Patent EA-005169-B1 (EA)
Title: RECOMBINANT ANTIBODY RECOGNIZING IOR C2, Fv FRAGMENT OF SAID ANTIBODY AND METHOD
Inventor: MATEO DE ACOSTA DEL RIO CRISTI
IPC: C07K16/30
Filed: 2000-11-16
Abstract: 1. Recombinant antibody interacting with the ior C2 antigen complex derived from murine monoclonal antibody IOR C5 produced by the hybridoma deposited under number ECCC 97061101, wherein the CDRs sequences of the light and heavy chains are the following: HEAVY CHAIN CDR1: S D Y N W H CDR2: Y I S Y N G T T S Y N P S L K S CDR3: N D E K A W F A Y LIGHT CHAIN CDR1: K S S Q S L L D S D G K T Y L N CDR2: L V S K L D S CDR3: W Q G T H F P H T 2. Recombinant antibody according to claim 1 which contains the CDRs and framework regions (FRs) of the antibody IOR C5 and human constant regions of the light and heavy chains of the antibody, wherein said framework amino acid sequences of the heavy and light chains are the following: HEAVY CHAIN FR1: D V Q L Q E S G P G L V K P S Q T L S L T C T V T G Y S I T FR2: W I R Q F P G K G L E W M G FR3: R I S I T R D T S K N Q F F L Q L N S V T T E D T A T Y Y C A R FR4: W G Q G T L V T V S A LIGHT CHAIN FR1: D V V M T Q T P L T L S V T L G Q P A S I S C FR2: W L L Q R P G Q S P R R L I Y FR3: G V P D R F S G S G S G T D F A L K I R R V E A E D L G V Y Y C FR4: F G G G T K L E I K R K S T L T G 3. Recombinant antibody according to claim 1 that additionally contains point mutations in the framework regions of the heavy and light chains for reducing its immunogenicity. 4. Recombinant antibody according to claim 3 which has in the framework regions of the heavy and light chains any of the following point mutations HEAVY CHAIN: Position 10 ASP by GLY Position 17 SER by THR Position 43 ASN by LYS Position 44 LYS by GLY LIGHT CHAIN: Position 15 ILE by LEU Position 45 LYS by ARG Position 63 THR by SER 5. Cellular line expressing the recombinant antibody of any one of claims 1 to 4. 6. Pharmaceutical composition for treating rectum and colon malignant tumours, its metastasis and recidives, comprising the recombinant antibody of any one of claims 1 to 4 and a suitable excipient. 7. Diagnostic composition for localisation and identification "in vivo" of rectum and colon malignant tumours, its metastasis and recidives, comprising the recombinant antibody of any one of claims 1 to 4. 8. Diagnostic composition according to claim 7 comprising additionally compounds for radiolabelling the antibody selected from the group consisting of technetium 99, rhenium 186 and rhenium 184. 9. Diagnostic method to identify "in vivo" rectum and colon malignant tumours, its metastasis and recidives, comprising administering to a subject a physiologically acceptable composition which contains the antibody of any one of claims 1-4, previously labelled with Tc-99m or any other radioactive analogue, and the monitoring of the biodistribution of this composition by immunogammagraphy methods. 10. Single chain Fv fragment of the recombinant antibody derived from murine monoclonal antibody IOR C5 produced by the hybridoma deposited under number ECCC 97061101, wherein the sequences of the CDRs and frameworks regions (FRs) of said fragment, being: HEAVY CHAIN FR1: D V Q L Q E S G P G L V K P S Q T L S L T C T V T G Y S I T FR2: W I R Q F P G K G L E W M G FR3: R I S I T R D T S K N Q F F L Q L N S V T T E D T A T Y Y C A R FR4: W G Q G T L V T V S A CDR1: S D Y N W H CDR2: Y I S Y N G T T S Y N P S L K S CDR3: N D E K A W F A Y LIGHT CHAIN FR1: D V V M T Q T P L T L S V T L G Q P A S I S C FR2: W L L Q R P G Q S P R R L I Y FR3: G V P D R F S G S G S G T D F A L K I R R V E A E D L G V Y Y C FR4: F G G G T K L E I K R K S T L T G CDR1: K S S Q S L L D S D G K T Y L N CDR2: L V S K L D S CDR3: W Q G T H F P H T 11. Single chain Fv fragment according to claim 10 wherein in the framework regions of the heavy and light chains it has any of the following point mutations for reducing its immunogenicity: HEAVY CHAIN: Position 10 ASP by GLY Position 17 SER by THR Position 43 ASN by LYS Position 44 LYS by GLY LIGHT CHAIN: Position 15 ILE by LEU Position 45 LYS by ARG Position 63 THR by SER. 12. A cellular line expressing the single chain Fv fragment of claim 10 or 11. 13. Pharmaceutical composition for treating rectum and colon malignant tumours, its metastasis and recidives, comprising the single chain Fv fragment of claims 10 or 11 and a suitable excipient. 14. Diagnostic composition for localisation and identification "in vivo" of rectum and colon malignant tumours, its metastasis and recidives, comprising the single chain Fv fragment of claims 10 or 11. 15. Diagnostic composition according to claim 14 comprising additionally compounds for radiolabelling the antibody selected from the group consisting of technetium 99, rhenium 186 and rhenium 184. 16. Diagnostic method to identify "in vivo" rectum and colon malignant tumours, its metastasis and recidives, comprising administering to a subject a physiologically acceptable composition which contains the single chain Fv fragment of any one of claims 10 or 11, previously labelled with Tc-99m or any other radioactive analogue, and the monitoring of the biodistribution of this composition by immunogammagraphy methods.