Separating agent for human serum-derived IgG polyclonal antibodies, and method for separating human serum-derived IgG polyclonal antibodies using same
View Patent ↗An object of the present invention is to provide a separation agent for separating a human serum-derived IgG polyclonal antibody. This object is achieved by a separation agent for separating a human serum-derived IgG polyclonal antibody, the separation agent including: a carrier; and a single-chain antibody which has a dissociation constant for a human serum-derived IgG polyclonal antibody of not more than 3.0×10 −8 M and which binds to the surface of the carrier via a chemical bond.
1. A separation agent for separating a human serum-derived IgG polyclonal antibody, the separation agent comprising: a carrier; and a single-chain antibody which has a dissociation constant for a human serum-derived IgG polyclonal antibody of not more than 3.0×10 −8 M and which binds to the surface of the carrier via a chemical bond,
wherein the single-chain antibody is selected from the following single-chain antibodies (a) to (e):
(a) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ATRYDSYGYAYNYWFGTLW (SEQ ID NO: 148);
wherein the single-chain antibody (a) further comprises the following amino acid sequence (a-1) or (a-2):
(a-1) the amino acid sequence of the heavy-chain CDR1: SGIDLSSNA (SEQ ID NO: 153),
the amino acid sequence of the heavy-chain CDR2: ISTVGKT (SEQ ID NO: 154),
the amino acid sequence of the light-chain CDR1: ENINSE (SEQ ID NO: 155),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QSTYYDGNYVYA (SEQ ID NO: 156); and
(a-2) the amino acid sequence of the heavy-chain CDR1: SGIDLSSNA (SEQ ID NO: 153),
the amino acid sequence of the heavy-chain CDR2: ISTGGST (SEQ ID NO: 157),
the amino acid sequence of the light-chain CDR1: QNINNE (SEQ ID NO: 158),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QSTYYDGNYVYA (SEQ ID NO: 156),
(b) the single-chain antibody having the amino acid sequence of the heavy-chain CDR3: GSYYDSHGYAYVSLW (SEQ ID NO: 149);
wherein the single-chain antibody (b) further comprises
the amino acid sequence of the heavy-chain CDR1: SGFSLSRYA (SEQ ID NO: 159),
the amino acid sequence of the heavy-chain CDR2: IGSGGST (SEQ ID NO: 160),
the amino acid sequence of the light-chain CDR1: QSISTA (SEQ ID NO: 161),
the amino acid sequence of the light-chain CDR2: SAS, and
the amino acid sequence of the light-chain CDR3: QSYYGSSSDNA (SEQ ID NO: 162);
(c) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ATDYGIYGYAYGHLW (SEQ ID NO: 150);
wherein the single-chain antibody (c) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGIDLSSYA (SEQ ID NO: 163),
the amino acid sequence of the heavy-chain CDR2: IGSGGGT (SEQ ID NO: 164),
the amino acid sequence of the light-chain CDR1: QSISTA (SEQ ID NO: 161),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QTYFGSDTDNA (SEQ ID NO: 165);
(d) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ARYSGDNGGALNLW (SEQ ID NO: 151);
wherein the single-chain antibody (d) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGFSLSSYA (SEQ ID NO: 166),
the amino acid sequence of the heavy-chain CDR2: ISSSGST (SEQ ID NO: 167),
the amino acid sequence of the light-chain CDR1: QHIRSY (SEQ ID NO: 168),
the amino acid sequence of the light-chain CDR2: AAS, and
the amino acid sequence of the light-chain CDR3: QRYYDIRNYGNG (SEQ ID NO: 169), and
(e) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ARYSGDNGGTLNLW (SEQ ID NO: 152);
wherein the single-chain antibody (e) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGIDLRRYA (SEQ ID NO: 170),
the amino acid sequence of the heavy-chain CDR2: IASGNTD (SEQ ID NO: 171),
the amino acid sequence of the light-chain CDR1: QSISSY (SEQ ID NO: 172),
the amino acid sequence of the light-chain CDR2: AAS, and
the amino acid sequence of the light-chain CDR3: QSYYSISSYGNT (SEQ ID NO: 173).
2. The separation agent according to claim 1 , wherein the single-chain antibody is also an antibody against a human serum-derived IgA polyclonal antibody.
3. The separation agent according to claim 2 , wherein the single-chain antibody has a dissociation rate constant for the human serum-derived IgA polyclonal antibody of not more than 1.0×10 −3 s −1 .
4. The separation agent according to claim 1 , wherein the single-chain antibody is an antibody which binds to L chain.
5. The separation agent according to claim 1 , wherein the single-chain antibody is an antibody which binds to kappa chain.
6. A method for separating a human serum-derived IgG polyclonal antibody from a mixed liquid of two or more kinds of substances, said mixed liquid comprising human serum-derived IgG polyclonal antibody, said method comprising the steps of: contacting said mixed liquid with a separation agent; and separating a human serum-derived IgG polyclonal antibody from said mixed liquid, the separation agent comprising: a carrier; and a single-chain antibody which has a dissociation constant for a human serum-derived IgG polyclonal antibody of not more than 3.0×10 −8 M and which binds to the surface of the carrier via a chemical bond,
wherein the single-chain antibody is selected from the following single-chain antibodies (a) to (e):
(a) the single-chain antibody having the amino acid sequence of the heavy-chain CDR3: ATRYDSYGYAYNYWFGTLW (SEQ ID NO: 148);
wherein the single-chain antibody (a) further comprises the following amino acid sequence (a-1) or (a-2):
(a-1) the amino acid sequence of the heavy-chain CDR1: SGIDLSSNA (SEQ ID NO: 153),
the amino acid sequence of the heavy-chain CDR2: ISTVGKT (SEQ ID NO: 154),
the amino acid sequence of the light-chain CDR1: ENINSE (SEQ ID NO: 155),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QSTYYDGNYVYA (SEQ ID NO: 156); and
(a-2) the amino acid sequence of the heavy-chain CDR1: SGIDLSSNA (SEQ ID NO: 153),
the amino acid sequence of the heavy-chain CDR2: ISTGGST (SEQ ID NO: 157),
the amino acid sequence of the light-chain CDR1: QNINNE (SEQ ID NO: 158),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QSTYYDGNYVYA (SEQ ID NO: 156),
(b) the single-chain antibody having the amino acid sequence of the heavy-chain CDR3: GSYYDSHGYAYVSLW (SEQ ID NO: 149);
wherein the single-chain antibody (b) further comprises
the amino acid sequence of the heavy-chain CDR1: SGFSLSRYA (SEQ ID NO: 159),
the amino acid sequence of the heavy-chain CDR2: IGSGGST (SEQ ID NO: 160),
the amino acid sequence of the light-chain CDR1: QSISTA (SEQ ID NO: 161),
the amino acid sequence of the light-chain CDR2: SAS, and
the amino acid sequence of the light-chain CDR3: QSYYGSSSDNA (SEQ ID NO: 162);
(c) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ATDYGIYGYAYGHLW (SEQ ID NO: 150);
wherein the single-chain antibody (c) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGIDLSSYA (SEQ ID NO: 163),
the amino acid sequence of the heavy-chain CDR2: IGSGGGT (SEQ ID NO: 164),
the amino acid sequence of the light-chain CDR1: QSISTA (SEQ ID NO: 161),
the amino acid sequence of the light-chain CDR2: DAS, and
the amino acid sequence of the light-chain CDR3: QTYFGSDTDNA (SEQ ID NO: 165);
(d) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ARYSGDNGGALNLW (SEQ ID NO: 151);
wherein the single-chain antibody (d) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGFSLSSYA (SEQ ID NO: 166),
the amino acid sequence of the heavy-chain CDR2: ISSSGST (SEQ ID NO: 167),
the amino acid sequence of the light-chain CDR1: QHIRSY (SEQ ID NO: 168),
the amino acid sequence of the light-chain CDR2: AAS, and
the amino acid sequence of the light-chain CDR3: QRYYDIRNYGNG (SEQ ID NO: 169), and
(e) the single-chain antibody having
the amino acid sequence of the heavy-chain CDR3: ARYSGDNGGTLNLW (SEQ ID NO: 152);
wherein the single-chain antibody (e) further comprises:
the amino acid sequence of the heavy-chain CDR1: SGIDLRRYA (SEQ ID NO: 170),
the amino acid sequence of the heavy-chain CDR2: IASGNTD (SEQ ID NO: 171),
the amino acid sequence of the light-chain CDR1: QSISSY (SEQ ID NO: 172),
the amino acid sequence of the light-chain CDR2: AAS, and
the amino acid sequence of the light-chain CDR3: QSYYSISSYGNT (SEQ ID NO: 173).
7. The method according to claim 6 , wherein the single-chain antibody is also an antibody against a human serum-derived IgA polyclonal antibody, and wherein the method further separates the human serum-derived IgA polyclonal antibody.
8. The method according to claim 7 , wherein the single-chain antibody has a dissociation rate constant for the human serum-derived IgA polyclonal antibody of not more than 1.0×10 −3 s −1 .
9. The method according to claim 6 , wherein the single-chain antibody is an antibody which binds to L chain.
10. The method according to claim 6 , wherein the single-chain antibody is an antibody which binds to kappa chain.