Carbohydrate deficient transferrin (CDT) -specific antibodies, their preparation and use
View Patent ↗The present invention relates to antibodies which, in agueous solution, selectively bind to a transferrin-homologous carbohydrate deficient transferrin (CDT) without the latter needing to be bound to a solid phase. CDT is characterized by at least one of the two oligosaccharide chains which are normally bound to Asn 413 and/or Asn 611 of transferrin being entirely or substantially entirely lacking.
1. An isolated antibody or an antigen binding fragment thereof which binds selectively to carbohydrate deficient transferrin, wherein the binding takes place in the region of the following four epitopes (1) to (4) of the carbohydrate deficient transferrin sequence:
SEQ ID NO: 1
VVARSMGGKEDLIWELL and
SEQ ID NO: 2
TTEDSIAKIMNGEADAMSLDGGF and
SEQ ID NO: 3
SKLSMGSGLNLSEPN and
SEQ ID NO: 4
YEKYLGEEYVKAV.
2. An isolated antibody which binds selectively to carbohydrate deficient transferrin, wherein the binding takes place in the region of three of the following four epitopes (1) to (4) of the carbohydrate deficient transferrin sequence:
SEQ ID NO: 1
VVARSMGGKEDLIWELL and
SEQ ID NO: 2
TTEDSIAKIMNGEADAMSLDGGF and
SEQ ID NO: 3
SKLSMGSGLNLSEPN and
SEQ ID NO: 4
YEKYLGEEYVKAV;
3. An isolated antibody which binds selectively to carbohydrate deficient transferrin, wherein the binding takes place in the region of two of the following four epitopes (1) to (4) of the carbohydrate deficient transferrin sequence:
SEQ ID NO:1
VVARSMGGKEDLIWELL
and
SEQ ID NO:2
TTEDSIAKIMNGEADAMSLDGGF
and
SEQ ID NO:3
SKLSMGSGLNLSEPN
and
SEQ ID NO:4
YEKYLGEEYVKAV.
4. The antibody as claimed in claim 1 , which is a monoclonal antibody.
5. A monoclonal antibody which is produced by the cell culture having the deposit number DSM ACC2540.
6. A monoclonal antibody which is produced by the cell culture having the deposit DSM ACC2541.
7. A process for preparing the antibody as claimed in claim 1 by immunizing a suitable experimental animal with unglycosylated transferrin, fusing the spleen cells of this experimental animal to myeloma cells, resulting in antibody-producing hybrid cells, cloning the hybrid cells and selecting a hybrid cell clone which produces an antibody whose binding according to the results of an epitope mapping takes place in the region of the following four epitopes (1) to (4) of the carbohydrate deficient transferrin sequence:
SEQ ID NO:1
VVARSMGGKEDLIWELL
and
SEQ ID NO:2
TTEDSIAKIMNGEADAMSLDGGF
and
SEQ ID NO:3
SKLSMGSGLNLSEPN
and
SEQ ID NO:4
YEKYLGEEYVKAV;
and obtaining antibody from the selected hybrid cell clone.
8. An immunoassay for detecting carbohydrate deficient transferrin in a sample, which comprises contacting the antibody as claimed in claim 1 with the sample, and detecting the formation of an immune complex consisting of carbohydrate deficient transferrin and said antibody.
9. A test kit for carrying out an immunoassay as claimed in claim 8 comprising the antibody which binds selectively to carbohydrate deficient transferrin, wherein the antibody binding takes place in the region of at least two of the following four epitopes (1) to (4) of the carbohydrate deficient transferrin sequence:
SEQ ID NO:1
VVARSMGGKEDLIWELL
and
SEQ ID NO:2
TTEDSIAKIMNGEADAMSLDGGF
and
SEQ ID NO:3
SKLSMGSGLNLSEPN
and
SEQ ID NO:4
YEKYLGEEYVKAV.
10. The test kit as claimed in claim 9 , wherein the antibody binding takes place in the region of three of the following four epitopes (1) to (4) of the carbohydrate transferrin sequence:
SEQ ID NO:1
VVARSMGGKEDLIWELL
and
SEQ ID NO:2
TTEDSIAKIMNGEADAMSLDGGF
and
SEQ ID NO:3
SKLSMGSGLNLSEPN
and
SEQ ID NO:4
YEKYLGEEYVKAV.
11. The test kit as claimed in claim 9 , wherein the antibody binding takes place in the region of all four of the following epitopes (1) to (4) of the carbohydrate transferrin sequence:
SEQ ID NO:1
VVARSMGGKEDLIWELL
and
SEQ ID NO:2
TTEDSIAKIMNGEADAMSLDGGF
and
SEQ ID NO:3
SKLSMGSGLNLSEPN
and
SEQ ID NO:4
YEKYLGEEYVKAV.