Chimeric antibodies directed against a human glycoprotein antigen

Claim 2

A recombinant DNA compound that comprises a first DNA sequence encoding the light chain variable region of a chimeric monoclonal antibody, the first DNA sequence coding for an amino acid sequence comprising: Asn-Ile-Val-Leu-Thr-Gln-Ser-Pro-Ala-Ser Leu-Ala-Val-Ser-Leu-Gly-Gln-Arg-Ala-Thr Ile-Ser-Cys-Arg-Ala-Ser-Glu-Ser-Val-Asp Ser-Tyr-Gly-Asn-Ser-Phe-Met-His-Trp-Tyr Gln-Gln-Lys-Pro-Gly-Gln-Pro-Pro-Lys-Leu Leu-Ile-Tyr-Leu-Ala-Ser-Asn-Leu-Glu-Ser Gly-Val-Pro-Ala-Arg-Phe-Ser-Gly-Ser-Gly Ser-Arg-Thr-Asp-Phe-Thr-Leu-Thr-Ile-Asp Pro-Val-Glu-Ala-Asp-Asp-Ala-Ala-Thr-Tyr Tyr-Cys-Gln-Gln-Asn-Asn-Glu-Asp-Pro-Leu Thr-Phe-Gly-Ser-Gly-Thr-Lys-Leu-Glu-Ile Lys-Arg.

Claim 3

The recombinant DNA compound of Claim 1 wherein the coding strand comprises: AAC-ATT-GTG-CTG-ACC-CAA-TCT-CCA-GCT-TCT TTC-GCT-GTG-TCT-CTA-GGG-CAG-AGG-GCC-ACC ATA-TCC-TGC-AGA-GCC-AGT-GAA-AGT-GTT-GAT AGT-TAT-GGC-AAT-AGT-TTT-ATG-CAC-TGG-TAC CAG-GAG-AAA-CCA-GGA-CAG-CCA-CCC-AAA-CTC CTC-ATC-TAT-CTT-GCA-TCC-AAC-CTA-GAA-TCT GGG-GTC-CCT-GCC-AGG-TTC-AGT-GGC-AGT-GGA TCT-AGG-ACA-GAC-TTC-ACC-CTC-ACC-ATT-GAT CCT-GTG-GAG-GCT-GAT-GAT-GCT-GCA-ACC-TAT TAC-TGT-CAA-CAA-AAT-AAT-GAG-GAT-CCT-CTC ACG-TTC-GGC-TCG-GGG-ACA-AAG-TTG-GAA-ATA AAA-CGG.

Claim 4

The recombinant DNA compound of Claim 1 wherein the first DNA sequence further comprises a DNA sequence encoding a eukaryotic signal peptide.

Claim 5

The recombinant DNA compound of Claim 3 wherein the DNA sequence encoding a leader peptide encodes a leader peptide with an amino acid sequence comprising: Met-Glu-Thr-Asp-Thr-Leu-Leu-Leu-Trp-Val Leu-Leu-Leu-Trp-Val-Pro-Gly-Ser-Thr-Gly.

Claim 6

The recombinant DNA compound of Claim 4 wherein the DNA encoding a leader peptide comprises: ATG-GAG-ACA-GAC-ACA-CTC-CTG-CTA-TGG-GTG CTG-CTG-CTC-TGG-GTT-CCA-GGT-TCC-ACA-GGT.

Claim 7

The recombinant DNA vector of Claim 5 that is plasmid pTZK910 (NRRL B-18478).

Claim 8

The recombinant DNA compound of Claim 1 that further comprises a second DNA sequence which encodes the light chain constant region of a chimeric monoclonal antibody.

Claim 9

A recombinant DNA vector that comprises the DNA compound of Claim 7.

Claim 10

The recombinant DNA vector of Claim 8 that is plasmid pG9.2.27K as shown in Figure 10.

Claim 11

A recombinant DNA compound that comprises a first DNA sequence encoding the heavy chain variable region of a chimeric monoclonal antibody, the first DNA sequence coding for an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 12

A recombinant DNA compound that comprises a first DNA sequence encoding the heavy chain variable region of a chimeric monoclonal antibody, the first DNA sequence coding for an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 13

The recombinant DNA compound of Claim 10 wherein the coding strand comprises: CAG-GTC-CAG-CTG-CAG-CAG-TCT-GGA-CCT-GAG CTG-GTG-AAG-CCT-GGG-GCC-TCA-GTG-AAG-ATT TCC-TGC-AAA-GCT-TCT-GGC-TAC-GCA-TTC-AGT AGG-TCT-TGG-ATG-AAC-TGG-GTG-AAG-CAG-AGG CCT-GGA-CAG-GGT-CTT-GAG-TGG-ATT-GGA-CGG ATT-TAT-CCT-GGA-GAT-GGA-GAT-ACT-AAC-TAC AAT-GGG-AAG-TTC-AAG-GGC-AAG-GCC-ACA-CTG ACT-GCA-GAC-AAA-TCC-TCC-AGC-ACA-GCC-TAC ATG-CAG-GTC-AGC-AGC-CTG-ACC-TCT-GTG-GAC TCT-GCG-GTC-TAT-TTC-TGT-GCA-AGA-GGG-AAT ACG-GTA-GTA-GTT-CCC-TAT-ACT-ATG-GAC-TAC TGG-GGT-CAA-GGA-ACC-TCA-GTC-ACC-GTC-TCC TCA.

Claim 14

The recombinant DNA compound of Claim 10 wherein the first DNA sequence further comprises a DNA sequence encoding a eukaryotic signal peptide.

Claim 15

The recombinant DNA compound of Claim 12 wherein the DNA sequence encoding a leader peptide encodes a leader peptide with an amino acid sequence comprising: Met-Gly-Trp-Ser-Arg-Ile-Phe-Leu-Phe-Leu Leu-Ser-Ile-Thr-Ala-Gly-Val-His-Cys.

Claim 16

The recombinant DNA compound of Claim 13 wherein the DNA encoding a leader peptide comprises: ATG-GGA-TGG-AGC-CGG-ATC-TTT-CTC-TTC-CTC CTG-TCA-ATA-ACT-GCA-GGT-GTC-CAT-TGC.

Claim 17

The recombinant DNA vector of Claim 14 that is plasmid pG4G21 (NRRL B-18479).

Claim 18

The recombinant DNA compound of Claim 15 that further comprises a second DNA sequence which encodes the heavy chain constant region of a chimeric monoclonal antibody.

Claim 19

A recombinant DNA vector that comprises the DNA compound of Claim 16.

Claim 20

The recombinant DNA vector of Claim 17 that is plasmid pN9.2.27G1 as shown in Figure 12.

Claim 21

A eukaryotic host cell capable of expression of a chimeric monoclonal antibody comprising at least one DNA vector which comprises a DNA sequence encoding for the light chain of the chimeric antibody, and transcriptional and translational DNA sequences positioned in relation to the light chain-encoding DNA sequence to direct expression of the light chain, wherein the DNA strand sequence encoding for the light chain variable region comprises: AAC-ATT-GTG-CTG-ACC-CAA-TCT-CCA-GCT-TCT TTC-GCT-GTG-TCT-CTA-GGG-CAG-AGG-GCC-ACC ATA-TCC-TGC-AGA-GCC-AGT-GAA-AGT-GTT-GAT AGT-TAT-GGC-AAT-AGT-TTT-ATG-CAC-TGG-TAC CAG-GAG-AAA-CCA-GGA-CAG-CCA-CCC-AAA-CTC CTC-ATC-TAT-CTT-GCA-TCC-AAC-CTA-GAA-TCT GGG-GTC-CCT-GCC-AGG-TTC-AGT-GGC-AGT-GGA TCT-AGG-ACA-GAC-TTC-ACC-CTC-ACC-ATT-GAT CCT-GTG-GAG-GCT-GAT-GAT-GCT-GCA-ACC-TAT TAC-TGT-CAA-CAA-AAT-AAT-GAG-GAT-CCT-CTC ACG-TTC-GGC-TCG-GGG-ACA-AAG-TTG-GAA-ATA AAA-CGG.

Claim 22

A eukaryotic host cell capable of expression of a chimeric monoclonal antibody comprising at least one DNA vector which comprises a DNA sequence encoding for the light chain of the chimeric antibody, and transcriptional and translational DNA sequences positioned in relation to the light chain-encoding DNA sequence to direct expression of the light chain, wherein the DNA strand sequence encoding for the light chain variable region comprises: AAC-ATT-GTG-CTG-ACC-CAA-TCT-CCA-GCT-TCT TTC-GCT-GTG-TCT-CTA-GGG-CAG-AGG-GCC-ACC ATA-TCC-TGC-AGA-GCC-AGT-GAA-AGT-GTT-GAT AGT-TAT-GGC-AAT-AGT-TTT-ATG-CAC-TGG-TAC CAG-GAG-AAA-CCA-GGA-CAG-CCA-CCC-AAA-CTC CTC-ATC-TAT-CTT-GCA-TCC-AAC-CTA-GAA-TCT GGG-GTC-CCT-GCC-AGG-TTC-AGT-GGC-AGT-GGA TCT-AGG-ACA-GAC-TTC-ACC-CTC-ACC-ATT-GAT CCT-GTG-GAG-GCT-GAT-GAT-GCT-GCA-ACC-TAT TAC-TGT-CAA-CAA-AAT-AAT-GAG-GAT-CCT-CTC ACG-TTC-GGC-TCG-GGG-ACA-AAG-TTG-GAA-ATA AAA-CGG.

Claim 23

The eukaryotic host cell according to Claim 19 wherein a second DNA vector comprises, or the first DNA construct fruther comprises, a DNA strand sequence encoding for the heavy chain of the chimeric antibody, and transcriptional and translational DNA sequences positioned in relation to the heavy chain-­encoding DNA strand sequence to direct expression of the heavy chain, wherein the DNA sequence encoding for the heavy chain variable region comprises: CAG-GTC-CAG-CTG-CAG-CAG-TCT-GGA-CCT-GAG CTG-GTG-AAG-CCT-GGG-GCC-TCA-GTG-AAG-ATT TCC-TGC-AAA-GCT-TCT-GGC-TAC-GCA-TTC-AGT AGG-TCT-TGG-ATG-AAC-TGG-GTG-AAG-CAG-AGG CCT-GGA-CAG-GGT-CTT-GAG-TGG-ATT-GGA-CGG ATT-TAT-CCT-GGA-GAT-GGA-GAT-ACT-AAC-TAC AAT-GGG-AAG-TTC-AAG-GGC-AAG-GCC-ACA-CTG ACT-GCA-GAC-AAA-TCC-TCC-AGC-ACA-GCC-TAC ATG-CAG-GTC-AGC-AGC-CTG-ACC-TCT-GTG-GAC TCT-GCG-GTC-TAT-TTC-TGT-GCA-AGA-GGG-AAT ACG-GTA-GTA-GTT-CCC-TAT-ACT-ATG-GAC-TAC TGG-GGT-CAA-GGA-ACC-TCA-GTC-ACC-GTC-TCC TCA.

Claim 24

The eukaryotic host cell of Claim 20 wherein the first DNA vector encodes the light chain of the chimeric monoclonal antibody and the second DNA vector encodes the heavy chain of the chimeric monoclonal antibody.

Claim 25

The eukaryotic host cell of Claim 21 that is SP2/0/pG9.2.27K/pN9.2.27G1.

Claim 26

A chimeric monoclonal antibody comprising a light chain variable region having an amino acid sequence comprising: Asn-Ile-Val-Leu-Thr-Gln-Ser-Pro-Ala-Ser Leu-Ala-Val-Ser-Leu-Gly-Gln-Arg-Ala-Thr Ile-Ser-Cys-Arg-Ala-Ser-Glu-Ser-Val-Asp Ser-Tyr-Gly-Asn-Ser-Phe-Met-His-Trp-Tyr Gln-Gln-Lys-Pro-Gly-Gln-Pro-Pro-Lys-Leu Leu-Ile-Tyr-Leu-Ala-Ser-Asn-Leu-Glu-Ser Gly-Val-Pro-Ala-Arg-Phe-Ser-Gly-Ser-Gly Ser-Arg-Thr-Asp-Phe-Thr-Leu-Thr-Ile-Asp Pro-Val-Glu-Ala-Asp-Asp-Ala-Ala-Thr-Tyr Tyr-Cys-Gln-Gln-Asn-Asn-Glu-Asp-Pro-Leu Thr-Phe-Gly-Ser-Gly-Thr-Lys-Leu-Glu-Ile Lys-Arg.

Claim 27

A chimeric monoclonal antibody comprising a light chain variable region having an amino acid sequence comprising: Asn-Ile-Val-Leu-Thr-Gln-Ser-Pro-Ala-Ser Leu-Ala-Val-Ser-Leu-Gly-Gln-Arg-Ala-Thr Ile-Ser-Cys-Arg-Ala-Ser-Glu-Ser-Val-Asp Ser-Tyr-Gly-Asn-Ser-Phe-Met-His-Trp-Tyr Gln-Gln-Lys-Pro-Gly-Gln-Pro-Pro-Lys-Leu Leu-Ile-Tyr-Leu-Ala-Ser-Asn-Leu-Glu-Ser Gly-Val-Pro-Ala-Arg-Phe-Ser-Gly-Ser-Gly Ser-Arg-Thr-Asp-Phe-Thr-Leu-Thr-Ile-Asp Pro-Val-Glu-Ala-Asp-Asp-Ala-Ala-Thr-Tyr Tyr-Cys-Gln-Gln-Asn-Asn-Glu-Asp-Pro-Leu Thr-Phe-Gly-Ser-Gly-Thr-Lys-Leu-Glu-Ile Lys-Arg.

Claim 28

The chimeric monoclonal antibody according to Claim 23 wherein the antibody is chimeric 9.2.27.

Claim 29

A chimeric monoclonal antibody comprising a heavy chain variable region having an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 30

A chimeric monoclonal antibody comprising a heavy chain variable region having an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 31

The chimeric monoclonal antibody according to Claim 25 wherein the antibody is chimeric 9.2.27.

Claim 32

A chimeric monoclonal antibody comprising a light chain variable region having an amino acid sequence comprising: Asn-Ile-Val-Leu-Thr-Gln-Ser-Pro-Ala-Ser Leu-Ala-Val-Ser-Leu-Gly-Gln-Arg-Ala-Thr Ile-Ser-Cys-Arg-Ala-Ser-Glu-Ser-Val-Asp Ser-Tyr-Gly-Asn-Ser-Phe-Met-His-Trp-Tyr Gln-Gln-Lys-Pro-Gly-Gln-Pro-Pro-Lys-Leu Leu-Ile-Tyr-Leu-Ala-Ser-Asn-Leu-Glu-Ser Gly-Val-Pro-Ala-Arg-Phe-Ser-Gly-Ser-Gly Ser-Arg-Thr-Asp-Phe-Thr-Leu-Thr-Ile-Asp Pro-Val-Glu-Ala-Asp-Asp-Ala-Ala-Thr-Tyr Tyr-Cys-Gln-Gln-Asn-Asn-Glu-Asp-Pro-Leu Thr-Phe-Gly-Ser-Gly-Thr-Lys-Leu-Glu-Ile Lys-Arg; and a heavy chain variable region having an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 33

A chimeric monoclonal antibody comprising a light chain variable region having an amino acid sequence comprising: Asn-Ile-Val-Leu-Thr-Gln-Ser-Pro-Ala-Ser Leu-Ala-Val-Ser-Leu-Gly-Gln-Arg-Ala-Thr Ile-Ser-Cys-Arg-Ala-Ser-Glu-Ser-Val-Asp Ser-Tyr-Gly-Asn-Ser-Phe-Met-His-Trp-Tyr Gln-Gln-Lys-Pro-Gly-Gln-Pro-Pro-Lys-Leu Leu-Ile-Tyr-Leu-Ala-Ser-Asn-Leu-Glu-Ser Gly-Val-Pro-Ala-Arg-Phe-Ser-Gly-Ser-Gly Ser-Arg-Thr-Asp-Phe-Thr-Leu-Thr-Ile-Asp Pro-Val-Glu-Ala-Asp-Asp-Ala-Ala-Thr-Tyr Tyr-Cys-Gln-Gln-Asn-Asn-Glu-Asp-Pro-Leu Thr-Phe-Gly-Ser-Gly-Thr-Lys-Leu-Glu-Ile Lys-Arg; and a heavy chain variable region having an amino acid sequence comprising: Gln-Val-Gln-Leu-Gln-Gln-Ser-Gly-Pro-Glu Leu-Val-Lys-Pro-Gly-Ala-Ser-Val-Lys-Ile Ser-Cys-Lys-Ala-Ser-Gly-Tyr-Ala-Phe-Ser Arg-Ser-Trp-Met-Asn-Trp-Val-Lys-Gln-Arg Pro-Gly-Gln-Gly-Leu-Glu-Trp-Ile-Gly-Arg Ile-Tyr-Pro-Gly-Asp-Gly-Asp-Thr-Asn-Tyr Asn-Gly-Lys-Phe-Lys-Gly-Lys-Ala-Thr-Leu Thr-Ala-Asp-Lys-Ser-Ser-Ser-Thr-Ala-Tyr Met-Gln-Val-Ser-Ser-Leu-Thr-Ser-Val-Asp Ser-Ala-Val-Tyr-Phe-Cys-Ala-Arg-Gly-Asn Thr-Val-Val-Val-Pro-Tyr-Thr-Met-Asp-Tyr Trp-Gly-Gln-Gly-Thr-Ser-Val-Thr-Val-Ser Ser.

Claim 34

The chimeric monoclonal antibody according to Claim 27 wherein the antibody is XCEM 449.