JPH07203974A - Gene fragment of antibody recognizing cancer-specific mucin - Google Patents

Abstract

PURPOSE:To provide a novel gene participating the production of an antibody recognizing cancer-specific mucins, thus being useful in analyses of physiological roles of cancer-specific mucins and production of therapeutic and diagnostic agents for cancers. CONSTITUTION:The gene encodes the V-domain in the H-chain of the antibody recognizing the cancer-specific mucin having the amino acid sequence of the formula and includes the base sequence of the formula. Clone NdH4 hybridizing to the JH probe is isolated from the genomic library of Nd 2 hybridoma producing an antibody Nd 2 which recognizes cancer-specific mucins and injected into pBLUEscript to prepare pBLUE/NdH4, then introduced into Escherichia coli MKV1184 strain and cultured to prepare a single strand DNA as a sequence template and determine the base sequence of the V-domain in the heavy strand of the Nd2 antibody. The gene can be also prepared by a DNA synthesizer.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to an antibody gene or the like that recognizes cancer-specific mucin. Specifically, culturing a V region part of an H chain or L chain of an antibody that recognizes cancer-specific mucin, a gene encoding the same, a vector containing this gene, and a host cell transformed with this vector The present invention also relates to a method for producing an antibody that recognizes the above-mentioned region portion or cancer-specific mucin, and further to a method for producing a chimeric antibody that recognizes cancer-specific mucin.

[0002]

2. Description of the Related Art It is known that an antibody that recognizes an antigen specifically expressed in cancer cells (cancer-specific antigen) is effective in the diagnosis and treatment of cancer. Glycoprotein in which N-acetylgalactosamine and serine or threonine are O-glycoside-linked is generically called mucin, but it is known that certain mucins are cancer-specific antigens.

Ho et al., Immunized mice with mucin extracted from a human pancreatic cancer cell line and established a hybridoma producing a mouse monoclonal antibody that specifically recognizes pancreatic cancer tissue (Ho et al., Cancer. Res., 51, p372, 1991).
Sawada et al. Demonstrated that Nd2, which is one of the antibodies produced by Ho et al., Specifically recognizes human pancreatic cancer by administering Nd2 to mice transplanted with human pancreatic cancer (Sawada et al., Antibody, Immunoconjugates, and Ra
diopharmaceuticals, vol. 4, p493, 1991).

Furthermore, Sawada et al. Administered Nd2 derived from mouse to pancreatic cancer patients, and suggested that Nd2 is useful for in vivo diagnosis of human pancreatic cancer (Sawada et al., Annual Meeting of the Japanese Cancer Society,
p3291991).

[0005]

In order to use a monoclonal antibody that recognizes cancer-specific mucin as an in-vivo diagnostic agent or therapeutic agent, it is necessary to administer the antibody in a large amount or repeatedly. is there. However, it is known that when a mouse-derived antibody such as Nd2 is administered to a human, an antibody that recognizes the mouse antibody appears in human serum (Levy et al., Annu. Rev. Med. 34, p107, 1983). Year). Therefore, administration of a large amount or repeated administration of a mouse-derived antibody to humans may lead to the risk of anaphylaxis due to an immune response, the administered antibody being rapidly degraded, and its original activity lost. There is. To solve this problem, human antibodies may be used, but it is difficult to isolate B cells producing the desired antibody from a population of human B cells, immortalize them, and culture them. .

In recent years, mouse-human chimeric antibodies have been proposed to solve these problems. A mouse-human chimeric antibody is an antibody (Oi and Morrison, Biotechnology) consisting of an antigen binding region derived from an animal species other than human, that is, a variable region (V region) and a human constant region (C region).
4, p214, 1986), the C region of human origin does not receive an immune response from human antibodies, so that the risk of anaphylaxis can be reduced.

More recently, humanized antibodies have been proposed. A humanized antibody is an antibody that comprises the minimum antigen-binding region derived from an animal species other than human, that is, the CDR region, and other human-derived V regions and C regions, and further reduces the possibility of receiving an immune response from human antibodies. You can

[0008] In order to enhance the function of the antibody as an in-vivo diagnostic agent or therapeutic agent, a fusion protein of the antibody and another protein, or an antibody consisting only of the V region has been proposed (Ward).
Et al., Nature, 341, p544, 1989).

As described above, it is possible to obtain more effective antibodies as in-vivo diagnostic agents and therapeutic agents by applying various techniques proposed in recent years to Nd2 and the like. In order to prepare Escherichia coli, a gene encoding the H chain and L chain of the antibody is required.

[0010]

In view of the above-mentioned problems, the present inventors have devised a mouse-derived anti-cancer-specific mucin antibody H chain or L chain.
As a result of intensive studies on chain genes, the inventors have found the genes of the V region and completed the present invention. That is, the present invention provides an H chain V region of an antibody that recognizes a cancer-specific mucin, which comprises the amino acid sequence shown in SEQ ID NO: 1, and a DNA encoding the H chain V region of an antibody that recognizes the cancer-specific mucin. A vector containing the DNA and capable of expressing the DNA in a host cell, and culturing a host cell transformed with the vector, which comprises the H chain V region of an antibody recognizing a cancer-specific mucin It is a manufacturing method.

The present invention also encodes the L chain V region of an antibody that recognizes a cancer-specific mucin and the L chain V region of an antibody that recognizes the cancer-specific mucin, which contains the amino acid sequence shown in SEQ ID NO: 2. L-chain V of an antibody that recognizes a cancer-specific mucin, which comprises culturing a host cell containing the DNA, the vector containing the DNA and capable of expressing the DNA in the host cell, and the host cell transformed with the vector It is a method of manufacturing a region.

Furthermore, the present invention provides a DNA encoding the H chain V region of the antibody recognizing the cancer-specific mucin and a DNA encoding the L chain V region of the antibody recognizing the cancer-specific mucin.
A method for producing a chimeric antibody that recognizes a cancer-specific mucin, which comprises culturing a vector containing the above vector and capable of expressing these DNAs in a host, and a host cell transformed with the vector. The present invention will be described in detail below.

The gene of the antibody which recognizes the cancer-specific mucin of the present invention is H which produces an antibody which recognizes the cancer-specific mucin.
Gene coding for V region of chain or V region of L chain
Is a gene to be cloned, and specifically has the nucleotide sequence shown in SEQ ID NO: 1 (FIG. 3) or SEQ ID NO: 2 (FIG. 4). The gene of the present invention can be isolated, for example, by the method shown in Example 1 using a hybridoma that produces an antibody that recognizes a cancer-specific mucin as a starting material. Hybridomas that produce Nd2 (Ho
, Cancer Res., 51, p372, 1991).

The gene of SEQ ID NO: 1 or SEQ ID NO: 2 can also be prepared by using, for example, a DNA synthesizer. In the gene of SEQ ID NO: 1 or SEQ ID NO: 2, one or more bases are added,
One or a plurality of bases may be deleted and one or a plurality of bases may be replaced.

The antibody (V region) recognizing the cancer-specific mucin of the present invention is encoded by the above-mentioned gene and has the sequence of SEQ ID NO: 1 (FIG. 3) or SEQ ID NO: 2 (FIG. 4). Is. In the antibody (V region) of SEQ ID NO: 1 or SEQ ID NO: 2, one or more amino acid residues are added, one or more amino acid residues are deleted, and one or more amino acids are The residue may be substituted.

In order to genetically engineer an antibody that recognizes a cancer-specific mucin, a vector containing a gene encoding it is required. The vector of the present invention contains, for example, all or part of the gene fragment of SEQ ID NO: 1 and / or SEQ ID NO: 2 and is capable of expressing the gene in a host cell. Therefore, the vector includes a promoter / operator for expressing (transcription) a gene other than the gene of the present invention, a terminator for terminating expression (transcription) of the gene, It may contain a known gene sequence such as a gene for replication in a host cell. In the present invention, the gene encoding the V region of the H chain and the V region of the L chain are used.
Include one that contains the gene that codes for the region. According to this vector, an antibody (Fv antibody) that does not include the C region and recognizes a cancer-specific mucin consisting of only the V region can be produced.

By further adding a gene fragment encoding the C region of human antibody to the vector, it can be used as a vector for producing the H chain or L chain of a mouse-human chimeric antibody. In this case, regarding the human antibody gene, the L chain C region is the L chain C region of the present invention so that the H chain C region of the human antibody gene is in a state of being in frame with the H chain V region of the present invention.
It is prepared so that the frame matches the chain V region. A gene fragment coding for the C region of H chain or L chain of human antibody has been publicly known (for example, Kameyama et al.
FEBS Lett., Volume 244, p301, 1989).

Among the vectors of the present invention, the vector for producing a mouse-human chimeric antibody that recognizes a cancer-specific mucin includes, for example, all or part of the gene of SEQ ID NO: 1 and the gene of SEQ ID NO: 2. All or part of human antibody H
Gene encoding the C region of the chain and C of the L chain of human antibody
It is a vector containing a gene coding for the region and capable of expressing the gene fragment in a host cell. Here, regarding the human antibody gene fragment, the H chain C region is in a state in which the frame matches the H chain V region.
The vector is constructed so that the L chain C region is in a state in which the frame matches the L chain V region.

Transformation of a host cell with the above vector may be carried out by a conventional method without any particular limitation. For example, if the vector is for E. coli, E. coli may be used as a host, and if the vector is for yeast, yeast may be used as a host.

By culturing the host cells thus transformed under appropriate conditions, various forms of antibody proteins can be produced. In the present invention, various forms of antibodies can be used to produce a protein depending on the vector used. However, if the vector of the present invention is selected and used, the V chain of the H chain of the antibody recognizing the cancer-specific mucin is selected. Region protein, V region protein of L chain of antibody that recognizes cancer-specific mucin, chimeric protein of V region of H chain of antibody that recognizes cancer-specific mucin and C region of H chain of human antibody, cancer-specific mucin For producing a chimeric protein of the V region of the antibody L chain and the C region of the human antibody L chain or a chimeric antibody of the V region of the antibody recognizing cancer-specific mucin and the C region of human antibody, etc. it can. In addition to this, an antibody protein comprising the V regions of the H chain and L chain of an antibody that recognizes cancer-specific mucin can be produced.

The present invention further provides the hypervariable region (CDR) of mouse Nd2. Specifically, it is an underlined sequence portion in the base sequence of the V region of the Nd2 · H chain or the V region of the Nd2 · L chain shown in FIG. 3 or FIG. By leaving only this CDR and converting the other part into, for example, the corresponding part of a human antibody, a humanized antibody having mucin-binding properties equivalent to mouse Nd2 can be obtained.

[0022]

[0023]

[Sequence Listing] SEQ ID NO: 1 Sequence length: 122 Sequence type: Amino acid Topology :: Linear Sequence type: Protein sequence 5 10 15 GAA GTG AAG CTG GTG GAG TCT GGG GGA GTC TTA GTG AAG TCT GGA GGG Glu Val Lys Leu Val Glu Ser Gly Gly Val Leu Val Lys Ser Gly Gly 20 25 30 TCC CTG AAA CTC TCC TGT GCA GTC TCT GGA TTC ACT TTC AGT AAC TAT Ser Leu Lys Leu Ser Cys Ala Val Ser Gly Phe Thr Phe Ser Asn Tyr 35 40 45 GGC ATG TCT TGG GTT CGC CAG ACT CCG GAG AAG AGG CTG GAG TGG GTC Gly Met Ser Trp Val Arg Gln Thr Pro Glu Lys Arg Leu Glu Trp Val 50 55 60 GCA ACC ATT AGC AAT AGT GGT AGA TAC ACC TAC TAC TTT CCA GAC AGT GTG Ala Thr Ile Ser Asn Ser Gly Arg Tyr Thr Tyr Phe Pro Asp Ser Val 65 70 75 80 AAG GGG CCG TTC GCC ATC TCC AGA GAC AAT GCC AAG AAC AAC CTG TAC Lys Gly Pro Phe Ala Ile Ser Arg Asp Asn Ala Lys Asn Asn Leu Tyr 85 90 95 CTG CAA ATG AGC AGT CTG AGG TCT GCG GAC ACG GCC TTG TAT TAC TGT Leu Gln Met Ser Ser Leu Arg Ser Ala Asp Thr Ala Leu Tyr Tyr Cys 100 105 110 ACA A GA CAT TTA GAC TAT GCT AAC TAC GAT GCT ATG GAC TAT TGG GGT Thr Arg His Leu Asp Tyr Ala Asn Tyr Asp Ala Met Asp Tyr Trp Gly 115 120 CAA GGA ACC TCT GTC ACC GTC TCC TCA GGT Gln Gly Thr Ser Val Thr Val Ser Ser Gly SEQ ID NO: 2 Sequence length: Sequence type: Amino acid Topology −: Linear Sequence type: Protein sequence 5 10 15 GAC ATT GTG CTG ACA CAG TCT CCT GCT TCC TTA GCT GTA TCT CTG GGG Asp Ile Val Leu Thr Gln Ser Pro Ala Ser Leu Ala Val Ser Leu Gly 20 25 30 CAG AGG GCC ACC ATC TCA TGC AGG GCC AGC AAA AGT GTC ACT ACA TCT Gln Arg Ala Thr Ile Ser Cys Arg Ala Ser Lys Ser Val Thr Thr Ser 35 40 45 GAC TTT AGT TAT ATG CAC TGG TAC CAA CAG AAA CCC GGA CAG CCA CCC Asp Phe Ser Tyr Met His Trp Tyr Gln Gln Lys Pro Gly Gln Pro Pro 50 55 60 AAA CTC CTC CTC TAT CTT GCA TCC AAC CTA GAA TCT GGG GTC CCT GAC Lys Leu Leu Leu Tyr Leu Ala Ser Asn Leu Glu Ser Gly Val Pro Asp 65 70 75 80 AGG TTC AGT GGC AGT GGG TCT GGG ACA GAC TTC ACC CTC AAC ATC CAT Arg Phe Ser Gly Ser Gly Ser G ly Tyr Asp Phe Tyr Leu Asn Ile His 85 90 95 CCT GTG GAG GAG GAG GAT GCT GCA ACC TAT TAC TGT CAG CAC AGT AGG Pro Val Glu Glu Glu Asp Ala Ala Thr Tyr Tyr Cys Gln His Ser Arg 100 105 110 GAG TTT CCG TGG ACG TTC GGT GGA GGC ACC AAA CTG GAA ATC AAA CGT Glu Phe Pro Trp Thr Phe Gly Gly Gly Thr Lys Leu Glu Ile Lys Arg

[0024]

EXAMPLES Examples will be shown below to explain the present invention in more detail, but the present invention is not limited to these examples.

Example 1. Isolation of Nd2 antibody gene First, Nd2 hybridoma DNA was prepared by the following method. 1 × 10 ⁸ Nd2 cultured in RPMI1640
Hybridoma (Ho et al., Cancer Res., Vol. 51, p372, 19
(1991) was washed with PBS, suspended in a guanidine isothiocyanate solution, layered on cesium chloride and subjected to ultracentrifugation to separate DNA (supernatant) and RNA (precipitate). After the supernatant was dialyzed, it was treated with proteinase and RNase, deproteinized and purified by a usual method to purify 10 mg of DNA. From the precipitate, deproteinization and purification are carried out by the usual method,
1 mg of RNA was purified.

Next, a genomic library was prepared by the following method. DN completely digested with restriction enzyme EcoRI
A (2 μg) and λD also completely digested with EcoRI
After ligating ASH II (manufactured by Toyobo Co., Ltd., 1 μg) at 4 ° C. for one day, it was mixed with 1 part tube of Fage-Extrast of in vitro packaging kit (manufactured by Toyobo Co., Ltd.). After incubating at 20 ° C. for 2 hours, 400 μl of SM buffer was added to prepare a genomic library.

The isolation of the antibody gene was performed as follows. 1
Escherichia coli SRB (P2) cultured overnight was inoculated into 100 ml of TB medium containing 0.2% maltose and 10 mM MgSO ₄ , and
After culturing until the D reaches 0.5, collect the bacteria and add 20 ml of 10 mM Mg.
Suspended in SO ₄ . Next, SM buffer containing a library corresponding to 1 × 10 ⁶ and P2392 were mixed in equal amounts, and the mixture was mixed at 37 ° C.
Incubate at room temperature for 30 minutes, and plate on 10 TB agarose plates (15 cm in diameter) containing 8 mM MgSO _4, and leave for 1 day 37 days.
It was kept warm at ℃. The next day, a nylon filter was placed on each plate and allowed to stand for 2 minutes, and the filter was removed. Next, the filter was immersed in 0.5 N NaOH and 1.5 M NaCl for alkali treatment, immersed in 0.5 M Tris-HCl (pH 8.0) and 1.5 M NaCl for neutralization, immersed in 0.2 X SSC, and then air-dried. Then J _H
Probe or Jκ probe (Kameyama et al., FEBS Let
. t, 244 Volume, p301, 1989 years) using a plug - Kuhaiburidaize - do Deployment, J _H pro - Breakfast hybridize to the black - down, NdH4, and Jκ professional - hybridize to the blanking black - down, NdK19 was isolated. As a result of isolating these insert DNAs and identifying them by Southern blot analysis, it was found that the fragment hybridizing to the _JH probe was 2.5 kbp and the fragment hybridizing to the Jκ probe was 6.2 kbp. .

Each of the above fragments was inserted into pBLUEscript to prepare pBLUE / NdH4 and pBLUE / NdK19. Furthermore, a restriction enzyme map of insert DNA was prepared, and it was confirmed that the isolated gene fragment was a rearranged antibody gene.
FIG. 1 shows a restriction map of the V region of the Nd2 · H chain, and FIG. 2 shows a restriction map of the V region of the Nd2 · L chain.

Example 2. Determination of Nd2 antibody gene nucleotide sequence The nucleotide sequences of the V region of the H chain and the V region of the L chain of the Nd2 antibody isolated by the method shown in Example 1 were determined by the following method.

Each of pBLUE / NdH4 and pBLUE / NdK19 was introduced into E. coli MV1184 strain, and then the E. coli was cultured,
Single-stranded DNA was prepared by a conventional method and used as a template for sequencing. Sequence is Sequenase Ve
Using the rsion2.0 DNA Sequencing Kit (manufactured by Toyobo Co., Ltd.), the nucleotide sequence of the V region gene of the H chain or L chain of the Nd2 antibody was determined. Results for pBLUE / NdH4 (V of H chain
Figure 3 shows the results for pBLUE / NdK19 (region V).
Area) is shown in FIG.

Example 3. Preparation of Nd2 mouse / human chimeric antibody pBLUE / NdH4 and pBLUE / NdK19 insert DNAs were used as mouse / human chimeric H chain expression vector pSV2-HG1gpt and mouse / human chimeric L chain expression vector pSV2-HCκneo (Ka
meyama et al., FEBS Lett., Vol. 244, p301, 1989), pSV2-HG1gpt / NdH4 and pSV2-HCκneo / Nd.
We made K19. Figure 5 shows a restriction map of pSV2-HG1gpt / NdH4, and Figure 6 shows a restriction map of pSV2-HCκneo / NdK19.
Shown respectively.

The expression vector was introduced into the cells as follows. 1 × 10 ⁷ SP2 / 0 cells cultured by the usual method in DMEM medium containing 10% bovine serum were added to PB.
PBS pre-cooled to 4 ° C after washing with S
Suspended at 0.4 ml. Add 50 μg each of pSV2-HG1gpt / NdH4 and pSV2-HCκneo / NdK19 to 0.4 ml of P
The solution dissolved in BS and cooled on ice was mixed, placed on ice for 10 minutes, and a Gene pulser (BioRad) was used.
Pulsed at 1100 V, 25 microfarads. After placing on ice for a further 10 minutes, the plate was washed with a medium and further cultured in a normal medium. After 3 days, the medium was replaced with a selective medium (normal medium containing 0.8 mg / ml of G418 (manufactured by Gibco)) and seeded on a 96-well plate. The medium was replaced after 7 and 14 days, and the chimeric antibody titer of the supernatant was measured after 21 days, and 15 positive clones were selected. Finally, clone HCNd8A with the highest expression level was selected. Binding of Nd2 Mouse / Human Chimeric Antibody to Mucin The assay of chimeric antibody titer was performed as follows. Anti-human Ig
The plate coated with G1 was blocked with 1% BSA and PBS. The culture supernatant was added to this, and it was left at room temperature for 2 hours or more. After washing the plate, HRP-conjugated anti-human Ig
κ was added and the mixture was allowed to stand at room temperature for 2 hours or longer. Finally, after washing the plate, a coloring reagent (manufactured by Dainippon Pharmaceutical Co., Ltd.) was added, and the degree of coloring was determined by using an immunoreader (MPR-A4, Tosoh Corporation).
(Manufactured by KK). As a result, it was found that HCNd8A produced about 10 μg / ml chimeric antibody in the culture supernatant.

The binding properties of the produced chimeric Nd2 antibody to mucin were determined as follows. Human pancreatic cancer cell line SW1
Mucin extracted from 990 (Ho et al., Cancer Res., 51
, P372, 1991) coated by a conventional method.
The sample was added to the autoclave and left at room temperature for 2 hours or more. After washing the plate, HRP-conjugated anti-human Igκ was added and the plate was allowed to stand at room temperature for 2 hours or longer. After washing the plate, a coloring reagent (manufactured by Dainippon Pharmaceutical Co., Ltd.) was added, and the degree of coloring was measured with an immunoreader (M
It was measured by PR-A4, manufactured by Tosoh Corporation.

The results are shown in FIG. As is clear from FIG. 7, the chimeric antibody contained in the culture supernatant of HCNd8A retained the binding property to mucin.

Example 5. Immunostaining of human pancreatic cancer tissue using Nd2 mouse / human chimeric antibody Immunostaining of human cancer tissue using chimeric Nd2 antibody
ABC method (Yuan et al., Cancer Res. 45, p6179, 1985).
Year)). First, the endogenous peroxidase activity was inhibited by 1% hydrogen peroxide. Next, the organization
The culture supernatant of HCNd8A diluted with 5% rabbit serum, HRP-labeled anti-human Igκ and ABC reagent (manufactured by Vector Laboratories) were added in that order in the usual way. As a comparison, immunostaining using a mouse Nd2 antibody was performed.

The results are shown in Table 1. As is clear from Table 1, the chimeric Nd2 antibody and the mouse Nd2 antibody showed similar staining patterns. This proves that the antigen-binding property of the chimeric Nd2 antibody is similar to that of the mouse Nd2 antibody.

[0037]

[Table 1]

[0038]

EFFECT OF THE INVENTION A gene encoding the V regions of the H chain and L chain of the antibody recognizing the cancer-specific mucin provided by the present invention, and further for producing the antibody using a genetic engineering technique. By the means and method described above, it is possible to mass-produce the antibody having various additional functions. They are,
It is important for the analysis of the physiological role of cancer-specific mucin and has great significance in the development of therapeutic and diagnostic agents for cancer.

[Brief description of drawings]

FIG. 1 is a diagram showing a restriction enzyme map of the V region of Nd2 · H chain, where E, H, X, S, and En are respectively Ec.
oRI site, HindIII site, XbaI site, SacI and enhancer are shown.

FIG. 2 is a diagram showing a restriction enzyme map of the V region of Nd2 · L chain, where E, B, H, and X indicate EcoRI site, BamHI site, HindIII site, and XbaI site, respectively.

FIG. 3 shows the nucleotide sequence of V region of Nd2 · H chain and the amino acid sequence of V region of H chain deduced from the sequence.
In the figure, the three underlined base sequences indicate the hypervariable regions (CDR1, CDR2 and CDR3 from the upstream side).

FIG. 4 shows the nucleotide sequence of V region of Nd2 L chain and the amino acid sequence of L region V region deduced from the sequence.
In the figure, the three underlined base sequences indicate the hypervariable regions (CDR1, CDR2 and CDR3 from the upstream side).

FIG. 5 is a view showing expression vector-pSV2-HG1gpt / NdH4, and symbols in the figure have the same meanings as symbols used in ordinary genetic engineering.

FIG. 6 is a diagram showing the expression vector pSV2-HCκneo / NdK19, and the symbols in the figure have the same meanings as those used in ordinary genetic engineering.

FIG. 5 shows the chimeric NDs obtained by the method shown in Example 4.
2 shows the color development of the plate when the binding of 2 antibody (○) and mouse type ND2 antibody (●) to mucin was examined, the vertical axis shows the absorbance at 500 nm, and the horizontal axis shows the absorbance. Co-
It shows the dilution of mucin at the time.

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Claims (13)

[Claims] 1. An H chain V region of an antibody that recognizes a cancer-specific mucin, which comprises the amino acid sequence shown in SEQ ID NO: 1. 2. A gene encoding the H chain V region of the antibody that recognizes the cancer-specific mucin according to claim 1. 3. A base sequence shown in SEQ ID NO: 1,
The gene according to claim 2. 4. A vector containing the DNA according to claim 2 and capable of expressing the gene in a host cell. 5. A method for producing an H chain V region of an antibody that recognizes a cancer-specific mucin, which comprises culturing a host cell transformed with the vector according to claim 4. 6. An L chain V region of an antibody that recognizes a cancer-specific mucin, which comprises the amino acid sequence shown in SEQ ID NO: 2. 7. A gene encoding the L chain V region of the antibody that recognizes the cancer-specific mucin according to claim 6. 8. A base sequence shown in SEQ ID NO: 2,
The gene according to claim 7. 9. A vector comprising the DNA according to claim 7 and capable of expressing the gene in a host cell. 10. A method for producing an L chain V region of an antibody which recognizes a cancer-specific mucin, which comprises culturing a host cell transformed with the vector according to claim 9. 11. A gene encoding the H chain V region of the antibody recognizing the cancer-specific mucin according to claim 2, and claim 7.
A vector containing a gene encoding the L chain V region of an antibody that recognizes the cancer-specific mucin described in 1 above, and capable of expressing these genes in a host. 12. The vector according to claim 11, which further comprises a gene encoding the H chain C region of a human-derived antibody and a gene encoding the L chain C region of a human-derived antibody. 13. A method for producing a chimeric antibody that recognizes a cancer-specific mucin, which comprises culturing a host cell transformed with the vector according to claim 12.