A sn38 antibody or antigen-binding fragment thereof and uses thereof

Abstract

The application discloses an SN38 antibody or an antigen binding fragment thereof and application thereof. The SN38 antibody or the antigen binding fragment thereof comprises a sequence shown in the application. The SN38 antibody of the application can recognize a small molecule SN38 with high specificity, has high affinity to the SN38, and can realize effective detection on the content of the SN38 in a sample.

Description

TECHNICAL FIELD

[0001] The present application relates to the field of biological medicine, in particular to a SN38 antibody or antigen-binding fragment thereof and application thereof. BACKGROUND

[0002] Antibody-drug conjugate (ADC) is a new generation of antibody targeted therapy drug, mainly used in the treatment of cancer. ADC drug is composed of small molecule cytotoxic drug (drug), antibody (antibody) and linker connecting antibody and cytotoxic drug (linker). Small molecule cytotoxic drug is combined with antibody protein by chemical coupling method. Clinical studies have proved that ADC drug has high efficacy, is relatively stable in blood, can effectively reduce the toxicity of small molecule cytotoxic drug (chemical drug) to circulatory system and healthy tissues, and is the research focus of international anticancer drugs.

[0003] Camptothecin (CPT) is a drug with a pentacyclic skeleton, which is composed of 1 (3, 4-β) quinoline group (A, B ring), 1 azaindole (C, D ring) and 1 α-hydroxy-δ-lactone ring (E ring), with 1 chiral carbon atom C-20.

[0004] SN38 (7-ethyl-10-hydroxy camptothecin) is a semi-synthetic natural anticancer alkaloid camptothecin analogue, and its molecular formula is as follows:

[0005]

[0006] SN38 regulates the topological structure of DNA by using supercoiled DNA to unwind and promote DNA replication, inhibits the synthesis of TOP. By forming a stable ternary complex with TOP1, DNA, it blocks the cell cycle to S phase, thereby preventing DNA replication and causing cell death. Therefore, SN38 can be used as a preferred small molecule cytotoxic drug for ADC (antibody-drug conjugate, antibody-drug conjugate).

[0007] In clinical trials, it is necessary to monitor the ADC concentration in vivo after administration of ADC constructed by SN38 to determine the administration dose. At present, there is no detection reagent that can well detect the content of ADC in the sample, so the market urgently needs an antibody that can effectively bind SN38 to quickly and accurately analyze SN38 ADC. SUMMARY

[0008] In view of the technical problem that there is no antibody for SN38 in the prior art and it is difficult to achieve good detection of SN38, the application provides an SN38 antibody or an antigen binding fragment thereof and application thereof.The SN38 antibody of the application can recognize small molecule SN38 with high specificity and has high affinity for SN38, and can achieve effective detection of the content of SN38 in a sample.

[0009] To solve the above technical problems, one of the technical solutions provided by the application is an SN38 antibody or an antigen binding fragment thereof, which comprises a heavy chain variable region and / or a light chain variable region.

[0010] The heavy chain variable region comprises HCDR1, HCDR2 and HCDR3, the HCDR1 comprises an amino acid sequence as shown in SEQ ID NO:1 or a variant 1 thereof; the HCDR2 comprises an amino acid sequence selected from the group consisting of SEQ ID NO:2, SEQ ID NO:5 and SEQ ID NO:8; and the HCDR3 comprises an amino acid sequence selected from the group consisting of SEQ ID NO:3, SEQ ID NO:6 and SEQ ID NO:9.

[0011] The light chain variable region comprises LCDR1, LCDR2 and LCDR3, the LCDR1 is an amino acid sequence as shown in SEQ ID NO:14 or a variant 2 thereof; the LCDR2 comprises an amino acid sequence as shown in GTN or GTY; and the amino acid sequence of the LCDR3 comprises an amino acid sequence as shown in SEQ ID NO:15 or a variant 3 thereof.

[0012] The variant at least has the function of the SN38 antibody or the antigen binding fragment thereof.

[0013] Preferably, the variant 1 is obtained by mutating the 6th, 7th and / or 8th positions of the amino acid sequence shown in SEQ ID NO:1, wherein the mutation of the 6th position is preferably T6G, the mutation of the 7th position is preferably Y7A, and the mutation of the 8th position is preferably W8G.

[0014] The variant 2 is obtained by mutating the 7th and / or 9th positions of the amino acid sequence shown in SEQ ID NO:14, wherein the mutation of the 7th position is preferably S7R, and the mutation of the 9th position is preferably F8Y.

[0015] The variant 3 is obtained by mutating the 4th, 5th and / or 6th positions of the amino acid sequence shown in SEQ ID NO:15, wherein the mutation of the 4th position is preferably Y4H, the mutation of the 5th position is preferably R5S or R5T, and the mutation of the 6th position is preferably N6T.

[0016] In a preferred embodiment,

[0017] the amino acid sequence of the variant 1 is set forth in SEQ ID NO: 4 or SEQ ID NO: 7;

[0018] the amino acid sequence of the variant 2 is set forth in SEQ ID NO: 10 or SEQ ID NO: 12;

[0019] the amino acid sequence of the variant 3 is set forth in SEQ ID NO: 11 or SEQ ID NO: 13.

[0020] Preferably, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 1-3, respectively; or, HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 4-6, respectively; or, HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 7-9, respectively;

[0021] and / or, the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 10, GTN, SEQ ID NO: 11, respectively; or, LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 12, GTY, SEQ ID NO: 13, respectively; or, LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 14, GTN, SEQ ID NO: 15, respectively.

[0022] More preferably, the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 1-3, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 10, GTN, SEQ ID NO: 11, respectively; or

[0023] the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 4-6, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 12, GTY, SEQ ID NO: 13, respectively;

[0024] The heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NOs: 7-9, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NOs: 14, GTN, 15, respectively.

[0025] In the technical solution, the heavy chain variable region can further comprise HFR1, HFR2, HFR3 and HFR4, the amino acid sequences of the HFR1, HFR2, HFR3 and HFR4 are set forth in SEQ ID NOs: 16-18 and SEQ ID NO: 19, respectively; or, the amino acid sequences of the HFR1, HFR2, HFR3 and HFR4 are set forth in SEQ ID NOs: 20-22 and SEQ ID NO: 19, respectively; or, the amino acid sequences of the HFR1, HFR2, HFR3 and HFR4 are set forth in SEQ ID NOs: 23-25 and SEQ ID NO: 19, respectively.

[0026] And / or, the light chain variable region comprises LFR1, LFR2, LFR3 and LFR4, the LFR1 comprises an amino acid sequence set forth in SEQ ID NO: 26 or a variant 4 thereof; the LFR2 comprises an amino acid sequence set forth in SEQ ID NO: 27 or a variant 5 thereof; the LFR3 comprises an amino acid sequence set forth in SEQ ID NO: 28 or a variant 6 thereof; and the LFR4 comprises an amino acid sequence set forth in SEQ ID NO: 29 or a variant 7 thereof.

[0027] Preferably:

[0028] The amino acid sequence of the variant 4 is set forth in SEQ ID NO: 30;

[0029] The amino acid sequence of the variant 5 is set forth in SEQ ID NO: 31;

[0030] The amino acid sequence of the variant 6 is set forth in SEQ ID NO: 32 or SEQ ID NO: 33;

[0031] The amino acid sequence of the variant 7 is set forth in SEQ ID NO: 34.

[0032] More preferably:

[0033] the heavy chain variable region comprises HFR1, HFR2, HFR3, and HFR4 having the amino acid sequences set forth in SEQ ID NO: 16-18 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3, and LFR4 having the amino acid sequences set forth in SEQ ID NO: 26-29, respectively; or, the heavy chain variable region comprises HFR1, HFR2, HFR3, and HFR4 having the amino acid sequences set forth in SEQ ID NO: 20-22 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3, and LFR4 having the amino acid sequences set forth in SEQ ID NO: 30-32 and SEQ ID NO: 29, respectively; the heavy chain variable region comprises HFR1, HFR2, HFR3, and HFR4 having the amino acid sequences set forth in SEQ ID NO: 23-25 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3, and LFR4 having the amino acid sequences set forth in SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 33, and SEQ ID NO: 34, respectively.

[0034] In a preferred embodiment, the heavy chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 35, SEQ ID NO: 37, or SEQ ID NO: 39, or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 35, SEQ ID NO: 37, or SEQ ID NO: 39; and the light chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 36, SEQ ID NO: 38, or SEQ ID NO: 40, or an amino acid sequence having at least 90%, 95%, 96%, 97%, 98%, or 99% identity to the amino acid sequence set forth in SEQ ID NO: 36, SEQ ID NO: 38, or SEQ ID NO: 40.

[0035] In a preferred embodiment, the heavy chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 35, and the light chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 36; or, the heavy chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 37, and the light chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 38; or, the heavy chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 39, and the light chain variable region comprises an amino acid sequence set forth in SEQ ID NO: 40.

[0036] Preferably, the SN38 antibody or antigen-binding fragment thereof comprises a murine or human antibody constant region or a mutation thereof; the murine antibody constant region comprises a heavy chain constant region of murine IgG1, IgG2a, IgG2b3 or IgG3 and a kappa or lambda type light chain constant region, and the human antibody constant region comprises a heavy chain constant region of human IgG1, IgG2, IgG3 or IgG4 and a kappa or lambda type light chain constant region; preferably, the mutation has at least 90%, 95%, 96%, 97%, 98% or 99% identity with the amino acid sequence of the murine or human antibody constant region.

[0037] In a preferred embodiment, the heavy chain constant region amino acid sequence of the SN38 antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 41, positions 124-459; and the light chain constant region amino acid sequence of the SN38 antibody or antigen-binding fragment thereof is shown in SEQ ID NO: 42, positions 110-209.

[0038] Preferably, the structure of the SN38 antibody or antigen-binding fragment thereof comprises an immunoglobulin, Fab, Fab', F(ab')2, Fv, such as a single-chain Fv fragment (scFv).

[0039] To solve the above technical problems, the second technical solution provided by the present application is an isolated nucleic acid encoding the SN38 antibody or antigen-binding fragment thereof as described in the first technical solution.

[0040] Preferably, in the nucleotide sequence encoding the SN38 antibody or antigen-binding fragment thereof, the nucleotide sequence encoding the heavy chain variable region is shown in SEQ ID NO: 43, positions 1-369, and the nucleotide sequence encoding the light chain variable region is shown in SEQ ID NO: 44, positions 1-327; or the nucleotide sequence encoding the heavy chain variable region is shown in SEQ ID NO: 47, positions 1-372, and the nucleotide sequence encoding the light chain variable region is shown in SEQ ID NO: 48, positions 1-327; or the nucleotide sequence encoding the heavy chain variable region is shown in SEQ ID NO: 51, positions 1-363, and the nucleotide sequence encoding the light chain variable region is shown in SEQ ID NO: 52, positions 1-327.

[0041] To solve the above technical problems, the third technical solution provided by the present application is an expression vector comprising the isolated nucleic acid as described in the second technical solution. Preferably, the starting vector of the expression vector is pcDNA3.1(+), more preferably pcDNA3.1(+) produced by Ingenium (Shanghai) Trading Co., Ltd.

[0042] To solve the above technical problems, the technical solution four provided by the present application is a transformant comprising the expression vector of the technical solution three. Preferably, the transformant is a genetically modified lentivirus or an immune effector cell. More preferably, the lentivirus is a lentivirus containing PLVX-EF1-CAR, lentiCRISPR v2-CAR or pELNS-CAR, and the immune effector cell is a T lymphocyte, an NK cell or an NKT cell.

[0043] To solve the above technical problems, the technical solution five provided by the present application is a detection composition comprising the SN38 antibody or antigen binding fragment thereof of the technical solution one, the isolated nucleic acid of the technical solution two, the expression vector of the technical solution three and / or the transformant of the technical solution four.

[0044] To solve the above technical problems, the technical solution six provided by the present application is a kit comprising the SN38 antibody or antigen binding fragment thereof of the technical solution one, the isolated nucleic acid of the technical solution two, the expression vector of the technical solution three, the transformant of the technical solution four and / or the detection composition of the technical solution five.

[0045] Preferably, the kit further comprises (i) a device for administering the antibody or antigen binding fragment thereof, the isolated nucleic acid, the expression vector, the transformant or the detection composition; and / or (ii) instructions for use.

[0046] To solve the above technical problems, the technical solution seven provided by the present application is a method for detecting SN38 in a sample, which uses the SN38 antibody or antigen binding fragment thereof of the technical solution one, the isolated nucleic acid of the technical solution two, the expression vector of the technical solution three, the transformant of the technical solution four, the detection composition of the technical solution five and / or the kit of the technical solution six to react with the sample.

[0047] To solve the above technical problems, the technical solution eight provided by the present application is the use of the SN38 antibody or antigen binding fragment thereof of the technical solution one, the isolated nucleic acid of the technical solution two, the expression vector of the technical solution three, the transformant of the technical solution four, the detection composition of the technical solution five or the kit of the technical solution six in detecting SN38.

[0048] In addition, the present application also provides the technical solution nine: a pharmaceutical composition comprising the SN38 antibody or antigen binding fragment thereof of the technical solution one, the isolated nucleic acid of the technical solution two, the expression vector of the technical solution three and / or the transformant of the technical solution four.

[0049] Preferably, the pharmaceutical composition comprises a pharmaceutically acceptable carrier.

[0050] In addition, the present application also provides a tenth technical solution: the use of the SN38 antibody or antigen binding fragment thereof of the first technical solution, the isolated nucleic acid of the second technical solution, the expression vector of the third technical solution, the transformant of the fourth technical solution, the pharmaceutical composition of the ninth technical solution, or the kit of the sixth technical solution in the preparation of a medicament for treating SN38-related diseases.

[0051] In addition, the present application also provides a technical solution: a method for treating SN38-related diseases (such as side effects), the method comprising administering to a patient in need thereof a therapeutically effective amount of the SN38 antibody or antigen binding fragment thereof of the first technical solution, the isolated nucleic acid of the second technical solution, the expression vector of the third technical solution, the transformant of the fourth technical solution, or the pharmaceutical composition of the ninth technical solution.

[0052] The term

[0053] In the present application, the term "CDR" refers to one of the six hypervariable regions within the variable domain of an antibody that primarily contribute to antigen binding, the definition of which six CDRs is provided by Kabat E. A. et al., (1991) Sequences of proteins of immunological interest. NIH Publication 91-3242. As used herein, the Kabat definition of CDRs applies to CDR1, CDR2, and CDR3 of the light chain variable domain (CDR L1, CDR L2, CDR L3 or L1, L2, L3), and CDR1, CDR2, and CDR3 of the heavy chain variable domain (CDR H1, CDR H2, CDR H3 or H1, H2, H3).

[0054] In the present invention, the term "antibody" is used interchangeably with "immunoglobulin" to refer to a glycoprotein comprising at least two heavy chains (HC) and two light chains (LC) interconnected by disulfide bonds. Each heavy chain is comprised of a heavy chain variable region (abbreviated herein as VH) and a heavy chain constant region. The heavy chain constant region is comprised of three domains, CH1, CH2 and CH3. Each light chain is comprised of a light chain variable region (abbreviated herein as VL) and a light chain constant region (abbreviated herein as CL). The light chain constant region is comprised of one domain, CL. Mammalian heavy chains are classified as alpha, delta, epsilon, gamma, and mu. Mammalian light chains are classified as lambda or kappa. Immunoglobulins comprising the heavy chains alpha, delta, epsilon, gamma, and mu are called IgA, IgD, IgE, IgG, and IgM, respectively. A complete antibody forms a "Y" shape. The stem of the Y is comprised of the second and third constant regions of the two heavy chains (and for IgE and IgM, the fourth constant region) bound together, and a disulfide bond (interchain) is formed in the hinge. The heavy chains gamma, alpha and delta have a constant region composed of three tandem (in line) Ig domains, and a hinge region for added flexibility; the heavy chains mu and epsilon have a constant region composed of four immunoglobulin domains. The second and third constant regions are referred to as "CH2 domain" and "CH3 domain", respectively. Each arm of the Y includes the variable and constant regions of a single light chain bound to the variable region and first constant region of a single heavy chain. The variable regions of the light and heavy chains are responsible for antigen binding.

[0055] In the present invention, "Fab" is comprised of one light chain and the CH1 and variable region of one heavy chain. The heavy chain of a Fab molecule cannot form a disulfide bond with another heavy chain molecule. The "Fc" region contains two heavy chain fragments comprising the CH2 and CH3 domains of an antibody. The two heavy chain fragments are held together by two or more disulfide bonds and by hydrophobic interactions of the CH3 domains. "Fab'" contains one light chain and a portion of one heavy chain comprising the VH domain and the CH1 domain and the region between the CH1 and CH2 domains, whereby interchain disulfide bonds can form between the two heavy chains of two Fab' to form a F(ab')2 molecule. "F(ab')2 fragments" contain two light chains and two heavy chains comprising a portion of the constant region between the CH1 and CH2 domains, whereby interchain disulfide bonds form between the two heavy chains. Thus F(ab')2 fragments are comprised of two Fab' held together by disulfide bonds between the two heavy chains. The term "Fv" means an antibody fragment consisting of the VL and VH domains of a single arm of an antibody, but lacking the constant regions.

[0056] In the present application, the scFv (single chain antibody fragment) can be a conventional single chain antibody in the art, which includes a heavy chain variable region, a light chain variable region, and a short peptide of 15-20 amino acids. Among them, the VL and VH domains are paired through a linker that enables them to be produced as a single polypeptide chain to form a monovalent molecule [see, for example, Bird et al., Science 242:423-426 (1988) and Huston et al., Proc. Natl. Acad. Sci. USA 85:5879-5883 (1988)]. Such scFv molecules can have a general structure: NH2-VL-linker-VH-COOH or NH2-VH-linker-VL-COOH. Suitable prior art linkers consist of repeating G4S amino acid sequences or variants thereof. For example, a linker with the amino acid sequence (G4S)4or (G4S)3may be used, but variants thereof can also be used.

[0057] As used herein, "pharmaceutically acceptable carrier" or "pharmaceutically acceptable carrier" includes any and all solvents, dispersion media, isotonic and absorption delaying agents, and the like that are physiologically compatible. A pharmaceutically acceptable carrier suitable for use in the application can be a sterile liquid such as water and oil, including those of either vegetable or animal origin, such as peanut oil, soybean oil, mineral oil, sesame oil and the like. Water is a preferred carrier when the pharmaceutical composition is administered intravenously. Saline solutions and aqueous dextrose and glycerol solutions can also be employed as liquid carriers, particularly for injectable solutions. Suitable excipients include starch, glucose, lactose, sucrose, gelatin, malt, rice, flour, chalk, silica gel, sodium stearate, glycerol monostearate, talc, sodium chloride, dried skim milk, glycerol, propylene, glycol, water, ethanol and the like. For use of excipients and their uses, see also "Handbook of Pharmaceutical Excipients", Fifth Edition, R.C. Rowe, P.J. Seskey and S.C. Owen, Pharmaceutical Press, London, Chicago. The compositions can also contain a small amount of wetting or emulsifying agents, or pH buffering agents, if desired. These compositions can take the form of solutions, suspensions, emulsions, tablets, pills, capsules, powders, sustained-release formulations and the like. Oral formulations can include standard pharmaceutical carriers and / or excipients, such as pharmaceutical grades of mannitol, lactose, starch, magnesium stearate, saccharin. A pharmaceutical formulation or pharmaceutical composition described herein can be prepared by mixing a antibody of the application or antigen-binding fragment thereof having the desired degree of purity with one or more optional pharmaceutical adjuvants (Remington's Pharmaceutical Sciences, 16thEdition, Osol, A. Ed. (1980)), preferably in the form of a lyophilized formulation or aqueous solution. The pharmaceutical compositions or formulations of the application can also contain more than one active ingredient, which are desirable for the particular indication being treated, preferably those having complementary activities that do not adversely affect each other. For example, it is desirable to provide other anti-infective active ingredients, such as other antibodies, anti-infective active agents, small molecule drugs or immunomodulators, etc. The active ingredients are suitably combined in amounts that are effective for the purpose intended. Sustained-release formulations can be prepared. Suitable examples of sustained-release formulations include semi-permeable matrices of solid hydrophobic polymers containing the antibody of the application or antigen-binding fragment thereof, which matrices are in the form of shaped articles, e.g., films, or microcapsules.

[0058] As known in the art, "nucleic acid" as used interchangeably herein in the application refers to a chain of nucleotides of any length, and includes DNA and RNA. Nucleotides can be deoxyribonucleotides, ribonucleotides, modified nucleotides or bases, and / or their analogs, or any substrate that can be incorporated into a chain by the action of a DNA or RNA polymerase.

[0059] Calculations of sequence identity between sequences are performed as follows. To determine the percent sequence identity of two amino acid sequences or of two nucleic acids sequences, the sequences are aligned for optimal comparison purposes (e.g., gaps can be introduced in the sequence of a first and a second amino acid or nucleic acid sequence for optimal alignment and non-homologous sequences can be disregarded for comparison purposes). In one preferred embodiment, the length of an aligned reference sequence for comparison purposes is at least 30%, preferably at least 40%, more preferably at least 50%, 60%, and even more preferably at least 70%, 80%, 90%, 100% of the length of the reference sequence. The amino acid residues or nucleotides at corresponding amino acid positions or nucleotide positions are then compared. When a position in the first sequence is occupied by the same amino acid residue or nucleotide as the corresponding position in the second sequence then the molecules are identical at that position. The comparison of sequences and the determination of the percent sequence identity between two sequences can be accomplished using a mathematical algorithm. In one preferred embodiment, the percent identity of two amino acid sequences is determined using the Needleman and Wunsch ((1970) J. Mol. Biol. 48: 444-453) algorithm which has been incorporated into the GAP program in the GCG software package (available at http: / / www.gcg.com), using either a Blossum 62 matrix or a PAM250 matrix, and a gap penalty of 16, 14, 12, 10, 8, 6, or 4, and a gap length penalty of 1, 2, 3, 4, 5, or 6. In another preferred embodiment, the percent identity of two nucleic acid sequences is determined using the GAP program (in the GCG software package; available at http: / / www.gcg.com), using a NWSgapdna.CMP matrix and gap penalty of 40, 50, 60, 70, or 80, and gap length penalty of 1, 2, 3, 4, 5, or 6. Especially preferred parameters for use in this program are Blossum 62 with gap penalty of 12, and gap extension penalty of 4. The percent identity between two deoxyribonucleotide sequences can also be determined using the algorithm of E. Meyers and W. Miller ((1989) CABIOS, 4: 11-17) which has been incorporated into the ALIGN program (version 2.0), using a PAM120 weight residue table, a gap length penalty of 12, and a gap penalty of 4. Additionally or alternatively, nucleic acid sequences and protein sequences described herein can be further used as a query sequence to perform a search against public databases to, for example, identify other family members or related sequences.

[0060] As used herein, "expression vector" means a construct that is capable of delivering one or more genes or sequences of interest into a host cell and preferably expressing the genes or sequences in the host cell. Examples of expression vectors include, but are not limited to, viral vectors, naked DNA or RNA expression vectors, plasmid, cosmid or bacteriophage vectors, DNA or RNA expression vectors associated with cationic condensing agents, DNA or RNA expression vectors encapsidated in liposomes, and certain eukaryotic cells, such as producer cells.

[0061] In the present application, the term "transformant" includes cells into which foreign nucleic acid has been introduced, including the progeny of those cells. Progeny can not be completely identical to the parent cell in nucleic acid content, but can contain mutations. The present application includes mutated progeny that have the same function or biological activity as the cells selected or screened in the originally transformed cells.

[0062] The numbers following the variants "variant 1", "variant 2" and the like in the present application have no special meaning and are only for reference of the same terms.

[0063] On the basis of common general knowledge in the art, the above-mentioned preferred conditions can be combined in any manner, i.e. to obtain the preferred embodiments of the present application.

[0064] The reagents and raw materials used in the present application are commercially available.

[0065] The positive progress effect of the present application is that:

[0066] The application discloses an SN38 antibody or an antigen binding fragment thereof and application thereof. Since there is no antibody for SN38 in the prior art, the SN38 antibody of the application is a brand-new sequence antibody. The SN38 antibody has the ability of specifically recognizing a small molecule SN38 and has high affinity, and has a broad prospect in clinical detection of SN38. DETAILED DESCRIPTION

[0067] The application will be further described in the following examples, but the application is not limited to the scope of the examples. The experimental methods in the following examples, for which no specific conditions are indicated, are selected according to conventional methods and conditions, or according to the instructions of the products.

[0068] Example 1 Preparation of SN38 immunogen

[0069] SN38 (Beijing Bailingwei Technology Co., Ltd.) is coupled with KLH (Sigma-Aldrich, item number H1757) and BSA (Sigma-Aldrich, item number A1933) respectively as immunogens.

[0070] Coupling KLH, first weigh 3 mg KLH, dissolve into 0.3 ml 10 mg / ml with 0.1 M sodium bicarbonate containing 5 mM ethylenediaminetetraacetic acid, pH 8.0. Then weigh a certain amount of 2-iminothiolane hydrochloride (Shanghai Genomics, item number C100211) and dissolve into 2 mg / ml (14 mM) with sterile water. Add 0.25 ml 2-iminothiolane hydrochloride to KLH, the molar ratio of KLH amino number and 2-iminothiolane hydrochloride is 1:2, and react for 2 h at room temperature. Dialyze for 2 h at room temperature with 0.1 M PBS containing 5 mM ethylenediaminetetraacetic acid, pH 6.5. Then weigh 1.2 mg SN38 and dissolve into 10 mg / ml with dimethyl sulfoxide. Take out KLH-SH after 2 h, add 1.2 mg SN38 to it, and react for 2 h at room temperature. Dialyze overnight at 4°C with 1x PBS, pH 7.4, and change the solution once.

[0071] Coupling BSA, weigh 3 mg BSA, dissolve into 0.3 ml 10 mg / ml with 0.1 M sodium bicarbonate containing 5 mM ethylenediaminetetraacetic acid, pH 8.0. Then weigh a certain amount of 2-iminothiolane hydrochloride and dissolve into 2 mg / ml (14 mM) with sterile water. Add 0.255 ml 2-iminothiolane hydrochloride to BSA. The molar ratio of BSA amino number and 2-iminothiolane hydrochloride is 1:2. React for 2 h at room temperature. Dialyze for 2 h at room temperature with 0.1 M PBS containing 5 mM ethylenediaminetetraacetic acid, pH 6.5. Weigh 1.3 mg SN38 and dissolve into 10 mg / ml with dimethyl sulfoxide. Take out BSA-SH after 2 h, add 1.3 mg SN38 to it, and react for 2 h at room temperature. Dialyze overnight at 4°C with 1x PBS, pH 7.4, and change the solution once.

[0072] Example 2 Animal immunization

[0073] The immunization is carried out in two groups, and different antigens are used for immunization in different groups. Five Balb / c mice (Vantianlihua) in group A are immunized with SN38-KLH, and five Balb / c mice in group B are immunized with SN38-BSA. Blood is collected 4 days before the start of immunization, and immunization is performed once every 14 days, for a total of 4 rounds of immunization. For each immunization, 50 μg of antigen is subcutaneously injected into each mouse. Blood is collected 7 days after the third round of immunization, and the serum of the immunized animals is detected by indirect ELISA to determine the level of immune response. Cell fusion is performed after the fourth round of immunization.

[0074] The serum titer was detected by indirect ELISA method 7 days after the third immunization. The enzyme-labeled plate (Nunc, item number 446442) with hydrophobic properties was coated with 1 μg / ml of SN38-BSA and SN38-KLH, respectively, to detect the SN38-KLH immunized group and the SN38-BSA immunized group. The coating solution was PBS (pH 7.4), and the serum was diluted 1000, 2000, 4000, 8000, 16000, 32000, 64000, 128000, 256000 and 512000 times, respectively, and then added to the enzyme-labeled plate to combine with the small molecules. The horseradish peroxidase-labeled goat anti-mouse Fc IgG (Bethyl Laboratories, item number A90-231P) was used as the secondary antibody for detection, and the OD 655nm The wavelength was the reference wavelength (Molecular Devices, model SpectraMax M2e), and the OD 450nm The absorbance was determined at 450 nm, and the serum titers of four mice numbered 46, 47, 50 and 54 were relatively strong. The results are shown in Table 1.

[0075] Table 1 Serum titer

[0076]

[0077] The serum of 10 mice was detected by competitive ELISA method, and the enzyme-labeled plate was coated with 0.1 μg / ml of SN38-BSA and SN38-KLH, respectively, to detect the serum of the SN38-KLH immunized group and the SN38-BSA immunized group. The coating solution was PBS (pH 7.4), and the serum was diluted 1000, 2000, 4000, 8000, 16000, 32000, 64000, 128000, 256000 and 512000 times, respectively, and then added to the enzyme-labeled plate with 0 μg / ml or 100 μg / ml of SN38. The horseradish peroxidase-labeled goat anti-mouse Fc IgG was used as the secondary antibody for detection, and the OD 655nm The wavelength was the reference wavelength, and the OD 450nm The absorbance was determined at 450 nm, and the results are shown in Table 2. The serum of the mice with strong competitive binding activity was screened by the excess SN38 small molecule, and two mice numbered 46 and 54 were selected for cell fusion experiment.

[0078] Table 2 ELISA detection value of mouse serum

[0079]

[0080] Example 3 Cell fusion and parent clone screening:

[0081] The cell fusion was performed by electrofusion method, and a total of 2 rounds were performed. The average fusion efficiency was about 2500 spleen cells to produce 1 hybridoma cell. All cells in each round of fusion were plated into 96-well plates for expansion culture. The supernatant of the mother clone produced by the fused cells was subjected to ELISA screening. The positive mother clone was screened by indirect ELISA method. The SN38-BSA immunized mouse hybridoma cell mother clone was coated with 1 μg / ml of SN38-KLH to coat the enzyme-labeled plate strip. The SN38-KLH binding supernatant of the SN38-BSA immunized mouse hybridoma cell mother clones 1C11, 1H3, 4F2, 10E9, 12E11, 13A1, 13H9, 14G7, 15B2, and 15G5 was added. The enzyme-labeled secondary antibody horseradish peroxidase labeled goat anti-mouse Fc IgG was added. The higher the OD value, the higher the antibody concentration.

[0082] The SN38-KLH immunized mouse hybridoma cell mother clone was coated with 1 μg / ml of SN38-BSA to coat the enzyme-labeled plate strip for detection. The SN38-KLH binding supernatant of the SN38-KLH immunized mouse hybridoma cell mother clones 17C5, 19G10, 20H4, 21C11, 25A4, 26D12, 29D11, 29F7, 30D9, and 30F6 was added. The enzyme-labeled secondary antibody horseradish peroxidase labeled goat anti-mouse Fc IgG was added. The ELISA detection results of the mouse hybridoma cell mother clone are shown in Table 3.

[0083] Table 3 ELISA detection results of the SN38 immunized mouse hybridoma cell mother clone

[0084]

[0085]

[0086] The supernatant of the positive mother clone detected by indirect ELISA was subjected to competitive ELISA analysis. 0.5 μg / ml of SN38-BSA was coated to detect the SN38-KLH immunized mother clone. 1:6 dilution of the cell supernatant and different concentrations of SN38 (0, 0.1, 1, and 10 μg / ml) were added. The SN38-BSA coated on the enzyme-labeled plate strip and the added SN38 competed with the antibody in the supernatant. The more SN38 added, the less antibody bound to the coated SN38. After washing the plate, the enzyme-labeled secondary antibody horseradish peroxidase labeled goat anti-mouse Fc IgG was added for color development reaction. The lower the OD value, the better the specificity of the antibody to SN38. The results are shown in Table 4. Similarly, the SN38-BSA immunized mother clone was detected, and the results are shown in Table 5.

[0087] Table 4 ELISA detection values of the supernatant of the positive SN38-KLH immunized mother clone

[0088]

[0089] Table 5. ELISA detection values ​​of SN38-BSA immunobrood clone supernatant.

[0090]

[0091] Clones 17C5, 25A4, 1H3, 13H9, and 14G7, when analyzed under identical conditions, exhibited relatively strong signal values. Furthermore, the addition of SN38 at concentrations of 0.01 μg / ml and 0.1 μg / ml resulted in better competition (lower signal values, i.e., OD). 450nm / OD 655nm If the value is less than 0.4%, cryopreservation of the selected strains will be carried out for subcloning.

[0092] Example 4: Subcloning Screening

[0093] Limiting dilution was used to subclone positive maternal clones to ensure the final production of single-clone cells. The subcloning stage was performed in three rounds using indirect ELISA and competitive ELISA methods for subclone screening. Two stable daughter clones were selected from each maternal clone for amplification and cryopreservation. Before cryopreservation, 5 ml of supernatant was collected from each daughter clone, and all subtypes were identified using a subtype kit (Sigma-Aldrich, catalog number: 11493027001).

[0094] Indirect ELISA was used to screen subclonal supernatants for positive results. For those immunized with SN38-BSA, 1 μg / ml SN38-KLH-coated ELISA strips were used; for those immunized with SN38-KLH (SN38-BSA), 1 μg / ml SN38-BSA (SN38-KLH)-coated strips were used. Serially diluted subclonal supernatants were added to the strips to bind with the small molecules coated on the ELISA strips. The reaction was visualized using horseradish peroxidase-labeled goat anti-mouse Fc IgG. Higher OD values ​​reflected higher antibody concentrations. Results are shown in Table 6.

[0095] Table 6. Detection of antibody concentration in subclonal cell supernatant

[0096]

[0097] Competitive ELISA analysis of subclone cell culture supernatant, 0.5 μg / ml SN38-BSA coated to detect SN38-KLH immunized subclone, add 300-fold dilution of subclone cell 17C5-1, 17C5-2, 25A4-1 and 25A4-2 supernatant and different concentrations of SN38 (0, 0.01, 0.1 μg / ml), add enzyme-linked secondary antibody horseradish peroxidase labeled goat anti-mouse Fc IgG for color development reaction, the lower the OD value, the better the specificity of the antibody to the small molecule. Similarly, test the subclone 1H3-1, 1H3-2, 13H9-1, 13H9-2, 14G7-1 and 14G7-2 immunized by SN38-BSA, the results are shown in Table 7.

[0098] Table 7 Specificity of antibodies in subclone cell supernatant

[0099]

[0100]

[0101] Example 5 Monoclonal antibody identification

[0102] Monoclonal antibodies 1H3-1, 13H9-2, 14G7-1 were produced, and the antibodies were purified by Protein A / G affinity chromatography method (Cytiva, item number 29049104). The purified antibodies were stored in phosphate buffer (PBS) by dialysis method. The titer was detected by polyacrylamide gel electrophoresis (SDS-PAGE), indirect ELISA, competitive ELISA, and the antibody concentration was determined by NanoDrop2000 (Thermo).

[0103] The titer of monoclonal antibody was detected by indirect ELISA method. The enzyme-labeled plate with hydrophobic properties was coated with 1 μg / ml SN38-KLH, and the coating solution was PBS (pH 7.4). The antibody was diluted 1000, 2000, 4000, 8000, 16000, 32000, 64000, 128000, 256000 and 512000 times, respectively, and then added to the enzyme-labeled plate to combine with SN38-KLH. The enzyme-linked secondary antibody horseradish peroxidase labeled goat anti-mouse Fc IgG was used for display reaction, and the OD 655nm The wavelength is the reference wavelength, and the OD 450nm The absorbance was determined at 1H3-1, 13H9-2, 14G7-1, and the titers were strong, reaching 512000, as shown in Table 8.

[0104] Table 8 Monoclonal antibody titer detection value of indirect ELISA

[0105]

[0106] Competitive ELISA assay: SN38-KLH was coated with 0.5 μg / ml of antibody. Purified 1H3-1, 13H9-2, and 14G7-1 antibodies (antibody concentration range: 5000–0.005 ng / ml) and different concentrations of SN38 (0, 0.01, and 0.1 μg / ml) were added, allowing the coated SN38-KLH to compete with the added SN38 for antibody binding. The more small the molecular weight of the added antibody, the less antibody would bind to the coated SN38. After washing the plate, horseradish peroxidase-labeled goat anti-mouse Fc IgG was added. The lower the OD value, the better the antibody specificity for SN38. The competitive binding results of SN38 monoclonal antibody 1H3-1 were as expected. As the SN38 dose increased, the binding affinity of the monoclonal antibody to the antigen decreased, indicating strong specificity of the monoclonal antibody for SN38.

[0107] Table 9. Specificity detection values ​​of 1H3-1 monoclonal antibody

[0108]

[0109]

[0110] Table 10 Specificity detection values ​​of 13H9-2 monoclonal antibody

[0111]

[0112] Table 11 Specificity detection values ​​of 14G7-1 monoclonal antibody

[0113]

[0114] Example 6: Monoclonal Antibody Sequence Identification

[0115] Hybridoma cells of types 1H3-1, 13H9-2, and 14G7-1 were revived, and the number of freshly collected hybridoma cell pellets was 5 × 10⁻⁶. 6 The hybridoma cell lysis buffer was thoroughly lysed at 5 × 10⁶ using TriZol Reagent. 6 Total RNA was extracted from freshly collected hybridoma cells, with a concentration of at least 100 ng / μl, a total volume of 20 μl, and a total weight of 5 μg. Agarose gel electrophoresis showed clear RNA bands with no significant degradation. Reverse transcription was performed to obtain cDNA, and the variable regions of the heavy and light chains were amplified, cloned into the target vector, and sequenced for both heavy and light chains. The results are shown in the tables below.

[0116] Table 12 HCDR amino acid sequence and number

[0117] HCDRs 1H3-1 13H9-2 14G7-1 SEQ ID NO: 1 4 7 HCDR1 GYTFTTYW GYTFTTAG GYTFTGYW SEQ ID NO: 2 5 8 HCDR2 INPSNGRT INTHSGVP INPGNGRT SEQ ID NO: 3 6 9 HCDR3 ATSGPYYSYDVAWFAY ARASSSLYRYDQAWFTY ARPLDGYYETWFSY

[0118] Table 13. Amino acid sequences and numbers of LCDR

[0119] LCDRs 1H3-1 13H9-2 14G7-1 SEQ ID NO: 10 12 14 LCDR1 TGAVTTRNF TGAVTTSNY TGAVTTSNF SEQ ID NO: LCDR2 GTN GTY GTN SEQ ID NO: 11 13 15 LCDR3 ALWYTTHWV ALWHSNHWV ALWYRNHWV

[0120] Table 14 HFR Amino Acid Sequences and Numbering

[0121]

[0122] Table 15 LFR Amino Acid Sequences and Numbering

[0123]

[0124]

[0125] Table 16 Light and Heavy Chain Variable Region Amino Acid Sequences and Numbering for Anti-SN38 Monoclonals

[0126]

[0127] 1H3-1 Heavy Chain Nucleic Acid Sequence (SEQ ID NO: 43):

[0128]

[0129] 1H3-1 light chain nucleic acid sequence (SEQ ID NO: 44):

[0130] CAGGCTGTTGTGACTCAGGAATCTGCACTCACCACATCACCTGGTGAAACAGTCACACTCACTTGTCGCTCAAGTACTGGGGCTGTTACAACCAGGAACTTTGCCAACTGGGTCCAAGAAAAACCAGATCATTTATTCACGGGTCTAATAGGTGGTACCAACAACCGAGCTCCAGGTGTTCCTGCCAGATTCTCAGGCTCCCTGATTGGAGACAAGGCTGCCCTCACCATCACAGGGGCACAGACTGAGGATGAGGCAATATATTTCTGTGCTCTATGGTACACCACCCATTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTAggtcagcccaagtccactcccactctcaccgtgtttccaccttcctctgaggagctcaaggaaaacaaagccacactggtgtgtctgatttccaacttttccccgagtggtgtgacagtggcctggaaggcaaatggtacacctatcacccagggtgtggacacttcaaatcccaccaaagagggcaacaagttcatggccagcagcttcctacatttgacatcggaccagtggagatctcacaacagttttacctgtcaagttacacatgaaggggacactgtggagaagagtctgtctcctgcagaatgtctc

[0131] 13H9-2 heavy chain nucleic acid sequence (SEQ ID NO: 47):

[0132]

[0133] 13H9-2 light chain nucleic acid sequence (SEQ ID NO: 48):

[0134] CAGGCTGTTGTGACTCAGGAATCTGCACTCACCACATCACCTGGTGAAACAGTCACACTCACTTGTCGCTCAAATACTGGGGCTGTTACAACTAGTAACTATGCCAACTGGGTCCAAGAAAAACCAGATCATTTATTCACTGCTCTAATAGGTGGTACCTACAACCGAGCTCCAGGTGTTCCTGCCAGATTCTCAGGCTCCCTGATTGGAGACAAGTCTGCCCTCACCATCACAGGGGCACAGACTGAGGATGAGGCAATATATTTCTGTGCTCTGTGGCACAGCAACCATTGGGTGTTCGGTGGAGGAACCAAACTGACTGTCCTAggtcagcccaagtccactcccactctcaccgtgtttccaccttcctctgaggagctcaaggaaaacaaagccacactggtgtgtctgatttccaacttttccccgagtggtgtgacagtggcctggaaggcaaatggtacacctatcacccagggtgtggacacttcaaatcccaccaaagagggcaacaagttcatggccagcagcttcctacatttgacatcggaccagtggagatctcacaacagttttacctgtcaagttacacatgaaggggacactgtggagaagagtctgtctcctgcagaatgtctc

[0135] 14G7-1 heavy chain nucleic acid sequence (SEQ ID NO: 51):

[0136]

[0137] 14G7-1 heavy chain nucleic acid sequence (SEQ ID NO: 51):

[0138] CAGGCTGTTGTGACTCAGGAATCTGCACTCACCACATCACCTGGTGAAACAGTCACACTCACTTGTCGCTCAAGTACTGGGGCTGTTACAACTAGTAACTTTGCCAACTGGGTCCAAGAAAAACCAGATCATTTATTCACTGGTCTAATAGGTGGTACCAACAACCGCGCTCCAGGTGTTCCTGCCAGATTCTCAGGCTCCCTGATTGGAGACAAGGCTGCCCTCACCATCACAGGGGCACAGACTGAGGATGAGGCAGTATATTTCTGTGCTCTCTGGTACAGAAACCACTGGGTGTTCGGTGGAGGAACCGAACTGACTGTCCTAggtcagcccaagtccactcccactctcaccgtgtttccaccttcctctgaggagctcaaggaaaacaaagccacactggtgtgtctgatttccaacttttccccgagtggtgtgacagtggcctggaaggcaaatggtacacctatcacccagggtgtggacacttcaaatcccaccaaagagggcaacaagttcatggccagcagcttcctacatttgacatcggaccagtggagatctcacaacagttttacctgtcaagttacacatgaaggggacactgtggagaagagtctgtctcctgcagaatgtctc

[0139] Note: In the nucleic acid sequence, upper case is variable region, lower case is constant region

[0140] Example 7 Antibody affinity assay

[0141] The affinity constants (KD) of antibodies 1H3-1, 13H9-2 and 14G7-1 to SN38 were detected by surface plasmon resonance (GE, model Biacore T200) analysis technology. SN38-KLH was covalently coupled to the surface of a CM5 chip by an amino group (GE Healthcare), and antibodies 1H3-1, 13H9-2 and 14G7-1 were diluted with buffer (10 mM HEPES, 150 mM NaCl, 3 mM EDTA, 0.05% Tween 20, pH 7.4) to different concentrations: 3.125 nM, 6.25 nM, 12.5 nM, 25 nM, 50 nM and 100 nM, respectively, and injected into the chip at a flow rate of 30 μL / min, with a sample binding time of 300 s and a dissociation time of 600 s. After each binding and dissociation, the chip surface was regenerated with 50 mM hydrochloric acid for 60 s at a flow rate of 100 μL / min.

[0142] Table 17 Affinity determination values

[0143] Clone ka(1 / Ms) kd(1 / s) KD(M) 1H3-1 1.28E+05 3.90E-04 3.06E-09 13H9-2 1.28E+05 1.35E-04 1.06E-09 14G7-1 2.78E+05 3.42E-03 1.23E-08

[0144] The results of detecting the affinity constants of antibodies 1H3-1, 13H9-2 and 14G7-1 to SN38 are shown in Table 17. SEQUENCE LISTING <110> Shanghai Fudan Zhangjiang Biomedicine Co., Ltd. <120> An SN38 antibody or antigen-binding fragment thereof and application thereof <130> P20016924C <160> 52 <170> PatentIn version 3.5 <210> 1 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 HCDR1 <400> 1 Gly Tyr Thr Phe Thr Thr Tyr Trp 1 5 <210> 2 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 HCDR2 <400> 2 Ile Asn Pro Ser Asn Gly Arg Thr 1 5 <210> 3 <211> 16 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 HCDR3 <400> 3 Ala Thr Ser Gly Pro Tyr Tyr Ser Tyr Asp Val Ala Trp Phe Ala Tyr 1 5 10 15 <210> 4 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 HCDR1 <400> 4 Gly Tyr Thr Phe Thr Thr Ala Gly 1 5 <210> 5 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 HCDR2 <400> 5 Ile Asn Thr His Ser Gly Val Pro 1 5 <210> 6 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 HCDR3 <400> 6 Ala Arg Ala Ser Ser Ser Leu Tyr Arg Tyr Asp Gln Ala Trp Phe Thr 1 5 10 15 Tyr <210> 7 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 HCDR1 <400> 7 Gly Tyr Thr Phe Thr Gly Tyr Trp 1 5 <210> 8 <211> 8 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 HCDR2 <400> 8 Ile Asn Pro Gly Asn Gly Arg Thr 1 5 <210> 9 <211> 14 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 HCDR3 <400> 9 Ala Arg Pro Leu Asp Gly Tyr Tyr Glu Thr Trp Phe Ser Tyr 1 5 10 <210> 10 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 LCDR1 <400> 10 Thr Gly Ala Val Thr Thr Arg Asn Phe 1 5 <210> 11 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 LCDR3 <400> 11 Ala Leu Trp Tyr Thr Thr His Trp Val 1 5 <210> 12 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 LCDR1 <400> 12 Thr Gly Ala Val Thr Thr Ser Asn Tyr 1 5 <210> 13 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 LCDR3 <400> 13 Ala Leu Trp His Ser Asn His Trp Val 1 5 <210> 14 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 LCDR1 <400> 14 Thr Gly Ala Val Thr Thr Ser Asn Phe 1 5 <210> 15 <211> 9 <212> PRT <213> Artificial Sequence <220> <223> 14G7‑1 LCDR3 <400> 15 Ala Leu Trp Tyr Arg Asn His Trp Val 1 5 <210> 16 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> 1H3‑1 VH FR1 <400> 16 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Asp Ser 20 25 <210> 17 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 1H3‑1 VH FR2 <400> 17 Met His Trp Val Met Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile Gly 1 5 10 15 Glu <210> 18 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 VH FR3 <400> 18 Asn Tyr Asn Glu Arg Phe Lys Asn Lys Ala Thr Val Thr Val Asp Lys 1 5 10 15 Ser Ser Thr Thr Ala Tyr Met Gln Leu Ser Ser Leu Thr Ser Glu Asp 20 25 30 Ser Ala Val Tyr Tyr Cys 35 <210> 19 <211> 11 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 VH FR4 <400> 19 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala 1 5 10 <210> 20 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VH FR1 <400> 20 Gln Ile His Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ala Val Arg Ile Ser Cys Lys Ala Ser 20 25 <210> 21 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VH FR2 <400> 21 Met Gin Trp Val Gin Lys Met Pro Gly Lys Gly Leu Arg Trp lie Gly 1 5 10 15 Trp <210> 22 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VH FR3 <400> 22 Arg Tyr Ala Glu Asp Phe Ala Gly Arg Phe Ala Phe Ser Leu Glu Thr 1 5 10 15 Ser Ala Thr lie Val Tyr Leu Gin lie Ser Asn Leu Lys Tyr Glu Asp 20 25 30 Thr Ala Thr Tyr Phe Cys 35 <210> 23 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VH FR1 <400> 23 Gln Val Arg Leu Gin Gin Pro Gly Ala Glu Leu Val Thr Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser 20 25 <210> 24 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VH FR2 <400> 24 Met His Trp Val Arg Gin Arg Pro Gly Gin Gly Leu Glu Trp He Gly 1 5 10 15 Glu <210> 25 <211> 38 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VH FR3 <400> 25 Asn Tyr He Glu Lys Phe Lys Thr Lys Ala Thr Leu Thr Val Asp Lys 1 5 10 15 Ser Ser Asn Thr Ala Tyr Met Gin Leu Ser Ser Leu Thr Ser Glu Asp 20 25 30 Ser Ala Val Tyr Tyr Cys 35 <210> 26 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 and 14G7-1 VL FR1 <400> 26 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Ser 20 25 <210> 27 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 and 14G7-1 VL FR2 <400> 27 Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly Leu Ile 1 5 10 15 Gly <210> 28 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 VL FR3 <400> 28 Asn Arg Ala Pro Gly Val Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly 1 5 10 15 Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala Gln Thr Glu Asp Glu Ala 20 25 30 Ile Tyr Phe Cys 35 <210> 29 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 and 13H9-2 VL FR4 <400> 29 Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 1 5 10 <210> 30 <211> 25 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VL FR1 <400> 30 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Asn 20 25 <210> 31 <211> 17 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VL FR2 <400> 31 Ala Asn Trp Val Gin Glu Lys Pro Asp His Leu Phe Thr Ala Leu Ile 1 5 10 15 Gly <210> 32 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VL FR3 <400> 32 Asn Arg Ala Pro Gly Val Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly 1 5 10 15 Asp Lys Ser Ala Leu Thr Ile Thr Gly Ala Gin Thr Glu Asp Glu Ala 20 25 30 Ile Tyr Phe Cys 35 <210> 33 <211> 36 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VL FR3 <400> 33 Asn Arg Ala Pro Gly Val Pro Ala Arg Phe Ser Gly Ser Leu Ile Gly 1 5 10 15 Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala Gln Thr Glu Asp Glu Ala 20 25 30 Val Tyr Phe Cys 35 <210> 34 <211> 10 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VL FR4 <400> 34 Phe Gly Gly Gly Thr Glu Leu Thr Val Leu 1 5 10 <210> 35 <211> 123 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 VH amino acid sequence <400> 35 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Asp Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Met Gin Arg Pro Gly Gin Gly Leu Glu Trp lie 35 40 45 Gly Glu lie Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Arg Phe 50 55 60 Lys Asn Lys Ala Thr Val Thr Val Asp Lys Ser Ser Thr Thr Ala Tyr 65 70 75 80 Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Ser Gly Pro Tyr Tyr Ser Tyr Asp Val Ala Trp Phe Ala Tyr 100 105 110 Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 36 <211> 109 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 VL amino acid sequence <400> 36 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Arg 20 25 30 Asn Phe Ala Asn Trp Val Gin Glu Lys Pro Asp His Leu Phe Thr Gly 35 40 45 Leu Ile Gly Gly Thr Asn Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Ala Leu Trp Tyr Thr Thr 85 90 95 His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <210> 37 <211> 124 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VH amino acid sequence <400> 37 Gln Ile His Leu Val Gin Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ala Val Arg Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Ala 20 25 30 Gly Met Gin Trp Val Gin Lys Met Pro Gly Lys Gly Leu Arg Trp Ile 35 40 45 Gly Trp lie Asn Thr His Ser Gly Val Pro Arg Tyr Ala Glu Asp Phe 50 55 60 Ala Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr lie Val Tyr 65 70 75 80 Leu Gin lie Ser Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Ala Ser Ser Ser Leu Tyr Arg Tyr Asp Gin Ala Trp Phe Thr 100 105 110 Tyr Trp Gly Gin Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 38 <211> 109 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 VL amino acid sequence <400> 38 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Asn Thr Gly Ala Val Thr Thr Ser 20 25 30 Asn Tyr Ala Asn Trp Val Gin Glu Lys Pro Asp His Leu Phe Thr Ala 35 40 45 Leu Ile Gly Gly Thr Tyr Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu Ile Gly Asp Lys Ser Ala Leu Thr Ile Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Ala Leu Trp His Ser Asn 85 90 95 His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu 100 105 <210> 39 <211> 121 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VH amino acid sequence <400> 39 Gln Val Arg Leu Gln Gln Pro Gly Ala Glu Leu Val Thr Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr 20 25 30 Trp Met His Trp Val Arg Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Gly Asn Gly Arg Thr Asn Tyr Ile Glu Lys Phe 50 55 60 Lys Thr Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Leu Asp Gly Tyr Tyr Glu Thr Trp Phe Ser Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala 115 120 <210> 40 <211> 109 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 VL amino acid sequence <400> 40 Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser 20 25 30 Asn Phe Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly 35 40 45 Leu Ile Gly Gly Thr Asn Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala Val Tyr Phe Cys Ala Leu Trp Tyr Arg Asn 85 90 95 His Trp Val Phe Gly Gly Gly Thr Glu Leu Thr Val Leu 100 105 <210> 41 <211> 459 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 heavy chain amino acid sequence <400> 41 Gln Val Gln Leu Gln Gln Pro Gly Ala Glu Leu Val Arg Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Asp Ser Gly Tyr Thr Phe Thr Thr Tyr 20 25 30 Trp Met His Trp Val Met Gln Arg Pro Gly Gln Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Ser Asn Gly Arg Thr Asn Tyr Asn Glu Arg Phe 50 55 60 Lys Asn Lys Ala Thr Val Thr Val Asp Lys Ser Ser Thr Thr Ala Tyr 65 70 75 80 Met Gln Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Thr Ser Gly Pro Tyr Tyr Ser Tyr Asp Val Ala Trp Phe Ala Tyr 100 105 110 Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro 115 120 125 Pro Ser Val Tyr Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly Ser 130 135 140 Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Ser Val 145 150 155 160 Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Ser Val His Thr Phe 165 170 175 Pro Ala Leu Leu Gln Ser Gly Leu Tyr Thr Met Ser Ser Ser Val Thr 180 185 190 Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Ser Val Ala 195 200 205 His Pro Ala Ser Ser Thr Thr Val Asp Lys Lys Leu Glu Pro Ser Gly 210 215 220 Pro Ile Ser Thr Ile Asn Pro Cys Pro Pro Cys Lys Glu Cys His Lys 225 230 235 240 Cys Pro Ala Pro Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe Pro 245 250 255 Pro Asn Ile Lys Asp Val Leu Met Ile Ser Leu Thr Pro Lys Val Thr 260 265 270 Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser 275 280 285 Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His 290 295 300 Arg Glu Asp Tyr Asn Ser Thr Ile Arg Val Val Ser Ala Leu Pro Ile 305 310 315 320 Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn 325 330 335 Asn Lys Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ile Lys 340 345 350 Gly Leu Val Arg Ala Pro Gln Val Tyr Ile Leu Pro Pro Pro Ala Glu 355 360 365 Gln Leu Ser Arg Glu Asp Val Ser Leu Thr Cys Leu Val Val Gly Phe 370 375 380 Asn Pro Gly Asp Ile Ser Val Glu Trp Thr Ser Asn Gly His Thr Glu 385 390 395 400 Glu Asn Tyr Lys Asp Thr Ala Pro Val Leu Asp Ser Asp Gly Ser Tyr 405 410 415 Phe Ile Tyr Ser Lys Leu Asp Ile Lys Thr Ser Lys Trp Glu Lys Thr 420 425 430 Asp Ser Phe Ser Cys Asn Val Arg His Glu Gly Leu Lys Asn Tyr Tyr 435 440 445 Leu Lys Lys Thr Ile Ser Arg Ser Pro Gly Lys 450 455 <210> 42 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> 1H3-1 light chain amino acid sequence <400> 42 Gln Ala Val Val Thr Gln Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Arg 20 25 30 Asn Phe Ala Asn Trp Val Gln Glu Lys Pro Asp His Leu Phe Thr Gly 35 40 45 Leu Ile Gly Gly Thr Asn Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu Ile Gly Asp Lys Ala Ala Leu Thr Ile Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala lie Tyr Phe Cys Ala Leu Trp Tyr Thr Thr 85 90 95 His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gin Pro 100 105 110 Lys Ser Thr Pro Thr Leu Thr Val Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125 Lys Glu Asn Lys Ala Thr Leu Val Cys Leu lie Ser Asn Phe Ser Pro 130 135 140 Ser Gly Val Thr Val Ala Trp Lys Ala Asn Gly Thr Pro lie Thr Gin 145 150 155 160 Gly Val Asp Thr Ser Asn Pro Thr Lys Glu Gly Asn Lys Phe Met Ala 165 170 175 Ser Ser Phe Leu His Leu Thr Ser Asp Gin Trp Arg Ser His Asn Ser 180 185 190 Phe Thr Cys Gin Val Thr His Glu Gly Asp Thr Val Glu Lys Ser Leu 195 200 205 Ser Pro Ala Glu Cys Leu 210 <210> 43 <211> 1377 <212> DNA <213> Artificial Sequence <220> <223> 1H3-1 heavy chain nucleic acid sequence <400> 43 CAGGTCCAAC TACAGCAGCC TGGGGCTGAA CTGGTGAGGC CTGGGGCTTC AGTGA AACTG 60 TCCTGCAAGG ATTCTGGCTA CACTTTTACC ACCTACTGGA TGC ACTGGGT GATGC AGAGG 120 CCTGGACAAG GTCTTGAGTG GATTGGAGAG ATTAATCCTA GCAACGGTCG GACTA ACTAC 180 AATGAGAGGT TCAAGAACAA GGCCACAGTG ACTGTA GACAATCCTCC ACCACAGCCT AC 240 ATGCAACTCA GCAGCCTGAC ATCTGAGGAC TCTGCGGTCT ATTACTGTGC AACATCAGGG 300 CCCTACTATA GTTACGACGT GGCCTGGTTT GCTTACTGGG GCCAAGGGAC TCTTGTCACT 360 GTCTCTGCAG CCAA AACAAC ACCCCCATC AGTCTATCCC ACTGGCCCCT GG GTTGGAGAT 420 ACA ACTGGTTCC TCTGTGACTC TGGGATGCCT GGTCAAGGGC TACTTCCCTG AGTCAGTG 480 ACTGTGACTT GGA ACTCTGG ATCCCTGTCC AGCAGTGTGC ACACCTTCCC AGCTCTCCTG 540 CAGTCTGGAC TCTACACTAT GAGCAGCTCA GTGACTGTCC CCTCCAGCAC CTGGCCAAGT 600 CAGACCCTCA CCTGCAGCGT TGCTCACCCA GCCAGCAGCA CCACG GTGGACA AAA AACTT 660 GAGCCCAGCG GGCCCATTTC AACAATCAAC CCCTGTCCTC CATGCAAGGA GTGTCACAAA 720 TGCCCAGCTC CTAACCTCGA GGGTGGACCA TCCGTCTTCA TCTTCCCTCC AAAATCAAG 780 GATGTACTCA TGATCTCCCT GACACCCAAG GTCACGTGTG TGGTGGTGGA TGTGAGCGAG 840 GATGACCCAG ACGTCCAGAT CAGCTGGTTT GTGAACAACG TGGAAGTACA CACAGCTCAG 900 ACACAACCAT AGAGAGGATT ACAACAGTAC TATCCGGGTG GTCAGTGCCC TCCTCCATC 960 CAGCACCAAG ACTGGATGAG TGGCAAGGAG TTCAAATGCA AGGTCAACAA CAAAGACCTC 1020 CCATCACCCA TCGAGAGACA TCTCAAAAAT TAAAGGGCT AGTCAGAGCT CCACAAGTA 1080 TACATCTTGC CGCCACCAGC AGAGCAGTTG TCCAGGGAAG ATGTCAGTCT CACTTGCCTG 1140 GTCGTGGGCT TCAACCCTGG AGACATCAGT GTGGAGTGGA CCAGCAATGG GCATAcAGAG 1200 GAGAACTACA AGGACACCAC CCAGTCCTGG ACTCTGACGG TTCTTACTT CATATAcAGC 1260 AAGCTCGATA TAAAAACAAG CAAGTGGGAG AAAACAGATT CCTTCTCATG CAACGTGAGA 1320 CACGAGGGTC TGAAAAATTA CTACCTGAAG AAGACCATCT CCCGCTTCCT GGGTAAG 1377 <210> 44 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> 1H3-1 Light Chain Nucleic Acid Sequence <400> 44 caggctgttg tgactcagga atctgcactc accacatcac ctggtgaaac agtcacactc 60 acttgtcgct caagtactgg ggctgttaca accaggaact ttgccaactg ggtccaagaa 120 aaaccagatc atttattcac gggtctaata ggtggtacca acaaccgagc tccaggtgtt 180 cctgccagat tctcaggctc cctgattgga gacaaggctg ccctcaccat cacaggggca 240 cagactgagg atgaggcaat atatttctgt gctctatggt acaccaccca ttgggtgttc 300 ggtggaggaa ccaaactgac tgtcctaggt cagcccaagt ccactcccac tctcaccgtg 360 tttccacctt cctctgagga gctcaaggaa aacaaagcca cactggtgtg tctgatttcc 420 aacttttccc cgagtggtgt gacagtggcc tggaaggcaa atggtacacc tatcacccag 480 ggtgtggaca cttcaaatcc caccaaagag ggcaacaagt tcatggccag cagcttccta 540 catttgacat cggaccagtg gagatctcac aacagtttta cctgtcaagt tacacatgaa 600 ggggacactg tggagaagag tctgtctcct gcagaatgtc tc 642 <210> 45 <211> 460 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 heavy chain amino acid sequence <400> 45 Gln Ile His Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly Glu 1 5 10 15 Ala Val Arg Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Thr Ala 20 25 30 Gly Met Gln Trp Val Gln Lys Met Pro Gly Lys Gly Leu Arg Trp Ile 35 40 45 Gly Trp Ile Asn Thr His Ser Gly Val Pro Arg Tyr Ala Glu Asp Phe 50 55 60 Ala Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser Ala Thr Ile Val Tyr 65 70 75 80 Leu Gln Ile Ser Asn Leu Lys Tyr Glu Asp Thr Ala Thr Tyr Phe Cys 85 90 95 Ala Arg Ala Ser Ser Ser Leu Tyr Arg Tyr Asp Gln Ala Trp Phe Thr 100 105 110 Tyr Trp Gly Gln Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr 115 120 125 Pro Pro Ser Val Tyr Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly 130 135 140 Ser Ser Val Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Ser 145 150 155 160 Val Thr Val Thr Trp Asn Ser Gly Ser Leu Ser Ser Ser Val His Thr 165 170 175 Phe Pro Ala Leu Leu Gln Ser Gly Leu Tyr Thr Met Ser Ser Ser Val 180 185 190 Thr Val Pro Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Ser Val 195 200 205 Ala His Pro Ala Ser Ser Thr Thr Val Asp Lys Lys Leu Glu Pro Ser 210 215 220 Gly Pro Ile Ser Thr Ile Asn Pro Cys Pro Pro Cys Lys Glu Cys His 225 230 235 240 Lys Cys Pro Ala Pro Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe 245 250 255 Pro Pro Asn Ile Lys Asp Val Leu Met Ile Ser Leu Thr Pro Lys Val 260 265 270 Thr Cys Val Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile 275 280 285 Ser Trp Phe Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr 290 295 300 His Arg Glu Asp Tyr Asn Ser Thr Ile Arg Val Val Ser Ala Leu Pro 305 310 315 320 Ile Gln His Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val 325 330 335 Asn Asn Lys Asp Leu Pro Ser Pro Ile Glu Arg Thr Ile Ser Lys Ile 340 345 350 Lys Gly Leu Val Arg Ala Pro Gln Val Tyr Ile Leu Pro Pro Pro Ala 355 360 365 Glu Gln Leu Ser Arg Glu Asp Val Ser Leu Thr Cys Leu Val Val Gly 370 375 380 Phe Asn Pro Gly Asp Ile Ser Val Glu Trp Thr Ser Asn Gly His Thr 385 390 395 400 Glu Glu Asn Tyr Lys Asp Thr Ala Pro Val Leu Asp Ser Asp Gly Ser 405 410 415 Tyr Phe Ile Tyr Ser Lys Leu Asp Ile Lys Thr Ser Lys Trp Glu Lys 420 425 430 Thr Asp Ser Phe Ser Cys Asn Val Arg His Glu Gly Leu Lys Asn Tyr 435 440 445 Tyr Leu Lys Lys Thr Ile Ser Arg Ser Pro Gly Lys 450 455 460 <210> 46 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> 13H9-2 Light Chain Amino Acid Sequence <400> 46 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Asn Thr Gly Ala Val Thr Thr Ser 20 25 30 Asn Tyr Ala Asn Trp Val Gin Glu Lys Pro Asp His Leu Phe Thr Ala 35 40 45 Leu Ile Gly Gly Thr Tyr Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu Ile Gly Asp Lys Ser Ala Leu Thr Ile Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala Ile Tyr Phe Cys Ala Leu Trp His Ser Asn 85 90 95 His Trp Val Phe Gly Gly Gly Thr Lys Leu Thr Val Leu Gly Gin Pro 100 105 110 Lys Ser Thr Pro Thr Leu Thr Val Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125 Lys Glu Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asn Phe Ser Pro 130 135 140 Ser Gly Val Thr Val Ala Trp Lys Ala Asn Gly Thr Pro Ile Thr Gin 145 150 155 160 Gly Val Asp Thr Ser Asn Pro Thr Lys Glu Gly Asn Lys Phe Met Ala 165 170 175 Ser Ser Phe Leu His Leu Thr Ser Asp Gin Trp Arg Ser His Asn Ser 180 185 190 Phe Thr Cys Gin Val Thr His Glu Gly Asp Thr Val Glu Lys Ser Leu 195 200 205 Ser Pro Ala Glu Cys Leu 210 <210> 47 <211> 1380 <212> DNA <213> Artificial Sequence <220> <223> 13H9-2 heavy chain nucleic acid sequence <400> 47 cagatccatc tggtgcagtc tggacctgag ctgaagaagc ctggagaggc agtcaggatc 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 ttgctgctga aaagctgctg gctgctgctg gctgctgctg gctgctgctg gctgctgctg 60 aaggatgtac tcatgatctc cctgacaccc aaggtcacgt gtgtggtggt ggatgtgagc 840 gaggatgacc cagacgtcca gatcagctgg tttgtgaaca acgtggaagt acacacagct 900 cagacacaaa cccatagaga ggattacaac agtactatcc gggtggtcag tgccctcccc 960 atccagcacc aggactggat gagtggcaag gagttcaaat gcaaggtcaa caacaaagac 1020 ctcccatcac ccatcgagag aaccatctca aaaattaaag ggctagtcag agctccacaa 1080 gtatacatct tgccgccacc agcagagcag ttgtccaggg aagatgtcag tctcacttgc 1140 ctggtcgtgg gcttcaaccc tggagacatc agtgtggagt ggaccagcaa tgggcataca 1200 gaggagaact acaaggacac cgcaccagtc ctggactctg acggttctta cttcatatac 1260 agcaagctcg atataaaaac aagcaagtgg gagaaaacag attccttctc atgcaacgtg 1320 agacacgagg gtctgaaaaa ttactacctg aagaagacca tctcccggtc tccgggtaaa 1380 <210> 48 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> 13H9-2 light chain nucleic acid sequence <400> 48 caggctgttg tgactcagga atctgcactc accacatcac ctggtgaaac agtcacactc 60 acttgtcgct caaatactgg ggctgttaca actagtaact atgccaactg ggtccaagaa 120 aaaccagatc atttattcac tgctctaata ggtggtacct acaaccgagc tccaggtgtt 180 cctgccagat tctcaggctc cctgattgga gacaagtctg ccctcaccat cacaggggca 240 cagactgagg atgaggcaat atatttctgt gctctgtggc acagcaacca ttgggtgttc 300 ggtggaggaa ccaaactgac tgtcctaggt cagcccaagt ccactcccac tctcaccgtg 360 tttccacctt cctctgagga gctcaaggaa aacaaagcca cactggtgtg tctgatttcc 420 aacttttccc cgagtggtgt gacagtggcc tggaaggcaa atggtacacc tatcacccag 480 ggtgtggaca cttcaaatcc caccaaagag ggcaacaagt tcatggccag cagcttccta 540 catttgacat cggaccagtg gagatctcac aacagtttta cctgtcaagt tacacatgaa 600 ggggacactg tggagaagag tctgtctcct gcagaatgtc tc 642 <210> 49 <211> 457 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 Heavy Chain Amino Acid Sequence <400> 49 Gln Val Arg Leu Gin Gin Pro Gly Ala Glu Leu Val Thr Pro Gly Ala 1 5 10 15 Ser Val Lys Leu Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr Gly Tyr 20 25 30 Trp Met His Trp Val Arg Gin Arg Pro Gly Gin Gly Leu Glu Trp Ile 35 40 45 Gly Glu Ile Asn Pro Gly Asn Gly Arg Thr Asn Tyr Ile Glu Lys Phe 50 55 60 Lys Thr Lys Ala Thr Leu Thr Val Asp Lys Ser Ser Asn Thr Ala Tyr 65 70 75 80 Met Gin Leu Ser Ser Leu Thr Ser Glu Asp Ser Ala Val Tyr Tyr Cys 85 90 95 Ala Arg Pro Leu Asp Gly Tyr Tyr Glu Thr Trp Phe Ser Tyr Trp Gly 100 105 110 Gln Gly Thr Leu Val Thr Val Ser Ala Ala Lys Thr Thr Pro Pro Ser 115 120 125 Val Tyr Pro Leu Ala Pro Gly Cys Gly Asp Thr Thr Gly Ser Ser Val 130 135 140 Thr Leu Gly Cys Leu Val Lys Gly Tyr Phe Pro Glu Ser Val Thr Val 145 150 155 160 Thr Trp Asn Ser Gly Ser Leu Ser Ser Ser Val His Thr Phe Pro Ala 165 170 175 Leu Leu Gln Ser Gly Leu Tyr Thr Met Ser Ser Ser Val Thr Val Pro 180 185 190 Ser Ser Thr Trp Pro Ser Gln Thr Val Thr Cys Ser Val Ala His Pro 195 200 205 Ala Ser Ser Thr Thr Val Asp Lys Lys Leu Glu Pro Ser Gly Pro Ile 210 215 220 Ser Thr Ile Asn Pro Cys Pro Pro Cys Lys Glu Cys His Lys Cys Pro 225 230 235 240 Ala Pro Asn Leu Glu Gly Gly Pro Ser Val Phe Ile Phe Pro Pro Asn 245 250 255 Ile Lys Asp Val Leu Met Ile Ser Leu Thr Pro Lys Val Thr Cys Val 260 265 270 Val Val Asp Val Ser Glu Asp Asp Pro Asp Val Gln Ile Ser Trp Phe 275 280 285 Val Asn Asn Val Glu Val His Thr Ala Gln Thr Gln Thr His Arg Glu 290 295 300 Asp Tyr Asn Ser Thr lie Arg Val Val Ser Ala Leu Pro lie Gin His 305 310 315 320 Gln Asp Trp Met Ser Gly Lys Glu Phe Lys Cys Lys Val Asn Asn Lys 325 330 335 Asp Leu Pro Ser Pro lie Glu Arg Thr lie Ser Lys lie Lys Gly Leu 340 345 350 Val Arg Ala Pro Gin Val Tyr lie Leu Pro Pro Pro Ala Glu Gin Leu 355 360 365 Ser Arg Glu Asp Val Ser Leu Thr Cys Leu Val Val Gly Phe Asn Pro 370 375 380 Gly Asp lie Ser Val Glu Trp Thr Ser Asn Gly His Thr Glu Glu Asn 385 390 395 400 Tyr Lys Asp Thr Ala Pro Val Leu Asp Ser Asp Gly Ser Tyr Phe lie 405 410 415 Tyr Ser Lys Leu Asp lie Lys Thr Ser Lys Trp Glu Lys Thr Asp Ser 420 425 430 Phe Ser Cys Asn Val Arg His Glu Gly Leu Lys Asn Tyr Tyr Leu Lys 435 440 445 Lys Thr lie Ser Arg Ser Pro Gly Lys 450 455 <210> 50 <211> 214 <212> PRT <213> Artificial Sequence <220> <223> 14G7-1 Heavy Chain Amino Acid Sequence <400> 50 Gln Ala Val Val Thr Gin Glu Ser Ala Leu Thr Thr Ser Pro Gly Glu 1 5 10 15 Thr Val Thr Leu Thr Cys Arg Ser Ser Thr Gly Ala Val Thr Thr Ser 20 25 30 Asn Phe Ala Asn Trp Val Gin Glu Lys Pro Asp His Leu Phe Thr Gly 35 40 45 Leu He Gly Gly Thr Asn Asn Arg Ala Pro Gly Val Pro Ala Arg Phe 50 55 60 Ser Gly Ser Leu He Gly Asp Lys Ala Ala Leu Thr He Thr Gly Ala 65 70 75 80 Gln Thr Glu Asp Glu Ala Val Tyr Phe Cys Ala Leu Trp Tyr Arg Asn 85 90 95 His Trp Val Phe Gly Gly Gly Thr Glu Leu Thr Val Leu Gly Gin Pro 100 105 110 Lys Ser Thr Pro Thr Leu Thr Val Phe Pro Pro Ser Ser Glu Glu Leu 115 120 125 Lys Glu Asn Lys Ala Thr Leu Val Cys Leu Ile Ser Asn Phe Ser Pro 130 135 140 Ser Gly Val Thr Val Ala Trp Lys Ala Asn Gly Thr Pro Ile Thr Gln 145 150 155 160 Gly Val Asp Thr Ser Asn Pro Thr Lys Glu Gly Asn Lys Phe Met Ala 165 170 175 Ser Ser Phe Leu His Leu Thr Ser Asp Gln Trp Arg Ser His Asn Ser 180 185 190 Phe Thr Cys Gln Val Thr His Glu Gly Asp Thr Val Glu Lys Ser Leu 195 200 205 Ser Pro Ala Glu Cys Leu 210 <210> 51 <211> 1371 <212> DNA <213> Artificial Sequence <220> <223> 14G7-1 heavy chain nucleic acid sequence <400> 51 caggtccgac tgcagcagcc tggggctgaa ctggtgacac ctggggcttc agtgaagctg 60 tcctgtaagg cttctggcta caccttcacc ggctactgga tgcactgggt gaggcagagg 120 cctggacaag gccttgagtg gattggagag attaatcctg gcaacggtcg tactaactac 180 attgagaagt tcaagaccaa ggccacactg actgtagaca aatcctccaa cacagcctac 240 atgcaactca gcagcctgac atctgaggac tctgcggtct attactgtgc aagacccctt 300 gatggttatt acgagacctg gttttcttac tggggccaag ggactctggt cactgtctct 360 gcagccaaaa caacaccccc atcagtctat ccactggccc ctgggtgtgg agatacaact 420 ggttcctctg tgactctggg atgcctggtc aagggctact tccctgagtc agtgactgtg 480 acttggaact ctggatccct gtccagcagt gtgcacacct tcccagctct cctgcagtct 540 ggactctaca ctatgagcag ctcagtgact gtcccctcca gcacctggcc aagtcagacc 600 gtcacctgca gcgttgctca cccagccagc agcaccacgg tggacaaaaa acttgagccc 660 agcgggccca tttcaacaat caacccctgt cctccatgca aggagtgtca caaatgccca 720 gctcctaacc tcgagggtgg accatccgtc ttcatcttcc ctccaaatat caaggatgta 780 ctcatgatct ccctgacacc caaggtcacg tgtgtggtgg tggatgtgag cgaggatgac 840 ccagacgtcc agatcagctg gtttgtgaac aacgtggaag tacacacagc tcagacacaa 900 acccatagag aggattacaa cagtactatc cgggtggtca gtgccctccc catccagcac 960 caggactgga tgagtggcaa ggagttcaaa tgcaaggtca acaacaaaga cctcccatca 1020 cccatcgaga gaaccatctc aaaaattaaa gggctagtca gagctccaca agtatacatc 1080 ttgccgccac cagcagagca gttgtccagg gaagatgtca gtctcacttg cctggtcgtg 1140 ggcttcaacc ctggagacat cagtgtggag tggaccagca atgggcatac agaggagaac 1200 tacaaggaca ccgcaccagt cctggactct gacggttctt acttcatata cagcaagctc 1260 gatataaaaa caagcaagtg ggagaaaaca gattccttct catgcaacgt gagacacgag 1320 ggtctgaaaa attactacct gaagaagacc atctcccggt ctccgggtaa a 1371 <210> 52 <211> 642 <212> DNA <213> Artificial Sequence <220> <223> 14G7-1 light chain nucleic acid sequence <400> 52 caggctgttg tgactcagga atctgcactc accacatcac ctggtgaaac agtcacactc 60 acttgtcgct caagtactgg ggctgttaca actagtaact ttgccaactg ggtccaagaa 120 aaaccagatc atttattcac tggtctaata ggtggtacca acaaccgcgc tccaggtgtt 180 cctgccagat tctcaggctc cctgattgga gacaaggctg ccctcaccat cacaggggca 240 cagactgagg atgaggcagt atatttctgt gctctctggt acagaaacca ctgggtgttc 300 ggtggaggaa ccgaactgac tgtcctaggt cagcccaagt ccactcccac tctcaccgtg 360 tttccacctt cctctgagga gctcaaggaa aacaaagcca cactggtgtg tctgatttcc 420 aacttttccc cgagtggtgt gacagtggcc tggaaggcaa atggtacacc tatcacccag 480 ggtgtggaca cttcaaatcc caccaaagag ggcaacaagt tcatggccag cagcttccta 540 catttgacat cggaccagtg gagatctcac aacagtttta cctgtcaagt tacacatgaa 600 ggggacactg tggagaagag tctgtctcct gcagaatgtc tc 642

Claims

1. An SN38 antibody or antigen-binding fragment thereof, comprising a heavy chain variable region and a light chain variable region; the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NO: 1-3, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 10, GTN, SEQ ID NO: 11, respectively; or the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NO: 4-6, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 12, GTY, SEQ ID NO: 13, respectively; the heavy chain variable region comprises HCDR1, HCDR2 and HCDR3 having the amino acid sequences set forth in SEQ ID NO: 7-9, respectively, and the light chain variable region comprises LCDR1, LCDR2 and LCDR3 having the amino acid sequences set forth in SEQ ID NO: 14, GTN, SEQ ID NO: 15, respectively. the heavy chain variable region comprises HFR1, HFR2, HFR3 and HFR4, and the light chain variable region comprises LFR1, LFR2, LFR3 and LFR4; the heavy chain variable region comprises HFR1, HFR2, HFR3 and HFR4 having the amino acid sequences set forth in SEQ ID NO: 16-18 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3 and LFR4 having the amino acid sequences set forth in SEQ ID NO: 26-29, respectively; or, the heavy chain variable region comprises HFR1, HFR2, HFR3 and HFR4 having the amino acid sequences set forth in SEQ ID NO: 20-22 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3 and LFR4 having the amino acid sequences set forth in SEQ ID NO: 30-32 and SEQ ID NO: 29, respectively; the heavy chain variable region comprises HFR1, HFR2, HFR3 and HFR4 having the amino acid sequences set forth in SEQ ID NO: 23-25 and SEQ ID NO: 19, respectively, and the light chain variable region comprises LFR1, LFR2, LFR3 and LFR4 having the amino acid sequences set forth in SEQ ID NO: 26, SEQ ID NO: 27, SEQ ID NO: 33 and SEQ ID NO: 34, respectively.

2. The SN38 antibody or antigen-binding fragment thereof of claim 1, wherein, ​ ​ 3. The SN38 antibody or antigen-binding fragment thereof of claim 1, wherein, the heavy chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 35 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 35, and the light chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 36 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 36; or, the heavy chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 37 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 37, and the light chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 38 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 38; or, the heavy chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 39 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO: 39, and the light chain variable region comprises an amino acid sequence as set forth in SEQ ID NO: 40 or that is at least 90% identical to the amino acid sequence set forth in SEQ ID NO:

40.

4. The SN38 antibody or antigen-binding fragment thereof of claim 1, wherein, The SN38 antibody or antigen-binding fragment thereof comprises a murine or human antibody constant region or a mutation thereof; the murine antibody constant region comprises a heavy chain constant region of murine IgGl, IgG2a, IgG2b3, or IgG3 and a kappa or lambda type light chain constant region, and the human antibody constant region comprises a heavy chain constant region of human IgGl, IgG2, IgG3, or IgG4 and a kappa or lambda type light chain constant region.

5. The SN38 antibody or antigen-binding fragment thereof of claim 4, wherein, The mutation has at least 90% identity to the amino acid sequence of the murine or human antibody constant region.

6. The SN38 antibody or antigen-binding fragment thereof of claim 5, wherein, The heavy chain constant region amino acid sequence of the SN38 antibody or antigen-binding fragment thereof is set forth in SEQ ID NO: 41, positions 124-459; and the light chain constant region amino acid sequence of the SN38 antibody or antigen-binding fragment thereof is set forth in SEQ ID NO: 42, positions 110-209.

7. The SN38 antibody or antigen-binding fragment thereof of any one of claims 1-6, wherein, The structure of the SN38 antibody or antigen-binding fragment thereof comprises an immunoglobulin, Fab, Fab', F(ab')2, Fv, and single-chain Fv fragment.

8. An isolated nucleic acid, comprising, The nucleic acid molecule encoding the SN38 antibody or antigen-binding fragment thereof as claimed in any one of claims 1-7.

9. The isolated nucleic acid of claim 8, wherein, In the nucleotide sequence encoding the SN38 antibody or the antigen-binding fragment thereof, the nucleotide sequence encoding the heavy chain variable region is set forth in positions 1-369 of SEQ ID NO: 43, the nucleotide sequence encoding the light chain variable region is set forth in positions 1-327 of SEQ ID NO: 44; or the nucleotide sequence encoding the heavy chain variable region is set forth in positions 1-372 of SEQ ID NO: 47, the nucleotide sequence encoding the light chain variable region is set forth in positions 1-327 of SEQ ID NO: 48; or the nucleotide sequence encoding the heavy chain variable region is set forth in positions 1-363 of SEQ ID NO: 51, the nucleotide sequence encoding the light chain variable region is set forth in positions 1-327 of SEQ ID NO:

52.

10. An expression vector, characterized in that, The expression vector comprises the isolated nucleic acid of claim 8 or 9.

11. The expression vector of claim 10, wherein, The expression vector is pcDNA3.1(+).

12. A transformant comprising the expression vector of claim 10 or 11.

13. The transformant of claim 12, wherein, The transformant is a genetically modified lentivirus or immune effector cell.

14. The transformant of claim 13, wherein, The lentivirus is a lentivirus containing PLVX-EF1-CAR, lentiCRISPR v2-CAR or pELNS-CAR, and the immune effector cell is a T lymphocyte, NK cell or NKT cell.

15. A test composition, characterized in that The detection composition comprises the SN38 antibody or the antigen-binding fragment thereof of any one of claims 1-7, the isolated nucleic acid of claim 8 or 9, the expression vector of claim 10 or 11, and / or the transformant of any one of claims 12-14.

16. A kit comprising, The kit comprises the SN38 antibody or the antigen-binding fragment thereof of any one of claims 1-7, the isolated nucleic acid of claim 8 or 9, the expression vector of claim 10 or 11, the transformant of any one of claims 12-14, and / or the detection composition of claim 15.

17. The kit of claim 16, wherein The kit further comprises (i) a device for administering the antibody or the antigen-binding fragment thereof, the isolated nucleic acid, the expression vector, the transformant or the detection composition; and / or (ii) instructions for use.

18. A method of detecting SN38 in a sample, characterized by, The method comprises reacting the sample with the SN38 antibody or the antigen-binding fragment thereof of any one of claims 1-7, the isolated nucleic acid of claim 8 or 9, the expression vector of claim 10 or 11, the transformant of any one of claims 12-14, the detection composition of claim 15, and / or the kit of claim 16 or 17.

19. Use of the SN38 antibody or the antigen-binding fragment thereof of any one of claims 1-7, the isolated nucleic acid of claim 8 or 9, the expression vector of claim 10 or 11, the transformant of any one of claims 12-14, the detection composition of claim 15, or the kit of claim 16 or 17 in detecting SN38.

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