Antibodies for diagnosis of acute myeloid leukemia

Patent metadata

Field	Value
Publication number	US-2016304618-A1
Application number	US-201414913843-A
Country	US
Kind code	A1
Filing date	Aug 27, 2014
Priority date	Aug 27, 2013
Publication date	Oct 20, 2016
Grant date	—

Abstract

A polypeptide in particular an antibody or antibody fragment is disclosed wherein the polypeptide is corresponding to certain complementarity determining regions CDR1, CDR2 and CDR3 of a heavy chain V H and a light chain V L of an antibody as well as a compound comprising the polypeptide, its use as diagnostic agent for acute myeloid leukemia subtype M2 and a kit comprising the compound.

First claim

1 . A polypeptide comprising an antibody or antibody fragment for AML subtype M2 specific diagnosis wherein the polypeptide corresponds to complementarity determining regions CDR1, CDR2 and CDR3 of a heavy chain V H and a light chain V L of an antibody, the complementarity determining regions comprising:
the CDR 1 region of the heavy chain V H defined by a sequence of 5 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
PO N /PO N /NP N /NP N /PO N

and the amino acids are linked via peptide bonds;
the CDR 2 region of the heavy chain V H defined by a sequence of 17 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
PO N /NP N /PO N /PO N /BP + or NP N /PO N /BP + or PO N /BP + /PO N /NP N /PO N /NP N /AP − /PO N /NP N /BP + /PO N

and the amino acids are linked via peptide bonds;
the CDR 3 region of the heavy chain V H defined by a sequence of 7 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
NP N /NP N or BP + /BP + or NP N /BP + or PO N /NP N /AP − /PO N

and the amino acids are linked via peptide bonds;
the CDR 1 region of the light chain V L defined by a sequence of 11 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
BP + /NP N /PO N /PO N /PO N /NP N /PO N /PO N /PO N /NP N /PO N

and the amino acids are linked via peptide bonds;
the CDR 2 region of the light chain V L defined by a sequence of 7 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
NP N or BP + /NP N /PO N /BP 0 or NP N /NP N /PO N /PO N

and the amino acids are linked via peptide bonds; and
the CDR 3 region of the light chain V L defined by a sequence of 9 amino acids wherein the amino acids have side chain polarities and charges at a pH of 7.4, and the amino acids of the amino acid sequence are symbolized by a symbol as represented by a formula
PO N /PO N /NP N or BP + /BP + or NP N /PO N or BP + /PO N /NP N /NP N /PO N

and the amino acids are linked via peptide bonds;
wherein the amino acids of the formulas are proteinogenic amino acids and the symbols have the meaning:
PO N represents an amino acid having a polar side chain polarity and a neutral side chain charge at pH 7.4,
NP N represents an amino acid having a non-polar side chain polarity and a neutral side chain charge at pH 7.4,
BP + represents an amino acid having a basic polar side chain polarity and a positive side chain charge at pH 7.4,
BP 0 represents an amino acid having a basic polar side chain polarity and a predominantly neutral side chain charge at pH 7.4, and
AP − represents an amino acid having an acidic polar side chain polarity and a negative side chain charge at pH 7.4.

2 . The polypeptide of claim 1 , wherein:
PO N represents an amino acid selected from the group consisting of asparagine, glutamine, serine, threonine, and tyrosine; NP N represents an amino acid selected from the group consisting of alanine, cysteine, glycine, isoleucine, leucine, methionine, phenylalanine, proline, tryptophane, and valine; BP + represents arginine or lysine; BP 0 represents histidine; and AP − represents aspartic acid or glutamic acid.

3 . The polypeptide of claim 1 , wherein:
the antibody or antibody fragment comprises in its heavy chain CDR 1 a peptide having at least 80% homology to the peptide of the amino acid sequence of SEQ ID NO: 1, the antibody or antibody fragment comprises in its heavy chain CDR 2 a peptide having at least 85% homology to the peptides of the amino acid sequences SEQ ID NO: 2 or SEQ ID NO: 3, and the antibody or antibody fragment comprises in its heavy chain CDR 3 a peptide having at least 85% homology to the peptides of the amino acid sequences SEQ ID NO: 4 or SEQ ID NO: 5.

4 . The polypeptide of claim 1 , wherein:
the antibody or antibody fragment comprises in its light chain CDR 1 a peptide having at least 80% homology to the peptide of the amino acid sequence of SEQ ID NO: 6, the antibody or antibody fragment comprises in its light chain CDR 2 a peptide having at least 70% homology to the peptides of the amino acid sequences of SEQ ID NO: 7 or SEQ ID NO: 8, and the antibody or antibody fragment comprises in its light chain CDR 3 a peptide having at least 50% homology to the peptides of the amino acid sequences of SEQ ID NO: 9 or SEQ ID NO: 10.

5 . The polypeptide of claim 1 , wherein:
the amino acid sequence of the heavy chain CDR 1 is the sequence of SEQ ID NO: 1, the amino acid sequence of the heavy chain CDR 2 is the sequence of SEQ ID NO: 2 or SEQ ID NO: 3, and the amino acid sequence of the heavy chain CDR 3 is the sequence of SEQ ID NO: 4 or SEQ ID NO: 5.

6 . The polypeptide of claim 1 , wherein:
the amino acid sequence of the light chain CDR 1 is the sequence of SEQ ID NO 6, the amino acid sequence of the light chain CDR 2 is the sequence of SEQ ID NO: 7 or SEQ ID NO: 8, and the amino acid sequence of the light chain CDR 3 is the sequence of SEQ ID NO: 9 or SEQ ID NO: 10.

7 . The polypeptide of claim 1 ,
wherein the CDR1, CDR2 and CDR3 of the heavy chain of the variable region of an antibody v H and CDR1, CDR2 and CDR3 of the light chain of the variable region of an antibody v L are linked with each other via a linker structure.

8 . The polypeptide of claim 1 ,
wherein the linker structure is (Gly4Ser)3.

9 . The polypeptide of claim 1 ,
wherein the polypeptide is an antibody or a recombinant antibody.

10 . A compound comprising the polypeptide of claim 1 comprising a detectable label.

11 . The compound of claim 10 ,
wherein the detectable label is selected from the group consisting of fluorescent dyes, gamma ray emitting radioisotopes, a quantum dot composed of heavy metals, noble metal nanoclusters composed of at least three, super paramagnetic iron oxid particles for MRI based molecular imaging, fluorescent proteins, and enzymes.

12 . The compound of claim 10 ,
wherein the polypeptide is linked with the detectable label by means of a chemical linking group.

13 . The compound according to claim 10 for use as a diagnostic for acute myeloid leukemia subtype M2.

14 . A method for diagnosing acute myeloid leukemia subtype M2 by applying the compound of claim 10 and detecting the detectable label.

15 . A diagnostic kit comprising the polypeptide of claim 1 for use in the diagnosis of acute myeloid leukemia subtype M2.

16 . The polypeptide of claim 9 ,
wherein the polypeptide is a single chain variable fragment (scFv).

17 . The compound of claim 11 , wherein:
the fluorescent dye is selected from the group consisting of fluorescein, rhodamine, coumarine, cyanine, and derivatives thereof; the radioisotopes are selected from the group consisting of iodine-131, lutetium-177, and yttrium-90; the quantum dot composed of heavy metals is selected from the group consisting of CdSe and InGaP; the noble metal nanoc

Assignees

• Fraunhofer Ges Forschung

Inventors

• Barth Stefan
• Tur Mehmet Kemal
• Fitting Jenny

Classifications

• G01N33/57505

  of the blood, e.g. leukaemia · CPC title

• C07K16/30 Primary

  from tumour cells · CPC title

• C07K2317/565

  Complementarity determining region [CDR] · CPC title

• C07K2317/622

  Single chain antibody (scFv) · CPC title

• G01N33/57426

  Physics · mapped topic