M, about 10 -2 M to about 10 -5 M, about 1×10 -2 M to about 5×10 -5 M, about 10 -2 M to about 10 -4 M, about 1×10 -2 M to about 5×10 -4 M, about 10 -2 M to about 10 -3 M, about 10 -3 M to about 10 -6 M, about 1×10 -3 M to about 5×10 -6 M, about 10 -3 M to about 10 -5 M, about 1×10 -3 M to about 5×10 -5 M, about 10 -3 M to about 10 -4 M, about 10 -4 M to about 10 -6 M, about 1×10 -4 M to about 5×10 -6 M, about 10 -4 M to about 10 -5 M, or about 10 -5 M to about 10 -6 M. In some embodiments, the K of the binding between the anti-nGPC3 antibody portion and soluble GPC3 d is about 10-1 M to about 10 -6 M. In some embodiments, the anti-nGPC3 antibody portion that specifically recognizes GPC3 bound to the cell surface does not bind to soluble GPC3.

[0160] In some embodiments, when referring to an anti-GPC3 antibody portion that specifically recognizes a target GPC3 (e.g., GPC3 bound to the cell surface) with high binding affinity and binds to a non-target (e.g., soluble GPC3) with low binding affinity, the K of the anti-GPC3 antibody portion binding to the target GPC3 (e.g., GPC3 bound to the cell surface) d is about 10 -7 M to about 10 -13 M (e.g., about 10 -7 M to about 10 -13 M, about 10 -9 M to about 10 -13 M, or about 10 -10 M to about 10 -12 M), and the K of binding to the non-target (e.g., soluble GPC3) d is about 10 -1 M to about 10 -6 M (e.g., about 10 -1 M to about 10 -6 M, about 10 -1 M to about 10 -5 M, or about 10 -2 M to about 10 -4 M).

[0161] In some embodiments, when referring to an anti-GPC3 antibody portion that specifically recognizes GPC3 bound to the cell surface (e.g., the binding affinity of the antibody portion for GPC3 bound to the cell surface is higher than that for soluble GPC3), or when referring to an anti-GPC3 antibody portion that specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity, the binding affinity of the anti-GPC3 antibody portion is compared to a control anti-GPC3 antibody (e.g., the monoclonal antibody GC33). In some embodiments, the K of the binding between GC33 (e.g., SEQ ID NO:510) and GPC3 bound to the cell surface d can be at least about 2 times the K of the binding between the anti-nGPC3 antibody portion described herein and GPC3 bound to the cell surface d e.g., about 2 times, about 3 times, about 4 times, about 5 times, about 6 times, about 7 times, about 8 times, about 9 times, about 10 times, about 10 - 100 times, about 100 - 1000 times, about 10 3 -10 4 times, about 10 4-10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11 -fold, or about 10 11 -10 12 -fold. In some embodiments, the K of the anti-GPC3 construct relative to GPC3 bound to the cell surface d is from about 0.1 nM to about 2 nM, such as from about 0.1 nM to about 0.5 nM, from about 0.5 nM to about 1 nM, or from about 1.5 to about 2 nM. In some embodiments, the K of the binding between the anti-nGPC3 antibody portion described herein and soluble GPC3 d can be at least about 2-fold of the K of the binding between GC33 (e.g., SEQ ID NO: 510) and soluble GPC3 d , such as about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 10 - 100-fold, about 100 - 1000-fold, about 10 3 -10 4 -fold, about 10 4 -10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11 -fold, or about 10 11 -10 12 -fold. In some embodiments, the K of the anti-GPC3 construct relative to GPC3 bound to the cell surface d is from about 10 nM to about 100 nM, such as from about 10 nM to about 20 nM, from about 20 nM to about 40 nM, from about 40 nM to about 80 nM, or from about 80 nM to about 100 nM. In some embodiments, the K of the binding between the control anti-GPC3 antibody and GPC3 bound to the cell surface dThe K for the binding between the anti-nGPC3 antibody portion described herein and GPC3 bound to the cell surface d is at least about 2-fold, such as about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 10-100 fold, about 100-1000 fold, about 10 3 -10 4 -fold, about 10 4 -10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11 -fold, or about 10 11 -10 12 -fold, where the control antibody includes: i) V comprising the amino acid sequence SEQ ID NO:503 H , or a variant thereof having at least about 95% sequence identity with SEQ ID NO:503; and ii) V comprising the amino acid sequence SEQ ID NO:504 L , or a variant thereof having at least about 95% sequence identity with SEQ ID NO:504. In some embodiments, the K for the binding between the anti-nGPC3 antibody portion described herein and soluble GPC3 d can be at least about 2-fold that of the K for the binding between the control anti-GPC3 antibody and soluble GPC3 d , such as about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 10-100 fold, about 100-1000 fold, about 10 3 -10 4 -fold, about 10 4 -10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11-fold, or about 10 11 -10 12 -fold, wherein the control antibody comprises: i) a V comprising the amino acid sequence SEQ ID NO:503 H , or a variant thereof having at least about 95% sequence identity to SEQ ID NO:503; and ii) a V comprising the amino acid sequence SEQ ID NO:504 L , or a variant thereof having at least about 95% sequence identity to SEQ ID NO:504. In some embodiments, the K of binding between the control anti-GPC3 antibody and GPC3 bound to the cell surface d can be at least about 2-fold of the K of binding between the anti-nGPC3 antibody moiety described herein and GPC3 bound to the cell surface d , such as about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 10 - 100-fold, about 100 - 1000-fold, about 10 3 -10 4 -fold, about 10 4 -10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11 -fold, or about 10 11 -10 12 -fold, wherein the control antibody comprises: i) a V H that comprises an HC-CDR1 having the amino acid sequence SEQ ID NO:497, an HC-CDR2 having the amino acid sequence SEQ ID NO:498, and an HC-CDR3 having the amino acid sequence SEQ ID NO:499; and ii) a V L that comprises an LC-CDR1 having the amino acid sequence SEQ ID NO:500, an LC-CDR2 having the amino acid sequence SEQ ID NO:501, and an LC-CDR3 having the amino acid sequence SEQ ID NO:502. In some embodiments, the K of binding between the anti-nGPC3 antibody moiety described herein and soluble GPC3 d can be the K of binding between the control anti-GPC3 antibody and soluble GPC3 dat least about 2-fold, such as about 2-fold, about 3-fold, about 4-fold, about 5-fold, about 6-fold, about 7-fold, about 8-fold, about 9-fold, about 10-fold, about 10 - 100-fold, about 100 - 1000-fold, about 10 3 -10 4 -fold, about 10 4 -10 5 -fold, about 10 5 -10 6 -fold, about 10 6 -10 7 -fold, about 10 7 -10 8 -fold, about 10 8 -10 9 -fold, about 10 9 -10 10 -fold, about 10 10 -10 11 -fold, or about 10 11 -10 12 -fold, wherein the control antibody comprises: i) V H , comprising an HC-CDR1 having the amino acid sequence of SEQ ID NO: 497, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 498, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 499; and ii) V L , comprising an LC-CDR1 having the amino acid sequence of SEQ ID NO: 500, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 501, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 502.

[0162] In some embodiments, the IC50 of soluble GPC3 in competing for binding between the anti-GPC3 construct and cell surface-bound GPC3 is from about 1 μg / ml to about 100 μg / ml, such as from about 1 μg / ml to about 5 μg / ml, 1 μg / ml to about 10 μg / ml, about 10 μg / ml to about 20 μg / ml, about 20 μg / ml to about 50 μg / ml, or 50 μg / ml to about 100 μg / ml. In some embodiments, the IC50 of soluble GPC3 in competing for binding between the anti-GPC3 construct and cell surface-bound GPC3 is at least about 2-fold, 3-fold, 4-fold, 5-fold, 10-fold, 20-fold, 50-fold, 100-fold, or more than 100-fold that of the IC50 of soluble GPC3 in competing for binding between a control anti-GPC3 antibody (such as GC33) and cell surface-bound GPC3.

[0163] Anti-GPC3 antibody partial form

[0164] The anti-GPC3 antibody portion described herein (against, for example, nGPC3 and / or sGPC3) can be in any form of an antibody or antigen-binding fragment.

[0165] In some embodiments, the anti-GPC3 antibody portion (against, for example, nGPC3 and / or sGPC3) is a full-length antibody or immunoglobulin derivative. In some embodiments, the anti-GPC3 antibody portion is an antigen-binding fragment, such as an antigen-binding fragment selected from the group consisting of: Fab, Fab', F(ab')2, Fv fragment, disulfide-stabilized Fv fragment (dsFv), or single-chain Fv (scFv). In some embodiments, the anti-GPC3 antibody portion is an scFv. In some embodiments, the anti-GPC3 antibody portion is a Fab or Fab'. In some embodiments, the anti-GPC3 antibody portion is chimeric, human, partially humanized, fully humanized, or semi-synthetic.

[0166] In some embodiments, the anti-GPC3 antibody portion (against nGPC3 and / or sGPC3) is a semi-synthetic antibody portion comprising a fully human sequence and one or more synthetic regions. In some embodiments, the anti-GPC3 antibody portion is a semi-synthetic antibody portion comprising a fully human light chain variable domain and a semi-synthetic heavy chain variable domain, the semi-synthetic heavy chain variable domain comprising fully human FR1, HC-CDR1, FR2, HC-CDR2, FR3, and FR4 regions and a synthetic HC-CDR3. In some embodiments, the semi-synthetic heavy chain variable domain comprises a fully synthetic HC-CDR3 having a sequence of about 5 to about 25 (such as any one of about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acids in length. In some embodiments, the semi-synthetic heavy chain variable domain or the synthetic HC-CDR3 is obtained from a semi-synthetic library (such as a semi-synthetic human library) comprising fully synthetic HC-CDR3s having a sequence of about 5 to about 25 (such as any one of about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, or 25) amino acids in length, wherein each amino acid in the sequence is randomly selected from the standard human amino acids other than cysteine. In some embodiments, the length of the synthetic HC-CDR3 is about 5 to about 19 (such as any one of about 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, or 19) amino acids. In some embodiments, the anti-GPC3 antibody portion is a semi-synthetic antibody portion comprising human CDRs and non-human framework sequences. In some embodiments, the non-human framework sequence comprises any sequence that can be used to generate synthetic heavy and / or light chain variable regions using one or more human CDR sequences as described herein, including, for example, mammals such as mice, rats, rabbits, pigs, bovines (such as cows, bulls, buffalo), deer, sheep, goats, chickens, cats, dogs, ferrets, primates (such as marmosets, rhesus monkeys), and the like. In some embodiments, the anti-GPC3 antibody portion is generated by grafting one or more human CDR sequences as described herein onto a non-human framework sequence (such as a mouse or chicken framework sequence).

[0167] Anti-GPC3 antibody portion sequence

[0168] In some embodiments, the anti-GPC3 antibody portion comprises a specific sequence or certain variants of such sequences. In some embodiments, the amino acid substitutions in the variant sequences generally do not reduce the ability of the anti-nGPC3 antibody portion to specifically recognize GPC3 bound to the cell surface, the ability of the anti-sGPC3 antibody portion to specifically recognize soluble GPC3, or the ability of the anti-GPC3 antibody portion to specifically recognize the target GPC3 (e.g., nGPC3 and / or sGPC3). For example, variants can be generated that generally do not reduce the binding affinity for the target GPC3 (e.g., nGPC3 and / or sGPC3). Also encompassed are variants that generally improve the target GPC3 binding affinity or affect some other property (e.g., specificity and / or cross-reactivity with related variants of the target GPC3).

[0169] Exemplary antibody sequences are shown in Tables 6 to 9. The exemplary CDR sequences in Tables 6 and 8 were predicted using the IgBLAST algorithm. See, e.g., Ye J et al., Nucleic Acids Research, 41:W34-W40 (2013), the entire disclosure of which is incorporated herein by reference. Those skilled in the art will recognize that a variety of algorithms are known for predicting CDR positions in the variable regions of antibody heavy and light chains, and antibody agents that include the CDRs from the antibodies described herein but are based on prediction algorithms other than IgBLAST are within the scope of the invention.

[0170] The exemplary antibody heavy and light chain variable region sequences in Tables 7 and 9 are defined pairs according to INTERNATIONAL IMMUNOGENETICS INFORMATION (IMGT). See, e.g., Lefranc, M.-P. et al., Nucleic Acids Res., 43:D413-422 (2015), the entire disclosure of which is incorporated herein by reference. Those skilled in the art will recognize that antibody agents that include the V H or V L sequences from the antibodies described herein but are based on algorithms other than IMGT are within the scope of the invention.

[0171] Anti - GPC3 antibody portion that specifically recognizes GPC3 bound to the cell surface

[0172] In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface (anti-nGPC3 antibody portion). In some embodiments, the anti-nGPC3 antibody portion has a higher binding affinity for GPC3 bound to the cell surface than for soluble GPC3. In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface with high binding affinity and binds to soluble GPC3 with low binding affinity.

[0173] In some embodiments, the anti-nGPC3 antibody portion comprises i) a heavy chain variable domain (V H ), which comprises an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain (V L ), which comprises an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions.

[0174] In some embodiments, the anti-nGPC3 antibody portion comprises i) an HC-CDR3 comprising an amino acid sequence of any one of SEQ ID NOs: 103-133; and ii) an LC-CDR3 comprising an amino acid sequence of any one of SEQ ID NOs: 256-286.

[0175] In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions.

[0176] In some embodiments, the anti-nGPC3 antibody portion comprises i) V H, which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31 or a variant thereof comprising up to about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82 or a variant thereof comprising up to about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184 or a variant thereof comprising up to about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235 or a variant thereof comprising up to about 3 (e.g., any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286.

[0177] In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions.

[0178] In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H, which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions occur in HC-CDR1 or HC-CDR2; and ii) V L , comprising an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions occur in LC-CDR1 or LC-CDR2.

[0179] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 1-31, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 52-82, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 103-133; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 154-184, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 205-235, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 256-286.

[0180] In some embodiments, the anti-nGPC3 antibody portion comprises: a) V H , which comprises the amino acid sequence of any one of SEQ ID NOs: 1-31, the amino acid sequence of any one of SEQ ID NOs: 52-82, and the amino acid sequence of any one of SEQ ID NOs: 103-133; and ii) V L , which comprises the amino acid sequence of any one of SEQ ID NOs: 154-184, the amino acid sequence of any one of SEQ ID NOs: 205-235, and the amino acid sequence of any one of SEQ ID NOs: 256-286.

[0181] In some embodiments, the anti-nGPC3 antibody portion comprises: a) V H, which comprises one, two or three CDRs of any one of SEQ ID NO: 307 - 337, and b) V L , which comprises one, two or three CDRs of any one of SEQ ID NO: 358 - 388. In some embodiments, the anti - nGPC3 antibody moiety comprises: a) V H , which comprises HC - CDR1, HC - CDR2 and HC - CDR3 of the heavy - chain variable domain of any one of SEQ ID NO: 307 - 337, and b) VL, which comprises LC - CDR1, LC - CDR2 and LC - CDR3 of the light - chain variable domain CDR of any one of SEQ ID NO: 358 - 388.

[0182] In some embodiments, the anti - nGPC3 antibody moiety comprises: a) V comprising the amino acid sequence of any one of SEQ ID NO: 307 - 337 H , or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with any one of SEQ ID NO: 307 - 337; and b) V comprising the amino acid sequence of any one of SEQ ID NO: 358 - 388 L , or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with any one of SEQ ID NO: 358 - 388.

[0183] In some embodiments, the anti - nGPC3 antibody moiety comprises: a) V comprising the amino acid sequence of any one of SEQ ID NO: 307 - 337 H ; and b) V comprising the amino acid sequence of any one of SEQ ID NO: 358 - 388 L .

[0184] In some embodiments, the anti - nGPC3 antibody moiety comprises the HC - CDR of V having an amino acid sequence of any one of SEQ ID NO: 307 - 337 H , and the LC - CDR of V having an amino acid sequence of any one of SEQ ID NO: 358 - 388 L .

[0185] The heavy - chain and light - chain variable domains can be combined in various paired combinations to produce a variety of anti - nGPC3 antibody moieties. Exemplary anti - nGPC3 antibodies are provided in Tables 6 and 7.

[0186] Table 6 Anti - GPC3 antibody moieties that specifically recognize GPC3 CDR sequences bound to the cell surface

[0187]

[0188]

[0189]

[0190] Table 7 Anti-GPC3 antibody portions that specifically recognize and bind to the cell surface GPC3 V H / V L sequences (CDR sequences Add Underline )

[0191]

[0192]

[0193]

[0194]

[0195]

[0196]

[0197] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:1 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:52 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:103 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ IDNO:154 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO:205 or a variant thereof having at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO:256 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:1, an HC-CDR2 having the amino acid sequence of SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:103, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQID NO:154, an LC-CDR2 having the amino acid sequence of SEQ ID NO:205, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:256, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:1, an HC-CDR2 having the amino acid sequence of SEQ ID NO:52, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:103; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:154, an LC-CDR2 having the amino acid sequence of SEQ ID NO:205, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:256.

[0198] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:2 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:53 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:104 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 155 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 206 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 257 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 2, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 53, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 104, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 155, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 206, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 257, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 2, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 53, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 104; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 155, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 206, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 257.

[0199] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:3 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:54 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:105 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:156 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:207 or a variant thereof comprising up to about 3 (such as any one of about 1, 2 or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:258 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:3, an HC-CDR2 having the amino acid sequence of SEQ ID NO:54 and an HC-CDR3 having the amino acid sequence of SEQ ID NO:105, or a variant thereof in which the HC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:156, an LC-CDR2 having the amino acid sequence of SEQ ID NO:207 and an LC-CDR3 having the amino acid sequence of SEQ ID NO:258, or a variant thereof in which the LC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:3, an HC-CDR2 having the amino acid sequence of SEQ ID NO:54 and an HC-CDR3 having the amino acid sequence of SEQ ID NO:105; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:156, an LC-CDR2 having the amino acid sequence of SEQ ID NO:207, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:258.

[0200] In some embodiments, the anti-nGPC3 antibody portion comprises: i)V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:4 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:55 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:106 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii)V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:157 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:208 or a variant thereof having at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:259 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1)i)V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:4, an HC-CDR2 having the amino acid sequence of SEQ ID NO:55, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:106, or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequence; and ii)V L , which comprises an LC-CDR1 having the amino acid sequence of SEQID NO:157, an LC-CDR2 having the amino acid sequence of SEQ ID NO:208, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:259, or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i)V H, which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:4, HC-CDR2 with the amino acid sequence of SEQ ID NO:55, and HC-CDR3 with the amino acid sequence of SEQ ID NO:106; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:157, LC-CDR2 with the amino acid sequence of SEQ ID NO:208, and LC-CDR3 with the amino acid sequence of SEQ ID NO:259.

[0201] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:5 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:56 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:107 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ IDNO:158 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:209 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:260 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:5, HC-CDR2 with the amino acid sequence of SEQ ID NO:56, and HC-CDR3 with the amino acid sequence of SEQ ID NO:107, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:158, an LC-CDR2 having the amino acid sequence of SEQ ID NO:209, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:260, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:5, an HC-CDR2 having the amino acid sequence of SEQ ID NO:56, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:107; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:158, an LC-CDR2 having the amino acid sequence of SEQ ID NO:209, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:260.

[0202] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:6 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:57 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:108 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:159 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:210 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:261 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H, which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:6, HC-CDR2 having the amino acid sequence of SEQ ID NO:57, and HC-CDR3 having the amino acid sequence of SEQ ID NO:108, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:159, LC-CDR2 having the amino acid sequence of SEQ ID NO:210, and LC-CDR3 having the amino acid sequence of SEQ ID NO:261, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:6, HC-CDR2 having the amino acid sequence of SEQ ID NO:57, and HC-CDR3 having the amino acid sequence of SEQ ID NO:108; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:159, LC-CDR2 having the amino acid sequence of SEQ ID NO:210, and LC-CDR3 having the amino acid sequence of SEQ ID NO:261.

[0203] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:7 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:58 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:109 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 160 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 211 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 262 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 7, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 58, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 109, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 160, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 211, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 262, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 7, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 58, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 109; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 160, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 211, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 262.

[0204] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:8 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:59 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:110 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:161 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:212 or a variant thereof comprising at most about 3 (such as any one of about 1, 2 or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:263 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:8, an HC-CDR2 having the amino acid sequence of SEQ ID NO:59 and an HC-CDR3 having the amino acid sequence of SEQ ID NO:110, or a variant thereof in which the HC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:161, an LC-CDR2 having the amino acid sequence of SEQ ID NO:212 and an LC-CDR3 having the amino acid sequence of SEQ ID NO:263, or a variant thereof in which the LC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:8, an HC-CDR2 having the amino acid sequence of SEQ ID NO:59 and an HC-CDR3 having the amino acid sequence of SEQ ID NO:110; and ii) V L, which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:161, LC-CDR2 with the amino acid sequence of SEQ ID NO:212, and LC-CDR3 with the amino acid sequence of SEQ ID NO:263.

[0205] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:9 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:60 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:111 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:162 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:213 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:264 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:9, HC-CDR2 with the amino acid sequence of SEQ ID NO:60, and HC-CDR3 with the amino acid sequence of SEQ ID NO:111, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:162, LC-CDR2 with the amino acid sequence of SEQ ID NO:213, and LC-CDR3 with the amino acid sequence of SEQ ID NO:264, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:9, an HC-CDR2 having the amino acid sequence of SEQ ID NO:60, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:111; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:162, an LC-CDR2 having the amino acid sequence of SEQ ID NO:213, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:264.

[0206] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:10 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:61 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:112 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:163 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:214 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:265 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:10, an HC-CDR2 having the amino acid sequence of SEQ ID NO:61, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:112, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:163, an LC-CDR2 having the amino acid sequence of SEQ ID NO:214, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:265, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:10, an HC-CDR2 having the amino acid sequence of SEQ ID NO:61, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:112; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:163, an LC-CDR2 having the amino acid sequence of SEQ ID NO:214, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:265.

[0207] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:11 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:62 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:113 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:164 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:215 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:266 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:11, an HC-CDR2 having the amino acid sequence of SEQ ID NO:62, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:113, or a variant thereof wherein the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:164, an LC-CDR2 having the amino acid sequence of SEQ ID NO:215, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:266, or a variant thereof wherein the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:11, an HC-CDR2 having the amino acid sequence of SEQ ID NO:62, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:113; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:164, an LC-CDR2 having the amino acid sequence of SEQ ID NO:215, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:266.

[0208] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:12 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:63 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:114 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 165 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 216 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 267 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 12, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 63, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 114, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 165, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 216, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 267, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 12, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 63, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 114; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 165, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 216, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 267.

[0209] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:13 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:64 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:115 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:166 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:217 or a variant thereof comprising at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:268 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:13, an HC-CDR2 having the amino acid sequence of SEQ ID NO:64, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:115, or a variant thereof in which the HC-CDR sequences comprise at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:166, an LC-CDR2 having the amino acid sequence of SEQ ID NO:217, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:268, or a variant thereof in which the LC-CDR sequences comprise at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:13, an HC-CDR2 having the amino acid sequence of SEQ ID NO:64, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:115; and ii) V L, which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:166, an LC-CDR2 with the amino acid sequence of SEQ ID NO:217, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:268.

[0210] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:14 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 with the amino acid sequence of SEQ ID NO:65 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:116 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:167 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 with the amino acid sequence of SEQ ID NO:218 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:269 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:14, an HC-CDR2 with the amino acid sequence of SEQ ID NO:65, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:116, or a variant thereof with up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequence; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:167, an LC-CDR2 with the amino acid sequence of SEQ ID NO:218, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:269, or a variant thereof with up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H, which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:14, HC-CDR2 having the amino acid sequence of SEQ ID NO:65, and HC-CDR3 having the amino acid sequence of SEQ ID NO:116; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:167, LC-CDR2 having the amino acid sequence of SEQ ID NO:218, and LC-CDR3 having the amino acid sequence of SEQ ID NO:269.

[0211] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:15 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:66 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:117 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ IDNO:168 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO:219 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO:270 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:15, HC-CDR2 having the amino acid sequence of SEQ ID NO:66, and HC-CDR3 having the amino acid sequence of SEQ ID NO:117, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:168, an LC-CDR2 having the amino acid sequence of SEQ ID NO:219, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:270, or a variant thereof wherein the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:15, an HC-CDR2 having the amino acid sequence of SEQ ID NO:66, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:117; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:168, an LC-CDR2 having the amino acid sequence of SEQ ID NO:219, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:270.

[0212] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:16 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:67 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:118 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:169 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:220 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:271 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:16, an HC-CDR2 having the amino acid sequence of SEQ ID NO:67, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:118, or a variant thereof wherein the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:169, an LC-CDR2 having the amino acid sequence of SEQ ID NO:220, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:271, or a variant thereof wherein the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:16, an HC-CDR2 having the amino acid sequence of SEQ ID NO:67, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:118; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:169, an LC-CDR2 having the amino acid sequence of SEQ ID NO:220, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:271.

[0213] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:17 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:68 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:119 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 170 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 221 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 272 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 17, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 68, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 119, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 170, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 221, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 272; or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 17, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 68, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 119; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 170, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 221, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 272.

[0214] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:18 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:69 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:120 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:171 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:222 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:273 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:18, an HC-CDR2 having the amino acid sequence of SEQ ID NO:69, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:120, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:171, an LC-CDR2 having the amino acid sequence of SEQ ID NO:222, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:273, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , comprising an HC-CDR1 having the amino acid sequence of SEQ ID NO:18, an HC-CDR2 having the amino acid sequence of SEQ ID NO:69, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:120; and ii) V L, which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:171, LC-CDR2 with the amino acid sequence of SEQ ID NO:222, and LC-CDR3 with the amino acid sequence of SEQ ID NO:273.

[0215] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:19 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:70 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:121, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:172 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:223 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:274, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:19, HC-CDR2 with the amino acid sequence of SEQ ID NO:70, and HC-CDR3 with the amino acid sequence of SEQ ID NO:121, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:172, LC-CDR2 with the amino acid sequence of SEQ ID NO:223, and LC-CDR3 with the amino acid sequence of SEQ ID NO:274, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H, which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:19, HC-CDR2 with the amino acid sequence of SEQ ID NO:70, and HC-CDR3 with the amino acid sequence of SEQ ID NO:121; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:172, LC-CDR2 with the amino acid sequence of SEQ ID NO:223, and LC-CDR3 with the amino acid sequence of SEQ ID NO:274.

[0216] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:20 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:71 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:122 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ IDNO:173 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:224 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:275 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , comprising HC-CDR1 with the amino acid sequence of SEQ ID NO:20, HC-CDR2 with the amino acid sequence of SEQ ID NO:71, and HC-CDR3 with the amino acid sequence of SEQ ID NO:122, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, comprising an LC-CDR1 with the amino acid sequence of SEQ ID NO:173, an LC-CDR2 with the amino acid sequence of SEQ ID NO:224, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:275, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:20, an HC-CDR2 with the amino acid sequence of SEQ ID NO:71, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:122; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:173, an LC-CDR2 with the amino acid sequence of SEQ ID NO:224, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:275.

[0217] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:21 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 with the amino acid sequence of SEQ ID NO:72 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:123 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ IDNO:174 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 with the amino acid sequence of SEQ ID NO:225 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:276 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:21, an HC-CDR2 having the amino acid sequence of SEQ ID NO:72, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:123, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:174, an LC-CDR2 having the amino acid sequence of SEQ ID NO:225, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:276, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:21, an HC-CDR2 having the amino acid sequence of SEQ ID NO:72, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:123; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:174, an LC-CDR2 having the amino acid sequence of SEQ ID NO:225, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:276.

[0218] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:22 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:73 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:124, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 175 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 226 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 277, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 22, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 73, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 124, or a variant thereof in the HC-CDR sequence comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 175, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 226, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 277, or a variant thereof in the LC-CDR sequence comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 22, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 73, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 124; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 175, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 226, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 277.

[0219] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:23 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:74 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:125 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:176 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:227 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:278 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:23, an HC-CDR2 having the amino acid sequence of SEQ ID NO:74, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:125, or a variant thereof in which the HC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:176, an LC-CDR2 having the amino acid sequence of SEQ ID NO:227, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:278, or a variant thereof in which the LC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:23, an HC-CDR2 having the amino acid sequence of SEQ ID NO:74, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:125; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:176, an LC-CDR2 having the amino acid sequence of SEQ ID NO:227, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:278.

[0220] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:24 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:75 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:126 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:177 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:228 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:279 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:24, an HC-CDR2 having the amino acid sequence of SEQ ID NO:75, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:126, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequence; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:177, an LC-CDR2 having the amino acid sequence of SEQ ID NO:228, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:279, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H, which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:24, HC-CDR2 with the amino acid sequence of SEQ ID NO:75, and HC-CDR3 with the amino acid sequence of SEQ ID NO:126; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:177, LC-CDR2 with the amino acid sequence of SEQ ID NO:228, and LC-CDR3 with the amino acid sequence of SEQ ID NO:279.

[0221] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:25 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:76 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:127 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ IDNO:178 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:229 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:280 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:25, HC-CDR2 with the amino acid sequence of SEQ ID NO:76, and HC-CDR3 with the amino acid sequence of SEQ ID NO:127, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:178, an LC-CDR2 having the amino acid sequence of SEQ ID NO:229, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:280, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:25, an HC-CDR2 having the amino acid sequence of SEQ ID NO:76, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:127; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:178, an LC-CDR2 having the amino acid sequence of SEQ ID NO:229, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:280.

[0222] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:26 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:77 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:128 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:179 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:230 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:281 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:26, an HC-CDR2 having the amino acid sequence of SEQ ID NO:77, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:128, or an HC-CDR sequence comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:179, an LC-CDR2 having the amino acid sequence of SEQ ID NO:230, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:281, or a variant thereof having an LC-CDR sequence comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:26, an HC-CDR2 having the amino acid sequence of SEQ ID NO:77, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:128; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:179, an LC-CDR2 having the amino acid sequence of SEQ ID NO:230, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:281.

[0223] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:27 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:78 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:129 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 180 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 231 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 282 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 27, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 78, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 129, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 180, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 231, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 282, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 27, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 78, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 129; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 180, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 231, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 282.

[0224] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:28 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:79 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:130 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:181 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:232 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:283 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:28, an HC-CDR2 having the amino acid sequence of SEQ ID NO:79, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:130, or a variant thereof in which the HC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:181, an LC-CDR2 having the amino acid sequence of SEQ ID NO:232, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:283, or a variant thereof in which the LC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:28, an HC-CDR2 having the amino acid sequence of SEQ ID NO:79, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:130; and ii) V L, which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:181, LC-CDR2 with the amino acid sequence of SEQ ID NO:232, and LC-CDR3 with the amino acid sequence of SEQ ID NO:283.

[0225] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:29 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:80 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:131 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ IDNO:182 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:233 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:284 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:29, HC-CDR2 with the amino acid sequence of SEQ ID NO:80, and HC-CDR3 with the amino acid sequence of SEQ ID NO:131, or a variant thereof with up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequence; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQID NO:182, LC-CDR2 with the amino acid sequence of SEQ ID NO:233, and LC-CDR3 with the amino acid sequence of SEQ ID NO:284, or a variant thereof with up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H, which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:29, HC-CDR2 having the amino acid sequence of SEQ ID NO:80, and HC-CDR3 having the amino acid sequence of SEQ ID NO:131; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:182, LC-CDR2 having the amino acid sequence of SEQ ID NO:233, and LC-CDR3 having the amino acid sequence of SEQ ID NO:284.

[0226] In some embodiments, the anti-nGPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:30 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:81 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:132 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ IDNO:183 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO:234 or a variant thereof having at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO:285 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:30, HC-CDR2 having the amino acid sequence of SEQ ID NO:81, and HC-CDR3 having the amino acid sequence of SEQ ID NO:132, or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:183, an LC-CDR2 having the amino acid sequence of SEQ ID NO:234, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:285, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:30, an HC-CDR2 having the amino acid sequence of SEQ ID NO:81, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:132; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:183, an LC-CDR2 having the amino acid sequence of SEQ ID NO:234, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:285.

[0227] In some embodiments, the anti-nGPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:31 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:82 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:133 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:184 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:235 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:286 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-nGPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:31, an HC-CDR2 having the amino acid sequence of SEQ ID NO:82, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:133, or a variant thereof wherein the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQID NO:184, an LC-CDR2 having the amino acid sequence of SEQ ID NO:235, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:286, or a variant thereof wherein the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-nGPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:31, an HC-CDR2 having the amino acid sequence of SEQ ID NO:82, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:133; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:184, an LC-CDR2 having the amino acid sequence of SEQ ID NO:235, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:286.

[0228] In some embodiments, the anti-nGPC3 antibody moiety comprises: a) a V comprising the amino acid sequence SEQ ID NO:307 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:307; and b) a V comprising the amino acid sequence SEQ ID NO:358 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:358. In some embodiments, the anti-nGPC3 antibody moiety comprises: a) a V comprising the amino acid sequence SEQ ID NO:307 H ; and b) a V comprising the amino acid sequence SEQ ID NO:358 L . In some embodiments, the anti-nGPC3 antibody moiety comprises a V containing the amino acid sequence SEQ ID NO:307H HC-CDR of, and V containing the amino acid sequence SEQ ID NO:358 L LC-CDR of.

[0229] In some embodiments, the anti-nGPC3 antibody portion comprises: a) V comprising the amino acid sequence SEQ ID NO:308 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:308; and b) V comprising the amino acid sequence SEQ ID NO:359 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:359. In some embodiments, the anti-nGPC3 antibody portion comprises: a) V comprising the amino acid sequence SEQ ID NO:308 H ; and b) V comprising the amino acid sequence SEQ ID NO:359 L . In some embodiments, the anti-nGPC3 antibody portion comprises HC-CDR of V containing the amino acid sequence SEQ ID NO:308 H and LC-CDR of V containing the amino acid sequence SEQ ID NO:359 L .

[0230] In some embodiments, the anti-nGPC3 antibody portion comprises: a) V comprising the amino acid sequence SEQ ID NO:309 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:309; and b) V comprising the amino acid sequence SEQ ID NO:360 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:360. In some embodiments, the anti-nGPC3 antibody portion comprises: a) V comprising the amino acid sequence SEQ ID NO:309 H ; and b) V comprising the amino acid sequence SEQ ID NO:360 L . In some embodiments, the anti-nGPC3 antibody portion comprises V containing the amino acid sequence SEQ ID NO:309 Hof the HC-CDR, and the V containing the amino acid sequence SEQ ID NO:360 L of the LC-CDR.

[0231] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:310 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:310; and b) a V comprising the amino acid sequence SEQ ID NO:361 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:361. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:310 H ; and b) a V comprising the amino acid sequence SEQ ID NO:361 L . In some embodiments, the anti-nGPC3 antibody portion comprises a HC-CDR containing the amino acid sequence SEQ ID NO:310 H and a LC-CDR containing the amino acid sequence SEQ ID NO:361 L .

[0232] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:311 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:311; and b) a V comprising the amino acid sequence SEQ ID NO:362 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:362. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:311 H ; and b) a V comprising the amino acid sequence SEQ ID NO:362 L . In some embodiments, the anti-nGPC3 antibody portion comprises a HC-CDR containing the amino acid sequence SEQ ID NO:311 H and a V containing the amino acid sequence SEQ ID NO:362L of LC-CDR.

[0233] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:312 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:312; and b) a V comprising the amino acid sequence SEQ ID NO:363 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:363. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:312 H ; and b) a V comprising the amino acid sequence SEQ ID NO:363 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:312 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:363 of V L .

[0234] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:313 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:313; and b) a V comprising the amino acid sequence SEQ ID NO:364 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:364. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:313 H ; and b) a V comprising the amino acid sequence SEQ ID NO:364 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:313 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:364 of V L .

[0235] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:314 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:314; and b) a V comprising the amino acid sequence SEQ ID NO:365 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:365. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:314 H ; and b) a V comprising the amino acid sequence SEQ ID NO:365 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:314 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:365 of V L .

[0236] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:315 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:315; and b) a V comprising the amino acid sequence SEQ ID NO:366 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:366. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:315 H ; and b) a V comprising the amino acid sequence SEQ ID NO:366 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:315 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:366 of V L .

[0237] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:316 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:316; and b) a V comprising the amino acid sequence SEQ ID NO:367 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:367. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:316 H ; and b) a V comprising the amino acid sequence SEQ ID NO:367 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:316 H and an LC-CDR containing the amino acid sequence SEQ ID NO:367 L .

[0238] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:317 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:317; and b) a V comprising the amino acid sequence SEQ ID NO:368 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:368. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:317 H ; and b) a V comprising the amino acid sequence SEQ ID NO:368 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:317 H and an LC-CDR containing the amino acid sequence SEQ ID NO:368 L .

[0239] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:318 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:318; and b) a V comprising the amino acid sequence SEQ ID NO:369 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:369. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:318 H ; and b) a V comprising the amino acid sequence SEQ ID NO:369 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:318 H and an LC-CDR containing the amino acid sequence SEQ ID NO:369 L .

[0240] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:319 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:319; and b) a V comprising the amino acid sequence SEQ ID NO:370 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:370. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:319 H ; and b) a V comprising the amino acid sequence SEQ ID NO:370 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:319 H and an LC-CDR containing the amino acid sequence SEQ ID NO:370 L .

[0241] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:320 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:320; and b) a V comprising the amino acid sequence SEQ ID NO:371 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:371. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:320 H ; and b) a V comprising the amino acid sequence SEQ ID NO:371 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:320 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:371 L of V.

[0242] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:321 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:321; and b) a V comprising the amino acid sequence SEQ ID NO:372 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:372. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:321 H ; and b) a V comprising the amino acid sequence SEQ ID NO:372 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:321 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:372 L of V.

[0243] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:322 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:322; and b) a V comprising the amino acid sequence SEQ ID NO:373 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:373. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:322 H ; and b) a V comprising the amino acid sequence SEQ ID NO:373 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:322 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:373 L of V.

[0244] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:323 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:323; and b) a V comprising the amino acid sequence SEQ ID NO:374 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:374. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:323 H ; and b) a V comprising the amino acid sequence SEQ ID NO:374 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:323 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:374 L of V.

[0245] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:324 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:324; and b) a V comprising the amino acid sequence SEQ ID NO:375 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:375. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:324 H ; and b) a V comprising the amino acid sequence SEQ ID NO:375 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:324 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:375 of V L .

[0246] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:325 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:325; and b) a V comprising the amino acid sequence SEQ ID NO:376 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:376. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:325 H ; and b) a V comprising the amino acid sequence SEQ ID NO:376 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:325 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:376 of V L .

[0247] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:326 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:326; and b) a V comprising the amino acid sequence SEQ ID NO:377 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:377. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:326 H ; and b) a V comprising the amino acid sequence SEQ ID NO:377 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:326 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:377 of V L .

[0248] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:327 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:327; and b) a V comprising the amino acid sequence SEQ ID NO:378 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:378. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:327 H ; and b) a V comprising the amino acid sequence SEQ ID NO:378 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:327 of V H , and an LC-CDR containing the amino acid sequence SEQ ID NO:378 of V L .

[0249] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:328 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:328; and b) a V comprising the amino acid sequence SEQ ID NO:379 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:379. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:328 H ; and b) a V comprising the amino acid sequence SEQ ID NO:379 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:328 H and an LC-CDR containing the amino acid sequence SEQ ID NO:379 L .

[0250] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:329 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:329; and b) a V comprising the amino acid sequence SEQ ID NO:380 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:380. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:329 H ; and b) a V comprising the amino acid sequence SEQ ID NO:380 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:329 H and an LC-CDR containing the amino acid sequence SEQ ID NO:380 L .

[0251] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:330 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:330; and b) a V comprising the amino acid sequence SEQ ID NO:381 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:381. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:330 H ; and b) a V comprising the amino acid sequence SEQ ID NO:381 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:330 H and an LC-CDR containing the amino acid sequence SEQ ID NO:381 L .

[0252] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:331 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:331; and b) a V comprising the amino acid sequence SEQ ID NO:382 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:382. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:331 H ; and b) a V comprising the amino acid sequence SEQ ID NO:382 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:331 H and an LC-CDR containing the amino acid sequence SEQ ID NO:382 L .

[0253] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:332 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:332; and b) a V comprising the amino acid sequence SEQ ID NO:383 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:383. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:332 H ; and b) a V comprising the amino acid sequence SEQ ID NO:383 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:332 H and an LC-CDR containing the amino acid sequence SEQ ID NO:383 L .

[0254] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:333 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:333; and b) a V comprising the amino acid sequence SEQ ID NO:384 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:384. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:333 H ; and b) a V comprising the amino acid sequence SEQ ID NO:384 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:333 H and an LC-CDR containing the amino acid sequence SEQ ID NO:384 L .

[0255] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:334 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:334; and b) a V comprising the amino acid sequence SEQ ID NO:385 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:385. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:334 H ; and b) a V comprising the amino acid sequence SEQ ID NO:385 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:334 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:385 L of V.

[0256] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:335 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:335; and b) a V comprising the amino acid sequence SEQ ID NO:386 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:386. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:335 H ; and b) a V comprising the amino acid sequence SEQ ID NO:386 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:335 H of V, and an LC-CDR containing the amino acid sequence SEQ ID NO:386 L of V.

[0257] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:336 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:336; and b) a V comprising the amino acid sequence SEQ ID NO:387 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:387. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:336 H ; and b) a V comprising the amino acid sequence SEQ ID NO:387 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:336 H and an LC-CDR containing the amino acid sequence SEQ ID NO:387 L .

[0258] In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:337 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:337; and b) a V comprising the amino acid sequence SEQ ID NO:388 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO:388. In some embodiments, the anti-nGPC3 antibody portion comprises: a) a V comprising the amino acid sequence SEQ ID NO:337 H ; and b) a V comprising the amino acid sequence SEQ ID NO:388 L . In some embodiments, the anti-nGPC3 antibody portion comprises an HC-CDR containing the amino acid sequence SEQ ID NO:337 H and an LC-CDR containing the amino acid sequence SEQ ID NO:388 L .

[0259] In some embodiments, the anti-nGPC3 antibody portion competes with a second anti-nGPC3 antibody portion for binding to GPC3 bound to the cell surface, where the second anti-nGPC3 antibody portion is any of the anti-nGPC3 antibody portions described herein. In some embodiments, the anti-nGPC3 antibody portion binds to the same or substantially the same epitope as the second anti-nGPC3 antibody portion. In some embodiments, binding of the anti-nGPC3 antibody portion to GPC3 bound to the cell surface inhibits binding of the second anti-nGPC3 antibody portion to the same GPC3 bound to the cell surface by at least about 70% (e.g., any of at least about 75%, 80%, 85%, 90%, 95%, 98%, or 99%) or vice versa. In some embodiments, the anti-nGPC3 antibody portion cross-competes with the second anti-nGPC3 antibody portion for binding to GPC3 bound to the cell surface, i.e., each of the anti-nGPC3 antibody portions competes with the other for binding to GPC3 bound to the cell surface.

[0260] Anti - GPC3 antibody portion that specifically recognizes GPC3

[0261] In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3. In some embodiments, the anti-GPC3 antibody portion specifically recognizes GPC3 bound to the cell surface. In some embodiments, the anti-GPC3 antibody portion specifically recognizes soluble GPC3. In some embodiments, the anti-sGPC3 antibody portion has a higher binding affinity for soluble GPC3 than for GPC3 bound to the cell surface. In some embodiments, the anti-sGPC3 antibody portion specifically recognizes an epitope within the amino acid sequence of SEQ ID NO: 461. In some embodiments, the anti-sGPC3 antibody portion specifically recognizes an epitope within amino acids 510 - 560 of SEQ ID NO: 460 (SEQ ID NO: 467). In some embodiments, the anti-GPC3 antibody portion specifically recognizes both GPC3 bound to the cell surface and soluble GPC3.

[0262] In some embodiments, the anti-GPC3 antibody portion comprises i) a heavy chain variable domain (V H ), which comprises an HC-CDR3 having an amino acid sequence of any of SEQ ID NOs: 134 - 153 or a variant thereof having at most about 5 (e.g., any of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) a light chain variable domain (V L ), which comprises an LC-CDR3 having an amino acid sequence of any of SEQ ID NOs: 287 - 306 or a variant thereof having at most about 5 (e.g., any of about 1, 2, 3, 4, or 5) amino acid substitutions.

[0263] In some embodiments, the anti-GPC3 antibody portion comprises i) VH which comprises an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153; and ii) V L which comprises an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306.

[0264] In some embodiments, the anti-GPC3 antibody moiety comprises i) V H which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 32-51 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 83-102 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 185-204 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 236-255 or a variant thereof having at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions.

[0265] In some embodiments, the anti-GPC3 antibody moiety comprises i) V H which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 32-51 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 83-102 or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153; and ii) V L, which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 185-204 or a variant thereof having at most about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 236-255 or a variant thereof having at most about 3 (e.g., any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 287-306.

[0266] In some embodiments, the anti-GPC3 antibody moiety comprises i) a V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 32-51, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 83-102, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 134-153, or a variant thereof having at most about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) a V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 185-204, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 236-255, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 287-306, or a variant thereof having at most about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences.

[0267] In some embodiments, the anti-GPC3 antibody moiety comprises i) a V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NOs: 32-51, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NOs: 83-102, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NOs: 134-153, or a variant thereof having at most about 5 (e.g., any one of about 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions occur in HC-CDR1 or HC-CDR2; and ii) a V L, which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 185-204, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 236-255, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306, or a variant thereof having at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, wherein the amino acid substitutions occur in LC-CDR1 or LC-CDR2.

[0268] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 32-51, an HC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 83-102, and an HC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 134-153; and ii) V L , which comprises an LC-CDR1 having an amino acid sequence of any one of SEQ ID NO: 185-204, an LC-CDR2 having an amino acid sequence of any one of SEQ ID NO: 236-255, and an LC-CDR3 having an amino acid sequence of any one of SEQ ID NO: 287-306.

[0269] In some embodiments, the anti-GPC3 antibody portion comprises: a) V H , which comprises the amino acid sequence of any one of SEQ ID NO: 32-51, the amino acid sequence of any one of SEQ ID NO: 83-102, and the amino acid sequence of any one of SEQ ID NO: 134-153; and ii) V L , which comprises the amino acid sequence of any one of SEQ ID NO: 185-204, the amino acid sequence of any one of SEQ ID NO: 236-255, and the amino acid sequence of any one of SEQ ID NO: 287-306.

[0270] In some embodiments, the anti-GPC3 antibody portion comprises: a) V H , which comprises one, two, or three CDRs of any one of SEQ ID NO: 338-357, and b) V L , which comprises one, two, or three CDRs of any one of SEQ ID NO: 389-408. In some embodiments, the anti-nGPC3 antibody portion comprises: a) V H, which comprises HC-CDR1, HC-CDR2 and HC-CDR3 of the heavy chain variable domain of any one of SEQ ID NO: 389-408, and b) VL, which comprises LC-CDR1, LC-CDR2 and LC-CDR3 of the light chain variable domain of any one of SEQ ID NO: 358-388.

[0271] In some embodiments, the anti-GPC3 antibody portion comprises: a) V H , which comprises the amino acid sequence of any one of SEQ ID NO: 338-357, or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with any one of SEQ ID NO: 338-357; and b) V L , which comprises the amino acid sequence of any one of SEQ ID NO: 389-408, or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with any one of SEQ ID NO: 389-408.

[0272] In some embodiments, the anti-GPC3 antibody portion comprises: a) V comprising the amino acid sequence of any one of SEQ ID NO: 338-357 H ; and b) V comprising the amino acid sequence of any one of SEQ ID NO: 389-408 L .

[0273] In some embodiments, the anti-GPC3 antibody portion comprises the HC-CDR of V having the amino acid sequence of any one of SEQ ID NO: 338-357 H , and the LC-CDR of V having the amino acid sequence of any one of SEQ ID NO: 389-408 L .

[0274] The heavy and light chain variable domains can be combined in various pairwise combinations to produce a variety of anti-GPC3 antibody portions. Exemplary anti-GPC3 antibody portions are provided in Tables 8 and 9.

[0275] Table 8 CDR Sequences of Anti-GPC3 Antibody Portions

[0276]

[0277]

[0278] Table 9 Anti-GPC3 Antibody Portion V H / V LSequence (CDR sequence Add underline )

[0279]

[0280]

[0281]

[0282]

[0283]

[0284] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO: 32 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO: 83 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 134 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO: 185 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO: 236 or a variant thereof comprising at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO: 287 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO: 32, HC-CDR2 having the amino acid sequence of SEQ ID NO: 83, and HC-CDR3 having the amino acid sequence of SEQ ID NO: 134, or a variant thereof in which the HC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:185, an LC-CDR2 having the amino acid sequence of SEQ ID NO:236, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:287, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:32, an HC-CDR2 having the amino acid sequence of SEQ ID NO:83, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:134; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:185, an LC-CDR2 having the amino acid sequence of SEQ ID NO:236, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:287.

[0285] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:33 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:84 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:135 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:186 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:237 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:288 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:33, an HC-CDR2 having the amino acid sequence of SEQ ID NO:84, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:135, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:186, an LC-CDR2 having the amino acid sequence of SEQ ID NO:237, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:288, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:33, an HC-CDR2 having the amino acid sequence of SEQ ID NO:84, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:135; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:186, an LC-CDR2 having the amino acid sequence of SEQ ID NO:237, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:288.

[0286] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:34 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:85 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:136 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 187 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 238 or a variant thereof comprising at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 289 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 85, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 136, or a variant thereof in which the HC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 187, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 238, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 289, or a variant thereof in which the LC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 34, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 85, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 136; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 187, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 238, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 289.

[0287] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:35 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:86 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:137 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:188 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:239 or a variant thereof comprising up to about 3 (such as any one of about 1, 2 or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:290 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:35, an HC-CDR2 having the amino acid sequence of SEQ ID NO:86, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:137, or a variant thereof in which the HC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:188, an LC-CDR2 having the amino acid sequence of SEQ ID NO:239, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:290, or a variant thereof in which the LC-CDR sequences comprise up to about 5 (such as any one of about 1, 2, 3, 4 or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:35, an HC-CDR2 having the amino acid sequence of SEQ ID NO:86, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:137; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:188, an LC-CDR2 having the amino acid sequence of SEQ ID NO:239, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:290.

[0288] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:36 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:87 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:138 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:189 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:240 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:291 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:36, an HC-CDR2 having the amino acid sequence of SEQ ID NO:87, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:138, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:189, an LC-CDR2 having the amino acid sequence of SEQ ID NO:240, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:291, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H, which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:36, HC-CDR2 having the amino acid sequence of SEQ ID NO:87, and HC-CDR3 having the amino acid sequence of SEQ ID NO:138; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:189, LC-CDR2 having the amino acid sequence of SEQ ID NO:240, and LC-CDR3 having the amino acid sequence of SEQ ID NO:291.

[0289] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:37 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:88 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:139 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ IDNO:190 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO:241 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO:292 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:37, HC-CDR2 having the amino acid sequence of SEQ ID NO:88, and HC-CDR3 having the amino acid sequence of SEQ ID NO:139, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:190, an LC-CDR2 having the amino acid sequence of SEQ ID NO:241, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:292, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:37, an HC-CDR2 having the amino acid sequence of SEQ ID NO:88, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:139; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:190, an LC-CDR2 having the amino acid sequence of SEQ ID NO:241, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:292.

[0290] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:38 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:89 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:140 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:191 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:242 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:293 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:38, an HC-CDR2 having the amino acid sequence of SEQ ID NO:89, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:140, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQID NO:191, an LC-CDR2 having the amino acid sequence of SEQ ID NO:242, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:293, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:38, an HC-CDR2 having the amino acid sequence of SEQ ID NO:89, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:140; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:191, an LC-CDR2 having the amino acid sequence of SEQ ID NO:242, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:293.

[0291] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:39 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:90 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:141 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 192 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 243 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 294 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 39, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 90, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 141, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 192, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 243, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 294, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 39, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 90, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 141; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 192, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 243, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 294.

[0292] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:40 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:91 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:142 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:193 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:244 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:295 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:40, an HC-CDR2 having the amino acid sequence of SEQ ID NO:91, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:142, or an HC-CDR sequence comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:193, an LC-CDR2 having the amino acid sequence of SEQ ID NO:244, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:295, or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequence. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:40, an HC-CDR2 having the amino acid sequence of SEQ ID NO:91, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:142; and ii) V L, which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:193, an LC-CDR2 with the amino acid sequence of SEQ ID NO:244, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:295.

[0293] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:41 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 with the amino acid sequence of SEQ ID NO:92 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:143 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:194 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 with the amino acid sequence of SEQ ID NO:245 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:296 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 with the amino acid sequence of SEQ ID NO:41, an HC-CDR2 with the amino acid sequence of SEQ ID NO:92, and an HC-CDR3 with the amino acid sequence of SEQ ID NO:143, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 with the amino acid sequence of SEQ ID NO:194, an LC-CDR2 with the amino acid sequence of SEQ ID NO:245, and an LC-CDR3 with the amino acid sequence of SEQ ID NO:296, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H, which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:41, HC-CDR2 having the amino acid sequence of SEQ ID NO:92, and HC-CDR3 having the amino acid sequence of SEQ ID NO:143; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ ID NO:194, LC-CDR2 having the amino acid sequence of SEQ ID NO:245, and LC-CDR3 having the amino acid sequence of SEQ ID NO:296.

[0294] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:42 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 having the amino acid sequence of SEQ ID NO:93 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 having the amino acid sequence of SEQ ID NO:144 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 having the amino acid sequence of SEQ IDNO:195 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 having the amino acid sequence of SEQ ID NO:246 or a variant thereof having up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 having the amino acid sequence of SEQ ID NO:297 or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises HC-CDR1 having the amino acid sequence of SEQ ID NO:42, HC-CDR2 having the amino acid sequence of SEQ ID NO:93, and HC-CDR3 having the amino acid sequence of SEQ ID NO:144, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:195, an LC-CDR2 having the amino acid sequence of SEQ ID NO:246, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:297, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:42, an HC-CDR2 having the amino acid sequence of SEQ ID NO:93, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:144; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:195, an LC-CDR2 having the amino acid sequence of SEQ ID NO:246, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:297.

[0295] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:43 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:94 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:145 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:196 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:247 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:298 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:43, an HC-CDR2 having the amino acid sequence of SEQ ID NO:94, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:145, or a variant thereof wherein the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:196, an LC-CDR2 having the amino acid sequence of SEQ ID NO:247, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:298, or a variant thereof wherein the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:43, an HC-CDR2 having the amino acid sequence of SEQ ID NO:94, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:145; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:196, an LC-CDR2 having the amino acid sequence of SEQ ID NO:247, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:298.

[0296] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:44 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:95 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:146 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 197 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 248 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 299 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 44, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 95, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 146, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 197, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 248, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 299, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 44, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 95, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 146; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 197, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 248, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 299.

[0297] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:45 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:96 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:147 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:198 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:249 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:300 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:45, an HC-CDR2 having the amino acid sequence of SEQ ID NO:96, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:147, or a variant thereof having up to about 5 (such as about 1, 2, 3, 4, or 5) amino acid substitutions in the HC-CDR sequences; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:198, an LC-CDR2 having the amino acid sequence of SEQ ID NO:249, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:300, or a variant thereof having up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions in the LC-CDR sequences. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:45, an HC-CDR2 having the amino acid sequence of SEQ ID NO:96, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:147; and ii) V L, which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:198, LC-CDR2 with the amino acid sequence of SEQ ID NO:249, and LC-CDR3 with the amino acid sequence of SEQ ID NO:300.

[0298] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:46 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:97 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:148 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ IDNO:199 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:250 or a variant thereof comprising at most about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:301 or a variant thereof comprising at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:46, HC-CDR2 with the amino acid sequence of SEQ ID NO:97, and HC-CDR3 with the amino acid sequence of SEQ ID NO:148, or a variant thereof in which the HC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQID NO:199, LC-CDR2 with the amino acid sequence of SEQ ID NO:250, and LC-CDR3 with the amino acid sequence of SEQ ID NO:301, or a variant thereof in which the LC-CDR sequence comprises at most about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:46, an HC-CDR2 having the amino acid sequence of SEQ ID NO:97, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:148; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:199, an LC-CDR2 having the amino acid sequence of SEQ ID NO:250, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:301.

[0299] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:47 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:98 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:149 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:200 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:251 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:302 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:47, an HC-CDR2 having the amino acid sequence of SEQ ID NO:98, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:149, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:200, an LC-CDR2 having the amino acid sequence of SEQ ID NO:251, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:302, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:47, an HC-CDR2 having the amino acid sequence of SEQ ID NO:98, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:149; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:200, an LC-CDR2 having the amino acid sequence of SEQ ID NO:251, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:302.

[0300] In some embodiments, the anti-GPC3 antibody portion comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:48 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:99 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:150 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ IDNO:201 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:252 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:303 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:48, an HC-CDR2 having the amino acid sequence of SEQ ID NO:99, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:150, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:201, an LC-CDR2 having the amino acid sequence of SEQ ID NO:252, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:303, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:48, an HC-CDR2 having the amino acid sequence of SEQ ID NO:99, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:150; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:201, an LC-CDR2 having the amino acid sequence of SEQ ID NO:252, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:303.

[0301] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:49 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:100 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:151 or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L, which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 202 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 253 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 304 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 49, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 100, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 151, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 202, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 253, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 304, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO: 49, an HC-CDR2 having the amino acid sequence of SEQ ID NO: 100, and an HC-CDR3 having the amino acid sequence of SEQ ID NO: 151; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO: 202, an LC-CDR2 having the amino acid sequence of SEQ ID NO: 253, and an LC-CDR3 having the amino acid sequence of SEQ ID NO: 304.

[0302] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H, which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:50 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an HC-CDR2 having the amino acid sequence of SEQ ID NO:101 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:152 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:203 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, an LC-CDR2 having the amino acid sequence of SEQ ID NO:254 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:305 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody portion comprises: (1) i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:50, an HC-CDR2 having the amino acid sequence of SEQ ID NO:101, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:152, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises an LC-CDR1 having the amino acid sequence of SEQ ID NO:203, an LC-CDR2 having the amino acid sequence of SEQ ID NO:254, and an LC-CDR3 having the amino acid sequence of SEQ ID NO:305, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitution occurs in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitution occurs in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody portion comprises i) V H , which comprises an HC-CDR1 having the amino acid sequence of SEQ ID NO:50, an HC-CDR2 having the amino acid sequence of SEQ ID NO:101, and an HC-CDR3 having the amino acid sequence of SEQ ID NO:152; and ii) V L, which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:203, LC-CDR2 with the amino acid sequence of SEQ ID NO:254, and LC-CDR3 with the amino acid sequence of SEQ ID NO:305.

[0303] In some embodiments, the anti-GPC3 antibody moiety comprises: i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:51 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, HC-CDR2 with the amino acid sequence of SEQ ID NO:102 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, and HC-CDR3 with the amino acid sequence of SEQ ID NO:153 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:204 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions, LC-CDR2 with the amino acid sequence of SEQ ID NO:255 or a variant thereof comprising up to about 3 (such as any one of about 1, 2, or 3) amino acid substitutions, and LC-CDR3 with the amino acid sequence of SEQ ID NO:306 or a variant thereof comprising up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the anti-GPC3 antibody moiety comprises: (1) i) V H , which comprises HC-CDR1 with the amino acid sequence of SEQ ID NO:51, HC-CDR2 with the amino acid sequence of SEQ ID NO:102, and HC-CDR3 with the amino acid sequence of SEQ ID NO:153, or a variant thereof in which the HC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions; and ii) V L , which comprises LC-CDR1 with the amino acid sequence of SEQ ID NO:204, LC-CDR2 with the amino acid sequence of SEQ ID NO:255, and LC-CDR3 with the amino acid sequence of SEQ ID NO:306, or a variant thereof in which the LC-CDR sequence comprises up to about 5 (such as any one of about 1, 2, 3, 4, or 5) amino acid substitutions. In some embodiments, the amino acid substitutions occur in HC-CDR1 or HC-CDR2. In some embodiments, the amino acid substitutions occur in LC-CDR1 or LC-CDR2. In some embodiments, the anti-GPC3 antibody moiety comprises i) V H, which includes HC-CDR1 with the amino acid sequence of SEQ ID NO:51, HC-CDR2 with the amino acid sequence of SEQ ID NO:102, and HC-CDR3 with the amino acid sequence of SEQ ID NO:153; and ii) V L , which includes LC-CDR1 with the amino acid sequence of SEQ ID NO:204, LC-CDR2 with the amino acid sequence of SEQ ID NO:255, and LC-CDR3 with the amino acid sequence of SEQ ID NO:306.

[0304] In some embodiments, the anti-GPC3 antibody portion includes: a) V including the amino acid sequence SEQ ID NO:338 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:338; and b) V including the amino acid sequence SEQ ID NO:389 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:389. In some embodiments, the anti-GPC3 antibody portion includes: a) V including the amino acid sequence SEQ ID NO:338 H ; and b) V including the amino acid sequence SEQ ID NO:389 L . In some embodiments, the anti-GPC3 antibody portion includes the HC-CDR of V with the amino acid sequence of SEQ ID NO:338 H and the LC-CDR of V with the amino acid sequence of SEQ ID NO:389 L .

[0305] In some embodiments, the anti-GPC3 antibody portion includes: a) V with the amino acid sequence of SEQ ID NO:339 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:339; and b) V with the amino acid sequence of SEQ ID NO:390 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO:390. In some embodiments, the anti-GPC3 antibody portion includes: a) V with the amino acid sequence of SEQ ID NO:339H ; and b) a V with the amino acid sequence of SEQ ID NO:390 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with the amino acid sequence of SEQ ID NO:339 H of HC-CDR, and a V with the amino acid sequence of SEQ IDNO:390 L of LC-CDR.

[0306] . In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:340 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO:340; and b) a V with the amino acid sequence of SEQ ID NO:391 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO:391. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:340 H ; and b) a V with the amino acid sequence of SEQ ID NO:391 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with the amino acid sequence of SEQ ID NO:340 H of HC-CDR, and a V with the amino acid sequence of SEQ IDNO:391 L of LC-CDR.

[0307] . In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:341 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO:341; and b) a V with the amino acid sequence of SEQ ID NO:392 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO:392. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:341 H ; and b) a V with the amino acid sequence of SEQ ID NO:392 L。In some embodiments, the anti-GPC3 antibody portion comprises a V with the amino acid sequence of SEQ ID NO:341 H of HC-CDR, and a V with the amino acid sequence of SEQ ID NO:392 L of LC-CDR.

[0308] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:342 H or a variant thereof having at least about 80% (including any one of, for example, at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:342; and b) a V with the amino acid sequence of SEQ ID NO:393 L or a variant thereof having at least about 80% (including any one of, for example, at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:393. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:342 H ; and b) a V with the amino acid sequence of SEQ ID NO:393 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with the amino acid sequence of SEQ ID NO:342 H of HC-CDR, and a V with the amino acid sequence of SEQ ID NO:393 L of LC-CDR.

[0309] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:343 H or a variant thereof having at least about 80% (including any one of, for example, at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:343; and b) a V comprising the amino acid sequence SEQ ID NO:394 L or a variant thereof having at least about 80% (including any one of, for example, at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO:394. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO:343 H ; and b) a V with the amino acid sequence of SEQ ID NO:394 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with the amino acid sequence of SEQ ID NO:343H HC-CDR with the amino acid sequence of SEQ ID NO: 394, and V with the amino acid sequence of SEQ ID NO: 394 L LC-CDR

[0310] In some embodiments, the anti-GPC3 antibody portion comprises: a) V with the amino acid sequence of SEQ ID NO: 344 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO: 344; and b) V with the amino acid sequence of SEQ ID NO: 395 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO: 395. In some embodiments, the anti-GPC3 antibody portion comprises: a) V with the amino acid sequence of SEQ ID NO: 344 H ; and b) V with the amino acid sequence of SEQ ID NO: 395 L . In some embodiments, the anti-GPC3 antibody portion comprises V with the amino acid sequence of SEQ ID NO: 344 H HC-CDR, and V with the amino acid sequence of SEQ ID NO: 395 L LC-CDR

[0311] In some embodiments, the anti-GPC3 antibody portion comprises: a) V with the amino acid sequence of SEQ ID NO: 345 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO: 345; and b) V with the amino acid sequence of SEQ ID NO: 396 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity with SEQ ID NO: 396. In some embodiments, the anti-GPC3 antibody portion comprises: a) V with the amino acid sequence of SEQ ID NO: 345 H ; and b) V with the amino acid sequence of SEQ ID NO: 396 L . In some embodiments, the anti-GPC3 antibody portion comprises V with the amino acid sequence of SEQ ID NO: 345 H HC-CDR, and V with the amino acid sequence of SEQ ID NO: 396 Lof LC-CDR.

[0312] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with an amino acid sequence of SEQ ID NO: 346 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 346; and b) a V with an amino acid sequence of SEQ ID NO: 397 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 397. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with an amino acid sequence of SEQ ID NO: 346 H ; and b) a V with an amino acid sequence of SEQ ID NO: 397 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with an amino acid sequence of SEQ ID NO: 346 H of HC-CDR, and a V with an amino acid sequence of SEQ ID NO: 397 L of LC-CDR.

[0313] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with an amino acid sequence of SEQ ID NO: 347 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 347; and b) a V with an amino acid sequence of SEQ ID NO: 398 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 398. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with an amino acid sequence of SEQ ID NO: 347 H ; and b) a V with an amino acid sequence of SEQ ID NO: 398 L . In some embodiments, the anti-GPC3 antibody portion comprises a V with an amino acid sequence of SEQ ID NO: 347 H of HC-CDR, and a V with an amino acid sequence of SEQ ID NO: 398 L of LC-CDR.

[0314] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO: 348 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 348; and b) a V with the amino acid sequence of SEQ ID NO: 399 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 399. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO: 348 H ; and b) a V with the amino acid sequence of SEQ ID NO: 399 L . In some embodiments, the anti-GPC3 antibody portion comprises the HC-CDR of the V with the amino acid sequence of SEQ ID NO: 348 H and the LC-CDR of the V with the amino acid sequence of SEQ ID NO: 399 L .

[0315] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO: 349 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 349; and b) a V with the amino acid sequence of SEQ ID NO: 400 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98%, or 99%) sequence identity to SEQ ID NO: 400. In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO: 349 H ; and b) a V with the amino acid sequence of SEQ ID NO: 400 L . In some embodiments, the anti-GPC3 antibody portion comprises the HC-CDR of the V with the amino acid sequence of SEQ ID NO: 349 H and the LC-CDR of the V with the amino acid sequence of SEQ ID NO: 400 L .

[0316] In some embodiments, the anti-GPC3 antibody portion comprises: a) a V with the amino acid sequence of SEQ ID NO: 350H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 350; and b) a V having the amino acid sequence of SEQ ID NO: 401 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 401. In some embodiments, the anti-GPC3 antibody moiety comprises: a) a V having the amino acid sequence of SEQ ID NO: 350 H ; and b) a V having the amino acid sequence of SEQ ID NO: 401 L . In some embodiments, the anti-GPC3 antibody moiety comprises a V having the amino acid sequence of SEQ ID NO: 350 H of the HC-CDR, and a V having the amino acid sequence of SEQ ID NO: 401 L of the LC-CDR.

[0317] In some embodiments, the anti-GPC3 antibody moiety comprises: a) a V having the amino acid sequence of SEQ ID NO: 351 H or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 351; and b) a V having the amino acid sequence of SEQ ID NO: 402 L or a variant thereof having at least about 80% (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) sequence identity to SEQ ID NO: 402. In some embodiments, the anti-GPC3 antibody moiety comprises: a) a V having the amino acid sequence of SEQ ID NO: 351 H ; and b) a V having the amino acid sequence of SEQ ID NO: 402 L . In some embodiments, the anti-GPC3 antibody moiety comprises a V having the amino acid sequence of SEQ ID NO: 351 H of the HC-CDR, and a V having the amino acid sequence of SEQ ID NO: 402 L of the LC-CDR.

[0318] In some embodiments, the anti-GPC3 antibody moiety comprises: a) a V having the amino acid sequence of SEQ ID NO: 352 Hor a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO: 352; and b) V having an amino acid sequence of SEQ ID NO: 403 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO: 403. In some embodiments, the anti-GPC3 antibody portion comprises: a) V having an amino acid sequence of SEQ ID NO: 352 H ; and b) V having an amino acid sequence of SEQ ID NO: 403 L . In some embodiments, the anti-GPC3 antibody portion comprises V having an amino acid sequence of SEQ ID NO: 352 H of the HC-CDR, and V having an amino acid sequence of SEQ ID NO: 403 L of the LC-CDR.

[0319] In some embodiments, the anti-GPC3 antibody portion comprises: a) V having an amino acid sequence of SEQ ID NO: 353 H or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO: 353; and b) V having an amino acid sequence of SEQ ID NO: 404 L or a variant thereof having at least about 80% sequence identity (including, for example, any one of at least about 80%, 85%, 90%, 95%, 96%, 97%, 98% or 99%) with SEQ ID NO: 404. In some embodiments, the anti-GPC3 antibody portion comprises: a) V having an amino acid sequence of SEQ ID NO: 353 H ; and b) V having an amino acid sequence of SEQ ID NO: 404 L . In some embodiments, the anti-GPC3 antibody portion comprises V having an amino acid sequence of SEQ ID NO: 353 H of the HC-CDR, and V having an amino acid sequence of SEQ ID NO: 404 L of the LC-CDR. ...

Claims

1. An isolated anti-GPC3 construct comprising an anti-GPC3 antibody portion that specifically recognizes cell surface-bound GPC3, wherein said antibody portion comprises: I.i) Heavy chain variable region V H , which comprises heavy chain complementarity determining regions HC-CDR1, HC-CDR2 and HC-CDR3, HC-CDR1 consists of the amino acid sequence of SEQ ID NO:3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:105; and ii) light chain variable region V L , which comprises light chain complementarity determining regions LC-CDR1, LC-CDR2 and LC-CDR3, LC-CDR1 consists of the amino acid sequence of SEQ ID NO:156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:258; II.i) V H , which includes HC - CDR1, HC - CDR2, and HC - CDR3. HC - CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC - CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii) V L , which includes LC - CDR1, LC - CDR2, and LC - CDR3. LC - CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC - CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO: 276; III.i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:24, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:75, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:126; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:177, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:228, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:279; IV.i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 1, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 52, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 103; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 154, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 205, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 256; V.i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 11, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 62, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 113; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 164, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 215, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 266; VI.i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:12, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:63, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:114; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:165, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:216, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:267; VII.i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 44, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 95, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 146; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 197, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 248, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 299; VIII.i)V H , which includes HC-CDR1, HC-CDR2 and HC-CDR3, and HC-CDR1 is composed of SEQ comprising the amino acid sequence of SEQ ID NO:28, HC-CDR2 comprising the amino acid sequence of SEQ ID NO:79, and HC-CDR3 comprising the amino acid sequence of SEQ ID NO:130; and ii) V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, LC-CDR1 comprising the amino acid sequence of SEQ ID NO:181, LC-CDR2 comprising the amino acid sequence of SEQ ID NO:232, and LC-CDR3 comprising the amino acid sequence of SEQ ID NO:283; IX.i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO:4, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:55, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:106; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO:157, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:208, and LC-CDR3 consists of SEQ the amino acid sequence of ID NO:259; X.i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:5, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:56, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:107; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:158, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:209, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:260; XI.i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3, where HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 30, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 81, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 132; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3, where LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 183, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 234, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 285; XII.i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 41, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 92, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 143; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 194, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 245, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 296; or XIII.i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 49, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 100, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 151; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 202, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 253, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 304; wherein the isolated anti-GPC3 construct is an anti-GPC3 single-chain Fv (scFv), an anti-GPC3 Fc fusion protein, a full-length anti-GPC3 antibody, an anti-GPC3 tandem bis-scFv bispecific T cell engager, an anti-GPC3 chimeric antigen receptor (CAR), an anti-GPC3 chimeric antibody-T cell receptor (TCR) (caTCR), or an anti-GPC3 costimulatory chimeric receptor (CSR); and wherein the anti-GPC3 tandem bis-scFv comprises two scFvs linked by a peptide linker, wherein the anti-GPC3 antibody portion is the first scFv, and wherein the second scFv is a second antibody portion that specifically recognizes CD3ε.

2. The isolated anti-GPC3 construct according to claim 1, wherein said anti-GPC3 antibody portion comprises: I.i)V H , which contains the amino acid sequence of SEQ ID NO: 327 and ii) V L , which contains SEQ ID NO: the amino acid sequence of 378; II.i)V H , which comprises the amino acid sequence of SEQ ID NO: 330 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 381; III.i)V H , which comprises the amino acid sequence of SEQ ID NO: 309 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 360; IV.i)V H , which comprises the amino acid sequence of SEQ ID NO: 307 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 358; V.i)V H , which comprises the amino acid sequence of SEQ ID NO: 317 and ii)V L , which comprises SEQ ID NO: the amino acid sequence of 368; VI.i)V H , which comprises the amino acid sequence of SEQ ID NO: 318 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 369; VII.i)V H , which comprises the amino acid sequence of SEQ ID NO: 350 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 401; VIII.i)V H , which comprises the amino acid sequence of SEQ ID NO: 334 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 385; IX.i)V H , which comprises the amino acid sequence of SEQ ID NO: 310 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 361; X.i)V H , which comprises the amino acid sequence of SEQ ID NO: 311 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 362; XI.i)V H , which comprises the amino acid sequence of SEQ ID NO: 336 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 387; XII.i)V H , which comprises the amino acid sequence of SEQ ID NO: 347 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 398; or XIII.i)V H , which comprises the amino acid sequence of SEQ ID NO: 355 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 406.

3. The isolated anti-GPC3 construct according to claim 1, wherein said anti-GPC3 antibody portion comprises: I.i)V H which comprises HC - CDR1, HC - CDR2 and HC - CDR3, wherein HC - CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC - CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii)V L which comprises LC - CDR1, LC - CDR2 and LC - CDR3, wherein LC - CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC - CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO: 276; II.i) V H , which includes HC - CDR1, HC - CDR2, and HC - CDR3. HC - CDR1 consists of the amino acid sequence of SEQ ID NO:24, HC - CDR2 consists of the amino acid sequence of SEQ ID NO:75, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO:126; and ii) V L , which includes LC - CDR1, LC - CDR2, and LC - CDR3. LC - CDR1 consists of the amino acid sequence of SEQ ID NO:177, LC - CDR2 consists of the amino acid sequence of SEQ ID NO:228, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO:279; 381; or III.i)V H , which includes HC-CDR1, HC-CDR2 and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO:3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:105; and ii)V L , which includes LC-CDR1, LC-CDR2 and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO:156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:

258.

4. The isolated anti-GPC3 construct according to claim 1, wherein said anti-GPC3 antibody portion comprises: I.i)V H , which comprises the amino acid sequence of SEQ ID NO: 309 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 360; II.i)V H , which contains the amino acid sequence of SEQ ID NO: 327 and ii) V L , which contains SEQ ID NO: the amino acid sequence of 378; or III.i)V H , which comprises the amino acid sequence of SEQ ID NO: 330 and ii) V L , which comprises SEQ ID NO: the amino acid sequence of 381.

5. The isolated anti-GPC3 construct according to claim 1, wherein: i. The anti-GPC3 antibody moiety is a full-length antibody, Fab, Fab', F(ab') 2 , Fv or single-chain Fv scFv; ii. the anti-GPC3 antibody portion is fused to an Fc fragment, optionally via a linker; and / or iii. the isolated anti-GPC3 construct is multispecific.

6. The isolated anti-GPC3 construct according to claim 1, wherein the isolated anti-GPC3 construct is an anti-GPC3 CAR, which comprises: (a) an extracellular domain comprising said anti-GPC3 antibody portion; (b) a transmembrane domain; and (c) an intracellular signaling domain.

7. The isolated anti-GPC3 construct according to claim 6, wherein the intracellular signaling domain comprises a CD3ζ intracellular signaling sequence and a CD28 intracellular signaling sequence.

8. The isolated anti-GPC3 construct according to claim 6, wherein the anti-GPC3 CAR comprises the amino acid sequence of any one of SEQ ID NO:491 and 516 - 521.

9. The isolated anti-GPC3 construct according to claim 1, wherein the isolated anti-GPC3 construct is an anti-GPC3 caTCR, which comprises: (a) an extracellular domain comprising said anti-GPC3 antibody portion; and (b) TCR module TCRm, where the TCRm includes a first TCR domain TCRD containing a first TCR transmembrane domain TCR-TM and a second TCRD containing a second TCR-TM, and where the TCRm promotes the recruitment of at least one TCR-related signaling molecule.

10. The isolated anti-GPC3 construct according to claim 9, wherein the TCR-related signaling molecule is selected from the group consisting of CD3δε, CD3γε, and ζζ.

11. The isolated anti-GPC3 construct according to claim 9, wherein: i. The first TCR-TM is derived from the transmembrane domain of TCRγ and the second TCR-TM is derived from the transmembrane domain of TCRδ; or ii. The first TCR-TM is derived from the transmembrane domain of TCRδ and the second TCR-TM is derived from the transmembrane domain of TCRγ.

12. The isolated anti-GPC3 construct according to claim 9, wherein: i. The first TCR-TM is derived from the transmembrane domain of TCRα and the second TCR-TM is derived from the transmembrane domain of TCRβ; or ii. The first TCR-TM is derived from the transmembrane domain of TCRβ and the second TCR-TM is derived from the transmembrane domain of TCRα.

13. The isolated anti-GPC3 construct according to claim 1, wherein the isolated anti-GPC3 construct is anti-GPC3 CSR, which comprises: (a) A ligand-binding module, which includes the anti-GPC3 antibody portion; (b) A transmembrane module; and (c) A co-stimulatory immune cell signaling module.

14. The isolated anti-GPC3 construct according to claim 13, wherein the anti-GPC3 CSR does not include a functional naive immune cell signaling sequence.

15. The isolated anti-GPC3 construct according to claim 13, wherein the anti-GPC3 CSR lacks any naive immune cell signaling sequence.

16. The isolated anti-GPC3 construct according to claim 13, wherein the co-stimulatory immune cell signaling module includes a portion of the intracellular domain of a co-stimulatory molecule selected from the group consisting of CD27, CD28, 4-1BB (CD137), OX40, CD30, CD40, PD-1, ICOS, LFA-1, CD2, CD7, LIGHT, KG2C, B7-H3, and a ligand that specifically binds CD83.

17. The isolated anti-GPC3 construct according to claim 13, wherein the anti-GPC3 CSR includes the amino acid sequence of SEQ ID NO:

531.

18. An isolated nucleic acid encoding a polypeptide component of the isolated anti-GPC3 construct according to any one of claims 1 to 17.

19. A vector comprising the isolated nucleic acid according to claim 18.

20. An isolated host cell comprising the anti-GPC3 construct according to any one of claims 1 to 17, the isolated nucleic acid according to claim 18, or the vector according to claim 19.

21. An effector cell that expresses an isolated anti-GPC3 construct according to any one of claims 1 to 17.

22. The effector cell according to claim 21, wherein the effector cell is a T cell.

23. The effector cell according to claim 22, wherein the T cell is selected from the group consisting of: cytotoxic T cells, helper T cells, and natural killer T cells.

24. The effector cell according to claim 21, wherein the expression of the anti-GPC3 construct is induced by activating the effector cell.

25. The effector cell according to claim 21, wherein the effector cell comprises an anti-GPC3 CSR and a caTCR.

26. The effector cell according to claim 25, wherein the caTCR comprises: (a) An extracellular domain, wherein the extracellular domain comprises: (i) An anti-GPC3 antibody portion; or (ii) An anti-AMC antibody portion that specifically binds to a complex comprising an alpha-fetoprotein AFP peptide and a major histocompatibility complex MHC class I protein (AFP / MHC class I complex, or AMC); and (b) A TCRM, wherein the TCRM comprises a first TCRD and a second TCRD, the first TCRD comprises a first TCR-TM, the second TCRD comprises a second TCR-TM, and wherein the TCRM promotes the recruitment of at least one TCR-associated signaling molecule.

27. The effector cell of claim 26, wherein the TCR-associated signaling molecule is selected from the group consisting of CD3δε, CD3γε, and ζζ.

28. The effector cell of claim 26, wherein the extracellular domain of the caTCR comprises an anti-GPC3 antibody portion, and wherein: (i) The anti-GPC3 antibody portion specifically recognizes cell surface-bound GPC3 with high binding affinity and binds soluble GPC3 with low binding affinity; and / or (ii) The IC of the soluble GPC3 competing for binding between the anti-GPC3 antibody portion and the GPC3 bound to the cell surface 50 is from about 1 μg / mL to about 100 μg / mL.

29. The effector cell of claim 26, wherein the caTCR is the anti-GPC3 caTCR of claim 9.

30. The effector cell of claim 21, wherein the effector cell comprises the anti-GPC3 caTCR, wherein the effector cell further comprises a CSR, and wherein the CSR comprises: (a) A ligand-binding module comprising an antibody portion; (b) A transmembrane module; and (c) A co-stimulatory immune cell signaling module.

31. The effector cell of claim 30, wherein the CSR is an anti-GPC3 CSR comprising an anti-GPC3 antibody portion.

32. The effector cell of claim 31, wherein: I. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:105; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:105; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; or II. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 105; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; or III. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 105; and ii) V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 24, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 75, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 126; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 177, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 228, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO: 381; or IV. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H , which includes HC - CDR1, HC - CDR2 and HC - CDR3, where HC - CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC - CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii) V L , which includes LC - CDR1, LC - CDR2 and LC - CDR3, where LC - CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC - CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO:3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO:54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO:105; and ii)V L which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO:156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO:207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO:258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; or V. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H , which comprises HC - CDR1, HC - CDR2 and HC - CDR3, wherein HC - CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC - CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii) V L , which comprises LC - CDR1, LC - CDR2 and LC - CDR3, wherein LC - CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC - CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; or VI. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H which comprises HC - CDR1, HC - CDR2 and HC - CDR3, wherein HC - CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC - CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii) V L which comprises LC - CDR1, LC - CDR2 and LC - CDR3, wherein LC - CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC - CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which includes HC-CDR1, HC-CDR2, and HC-CDR3. HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 24, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 75, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 126; and ii)V L , which includes LC-CDR1, LC-CDR2, and LC-CDR3. LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 177, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 228, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO: 381; or VII. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H which comprises HC - CDR1, HC - CDR2 and HC - CDR3, wherein HC - CDR1 consists of the amino acid sequence of SEQ ID NO:24, HC - CDR2 consists of the amino acid sequence of SEQ ID NO:75, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO:126; and ii) V L which comprises LC - CDR1, LC - CDR2 and LC - CDR3, wherein LC - CDR1 consists of the amino acid sequence of SEQ ID NO:177, LC - CDR2 consists of the amino acid sequence of SEQ ID NO:228, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO:279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO: 381; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 3, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 54, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 105; and ii) V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 156, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 207, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 258; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 309 and a VL comprising the amino acid sequence of SEQ ID NO: 360; or VIII. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a) i) V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 24, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 75, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 126; and ii) V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 177, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 228, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO: 381; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 21, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 72, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 123; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 174, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 225, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 276; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 327 and a VL comprising the amino acid sequence of SEQ ID NO: 378; or IX. (1) The anti-GPC3 caTCR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC - CDR1, HC - CDR2 and HC - CDR3, wherein HC - CDR1 consists of the amino acid sequence of SEQ ID NO:24, HC - CDR2 consists of the amino acid sequence of SEQ ID NO:75, and HC - CDR3 consists of the amino acid sequence of SEQ ID NO:126; and ii)V L , which comprises LC - CDR1, LC - CDR2 and LC - CDR3, wherein LC - CDR1 consists of the amino acid sequence of SEQ ID NO:177, LC - CDR2 consists of the amino acid sequence of SEQ ID NO:228, and LC - CDR3 consists of the amino acid sequence of SEQ ID NO:279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO: 381; and (2) The anti-GPC3 CSR comprises an anti-GPC3 antibody portion, and the anti-GPC3 antibody portion comprises: (a)i)V H , which comprises HC-CDR1, HC-CDR2 and HC-CDR3, wherein HC-CDR1 consists of the amino acid sequence of SEQ ID NO: 24, HC-CDR2 consists of the amino acid sequence of SEQ ID NO: 75, and HC-CDR3 consists of the amino acid sequence of SEQ ID NO: 126; and ii)V L , which comprises LC-CDR1, LC-CDR2 and LC-CDR3, wherein LC-CDR1 consists of the amino acid sequence of SEQ ID NO: 177, LC-CDR2 consists of the amino acid sequence of SEQ ID NO: 228, and LC-CDR3 consists of the amino acid sequence of SEQ ID NO: 279; and / or (b) A VH comprising the amino acid sequence of SEQ ID NO: 330 and a VL comprising the amino acid sequence of SEQ ID NO:

381.

33. A pharmaceutical composition comprising the isolated anti-GPC3 construct according to any one of claims 1 to 5 and a pharmaceutically acceptable carrier.

34. A pharmaceutical composition comprising the effector cell according to any one of claims 21 to 32 and a pharmaceutically acceptable carrier.

35. A kit comprising the isolated anti-GPC3 construct according to any one of claims 1 to 5.

36. Use of the pharmaceutical composition according to claim 33 in the preparation of a medicament for treating an individual suffering from a GPC3-positive disease, wherein the GPC3-positive disease is hepatocellular carcinoma (HCC).

37. Use of the pharmaceutical composition according to claim 34 in the preparation of a medicament for treating an individual suffering from a GPC3-positive disease, wherein the GPC3-positive disease is hepatocellular carcinoma (HCC).

38. A method for producing an isolated anti-GPC3 construct, which comprises: (a) Culturing a isolated host cell comprising the isolated nucleic acid according to claim 18 under conditions effective to express the anti-GPC3 construct; and (b) Obtaining the expressed anti-GPC3 construct from the host cell.

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