Biotin-specific monoclonal antibody and use thereof

Patent metadata

Field	Value
Publication number	US-12025613-B2
Application number	US-202117551723-A
Country	US
Kind code	B2
Filing date	Dec 15, 2021
Priority date	Dec 27, 2016
Publication date	Jul 2, 2024
Grant date	Jul 2, 2024

Abstract

The present invention relates to a monoclonal antibody capable of binding to biotin. In one embodiment the monoclonal antibody according to the invention also does not bind to a biotin moiety on a biotinylated molecule, wherein the biotin moiety is attached to the molecule via the carbon atom of the carboxyl function of the valeric acid moiety of biotin. Also disclosed is a method for generation of an antibody as discloed herein. The monoclonal antibody according to the invention is of specific use in a method for measuring an analyte in a sample, wherein a (strept)avidin/biotin pair is used to bind a biotinylated analyte specific binding agent to a (strept)avidin coated solid phase.

First claim

The invention claimed is:

1. A compound having a structure according to the following formula:

wherein M is selected from (i) a hapten which does not contain a biotin moiety, or (ii) a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH).

2. The compound of claim 1 , wherein said compound has a structure according to Formula [28]:

3. A compound having a structure according to the following formula:

wherein M is selected from (i) a hapten which does not contain a biotin moiety, or (ii) a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH).

4. The compound of claim 3 , wherein said compound has a structure according to Formula [22]:

5. A method for producing an antibody, the method comprising the steps of
a) immunizing an experimental animal with a compound according to claim 1 , wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH), thereby inducing B-cells producing antibodies binding to the compound,
b) obtaining a monoclonal antibody binding to the compound produced by the B-cell of step (a), either via hybridoma technology or by B-cell PCR technology,
c) further selecting the antibody of step (b) for binding to biotin
thereby obtaining the antibody.

6. The method of claim 5 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to a compound of claim 1 .

7. The method of claim 5 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to the compound of claim 1 , wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH).

8. The method of claim 5 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to a compound of Formula III,

9. The method of claim 5 , wherein said method comprises further step d), selecting those antibodies which do not bind to the compound of Formula II,

10. A method for producing an antibody specifically binding to biotin and to a compound of claim 1 ,
but not to a compound of Formula II, n M

the method comprising the steps of
a) immunizing an experimental animal with a compound according to claim 1 , wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH), thereby inducing B-cells producing antibodies binding to the compound,
b) obtaining a monoclonal antibody binding to the compound produced by the B-cell of step (a), either via hybridoma technology or by B-cell PCR technology,
c) selecting the antibody of step (b) for binding to biotin, and
d) selecting those antibodies which do not bind to the compound of Formula II,

thereby obtaining the antibody.

11. A method for producing an antibody, the method comprising the steps of
a) immunizing an experimental animal with a compound according to claim 3 , wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH), thereby inducing B-cells producing antibodies binding to the compound,
b) obtaining a monoclonal antibody binding to the compound produced by the B-cell of step (a), either via hybridoma technology or by B-cell PCR technology,
c) further selecting the antibody of step (b) for binding to biotin
thereby obtaining the antibody.

12. The method of claim 11 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to the compound of claim 3 wherein M is a hapten which does not contain a biotin moiety.

13. The method of claim 11 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to the compound of claim 3 wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH).

14. The method of claim 11 wherein in step (c) the selection is performed in a competitive assay using biotin as a competitor for binding of the antibody to a compound of Formula III:

15. The method of claim 11 , wherein said method comprises further step d), selecting those antibodies which do not bind to the compound of Formula II,

16. A method for producing an antibody specifically binding to biotin and to a compound of claim 3 , but not to a compound of Formula II,

the method comprising the steps of
a) immunizing an experimental animal with a compound according to claim 3 , wherein M is a polypeptide selected from the group consisting of rat, rabbit, mouse, porcine or bovine serum albumin, bovine or porcine thyroglobulin, ovalbumin, tetanus toxoid, gelatin, soybean trypsin inhibitor and keyhole limpet hemocyanin (KLH), thereby inducing B-cells producing antibodies binding to the compound,
b) obtaining a monoclonal antibody binding to the compound produced by the B-cell of step (a), either via hybridoma technology or by B-cell PCR technology,
c) selecting the antibody of step (b) for binding to biotin, and
d) selecting those antibodies which do not bind to the compound of Formula II,

thereby obtaining the antibody.

Assignees

• Roche Diagnostics Operations Inc

Inventors

• Gerg Michael
• Heindl Dieter
• Hillringhaus Lars
• Hirzel Klaus
• Hojer Caroline Dorothea
• Huber Florian
• Josel Hans-Peter
• Meier Thomas
• Schraeml Michael
• Voss Edgar

Classifications

• C07K16/44 Primary

  against material not provided for elsewhere {, e.g. haptens, metals, DNA, RNA, amino acids} · CPC title

• C07K2319/92

  containing an intein ("protein splicing")domain · CPC title

• C07K2317/34

  Identification of a linear epitope shorter than 20 amino acid residues or of a conformational epitope defined by amino acid residues · CPC title

• C07D495/04

  Ortho-condensed systems · CPC title

• G01N33/82

  involving vitamins {or their receptors} · CPC title