JPH04346792A - Amino acid sequence of anti-cancer human monoclonal antibody and dna base sequence coding the same antibody - Google Patents

Abstract

PURPOSE:To provide the subject amino acid sequence constituting the heavy chain and light chain of an anti-cancer human monoclonal antibody produced by the fused cell of the B cell of a human uterine cancer to a human lymphoblast cell and useful for the prevention, treatment, disgnosis, etc., of cancers, etc. CONSTITUTION:The production of the objective variable region fragment containing at least one of the super-variable regions Hv1, Hv2, and Hv3 (formulas I, II and III) of an anti-cancer human monoclonal antibody comprises extracting all RNAs by the guanidine thiocyanate method from a human/human hydridoma CLN/SUZ H11 produced by fusing the B lymphocyte of a human uterine cancer patient to a human lymphoblast cell, separating mRNA from the extracted RNA through an oligo (dT) cellulose column, synthesizing cDNAs with the separated mRNA as a template by a conventional method to produce a cDNA library, screening a clone containing a DNA coding the heavy chain and light chain variable region of a cancer cell antigen specific human immune globulin from the cDNA library, combining the DNA with a vector, transforming a host cell with the combined vector and subsequently culturing the transformed cell.

Description

[Detailed description of the invention]

0001

[Industrial Field of Application] The present invention is applicable to, for example, the medical and pharmaceutical fields such as the prevention, treatment, and diagnosis of human diseases, and the pharmacological and biochemical fields such as biochemical reagents and reagents for purifying biopolymers. The present invention relates to structures of variable regions of antigen-specific human immunoglobulins useful in a wide range of fields. For more details,
The present invention provides a human/human fusion cell line CLN/SUZ H1 of B cells of a human uterine cancer patient and a human lymphoblastoid cell line.
The present invention relates to the amino acid sequences of the heavy chain and light chain variable regions of the cancer cell antigen-specific human immunoglobulin produced by No. 1 and the base sequence of its gene.

0002

BACKGROUND OF THE INVENTION Since the development of techniques for producing monoclonal antibodies by cell fusion or cell immortalization, many useful antibodies have been obtained mainly using mice and the like. Among these, monoclonal antibodies against malignant tumor cells have extremely high utility value, as they are used not only for basic research such as tumor antigen analysis, but also for serum diagnosis and tumor image diagnosis using labeled antibodies. However, foreign antibodies from mice and the like are foreign to humans, and frequent administration to humans induces an immune response against the administered antibodies, resulting in side effects and a decrease in the therapeutic or preventive effects of the antibodies. From the above points, considering clinical fields in which antibodies are actually administered to humans, such as human cancer prevention, treatment, and in-vivo diagnosis, it is desirable to use human antibodies. However, human monoclonal antibodies are difficult to produce, so they are hardly put to practical use at present.

Under these circumstances, one of the inventors of the present invention
Japanese Patent Application Publication No. 58-201994 (Patent Publication No. 01-598
As disclosed in detail in JP-A-59-135898 and JP-A-59-137497, the cell line CLN/SUZ produces human monoclonal antibodies with high reactivity to human cancer cells. H1
1 (ATCC No. HB8307) was established. The antibody produced by this cell line (named CLN-IgG) is
The antibody class is IgG, the isotype is γ1 type and κ type, and an interesting finding has been obtained that immunohistologically, it binds to cancer antigens present on the surface of cancer cells and has the effect of suppressing the proliferation of cancer cells. It is being

0004

[Problems to be Solved by the Invention] Against this technical background, there are problems to be solved as described below.

[0005] 1) It is known that monoclonal antibody-producing cell lines generally lose their antibody-producing ability with passage. Generally speaking, human hybridomas produce lower amounts of antibodies than mouse hybridomas. When using human monoclonal antibodies for cancer treatment or diagnosis, a large amount of antibodies are required, and it is essential to solve this problem.

2) According to current immunological knowledge, there is no known mechanism by which monoclonal antibodies bind to human cancer cells and suppress cancer cell proliferation or kill cancer cells through the action of the antibody itself. There is. Furthermore, complement or K-
It is known to suppress the proliferation of cancer cells with the help of cells and macrophages, causing their death. However, these effects are actually not as strong as expected, and therefore further attempts are needed to increase the anticancer activity of antibodies.

[0007] One means of solving the above-mentioned problems, ie, improving the amount of antibody produced and increasing the anticancer activity of the antibody, is a method using genetic manipulation. For example, in the case of problem 1), after cloning the antibody gene,
A possible solution would be to introduce a gene into a host cell such as an animal cell or E. coli, express the antibody gene, and obtain a large amount of antibody.Also, in the case of problem 2), it would be possible to artificially change the antibody gene. By modifying various functions of antibodies, such as increasing binding affinity with antigens, anti-cancer activity or tissue infiltration through immunocompetent cells, and even modifying functions that antibodies do not originally have, such as increasing cell invasiveness. It is possible to design molecules with higher anticancer activity by adding toxicity, enzymatic activity, immune-inducing activity, etc. to antibody molecules or fragments thereof.

[0008] In order to achieve these objectives, it is important to isolate and elucidate the structure of antibody genes. However, the structure of the light chain and heavy chain constituting the CLN-lgG monoclonal antibody, as well as the gene structure of the variable region that has the function of specifically binding to an antigen, are completely unknown so far.

[0009] Therefore, the main object of the present invention is to
The purpose of this project is to elucidate the genetic structure of the light chain and heavy chain of IgG monoclonal antibodies.

0010

[Means for solving the problems] The present inventors have discovered that the CL
The cDNA encoding the light chain and heavy chain of the N-lgG monoclonal antibody was isolated, the DNA base sequence was elucidated, and the amino acid sequences of the light chain and heavy chain variable regions of the antibody were determined from the sequence. It was completed.

Specifically, mRNA was prepared from CLN/SUZ H11, and a cDNA lambda phage library created therefrom was screened by the plaque hybridization method, and the bases of the inserted DNA of the isolated phage clone were determined. This was achieved by determining the sequence. The present invention will be explained in more detail below.

According to the present invention, the DNA base sequences of the light chain variable region and the heavy chain variable region of a CLN-lgG monoclonal antibody are determined using a human antibody gene fragment amplified by the PCR method (polymerase chain reaction method) as a probe.
It was determined by cloning the cDNA of the IgG monoclonal antibody light chain and heavy chain and analyzing the DNA base sequence. These steps will be explained in more detail below.

[1] mRNA isolation and purification The cell line used in the present invention is a human/human hybridoma obtained by fusion of human uterine cancer patient lymphocytes and human lymphoblasts, and specifically, the cell line used in the present invention is a human/human hybridoma obtained by fusion of human uterine cancer patient lymphocytes and human lymphoblasts. 201994
The method is disclosed in detail in Japanese Patent Publication No. 01-59878, and produces human monoclonal antibodies that specifically react with cell surface antigens of cancer cells such as human brain tumors, lung cancer, gastric cancer, malignant melanoma, etc. Hybridoma CLN-
It is SUZH11. This hybridoma is ATCC (
American TypeCulture Coll.
registration number HB8307.

[0014] The cells are cultured and grown under appropriate conditions, for example, at 37° C. and a carbon dioxide concentration of 5%, in a culture medium containing 5% fetal bovine serum, such as RDF medium, and the resulting cells are collected by centrifugation. After that, the cells are removed using a conventional method such as H
The guanidium thiocyanate method of Han et al. [Han, J.;
H. , Stratowa, C. , & Rutte
r, W. J. (1987) Biochemistry
, 26, 1617-1625], and then the poly(A) + RNA fraction is separated and purified using a conventional method, such as adsorption column chromatography using oligo dT cellulose or a batch method.

[0015] The poly(A)+RNA obtained is further cD
It can be used to create a NA library. In the present invention, specifically, total RNA is extracted from CLN-SUZ H11 hybridoma cells, and poly(A)+R is extracted from this extract using an oligo dT cellulose column.
NA was purified and used for the construction of the following cDNA library.

[2] Preparation of cDNA library [1]
Using the poly(A)+RNA obtained in the step as a template and using oligo dT corresponding to polyA or a synthetic nucleotide having a base sequence thought to correspond to the constant region of an antibody as a primer, dATP, dGTP, dTTP,
Single-stranded DNA complementary to mRNA is synthesized by reverse transcriptase in the presence of dCTP. Next, after fragmenting the RNA with E. coli RNAase H, using this single-stranded DNA as a template, double-stranded DNA was fragmented using E. coli DNA polymerase I.
Synthesize DNA. A sticky end can be introduced into the thus obtained cDNA by, for example, ligating an EcoRI linker and then digesting it with EcoRI. The resulting fragments were inserted into a suitable phage vector, such as λgt10.
After ligation to the EcoRI site of a vector, λgt11 vector, etc., in vitro packaging is performed to generate cDNA.
A library can be created.

In the specific operation of the present invention, poly(A)+RNA obtained in step [1] is used as a template to generate cDNA.
was synthesized, and this cDNA was ligated to a λgt10 vector to create a cDNA library.

[3] Preparation of probes The probes include human immunoglobulin light chain or heavy chain constant region or variable region genes or fragments thereof;
Alternatively, a chemically synthesized oligonucleotide having a base sequence corresponding to the amino acid sequence of that portion and labeled with 32P, biotin, etc. by the nick translation method can be used.

In the present invention, preferably, a fragment amplified by PCR using cDNA as a template and sequences corresponding to parts of the light chain and heavy chain of an antibody as primers is biotinylated by a nick translation method. The probe can be used as a probe.

[4] Cloning of cDNA A desired clone is selected from the cDNA library obtained in step [2] using the probe obtained in step [3]. For example, the cDNA obtained in step [2]
A library λgt10 phage was transferred to E. coli strain (C6
00Hfl-) to form plaques, and positive clones are further selected by plaque hybridization. As a result, λCLN-G111 was identified as a CLN-lgG heavy chain cDNA clone.
λCLN-K4 as N-lgG light chain cDNA clone
11 were selected and subjected to nucleotide sequencing.

[5] Base sequence determination [4] The cDNA clone obtained in the step [4] can be used, for example, as a plasmid vector such as pUC18, a phage vector such as M13 phage, or a plasmid vector such as pUC18, pUC118, or
The DNA base sequence of the inserted portion of the resulting subclone can be determined by recloning into a Phagemid vector such as Bluescript SK+ using the Maxam, Gilbert method or Sanger method.

In the specific operation of the present invention, λCLN-G111 and λCLN-K obtained in step [4]
The EcoRI fragment of 411 was converted into pBluescript
After recloning into SK+, single-stranded DNA was prepared by infection with helper phage R408, and its nucleotide sequence was determined by Sanger method.

[0023] The present invention will now be explained in more detail with reference to Examples.

[0024]

[Example] Example 1: Hybridoma CLN/SUZ
Isolation of mRNA (Poly(A) + RNA) from H11 The method for obtaining the mRNA fraction from purified human hybridoma CLN/SUZ H11 is as follows. C.L.N.
/SUZ H11 cells using the guanidium thiocyanate method [Han, J.; H. , Stratowa, C.
．． , & Rutter, W. J. (1987).
Biochemistry, 26, 1617-162
5], total RNA was prepared. 109 cultured cells were collected by centrifugation and washed with physiological saline. 20 ml of 5M guanidinium thiocyanate containing 8% 2-mercaptoethanol, which has been ice-cooled in advance, is added to the cell pellet collected by centrifugation at 800 rpm, and homogenized immediately. Prepare the cell disruption solution for 7 hours in advance.
．． Add 5 ml of ethanol to a polypropylene centrifuge tube that has been chilled at -20°C, mix, and immediately
Centrifuge at 0,000 rpm for 5 minutes. 8% 2-mercaptoet, which had been ice-cooled in advance, was added to the resulting precipitate.
5M guanidium thiocyanate containing hanol
After adding 0ml and homogenizing, add 0.0ml of 1M acetic acid.
Add 25 ml and 7.5 ml of cold ethanol and heat to -20°C.
Leave it overnight. The precipitate obtained by centrifugation at 10,000 rpm for 10 minutes was mixed with 10 mM 2-mercaptoet.
Dissolve in 10 ml of 6M guanidine hydrochloride containing hanol, add 0.25 ml of 1M acetic acid and 5 ml of cold ethanol, and leave at -20°C for 3 hours. 10,000rpm
The precipitate obtained by centrifugation at
Add cold ethanol and leave at -20°C for an additional 3 hours. The precipitate obtained by centrifugation at 10,000 rpm for 10 minutes was dissolved in 5 ml of sterile pure water, and 21 M sodium acetate 0
．． Add 5 ml and 12.5 ml of cold ethanol at -20°C.
and save it as a total RNA fraction.

Poly(A)+RNA was obtained by the Chirgwin method [Chirgwin's method] from the total RNA fraction obtained by the above method.
rgwin, J. M. , Przybyla, A. E.
, MacDonald, R. J. , & Rutter
, W. J. (1979) Biochemistry,
18, 5294-5299]. First, C
9 mg of total RNA from LN/SUZH11 cells is dissolved in pure water to give a concentration of 2.5 mg/ml. After heat treatment at 100°C for 5 minutes and quenching on ice, 5M lithium chloride, 10% S
Add DS and 1M triethanolamine hydrochloric acid pH 7.4 to final concentrations of 0.5M, 0.2%, and 10mM, respectively. The solution was mixed in advance with binding buffer (0.5
Apply to an oligo(dT) cellulose column equilibrated with M lithium chloride, 0.2% SDS, 10mM triethanolamine hydrochloride pH 7.4). Further wash with 10 column volumes of binding buffer. The poly(A)+RNA bound to the column is eluted with an elution buffer (10mM triethanolamine hydrochloride pH 7.4) and the RNA fraction is collected. The obtained poly(A)+RNA solution is heat-treated at 100° C. for 5 minutes, and then the above-described chromatography using the oligo(dT) cellulose column is repeated once again using a new column. For RNA, add 2.5 times the volume of ethanol and 1/10 volume of 2M sodium acetate to the eluate.
Collect as a precipitate after centrifugation at 10,000xg. Dissolve the purified poly(A) + RNA precipitate in sterile pure water.
Store at -70°C at a concentration of μg/μl.

Example 2: Creation of CLN/SUZ H11 library Using 4 μg of poly(A)+RNA obtained in Example 1,
According to the protocol of Amersham's cDNA synthesis kit, 3.2 μg of cDNA synthesized was recovered. This cD
After treating the NA fragment with EcoRI methylase, EcoRI linkers were ligated using T4 DNA ligase. After EcoRI digestion, a cDNA fraction having EcoRI ends was collected using an Amersham column. After ligating 100 ng of this cDNA to 1 μg of λgt10 vector (Stratagene), in vitro packaging was performed using a packaging kit (GIGAPACK).
CLN/
SUZ H11 cDNA library 7.8×106
pfu/μg DNA was prepared.

Example 3: Determination of the amino acid sequence of CLN-lgG using a protein sequencer After reducing the purified CLN-lgG, 30 μg each of the heavy chain and light chain purified by gel filtration were added to a protein sequencer 477A (manufactured by Aplite Biosystems). The amino acid sequence of about 30 residues from the N-terminus was determined. In addition, the heavy chain was cleaved specifically for methionine with cyanogen bromide, the fragments were separated and purified by reversed-phase liquid chromatography, and then a portion of the amino acid sequence was similarly determined.

Example 4: Probe preparation method (1) Heavy chain probe A sequence with homology to the partial amino acid sequence of the CLN-lgG heavy chain determined in Example 3 was searched in the NBRF protein database (NBRF-PDB; National Biom.
edical Research Foundation
Human immunoglobulin germ line VH
26 (Entry name H3HU26, Accession
It became clear that the number A02047) had the highest homology. So EMBL D
NA database (EMBL-GDB; Europe
n Molecular Biology Labora
Tory Gene data Base)
26 DNA sequences (ID name HSIGHAU, Access
sion number M17747)
Thirty nucleotides (primer No. 1) corresponding to the terminal ten amino acid residues were synthesized. In addition, the DNA sequence of the heavy chain γ1CH1 domain (EMBL-GDB; ID name HSI
GCC4, Accession number J00
30 nucleotides (primer No. 2) corresponding to the 10 C-terminal amino acid residues of 228) were synthesized.

[0029]

[Chemical formula 9]

Example 2 using these two types of primers
PCR (polymerase chain reaction) was performed using 4 ng of CLN/SUZ H11 cDNA prepared in 1 as a template. As a result, a fragment of about 660 base pairs (PCRγC3) was amplified, and the base sequence was determined to be a fragment of antibody heavy chain γ1 CH1.
It became clear that these correspond to domains and variable regions. This γC3 was biotinylated by a nick translation method to obtain a probe (biotinylated PCR γC3).

(2) Light chain probe As a result of searching the NBRF protein database for a sequence that has homology to the partial amino acid sequence of the CLN-lgG light chain determined in Example 3, it was found that human immunoglobulin derived from Daudi cells (entry name K1HUDI) was found. , Access
ion number A01884). Therefore, the DNA sequence of the Daudi antibody light chain in the EMBL DNA database (ID name HSVK02, Accession number
00966), 30 nucleotides (primer No. 3) corresponding to the 10 N-terminal amino acid residues were synthesized. In addition, the DNA sequence of the light chain (κ chain) C domain (EMB
L-GDB; ID name HSIGK1, Accession
number V00557) C-terminal amino acid 1
30 nucleotides corresponding to residue 0 (primer No.
4) was synthesized.

[0032]

[Chemical formula 10]

Example 2 using these two types of primers
PCR was performed using 4 ng of CLN/SUZ H11 cDNA prepared in 1 as a template. As a result, a fragment of approximately 660 base pairs (PCRκA4) was amplified, and base sequencing revealed that it corresponded to the antibody light chain (κ chain) C domain and variable region. This PCRκ4 was biotinylated by nick translation method and probe (
Biotinylated PCRκA4) was obtained.

Example 5: Cloning of cDNA (1)
Cloning of heavy chain cDNA CLN/SUZ H11cD obtained in Example 2 above
Plaque hybridization was performed on the NA library using the biotinylated probe obtained in Example 4, and 13 positive clones were obtained. One of these clones has an inserted DNA of approximately 1.6K base pairs,
This phage was named λCLN-G111.

(2) Cloning of light chain cDNA Plaque hybridization was performed on the CLN/SUZ H11 cDNA library obtained in Example 2 using the biotinylated probe obtained in Example 4.
Seven positive clones were obtained. Among them, clone 411 had an inserted DNA of approximately 1.0 K base pairs, and this phage was named λCLN-K411.

Example 6: Determination of base sequence λCLN-G111 and λCLN- cloned in Example 5
Phagemid Blues uses the EcoRI fragment of K411.
Re-cloned into crypt SK+. E. coli KL1-Blue transformed with this phagemid was infected with helper phage R408, single-stranded DNA was prepared, and the Sanger method (Sanger, F. et al.
roc. Natl. Acad. Sci. USA
．． 74; 5463 (1977)). The base sequence of the variable region of the resulting CLN-lgG light chain cDNA clone and the amino acid sequence predicted therefrom are shown below.

[0037]

[Table 1]

[0038]

[Table 2]

Furthermore, in order to determine the subgroups of the CLN-lgG heavy chain and light chain, we performed a computer search and examined genes with high homology. As a result, the CLN-lgG heavy chain variable region is in subgroup 3, and the light chain ( It was revealed that the κ chain) variable region belongs to subgroup 1.

Example 7: Determination of hypervariable regions Comparing the amino acid sequences of the heavy and light chains of various antibodies reveals that there are regions with constant sequences (constant regions) and regions with different sequences (variable regions). I understand that. Parts of the variable region that are particularly highly variable are called hypervariable regions (hyper-variable regions).
There are three regions in each of the heavy chain and light chain. From the amino terminus, they are called Hv1, Hv2, and Hv3, respectively. These hypervariable regions are thought to form the antigen binding site and contain amino acid residues that make direct contact with antigenic determinants. Therefore, hypervariable regions are complementarity determining regions (CDRs)
;Complementality determination
This region is also called a region that controls the antigen specificity of an antibody.

Kabat & Wu plots were used to determine the hypervariable regions of CLN-IgG. [Kab
at, E. A. , Wu, T. T. , Bilofsky
,H. Variable Regions of Imm
Unoglobulin Chains (Medica
l Compute. Systems, Bolt,
Beranek & Newman, Cambridge
E, 1976)] This involves arranging the sequences of various antibodies and calculating and plotting the degree of variation for each position. The degree of variation here is the ratio between "the number of types of amino acids that appear at a given position" and "the frequency of the amino acid that appears most frequently at that position", and theoretically takes a value from 1 to 400. A region with a large degree of variation is a hypervariable region.

(1) Determination of the hypervariable region of CLN-lgG light chain The amino acid sequence of CLN-lgG light chain revealed that it belongs to kappa chain subgroup 1. So NBR
The 24 sequences belonging to subgroup 1 included in F-PDB (rel. 26) were aligned with the CLN-lgG light chain, and the degree of variation was calculated at each position to create a Kabat & Wu plot (FIG. 1). From the results, Hv1, Hv
2, Hv3 was determined to have residue numbers 28 to 34, 50 to 56, and 91 to 96, respectively.

(2) Determination of the hypervariable region of CLN-lgG heavy chain The amino acid sequence of CLN-lgG heavy chain revealed that it belongs to Hv subgroup 3. Therefore, NBRF-
Subgroup 3 included in PDB (rel.26)
21 sequences belonging to the CLN-lgG heavy chain are aligned,
The degree of variation was calculated at each position and a Kabat & Wu plot was created (Figure 2, residue numbers up to 96 are shown). As a result, Hv1 and Hv2 were determined from residue numbers 31 to 35 and 4, respectively.
It was decided to be 9 to 59. Regarding Hv3, in the case of the heavy chain, it is difficult to precisely match the positions because the chain lengths are significantly different between each sequence, as shown below. Calculating the degree of mutation without considering gaps, residue number 96 cysteine is 1.0, 97 glycine is 2.1, 98 arginine is 3.9, 99 valine is 29.3, and 109 tyrosine is 18
．． 4, 110 tryptophan is 3.5, and 111 glycine is 1.0. The degree of mutation is clearly high at residue numbers 99 to 109, and this region corresponds to Hv3.

[0044]

[Table 3]

[0045]

EFFECT OF THE INVENTION Now that the CLN-IgG antibody and its gene structure have been clarified, it has become possible to introduce and express this gene into host cells such as animal cells and Escherichia coli to obtain large quantities of the antibody. Furthermore, not only complete antibodies but also certain antibody fragments, such as heavy chain only, light chain only, Fab
fragment, F(ab)'2 fragment, Fv fragment, domain fragment (
It becomes possible to obtain various antibody-derived fragments such as dAb) and CDR fragments. Furthermore, by artificially causing mutations in antibody genes, complete antibodies or fragments derived from various antibodies with partially different amino acid sequences can be obtained.

[0046] As a result of the research to date, CLN-lgG acts on human cancer cells such as human gastric cancer, lung cancer, brain tumor, malignant melanoma, etc., and suppresses the proliferation of these cancer cells by its own action. Alternatively, it is expected to kill cancer cells, and further suppress the proliferation of cancer cells with the help of complement, K-cells, macrophages, etc., and cause death of cancer cells. However, by modifying the CLN-IgG gene and substituting some amino acids in the antibody, it is possible to further increase the activity of the antibody. For example, it can be modified to increase binding affinity with antigens, anticancer activity via immunocompetent cells, or tissue invasiveness. Furthermore, by adding cytotoxicity, enzymatic activity, immunity-inducing activity, etc. to antibody molecules or fragments thereof at the genetic level,
One possibility is to design molecules with higher anticancer activity. To give specific examples, using a cancer-specific antibody as a carrier, for example, chemotherapeutic agent-conjugated human monoclonal antibody, interferon-conjugated human monoclonal antibody, polymer toxin-conjugated human monoclonal antibody, drug-containing liposome-conjugated human monoclonal antibody, etc. It is useful as a drug in the form of a monoclonal antibody that inhibits the proliferation or induces the death of cancer cells. It is also conceivable to bind a radiosensitive substance to an antibody and administer it to a patient so that the antibody selectively accumulates in cancer cells, thereby increasing the effectiveness of treatment and diagnosis. When used against such cancers, a complete human monoclonal antibody may be used, or as mentioned above, for example, heavy chain only, light chain only, Fab fragment, F(ab)'2 fragment, Fv fragment, Smaller fragments containing specific antigen recognition sites such as domain fragments (dAbs), CDR fragments, etc. can also be used.

[Brief explanation of drawings]

[Figure 1] Figure 1 shows 24 proteins belonging to human κ chain subgroup 1.
Figure 3 shows a Kabat & Wu plot of the variable region sequences of .

[Figure 2] Figure 2 shows 21 cells belonging to human heavy chain subgroup 3.
Figure 3 shows a Kabat & Wu plot of the variable region sequences of .

Claims (10)

[Claims] 1. An immunoglobulin heavy chain variable region fragment comprising at least one hypervariable region selected from hypervariable regions Hv1, Hv2 and Hv3 having the following amino acid sequence: [Claim 2] The following amino acid sequence [Chemical 2] An immunoglobulin heavy chain variable region fragment having the following. 3. An immunoglobulin heavy chain variable region comprising a nucleotide sequence encoding at least one hypervariable region selected from hypervariable regions Hv1, Hv2, and Hv3 having the following amino acid sequence: DNA and RNA fragments encoding at least a portion of 4. DNA and RNA base sequences encoding the amino acid sequence according to claim 2. 5. The DNA base sequence of claim 4 and the corresponding RNA base sequence having the following base sequence: 6. An immunoglobulin light chain variable region fragment comprising at least one hypervariable region selected from hypervariable regions Hv1, Hv2 and Hv3 having the following amino acid sequence: [Claim 7] The following amino acid sequence [C6] An immunoglobulin light chain variable region fragment having the following. 8. An immunoglobulin light chain variable region comprising a nucleotide sequence encoding at least one hypervariable region selected from hypervariable regions Hv1, Hv2, and Hv3 having the following amino acid sequence: DNA and RNA fragments encoding at least a portion of 9. DNA and RNA base sequences encoding the amino acid sequence according to claim 7. 10. The DNA base sequence of claim 9, which has the following base sequence: [Claim 10] The DNA base sequence of claim 9 and its corresponding RNA base sequence.