Antibody conjugates

Patent metadata

Field	Value
Publication number	US-9315789-B2
Application number	US-201414146389-A
Country	US
Kind code	B2
Filing date	Jan 2, 2014
Priority date	Apr 28, 2005
Publication date	Apr 19, 2016
Grant date	Apr 19, 2016

Abstract

Antibody/signal-generating moiety conjugates are disclosed that include an antibody covalently linked to a signal-generating moiety through a heterobifunctional polyalkyleneglycol linker. The disclosed conjugates show exceptional signal-generation in immunohistochemical and in situ hybridization assays on tissue sections and cytology samples. In one embodiment, enzyme-metallographic detection of nucleic acid sequences with hapten-labeled probes can be accomplished using the disclosed conjugates as a primary antibody without amplification.

First claim

We claim:

1. An antibody-PEG linker-enzyme signal-generating moiety conjugate, comprising:
an antibody;
a plurality of enzyme signal-generating moieties; and
a plurality of monodisperse, heterobifunctional PEG linkers, wherein each enzyme signal-generating moiety is linked to the antibody through at least one of the plurality of monodisperse, heterobifunctional PEG linker to provide the antibody-PEG linker-enzyme signal-generating moiety conjugate having the formula

wherein Ab is the antibody, SM is the enzyme signal-generating moiety, n=4 to 12 and s=3 to 10.

2. The conjugate of claim 1 , wherein SM is an oxido-reductase enzyme.

3. The conjugate of claim 1 , wherein each SM independently is horseradish peroxidase, alkaline phosphatase, acid phosphatase, glucose oxidase, β-galactosidase, β-glucuronidase or β-lactamase.

4. The conjugate of claim 1 , wherein each SM independently is horseradish peroxidase or alkaline phosphatase.

5. The conjugate of claim 1 , wherein SM is alkaline phosphatase.

6. The conjugate of claim 1 , wherein SM is horseradish peroxidase.

7. The conjugate of claim 1 , wherein the antibody comprises an anti-antibody antibody.

8. The conjugate of claim 7 , wherein the anti-antibody antibody is an anti-IgG antibody.

9. The conjugate of claim 1 , wherein the antibody comprises an anti-hapten antibody.

10. The conjugate of claim 1 , wherein s=3 to 6.

11. The conjugate of claim 10 , wherein s=3 to 4.

12. The conjugate of claim 11 , wherein SM is horseradish peroxidase or alkaline phosphatase.

Assignees

• Ventana Med Syst Inc

Inventors

• Bieniarz Christopher
• Wong Jennifer
• Lefever Mark
• Kosmeder Ii Jerome W
• Ashworth-Sharpe Julia
• Kernag Casey A

Classifications

• G01N33/535

  with enzyme label {or co-enzymes, co-factors, enzyme inhibitors or enzyme substrates} · CPC title

• C07K16/44

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

• C12N9/16 Primary

  acting on ester bonds (3.1) · CPC title

• C12N9/0065

  acting on hydrogen peroxide as acceptor (1.11) · CPC title

• C07K16/42

  against immunoglobulins · CPC title