Method for assaying soluble GPC3 protein
The present invention relates to a method for assaying soluble GPC3 protein in a test sample, comprising using two different antibodies binding to different epitopes present in the N-terminal region of GPC3 protein.
1. An immunoassay method for assaying soluble Glypican-3 (GPC3) protein in a test sample, comprising contacting the sample with a first and a second antibody that each specifically bind to different epitopes of soluble GPC3 protein to form a complex; and detecting the complex to assay soluble GPC3 in the sample,
wherein the first antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 38 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 39,
and wherein the second antibody comprises a heavy chain variable region comprising the amino acid sequence of SEQ ID NO: 40 and a light chain variable region comprising the amino acid sequence of SEQ ID NO: 41,
and wherein the soluble GPC3 protein comprises amino acids 128-357 of SEQ ID NO: 70.
2. The method according to claim 1 , wherein any one of the two different antibodies is bound with a magnetic particle.
3. The method according to claim 1 , wherein the test sample is a whole blood sample, a plasma sample, or a serum sample isolated from a human.
4. An immunoassay method for assaying soluble Glypican-3 (GPC3) protein in a test sample, comprising contacting the sample with a first and a second antibody that each specifically bind to different epitopes of soluble GPC3 protein to form a complex; and detecting the complex to assay soluble GPC3 in the sample,
wherein the first antibody comprises three light chain complementarity determining regions (CDRs) in which CDR1 comprises the amino acid sequence of SEQ ID NO: 49, CDR2 comprises the amino acid sequence of SEQ ID NO: 50 and CDR3 comprises the amino acid sequence of SEQ ID NO: 51, and said first antibody further comprises three heavy chain complementarity determining regions (CDRs) in which CDR1 comprises the amino acid sequence of SEQ ID NO: 46, CDR2 comprises the amino acid sequence of SEQ ID NO: 47 and CDR3 comprises the amino acid sequence of SEQ ID NO: 48, and
wherein the second antibody comprises three light chain complementarity determining regions (CDRs) in which CDR1 comprises the amino acid sequence of SEQ ID NO: 55, CDR2 comprises the amino acid sequence of SEQ ID NO: 56 and CDR3 comprises the amino acid sequence of SEQ ID NO: 57, and said second antibody further comprises three heavy chain complementarity determining regions (CDRs) in which CDR1 comprises the amino acid sequence of SEQ ID NO: 52, CDR2 comprises the amino acid sequence of SEQ ID NO: 53 and CDR3 comprises the amino acid sequence of SEQ ID NO: 54,
and wherein the soluble GPC3 protein comprises amino acids 128-357 of SEQ ID NO: 70.
5. The method according to claim 4 , wherein any one of the two different antibodies is bound with a magnetic particle.
6. The method according to claim 4 , wherein the test sample is a whole blood sample, a plasma sample, or a serum sample isolated from a human.