Immunosuppressant binding antibodies and methods of obtaining and using same
AssigneeAbbott Laboratories
InventorsRobert W. Siegel, Joan D. Tyner, Terry Y. Nakagawa
Life of this patent
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Abstract
The present invention relates among other things to antibodies that immunospecifically bind to at least one agent of interest (e.g., an immunosuppressive agent), methods for producing such antibodies, and immunoassays that employ said antibodies. Additionally, the present invention also relates to methods for selecting an antibody for use in a diagnostic immunoassay and methods for selecting an antigen for use in a diagnostic immunoassay. The present invention further relates to the improvement of antibody recognition of an active parent drug in the presence of one or more of its major metabolites.
Claims41
An isolated antibody which specifically binds to an immunosuppressive agent with an equilibrium dissociation constant (KD) less than 1.9×10−11 M when said antibody has not been exposed to or incubated with at least one selection diluent.
An isolated antibody which specifically binds to an immunosuppressive agent with an equilibrium dissociation constant (KD) less than 1.52×10−10 M when said antibody is incubated with, exposed to, or is in the presence of at least one selection diluent.
A Chinese Hamster Ovary cell line 1-60-46 AM2 CHO 2-577 having A.T.C.C. Accession No. PTA-7436.
An antibody made from DNA extracted from a Chinese Hamster Ovary cell line 1-60-46 AM2 CHO 2-577 having A.T.C.C. Accession No. PTA-7436.
A chimeric antibody or a tacrolimus binding fragment thereof produced by a Chinese Hamster Ovary cell line designated tacrolimus 1-60-46 AM2 CHO 2-577, wherein said cell line has A.T.C.C. Accession No. PTA-7436.
A Chinese Hamster Ovary cell line designated tacrolimus 1-60-46 AM2 CHO 1-1157 having A.T.C.C. Accession No. PTA-7446.
An antibody made from DNA extracted from a Chinese Hamster Ovary cell line designated tacrolimus 1-60-46 AM2 CHO 1-1157 having A.T.C.C. Accession No. PTA-7446.
A chimeric antibody or a tacrolimus binding fragment thereof produced by a Chinese Hamster Ovary cell line designated tacrolimus 1-60-46 AM2 CHO 1-1157, wherein said cell line has A.T.C.C. Accession No. PTA-7446.
An isolated antibody which specifically binds to tacrolimus, wherein said antibody has a variable heavy domain and a variable light domain, the variable heavy domain comprising a heavy chain complementary determining region (“CDR”) 1, a heavy chain CDR 2 and a heavy chain CDR 3, the variable light domain comprising a light chain CDR 1, a light chain CDR 2 and a light chain CDR 3, wherein (a) Heavy Chain CDR 1 has an amino acid sequence of:
- Gly-Phe-Thr-Phe-Ser-Ser-Tyr-Gly-Met-Ser (SEQ ID NO:2);
- (b) Heavy Chain CDR 2 has an amino acid sequence having a formula of:
- (SEQ ID NO:33)
- Thr-Ile-Ser-Ser-Gly-Gly-Xaa1-Xaa2-Xaa3-Phe
- wherein Xaa1 is selected from the group consisting of threonine (Thr), alanine (Ala), lysine (Lys) and glutamic acid (Glu);
- wherein Xaa2 is selected from the group consisting of tyrosine (Tyr) and tryptophan (Trp);
- and
- wherein Xaa3 is selected from the group consisting of threonine (Thr) and valine (Val);
- (c) Heavy Chain CDR 3 has an amino acid sequence of:
- Gln-Thr-Asp-Gly-Tyr-Ser-Trp-Phe-Pro-Tyr (SEQ ID NO:6);
- (d) Light Chain CDR 1 has an amino acid sequence having a formula of:
- (SEQ ID NO:34)
- Lys-Ser-Ser-Xaa4-Xaa5-Xaa6-Val-His-Ser-Thr-Gly-
- Asn-Thr-Phe-Leu-Glu
- wherein Xaa4 is selected from the group consisting of:
- glutamine (Gln), alanine (Ala) and glycine (Gly);
- wherein Xaa5 is selected from the group consisting of:
- serine (Ser) and glycine (Gly);
- and
- wherein Xaa6 is selected from the group consisting of:
- isoleucine (Ile) and leucine (Leu);
- (e) Light Chain CDR 2 has an amino acid sequence having the formula of:
- Lys-Ile-Ser-Asn-Arg-Phe-Ser
- (SEQ ID NO:11)
- (f) Light Chain CDR 3 has an amino acid sequence having a formula of:
- (SEQ ID NO:35)
- Phe-Gln-Gly-Xaa7-Xaa8-Xaa9-Pro-Leu-Thr,
- wherein Xaa7 is selected from the group consisting of:
- Serine (Ser) and Glycine (Gly);
- wherein Xaa8 is selected from the group consisting of:
- histidine (His), arginine (Arg), valine (Val), threonine (Thr), lysine (Lys) and serine (Ser);
- and
- wherein Xaa9 is selected from the group consisting of:
- valine (Val), alanine (Ala), aspartic acid (Asp), cysteine (Cys) and Serine (Ser);
- with the proviso that if in heavy chain CDR 2 Xaa1 is Thr, Xaa2 is Tyr and Xaa3 is Thr and in the light chain CDR 1 Xaa4 is Gln, Xaa5 is Ser and Xaa6 is Ile, then in light chain CDR 3 Xaa9 is other than Val if Xaa7 is Ser and Xaa8 is His, or Xaa8 is other than His if Xaa7 is Ser and Xaa9 is Val or Xaa7 is other than Ser if Xaa8 is His and Xaa9 is Val.
A method for selecting an antibody for use in a diagnostic immunoassay, wherein said antibody binds to an epitope of interest, the method comprising the steps of:
- a) contacting at least one antibody with a sample in the presence of at least one selection diluent, wherein said sample comprises an epitope of interest to which said antibody is believed to bind and further wherein said antibody is present in a bio-display format;
- b) determining the equilibrium dissociation constant (KD), disassociation rate constant (kd), association rate constant (ka) or functional activity of the antibody;
- and
- c) selecting an antibody based on the equilibrium dissociation constant, dissociation rate constant, association rate constant or functional activity determined in step b).
A method for selecting an antibody for use in a diagnostic immunoassay, wherein said antibody binds to an epitope of interest, the method comprising the steps of:
- a) incubating at least one antibody in the presence of at least one selection diluent, wherein said antibody is present in a bio-display format;
- b) contacting said antibody with a sample, wherein said sample contains an epitope of interest to which said antibody is believed to bind;
- c) determining the equilibrium dissociation constant (KD), disassociation rate constant (kd), association rate constant (ka) or functional activity of the antibody;
- and
- d) selecting an antibody based on the equilibrium dissociation constant, disassociation rate constant, association rate constant or functional activity determined in step c).
A method for selecting a specific binding partner for detecting an analyte of interest in test sample for use in a diagnostic immunoassay, the method comprising the steps of:
- a) contacting a specific binding partner with a sample in the presence of at least one selection diluent, wherein said sample contains the epitope of interest and the specific binding partner binds to the epitope of interest, and further wherein said specific binding partner is present in a bio-display format;
- b) determining the equilibrium dissociation constant (KD), disassociation rate constant (kd), association rate constant (ka) or functional activity of the specific binding partner;
- and
- c) selecting a specific binding partner based on the equilibrium dissociation constant, dissociation rate constant, association rate constant or functional activity of the specific binding partner determined in step b).
A method of screening for antibodies having improved specificity for an agent using yeast display wherein said method comprises:
- (a) obtaining a yeast display library comprising antibodies present on the surface of yeast cells;
- (b) contacting said yeast cells with said agent in the presence of a binding competitor;
- (c) identifying yeast cells having antibodies displayed thereon which exhibit binding to said agent in the presence of said binding competitor, wherein such binding indicates an improved specificity for said agent.
A method of screening for antibodies having improved affinity for an epitope of interest, said method comprising the steps of:
- (a) obtaining a library comprising antibodies present in a bio-display format wherein said antibodies comprise mutations;
- (b) contacting said antibodies with sample comprising said epitope in the presence of at least one selection diluent;
- and
- (c) identifying antibodies present in said bio-display format which exhibit reduced dissociation rates in the presence of said selection diluent as compared to the dissociation rate of comparable antibody not comprising mutations, wherein such reduced dissociation rates indicate an improved affinity for said epitope.
A method of screening for a specific binding partner having improved affinity for an epitope of interest, said method comprising the steps of:
- (a) obtaining a library comprising specific binding partners present in a bio-display format wherein said specific binding partners comprise mutations;
- (b) contacting said specific binding partners with sample comprising said epitope in the presence of at least one selection diluent;
- and
- (c) identifying specific binding partners present in said bio-display format which exhibit reduced dissociation rates in the presence of said selection diluent as compared to the dissociation rate of a comparable specific binding partner not comprising mutations, wherein such reduced dissociation rates indicate an improved affinity for said epitope.
Dependencies are inferred from the claim text.