Design, preparation and application of innate immune super connector and complete immune antibody

Abstract

The invention provides the design of an Innate Super Enager (ISE) and a Complete Immune Antibody (CIA) as well as a preparation method and application of the Innate Super Enager (ISE) and the Complete Immune Antibody (CIA). The innate immune super connector contains an antibody or a ligand which connects and activates innate immune cells, or a cytokine which stimulates proliferation of the innate immune cells, or a pattern recognition receptor (PRR) agonist. The innate immune super connector can be used independently, can also be combined with other immune antibodies, or can be further connected with one or more postnatal immune antibodies or ligands to form a complete immune antibody. The complete immune antibody has two forms: (1) an end N of the innate immune super connector is connected with an acquired immune antibody or a tumor antigen antibody; and (2) connecting the C end or the N end of the acquired immune antibody or the tumor antigen antibody to an innate immune super connector.

Description

TECHNICAL FIELD

[0001] The present application belongs to the technical field of biological medicine, and more particularly relates to the preparation and application of cell engager or multispecific antibody. BACKGROUND

[0002] Immunity is divided into innate immunity and acquired (acquired) immunity. Innate immunity is born with, and acquired immunity is gradually acquired by the body during life. Innate immunity, also known as non-specific immunity, is a function of maintaining health from birth. This immunity has the function of rejection and phagocytosis to all foreign substances, bacteria, foreign bodies, etc. It includes body surface barrier, blood-brain barrier, blood-placenta barrier, cell phagocytosis, and antibacterial substances in normal body fluids and tissues. The innate immune system constitutes the first line of defense against microbial pathogens, and relies on pattern recognition receptors (PRRs). PRRs are host sensors encoded by the germline, which can detect typical molecules belonging to pathogens, and play a crucial role in the normal function of the innate immune system [1] . They are proteins expressed mainly by innate immune cells (such as dendritic cells, macrophages, monocytes, neutrophils, NK cells) and epithelial cells, which are used to recognize two types of molecules: pathogen-associated molecular patterns (PAMPs) associated with microbial pathogens; damage-associated molecular patterns (DAMPs) associated with host cell components released during cell damage or death. According to the protein domain, PRRs in the vertebrate innate immune system can be divided into the following five types: Toll-like receptors (TLRs), nucleotide oligomerization domain (NOD)-like receptors (NLRs), retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs), C-type lectin receptors (CLRs), and melanoma deficiency factor 2 (AIM2)-like receptors (ALRs) [1] . TLRs are the most widely distributed and most important PRRs. There are 13 types of TLRs found so far: humans have TLR1-10, and mice have 12 TLRs (TLR1-9, TLR11, 12, 13). PRRs are activated to produce innate immune cytokines such as type I interferons, which can directly inhibit viral replication; and by activating other innate immune cells and acquired immune cells, bridging acquired immunity to clear pathogenic microorganisms and regulate immune function.

[0003] pDC is a very basic and key innate immune cell. The endosomal TLR-7 or TLR-9 receptor of pDC is activated by ligands to produce a large amount of type I interferon, which activates a large number of interferon stimulated genes (ISGs) to directly inhibit the proliferation of viruses. Type I interferon is also a key cytokine for immune regulation of the body, which can activate all immune cells: dendritic cells, macrophages, T cells, B cells, to achieve immune amplification and immune regulation; can inhibit the proliferation of tumor cells and cause tumor cell apoptosis [2] , and can regulate a series of downstream immune responses to achieve the re-immune balance of the body. Non-methylated DNA ODN in pathogenic microorganisms contains CpG structure pattern: its sequence contains a cytosine deoxyribonucleotide (Cytosine), followed by a guanine deoxyribonucleotide (Guanine), and "p" refers to the phosphodiester bond between the two. CPG ODN is an agonist of TLR-9, which can stimulate pDC to produce a large amount of type I interferon. CpG ODN has been clinically shown to convert "cold tumors" into "hot tumors" and has a good synergistic effect in improving the clinical trials of PD-1 therapy [3] .

[0004] PRR-activated macrophages can phagocytose pathogenic microorganisms and tumor cells. Macrophages are the main component of immune cell infiltration in the tumor microenvironment, which can account for 50% of solid tumor mass. Although macrophages can directly phagocytose and kill tumor cells, they are immunosuppressed in the tumor microenvironment due to the expression of CD47 by tumor cells. The signal regulatory protein-alpha (SIRPα) expressed by macrophages is a major myeloid inhibitory receptor, which is a regulatory mechanism to prevent excessive phagocytosis [4] . Its ligand is the CD47 molecule on the surface of some cells, such as the CD47 expressed by red blood cells, which binds to the SIRPα expressed by macrophages to convey a "don't eat me" signal, thereby preventing damage to red blood cells. The SIRPα of aged red blood cells is significantly reduced, making macrophages lack the "don't eat me" signal, and actively phagocytose aged red blood cells, so that red blood cells achieve homeostasis, which is a self-regulatory mechanism of the body. Tumor cells also convey a "don't eat me" signal by overexpressing CD47, thereby achieving phagocytosis escape. There are many monoclonal antibodies and Fc-SIRPα fusion molecules designed based on this mechanism to re-activate the phagocytosis function of macrophages to tumor cells by blocking the "don't eat me" signal. Such products include Magrolimab (anti-CD47) and TTI-622 (Fc-SIRPα fusion molecule). Such products have shown some efficacy in clinical trials for hematological tumors such as acute myeloid leukemia (AML), but the efficacy is not significant [5]The reason for this is that the immune system of different tumors or the same tumor at different stages is extremely heterogeneous, showing a lack of innate immune components such as macrophages, NK cells, and some parts of innate immune factors, and a depletion of some subgroups of adaptive immune functions such as antigen-specific T cells and B cells. Especially in patients with advanced tumors, the immune function is generally low or extremely heterogeneous, and multiple immunosuppressive functions are at work, so simply improving the phagocytic function of macrophages cannot translate into clinical efficacy. At the same time, if the number of macrophages in the tumor population in the clinical trial is low and the M2 subtype is high, it will result in no target cells to correct for the CD47 "don't eat me" drug, which also leads to the failure of the clinical trial. However, if the number of macrophages and the number of M1 cells in the selected tumor patients reach a certain level through screening, it can be predicted that the effect of CD47 target therapy will be significant.

[0005] Natural killer cells, also known as NK cells or large granular lymphocytes (LGL), are a type of cytotoxic lymphocyte that is critical to the innate immune system [6] . They belong to the rapidly expanding family of known innate lymphoid cells (ILCs) and account for 5-20% of all circulating lymphocytes in humans. NK cells function similarly to cytotoxic T cells in the adaptive immune response of vertebrates. NK cells act on virus-infected cells and other intracellular pathogens about 3 days after infection, providing a rapid response and monitoring the body's tumor formation; they can respond and kill tumors at the early stage of tumor formation. Most immune cells recognize and destroy infected cells or cancer cells by sensing the antigens presented on the surface of the major histocompatibility complex (MHC) of infected cells or cancer cells, but NK cells can recognize and kill infected cells or tumor cells without antibodies and MHC, thereby achieving a faster immune response. Because some tumor cells escape the killing of specific cytotoxic T cells (Cytotoxic T Lymphocyte, CTL) belonging to the adaptive immune system by not expressing MHC I, the natural killer function of NK cells becomes particularly important in the body's tumor monitoring and killing. In addition to being able to directly kill cancer cells or infected cells, NK cells are also effector cells of the adaptive immune system, and antibodies against tumor or microbial antigens can activate NK cells by binding to CD16a on the surface of NK cells, achieving the function of killing tumors and infected cells, which is also known as antibody-dependent cell-mediated cytotoxicity (ADCC). AFM-13 is a CD16a and CD30 bispecific antibody that has entered clinical trials and has shown good results in the treatment of Hodgkin's lymphoma expressing CD30. The combination therapy of AFM-13 and AlloNK has entered the fast track of clinical development by the US FDA [7] .

[0006] In summary, by activating PRR receptors such as TLR9, pDC can be activated to produce a large amount of interferon, directly inhibiting the proliferation of tumor cells and inducing the growth of tumor cells in the acquired immune system; by preventing the "don't eat me" signal, macrophages can be activated to phagocytose and kill cancer cells; by connecting and activating NK cells, general tumor cells and tumor cells without markers (MHCI deficiency) can be quickly killed. These mechanisms can fully exert the functions of innate immunity to effectively kill tumor cells.

[0007] The connector formed by connecting the innate immune mechanisms can directly or indirectly bridge various innate immune effector cells and cancer cells, fully exerting the tumor cell killing functions of these different functional cells, not only achieving a strong tumor cell killing effect in the case of normal innate immune cells, but also achieving a certain therapeutic effect in the case of weak immune cells (such as macrophage dysfunction). Since the innate immune status of tumor patients is different, there may be a situation where one cell-mediated innate immunity is strong and another cell-mediated innate immunity is weak, so the cell connector recruiting a single innate immune cell may not be effective. To solve this problem, the Innate Super Engager (ISE) disclosed in the present application can activate pDC, macrophages and natural killer cells at the same time, and can effectively kill tumor cells and treat cancer under various different immune states.

[0008] In a normal body, both innate immunity and acquired immunity play a key role in killing and controlling tumor cells. In the early stages of tumor formation, the number of tumor cells is small, and the inhibition of immune mechanisms is absent or very low, so any part of the human immune system can suppress the growth of tumor cells, such as NK cells, which can quickly kill tumor cells regardless of whether the tumor cells contain MHCI; activated macrophages can directly phagocytose tumor cells regardless of whether they contain MHCI. After pDC is activated (such as by viruses or bacteria containing CpG fragments), a large amount of interferon is produced, which can directly inhibit the proliferation of cancer cells and achieve the effect of controlling tumor growth by activating a large number of innate and acquired immune cells. After the formation of acquired immunity against tumor cells, the strong tumor antigen-specific cytotoxic cells can directly and effectively kill tumor cells; the tumor antigen-specific B cells induced can produce anti-tumor antibodies, which can kill tumor cells through the ADCC function of recruiting NK cells.

[0009] But after the tumor cells achieve uncontrolled proliferation through numerous immune escape mechanisms, the body's immune cells and functions are greatly damaged, especially in patients with advanced cancer or metastasis, whose immune systems, including innate immunity and acquired immunity, have been damaged to varying degrees. Moreover, the immune status of different tumor patients also varies greatly, some may have a relatively sound acquired immune system, but the innate immune function is low; some have low acquired immunity but relatively strong innate immunity. In this case, affecting a single immune system or a single immune cell, such as only regulating the function of innate immune cells, such as only regulating a single innate immune effector cell such as NK cells or macrophages to kill and control tumors will only achieve limited effect. At the same time, in this case, it is difficult to determine which immune system or cell can be activated to kill tumor cells. Therefore, if the main innate immune regulation strategy and the main acquired immune regulation strategy can be integrated together, effective immunotherapy can be achieved in the case of immune system damage in tumor patients, and the nature and extent of immune damage cannot be determined. The Complete Immune Antibody (CIA) disclosed in the present application can simultaneously integrate innate immune drugs such as the Innate Super Connector (ISE) disclosed in the present application and acquired immune drugs such as T cell function regulating drugs (anti-PD-1 / PD-L1 antibodies) or anti-tumor antigen antibodies (such as anti-EGFR / Her2 antibodies) to achieve the best effect of killing and controlling tumors. SUMMARY

[0010] The present disclosure discloses a kind of innate immunity super connector, it includes at least two components of innate immune system selected from the following: pattern recognition receptors (PRR) agonist, macrophage activator, NK cell connection and activator, immune cytokine.More specifically, the innate immunity super connector disclosed in the present disclosure uses conventional antibody as basic structure, removes the variable region of antibody, or uses no targeting antibody in human body, and is connected with the component of innate immune system selected from the following at the N terminal or C terminal of antibody heavy chain constant region, or the N terminal or C terminal of antibody light chain constant region: PRR agonist, macrophage activator, NK cell connection and activator, immune cytokine.Two or more of PRR agonist, macrophage activator, NK cell connection and activator and immune cytokine can be combined with the skeleton antibody by linker, to form the innate immunity super connector with the required configuration.Various PRR agonists, macrophage activators, NK cell connection and activators and immune cytokines known in the art can be used.PRR agonist includes but is not limited to Toll-like receptor (TLR) agonist, NOD-like receptor (NLR) agonist, RIG-I-like receptor (RLR) agonist, C-type lectin receptor (CLR) agonist and melanoma deficiency factor 2 (AIM2) like receptor (ALR) agonist.Macrophage activator includes but is not limited to SIRP alpha D1 protein, CD40 antibody and CD24 antibody.NK cell connection and activator include but are not limited to anti-CD16a antibody.Immune cytokine includes but is not limited to IL-15, IL-18, IL-2, IL-7, IL-10 and IL-12.

[0011] The innate immune superconnector disclosed in the present invention is capable of recruiting or activating innate immune cells. As an example, the ISE disclosed in the present invention may have the following immune components: (1) ISE-A, containing SIRPα protein and anti-CD16a single-chain antibody; (2) ISE-B, containing SIRPα protein, anti-CD16a nanoantibody and immune cytokines such as IL-15; (3) ISE-C, containing SIRPα protein, anti-CD16a nanoantibody and immune cytokines and their receptors such as IL-15 and IL-15Rα; (4) ISE-D, containing SIRPα protein, anti-CD16a single-chain antibody and immune cytokines such as IL-15 or IL-2; (5) ISE-E, a PRR agonist is connected to the long chain N-terminus of ISE-A; (6) ISE-F, a PRR agonist is connected to the short chain N-terminus or C-terminus of ISE-A; (7) ISE-G, a PRR agonist is connected to the long chain C-terminus of ISE-A; (8) ISE-H, an IL-2 mutant or IFNα mutant is connected to the long chain N-terminus of ISE-A; (9) ISE-I, an IL-2 mutant or IFNα mutant is connected to the long chain N-terminus of ISE-B; (10) ISE-J, an IL-2 mutant or IFNα mutant is connected to the long chain N-terminus of ISE-D. SIRPα protein can block the "Don't eat me" signal of macrophages, and Anti-CD16a antibodies can specifically connect to and activate NK cells; cytokines such as IL-15 can cause NK cells to proliferate, so that the ability of NK cells to kill tumors will not be limited by the small number of cells. PRR agonists can activate the interferon production mechanism, IL-2 mutants or IFNα mutants, by introducing mutations, improve the binding ability to NK cells and CD8+T cell receptors.

[0012] The present invention also discloses a complete immune antibody (CIA), which comprises an ISE and an acquired immune antibody. The acquired immune antibody includes antibodies that regulate T cell immunity such as anti-PD-1 antibodies and PD-L1 antibodies, anti-tumor antigen antibodies such as anti-HER2 antibodies and anti-EGFR antibodies, and antibodies that regulate B cell function, such as anti-B cell surface receptor antibodies such as anti-CD40 antibodies or anti-CD40L antibodies. T cell regulators can relieve T cell immunosuppression by antagonizing immune checkpoints; anti-tumor antigen antibodies can target tumor cells and inhibit the growth of tumor cells; antibodies against B cell surface antigens can increase the ability of B cells to present tumor antigens. At least one ISE and one acquired immune antibody constitute a series of complete immune antibodies (CIA), and the possible composition of CIA antibodies is shown in the following table:

[0013] Table 1. Complete immune antibody combination table

[0014]

[0015] The key technical problems focused on by the present application are as follows:

[0016] (1) By innovatively designing the structure of ISE, NK cells are recruited while the "Don't eat me" signal of macrophages is blocked, thereby exerting the maximum innate immune effect.

[0017] (2) The immune cytokine and its receptor introduced in the structure, such as IL-15 or IL-15Rα fragment, can stimulate the proliferation of NK cells, thereby improving the killing function on tumor cells.

[0018] (3) After the activation of PRR, innate immune cytokines such as type I interferon are produced, which can directly inhibit the propagation of viruses; and by activating other innate immune cells and adaptive immune cells, the purpose of regulating immune function is achieved.

[0019] (4) Human CD16 protein has two subtypes, CD16a and CD16b, CD16a is mainly expressed in NK cells, and also expressed in monocytes and macrophages, which belongs to an activated receptor; CD16b is expressed in neutrophils, and the two subtypes are highly homologous, the present application ensures the specificity of anti-CD16a and does not bind to CD16b on the surface of neutrophils, thereby avoiding the side effects caused by non-specific binding [8] .

[0020] (5) ISE alone has the effect of killing tumor cells, and due to the non-specificity of ISE, it can be widely used in different tumors.

[0021] (6) ISE can combine innate immunity and adaptive immunity by connecting adaptive immune antibodies into complete immune antibodies (CIA), thereby fully improving the killing effect on tumor cells.

[0022] (7) CIA can replace the targeting antibody of adaptive immunity, thereby realizing specific targeted killing of different tumors.

[0023] (8) By innovatively designing the structure of ISE and CIA, the toxic and side effects of cytokines are reduced.

[0024] In order to achieve the above purposes, the technical solutions of the present application are as follows.

[0025] One object of the present application is to provide a kind of innate immune super connector and complete immune antibody, the innate immune super connector includes following two or more components: (a) anti-CD16a antibody, (b) SIRP alpha D1 protein, (c) immune cytokine, (d) pattern recognition receptor agonist.The complete immune antibody is based on the structure of ISE increases the antibody of adaptive immunity or tumor antigen.

[0026] In a particular embodiment, the traditional antibody is used as a basic structure, the variable region of the antibody is removed, or a targeting antibody is not used in the human body, and the anti-CD16a single-chain antibody or the SIRPaDl protein is connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker, and the SIRPaDl protein or the anti-CD16a single-chain antibody is connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker.

[0027] In a particular embodiment, the traditional antibody is used as a basic structure, the variable region of the antibody is removed, or a targeting antibody is not used in the human body, and the human immune cytokine and / or human immune cytokine receptor and anti-CD16a VHH are connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker, and the SIRPaDl protein is connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker, or the human immune cytokine and / or human immune cytokine receptor and anti-CD16a VHH are connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker, and the SIRPaDl protein is connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker.

[0028] In a particular embodiment, the traditional antibody is used as a basic structure, the variable region of the antibody is removed, or a targeting antibody is not used in the human body, and the human immune cytokine such as IL-15 and the anti-CD16a single-chain antibody are connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker, and the SIRPaDl protein is connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker, or the human immune cytokine such as IL-15 and the anti-CD16a single-chain antibody are connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker, and the SIRPaDl protein is connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker.

[0029] In a further embodiment, a pattern recognition receptor agonist is connected to the N-terminus or C-terminus of the ISE through a linker.

[0030] In a further embodiment, human IL-2 or IFNa or a mutant thereof is connected to the N-terminus or C-terminus of the ISE through a linker.

[0031] In a particular embodiment, the immune cytokine in the ISE includes but is not limited to IL-15, IL-18, IL-2, IL-7, IL-10, IL-12, etc.

[0032] In a particular embodiment, the ISE is used as a basic structure, and the acquired immune antibody variable region or the tumor-associated antigen antibody variable region is added to the N-terminus of the ISE to form a complete immune antibody.

[0033] In a particular embodiment, the fully immune antibody targeting moiety can be a tumor associated antigen antibody, including but not limited to anti-PD-L1, Her2, VEGF, EGFR, CD38, Claudin18.2, Claudin6, CD19, MSLN, Trop-2, CD20, Nectin-4, c-Met, MUC16, MUC17, 5T4, DLL3 or CD30, etc.

[0034] In a particular embodiment, the fully immune antibody targeting moiety can be a tumor associated antigen antibody, including but not limited to anti-PD-L1, Her2, VEGF, EGFR, CD38, Claudin18.2, Claudin6, CD19, MSLN, Trop-2, CD20, Nectin-4, c-Met, MUC16, MUC17, 5T4, DLL3 or CD30, etc.

[0035] In a particular embodiment, the fully immune antibody targeting moiety can be a tumor associated antigen antibody, including but not limited to anti-PD-L1, Her2, VEGF, EGFR, CD38, Claudin18.2, Claudin6, CD19, MSLN, Trop-2, CD20, Nectin-4, c-Met, MUC16, MUC17, 5T4, DLL3 or CD30, etc.

[0036] In one embodiment, the basic scaffold is an anti-hen egg lysozyme antibody (anti-hen egg lysozyme HEL antibody). In particular embodiments, the amino acid sequence of the long chain of the monoclonal antibody HELAB01 is SEQ ID NO: 20, and the amino acid sequence of the short chain is SEQ ID NO: 21. The amino acid sequence of the long chain of the bispecific antibody HELAB02 used as a mechanism verification is SEQ ID NO: 22, and the amino acid sequence of the short chain is SEQ ID NO: 23. The amino acid sequence of the long chain of the bispecific antibody HELAB03 used as a mechanism verification is SEQ ID NO: 24, and the amino acid sequence of the short chain is SEQ ID NO: 25. The amino acid sequence of the long chain of the trispecific antibody HELAB04 used as a mechanism verification is SEQ ID NO: 26, and the amino acid sequence of the short chain is SEQ ID NO: 27. The amino acid sequence of the long chain of the trispecific antibody HELAB05 used as a mechanism verification is SEQ ID NO: 28, and the amino acid sequence of the short chain is SEQ ID NO: 29.

[0037] In some embodiments, the Fc fragment of the antibody introduces L234A and L235A mutations, or further introduces M252Y, S254T and / or T256E mutations in the Fc fragment.

[0038] Another object of the present application is to provide a method for preparing the innate immune super connector and the complete immune antibody as described above, comprising the following steps:

[0039] (1) The structure of the present application is shown in Figures 1-3 , which is a symmetrical structure. The two chains of ISE or CIA are amplified respectively by PCR technology; M252Y / S254T / T256E mutations are introduced into the Fc segment of the antibody to prolong the half-life of the protein;

[0040] (2) The DNA fragment of one chain obtained in step (1) is cloned into a pcDNA series vector or other vector for a mammalian cell expression system at an appropriate position by conventional molecular biology means, and the DNA fragment of the other chain obtained in step (1) is cloned into another pcDNA series vector or other vector for a mammalian cell expression system at an appropriate position, including a fusion DNA sequence connected with appropriate transcription and translation regulatory sequences in the vector of the expression system;

[0041] (3) The recombinant vector obtained in step (2) is transfected into mammalian cells for expression of the fusion protein, and the purified ISE or CIA is obtained after one step of affinity chromatography. The transfection method can be chemical reagent transfection or electroporation transfection, and the mammalian cells can be HEK293 cells or CHO (Chinese Hamster Ovary) cells or derivative cells of the above-mentioned cells or other expression systems (such as E. coli and yeast). DETAILED DESCRIPTION

[0042] I. Innate immune super connector

[0043] The innate immunity superconnectors described herein comprise components of the innate immune system, including but not limited to pattern recognition receptor (PRR) agonists, macrophage activators, NK cell connecting and activators, and immune cytokines. Various PRR agonists, macrophage activators, NK cell connecting and activators, and immune cytokines known in the art can be employed. The pattern recognition receptors include Toll-like receptors (TLRs), nucleotide oligomerization domain (NOD)-like receptors (NLRs), retinoic acid-inducible gene-I (RIG-I)-like receptors (RLRs), C-type lectin receptors (CLRs), and absent in melanoma 2 (AIM2)-like receptors (ALRs). The PRR agonists used herein include, but are not limited to, Toll-like receptor (TLRs) agonists, NOD-like receptor (NLRs) agonists, RIG-I-like receptor (RLRs) agonists, C-type lectin receptor (CLRs) agonists, and absent in melanoma 2 (AIM2)-like receptor (ALRs) agonists. The macrophage activators used herein include, but are not limited to, SIRPaD1 protein, CD40 antibody, and CD24 antibody. The NK cell connecting and activators used herein include, but are not limited to, CD16a antibody. The immune cytokines used herein include, but are not limited to, IL-15, IL-18, IL-2, IL-7, IL-10, and IL-12.

[0044] The innate immunity superconnectors described herein employ a suitable basic scaffold to connect components of the innate immune system. The basic scaffold described herein can include conventional antibodies with variable regions removed, such as IgG, IgA, IgM, IgE, and IgD with variable regions removed. The basic scaffold described herein can include off-target antibodies in the human body, such as anti-chicken egg white lysozyme antibodies, anti-pathogenic microorganism antibodies such as anti-viral, bacterial antibodies, and the like.

[0045] In some embodiments, the innate immunity superconnectors of the present application comprise one or more of a macrophage activator, an NK cell connecting and activator, a PRR agonist, and an immune cytokine to activate macrophages, natural killer cells, and / or DC cells. In some embodiments, the innate immunity superconnectors of the present application comprise a macrophage activator, preferably a SIRPaD1 protein, and an NK cell connecting and activator, preferably an anti-CD16a antibody. The anti-CD16a antibody can be an anti-CD16a single chain antibody or an anti-CD16a nanobody.

[0046] In some embodiments, the innate immunity super-connector of the present application comprises two or more of the following components: (a) an anti-CD16a antibody, (b) a SIRPaDl protein, (c) an immune cytokine, (d) a pattern recognition receptor agonist to activate pDCs, macrophages and / or natural killer cells.

[0047] In some embodiments, the innate immunity super-connector of the present application comprises two or more of the following components: (a) an anti-CD16a antibody, (b) a SIRPaDl protein, (c) an immune cytokine, (d) a pattern recognition receptor agonist to activate pDCs, macrophages and / or natural killer cells. Figure 1 In some embodiments, the innate immunity super-connector of the present application comprises two or more of the following components: (a) an anti-CD16a antibody, (b) a SIRPaDl protein, (c) an immune cytokine, (d) a pattern recognition receptor agonist to activate pDCs, macrophages and / or natural killer cells. Figure 1In some embodiments, a conventional antibody is used as the basic structure, the variable region of the antibody is removed, or an antibody without a target in the human body is used as the basic structure, and a human immunocytokine and / or human immunocytokine receptor, an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH), and a SIRPαD1 protein are connected at the N-terminus and / or C-terminus of the constant region of the heavy chain of the antibody via a linker. In some embodiments, a conventional antibody is used as the basic structure, the variable region of the antibody is removed, or an antibody without a target in the human body is used as the basic structure, and a human immunocytokine and / or human immunocytokine receptor, an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH), and a SIRPαD1 protein are connected at the N-terminus and / or C-terminus of the constant region of the light chain of the antibody via a linker. In some embodiments, a traditional antibody is used as the basic structure, the variable region of the antibody is removed, or a non-targeted antibody in the human body is used as the basic structure, (1) a human immunocytokine and / or human immunocytokine receptor and an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) are connected to the N-terminus and / or C-terminus of the antibody heavy chain constant region through a linker, and (2) a SIRPαD1 protein is connected to the N-terminus or C-terminus of the antibody light chain constant region through a linker. In the above embodiments, human immunocytokines include, but are not limited to, IL-15, IL-18, IL-2, IL-7, IL-10, and IL-12. In the above embodiments, human immunocytokine receptors include, but are not limited to, IL-15 receptor, IL-18 receptor, IL-2 receptor, IL-7 receptor, IL-10 receptor, and IL-12 receptor.

[0048] In a specific embodiment, a traditional antibody is used as the basic structure, the variable region of the antibody is removed or a human non-targeted antibody is used, and a human immune cytokine (e.g., human IL-15) and an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) are connected to the C-terminus of the heavy chain constant region of the antibody through a linker, and a SIRPαD1 protein is connected to the N-terminus of the heavy chain constant region of the antibody through a linker, for example, see Figure 1 In a specific embodiment, a traditional antibody is used as the basic structure, the variable region of the antibody is removed or a non-targeted antibody in the human body is used, and a human immune cytokine (e.g., human IL-15), a human immune cytokine receptor (e.g., human IL-15 receptor), and an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) are connected to the C-terminus of the antibody heavy chain constant region via a linker, and a SIRPαD1 protein is connected to the N-terminus of the antibody heavy chain constant region via a linker, for example, seeFigure 1 C1.

[0049] In a specific embodiment, a traditional antibody is used as the basic structure, the variable region of the antibody is removed or a non-targeted antibody in the human body is used, and a human immune cytokine (e.g., human IL-15) and an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) are connected to the C-terminus of the heavy chain constant region of the antibody through a linker, and a SIRPαD1 protein is connected to the C-terminus of the light chain constant region of the antibody through a linker, for example, see Figure 1 In a specific embodiment, a traditional antibody is used as the basic structure, the variable region of the antibody is removed or a non-targeted antibody in the human body is used, and a human immune cytokine (e.g., human IL-15), a human immune cytokine receptor (e.g., human IL-15 receptor), and an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) are connected to the C-terminus of the antibody heavy chain constant region via a linker, and a SIRPαD1 protein is connected to the C-terminus of the antibody light chain constant region via a linker, for example, see Figure 1 In any of the above embodiments, the macrophage activator, the NK cell ligation and activator, the human immunocytokine, and the human immunocytokine receptor can be linked in any order. In any of the above embodiments, the immunocytokine, the immunocytokine receptor, the anti-CD16a antibody, and the SIRPαD1 protein can be linked in any order.

[0050] In some embodiments, the innate immune super connector of the present application is based on a traditional antibody with a variable region removed or an antibody without a target in the human body as a basic skeleton, connecting a macrophage activator, a NK cell connection and activator and a PRR agonist. In a preferred embodiment, the innate immune super connector of the present application is based on a traditional antibody with a variable region removed or an antibody without a target in the human body as a basic skeleton, connecting an anti-CD16a antibody (such as an anti-CD16a single-chain antibody or an anti-CD16aVHH), a SIRPαD1 protein and a PRR agonist, for example, see Figure 1E1 / E2, F1 / F2, and G1 / G2 of ISE-E, ISE-F, and ISE-G configurations. In some embodiments, based on a traditional antibody as a basic structure, removing the variable region of the antibody or using a human body without a targeting antibody, an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH), a PRR agonist, and / or a SIRPaD1 protein are connected by a linker at the N-terminus or C-terminus of the heavy chain constant region of the antibody, and a SIRPaD1 protein and / or an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH), a PRR agonist are connected by a linker at the N-terminus or C-terminus of the light chain constant region of the antibody.

[0051] In a particular embodiment, based on a traditional antibody as a basic structure, removing the variable region of the antibody or using a human body without a targeting antibody, an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) and / or a PRR agonist are connected by a linker at the N-terminus and / or C-terminus of the heavy chain constant region of the antibody, and a SIRPaD1 protein is connected by a linker at the N-terminus and / or C-terminus of the light chain constant region of the antibody, for example, see Figure 1 E1 / E2 and G1 / G2 of ISE-E, ISE-F, and ISE-G configurations. In a particular embodiment, based on a traditional antibody as a basic structure, removing the variable region of the antibody or using a human body without a targeting antibody, an anti-CD16a antibody (e.g., an anti-CD16a single-chain antibody or an anti-CD16a VHH) is connected by a linker at the N-terminus and / or C-terminus of the heavy chain constant region of the antibody, and a SIRPaD1 protein and / or a PRR agonist is connected by a linker at the N-terminus and / or C-terminus of the light chain constant region of the antibody, for example, see Figure 1 F1 and F2 of ISE-E, ISE-F, and ISE-G configurations. In any of the above embodiments, the macrophage activator, the NK cell linking and activator, and the PRR agonist can be connected in any order. In any of the above embodiments, the anti-CD16a antibody, the PRR agonist, and the SIRPaD1 protein can be connected in any order.

[0052] Based on the above ISE-A, ISE-B, ISE-C, and ISE-D configurations, the innate immune super connector described herein can further connect IL-2 or IFNa or a mutant thereof.

[0053] In some embodiments, based on the ISE-A configuration, IL-2 or IFNa or a mutant thereof is connected in ISE-A, for example, see Figure 1the ISE-H configuration of H1 / H2. For example, based on the ISE-A configuration, an IL-2 mutant or an IFNα mutant is linked at the N-terminus of the antibody heavy chain constant region, wherein the SIRPαD1 protein can be linked at the N-terminus and / or C-terminus of the antibody light chain constant region via a linker.

[0054] In some embodiments, based on the ISE-B configuration, an IL-2 or an IFNα or a mutant thereof is linked in ISE-B, for example, see Figure 1 the ISE-I configuration of I1 / I2. For example, based on the ISE-B configuration, an IL-2 mutant or an IFNα mutant is linked at the N-terminus of the antibody heavy chain constant region, wherein the SIRPαD1 protein can be linked at the N-terminus and / or C-terminus of the antibody light chain constant region via a linker.

[0055] In some embodiments, based on the ISE-D configuration, an IL-2 or an IFNα or a mutant thereof is linked in ISE-D, for example, see Figure 1 the ISE-J configuration of J1 / J2. For example, based on the ISE-D configuration, an IL-2 mutant or an IFNα mutant is linked at the N-terminus of the antibody heavy chain constant region, wherein the SIRPαD1 protein can be linked at the N-terminus and / or C-terminus of the antibody light chain constant region; further, the N-terminus and / or C-terminus of the antibody heavy chain constant region can also be linked to a cytokine receptor. In some embodiments, the heavy chain variable region of the anti-CD16a single chain antibody comprises: CDR1 as shown in SEQ ID NO: 1, CDR2 as shown in SEQ ID NO: 2, and CDR3 as shown in SEQ ID NO: 3; the light chain variable region comprises: CDR1 as shown in SEQ ID NO: 4, CDR2 as shown in SEQ ID NO: 5, and CDR3 as shown in SEQ ID NO: 6.

[0056] In further embodiments, the heavy chain variable region of the anti-CD16a single chain antibody comprises an amino acid sequence as shown in SEQ ID NO: 7, and the light chain variable region comprises an amino acid sequence as shown in SEQ ID NO: 8.

[0057] In further embodiments, the anti-CD16a single chain antibody comprises an amino acid sequence as shown in SEQ ID NO: 9.

[0058] In one embodiment, the anti-CD16a nanobody comprises: CDR1 as shown in SEQ ID NO: 10, CDR2 as shown in SEQ ID NO: 11, and CDR3 as shown in SEQ ID NO: 12.

[0059] In a further embodiment, the anti-CD16a nanobody comprises the amino acid sequence set forth in SEQ ID NO: 13.

[0060] In one embodiment, the SIRPaD1 protein comprises the amino acid sequence set forth in SEQ ID NO: 14.

[0061] In one embodiment, the human IL-15 protein fragment comprises the amino acid sequence set forth in SEQ ID NO: 15 or SEQ ID NO: 16.

[0062] In one embodiment, the human IL-15Ra protein fragment comprises the amino acid sequence set forth in SEQ ID NO: 17.

[0063] The present application has prepared different structures of innate immune connectors, such as Figure 1 A-J and Figure 3 as shown in ISE-A, ISE-B, ISE-C, ISE-D, ISE-E, ISE-F, ISE-G, ISE-H, ISE-I and ISE-J, HELAB01, HELAB02, HELAB03, HELAB04 and HELAB05, respectively.

[0064] In some embodiments, the long chain of the innate immune connector ISE-A has the structure of SIRPaD1 protein-linker-antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the short chain has the structure of antibody kappa chain constant region (see, e.g., A1 of Figure 1 In preferred embodiments, the amino acid sequence of the long chain of ISE-A1 comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 30, and the amino acid sequence of the short chain of ISE-A1 comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 31. In more preferred embodiments, the amino acid sequence of the long chain of ISE-A1 comprises the sequence set forth in SEQ ID NO: 30, and the amino acid sequence of the short chain of ISE-A1 comprises the sequence set forth in SEQ ID NO: 31. In one particular embodiment, the amino acid sequence of the long chain of the innate immune connector ISE-A1 is SEQ ID NO: 30, and the amino acid sequence of the short chain is SEQ ID NO: 31.

[0065] In some embodiments, the long chain of the innate immune connector ISE-A has the structure of antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the short chain has the structure of antibody kappa chain constant region-linker-SIRPaD1 protein (see, e.g., A2 of Figure 1of A2). In preferred embodiments, the ISE-A2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 32, and the ISE-A2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 33. In more preferred embodiments, the ISE-A2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 32, and the ISE-A2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 33. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-A2 is SEQ ID NO: 32, and the amino acid sequence of the short chain is SEQ ID NO: 33.

[0066] In some embodiments, the long chain of the innate immune linker ISE-B has the structure: SIRPaD1 protein-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region (see, e.g., Figure 1 of B1). In preferred embodiments, the ISE-B1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 34, and the ISE-B1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 35. In more preferred embodiments, the ISE-B1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 34, and the ISE-B1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 35. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-B1 is SEQ ID NO: 34, and the amino acid sequence of the short chain is SEQ ID NO: 35.

[0067] In some embodiments, the long chain of the innate immune linker ISE-B has the structure: antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region-linker-SIRPaD1 protein (see, e.g., Figure 1B2) of WO 2018 / 231 158. In preferred embodiments, the ISE-B2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 36, and the ISE-B2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 37. In more preferred embodiments, the ISE-B2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 36, and the ISE-B2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 37. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-B2 is SEQ ID NO: 36, and the amino acid sequence of the short chain is SEQ ID NO: 37.

[0068] In some embodiments, the long chain of the innate immune linker ISE-C has the structure: SIRPaD1 protein-linker-antibody heavy chain constant region-linker-human IL-15Ra- linker-human IL-15-linker-anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region (see, e.g., WO 2018 / 231 158, C1). In preferred embodiments, the ISE-C1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 39. In more preferred embodiments, the ISE-C1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 39. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-C1 is SEQ ID NO: 38, and the amino acid sequence of the short chain is SEQ ID NO: 39. Figure 1

[0069] In some embodiments, the long chain of the innate immune linker ISE-C has the structure: SIRPaD1 protein-linker-antibody heavy chain constant region-linker-human IL-15Ra- linker-human IL-15-linker-anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region (see, e.g., WO 2018 / 231 158, C1). In preferred embodiments, the ISE-C1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 39. In more preferred embodiments, the ISE-C1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 39. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-C1 is SEQ ID NO: 38, and the amino acid sequence of the short chain is SEQ ID NO: 39.

[0069] In some embodiments, the long chain of the innate immune linker ISE-C has the structure: SIRPaD1 protein-linker-antibody heavy chain constant region-linker-human IL-15Ra- linker-human IL-15-linker-anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region (see, e.g., WO 2018 / 231 158, C1). In preferred embodiments, the ISE-C1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 39. In more preferred embodiments, the ISE-C1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 38, and the ISE-C1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 39. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-C1 is SEQ ID NO: 38, and the amino acid sequence of the short chain is SEQ ID NO: 39. Figure 1In preferred embodiments, the ISE-C2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 40, and the ISE-C2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 41. In more preferred embodiments, the ISE-C2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 40, and the ISE-C2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 41. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-C2 is SEQ ID NO: 40, and the amino acid sequence of the short chain is SEQ ID NO: 41.

[0070] In some embodiments, the long chain of the innate immune linker ISE-D has the structure: SIRPaD1 protein-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain has the structure: antibody kappa chain constant region (see, e.g., Figure 1 In preferred embodiments, the ISE-D1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 42, and the ISE-D1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 43. In more preferred embodiments, the ISE-D1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 42, and the ISE-D1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 43. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-D1 is SEQ ID NO: 42, and the amino acid sequence of the short chain is SEQ ID NO: 43.

[0071] In some embodiments, the long chain of the innate immune linker ISE-D has the structure: antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain has the structure: antibody kappa chain constant region-linker-SIRPaD1 protein (see, e.g., Figure 1In a preferred embodiment, the ISE-D2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 44, and the ISE-D2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 45. In a more preferred embodiment, the ISE-D2 long chain amino acid sequence comprises the sequence shown in SEQ ID NO: 44, and the ISE-D2 short chain amino acid sequence comprises the sequence shown in SEQ ID NO: 45. In a specific embodiment, the amino acid sequence of the long chain of the innate immunity connector ISE-D2 is SEQ ID NO: 44, and the amino acid sequence of the short chain is SEQ ID NO: 45.

[0072] In some embodiments, the structure of the long chain of the innate immune connector ISE-E is: CpG ODN-linker-antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the structure of the short chain is: antibody kappa chain constant region-linker-SIRPαD1 protein (see, for example Figure 1 In a preferred embodiment, the ISE-E1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 46, and the ISE-E1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 47. In a more preferred embodiment, the ISE-E1 long chain amino acid sequence comprises the sequence shown in SEQ ID NO: 46, and the ISE-E1 short chain amino acid sequence comprises the sequence shown in SEQ ID NO: 47. In a specific embodiment, the amino acid sequence of the long chain of the innate immunity connector ISE-E1 is SEQ ID NO: 46, and the amino acid sequence of the short chain is SEQ ID NO: 47.

[0073] In some embodiments, the structure of the long chain of the innate immune connector ISE-E is: CpG ODN-linker-antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the structure of the short chain is: SIRPαD1 protein-linker-antibody kappa chain constant region (see, for example Figure 1In a preferred embodiment, the ISE-E2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 48, and the ISE-E2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 49. In a more preferred embodiment, the ISE-E2 long chain amino acid sequence comprises the sequence shown in SEQ ID NO: 48, and the ISE-E2 short chain amino acid sequence comprises the sequence shown in SEQ ID NO: 49. In a specific embodiment, the amino acid sequence of the long chain of the innate immunity connector ISE-E2 is SEQ ID NO: 48, and the amino acid sequence of the short chain is SEQ ID NO: 49.

[0074] In some embodiments, the structure of the long chain of the innate immune connector ISE-F is: antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the structure of the short chain is: CpG ODN-linker-antibody kappa chain constant region-linker-SIRPαD1 protein (see, for example Figure 1 In a preferred embodiment, the ISE-F1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 50, and the ISE-F1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97% or at least 99% sequence identity to SEQ ID NO: 51. In a more preferred embodiment, the ISE-F1 long chain amino acid sequence comprises the sequence shown in SEQ ID NO: 50, and the ISE-F1 short chain amino acid sequence comprises the sequence shown in SEQ ID NO: 51. In a specific embodiment, the amino acid sequence of the long chain of the innate immunity connector ISE-F1 is SEQ ID NO: 50, and the amino acid sequence of the short chain is SEQ ID NO: 51.

[0075] In some embodiments, the structure of the long chain of the innate immune connector ISE-F is: antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the structure of the short chain is: SIRPαD1 protein-linker-antibody kappa chain constant region-linker-CpG ODN (see, for example Figure 1In preferred embodiments, the ISE-F2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 52, and the ISE-F2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 53. In more preferred embodiments, the ISE-F2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 52, and the ISE-F2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 53. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-F2 is SEQ ID NO: 52, and the amino acid sequence of the short chain is SEQ ID NO: 53.

[0076] In some embodiments, the long chain of the innate immune linker ISE-G has the structure: antibody heavy chain constant region - linker - anti-CD16a single chain antibody - linker - CpG ODN, and the short chain has the structure: antibody kappa chain constant region - linker - SIRPaD1 protein (see, e.g., Figure 1 In preferred embodiments, the ISE-G1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 54, and the ISE-G1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 55. In more preferred embodiments, the ISE-G1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 54, and the ISE-G1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 55. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-G1 is SEQ ID NO: 54, and the amino acid sequence of the short chain is SEQ ID NO: 55.

[0077] In some embodiments, the long chain of the innate immune linker ISE-G has the structure: antibody heavy chain constant region - linker - anti-CD16a single chain antibody - linker - CpG ODN, and the short chain has the structure: SIRPaD1 protein - linker - antibody kappa chain constant region (see, e.g., Figure 1In preferred embodiments, the ISE-G2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 56, and the ISE-G2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 57. In more preferred embodiments, the ISE-G2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 56, and the ISE-G2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 57. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-G2 is SEQ ID NO: 56, and the amino acid sequence of the short chain is SEQ ID NO: 57.

[0078] In some embodiments, the long chain of the innate immune linker ISE-H has the structure: human IL-2 mutant-linker-antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the short chain has the structure: antibody kappa chain constant region-linker-SIRPaD1 protein (see, e.g., Figure 1 In preferred embodiments, the ISE-H1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 58, and the ISE-H1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 59. In more preferred embodiments, the ISE-H1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 58, and the ISE-H1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 59. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-H1 is SEQ ID NO: 58, and the amino acid sequence of the short chain is SEQ ID NO: 59.

[0079] In some embodiments, the long chain of the innate immune linker ISE-H has the structure: human IL-2 mutant-linker-antibody heavy chain constant region-linker-anti-CD16a single chain antibody, and the short chain has the structure: SIRPaD1 protein-linker-antibody kappa chain constant region (see, e.g., Figure 1In preferred embodiments, the ISE-H2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 58, and the ISE-H2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 60. In more preferred embodiments, the ISE-H2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 58, and the ISE-H2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 60. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-H2 is SEQ ID NO: 58, and the amino acid sequence of the short chain is SEQ ID NO: 60.

[0080] In some embodiments, the long chain of the innate immune linker ISE-I has the structure: human IL-2 mutant - linker - antibody heavy chain constant region - linker - human IL-15 - linker - anti-CD16a nanobody, and the short chain has the structure: antibody kappa chain constant region - linker - SIRPaD1 protein (see, e.g., Figure 1 In preferred embodiments, the ISE-I1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 61, and the ISE-I1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 62. In more preferred embodiments, the ISE-I1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 61, and the ISE-I1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 62. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-I1 is SEQ ID NO: 61, and the amino acid sequence of the short chain is SEQ ID NO: 62.

[0081] In some embodiments, the long chain of the innate immune linker ISE-I has the structure: human IL-2 mutant - linker - antibody heavy chain constant region - linker - human IL-15 - linker - anti-CD16a nanobody, and the short chain has the structure: SIRPaD1 protein - linker - antibody kappa chain constant region (see, e.g., Figure 1In preferred embodiments, the ISE-I2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 63, and the ISE-I2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 64. In more preferred embodiments, the ISE-I2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 63, and the ISE-I2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 64. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-I2 is SEQ ID NO: 63, and the amino acid sequence of the short chain is SEQ ID NO: 64.

[0082] In some embodiments, the long chain of the innate immune linker ISE-J has the structure: human IL-2 mutant-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain has the structure: antibody kappa chain constant region-linker-SIRPaD1 protein (see, e.g., Figure 1 In preferred embodiments, the ISE-J1 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 65, and the ISE-J1 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 66. In more preferred embodiments, the ISE-J1 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 65, and the ISE-J1 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 66. In one specific embodiment, the amino acid sequence of the long chain of the innate immune linker ISE-J1 is SEQ ID NO: 65, and the amino acid sequence of the short chain is SEQ ID NO: 66.

[0083] In some embodiments, the long chain of the innate immune linker ISE-J has the structure: human IL-2 mutant-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain has the structure: SIRPaD1 protein-linker-antibody kappa chain constant region (see, e.g., Figure 2In preferred embodiments, the ISE-J2 long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 65, and the ISE-J2 short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 67. In more preferred embodiments, the ISE-J2 long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 65, and the ISE-J2 short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 67. In one specific embodiment, the ISE-J2 long chain has an amino acid sequence of SEQ ID NO: 65, and the short chain has an amino acid sequence of SEQ ID NO: 67.

[0084] In some embodiments, the ISE-Negative Control long chain has the structure of an antibody heavy chain constant region, and the short chain has the structure of an antibody kappa chain constant region (see, e.g., Figure 2 In preferred embodiments, the ISE-Negative Control long chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 116, and the ISE-Negative Control short chain amino acid sequence comprises a sequence having at least 90%, at least 95%, at least 97%, or at least 99% sequence identity to SEQ ID NO: 31. In more preferred embodiments, the ISE-Negative Control long chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 116, and the ISE-Negative Control short chain amino acid sequence comprises the sequence set forth in SEQ ID NO: 31. In one specific embodiment, the ISE-Negative Control long chain has an amino acid sequence of SEQ ID NO: 116, and the short chain has an amino acid sequence of SEQ ID NO: 31.

[0085] II. HEL antibody

[0086] To demonstrate the possibility of the ISE concept, the present application prepared five antibodies with the main structure of ISE, which are HE LAB01, HE LAB02, HE LAB03, HE LAB04 and HE LAB05, by adding the variable region of an antibody against hen egg lysozyme (anti-hen egg lysozyme HEL antibody) to the ISE, which has no target in the human body, in order to show the superposition function of different parts of ISE and the function of cytokines.

[0087] Monoclonal antibody HELAB01 consists of an anti-HEL antibody heavy chain and an anti-HEL antibody light chain. Preferably, the amino acid sequence of the long chain of HELAB01 (i.e. the anti-HEL antibody heavy chain) is SEQ ID NO: 20 and the amino acid sequence of the short chain (i.e. the anti-HEL antibody light chain) is SEQ ID NO: 21.

[0088] Bispecific antibody HELAB02 is based on an anti-HEL antibody with a SIRPaD1 protein connected by a linker at the N- or C-terminus of the antibody heavy chain or at the N- or C-terminus of the antibody light chain. In one embodiment, the long chain structure of bispecific antibody HELAB02 is: an anti-HEL antibody heavy chain and the short chain structure is: an anti-HEL antibody light chain-linker-SIRPaD1 protein. In one specific embodiment, the amino acid sequence of the long chain of bispecific antibody HELAB02 used for mechanism validation is SEQ ID NO: 22 and the amino acid sequence of the short chain is SEQ ID NO: 23. In an alternative embodiment, the short chain structure of bispecific antibody HELAB02 is replaced by: SIRPaD1 protein-linker-anti-HEL antibody light chain, and preferably the amino acid sequence of the replaced short chain is SEQ ID NO: 85.

[0089] Bispecific antibody HELAB03 is based on an anti-HEL antibody with an anti-CD16a single chain antibody connected by a linker at the N- or C-terminus of the antibody heavy chain or at the N- or C-terminus of the antibody light chain. In one embodiment, the long chain structure of bispecific antibody HELAB03 is: an anti-HEL antibody heavy chain-linker-anti-CD16a single chain antibody and the short chain structure is: an anti-HEL antibody light chain. In one specific embodiment, the amino acid sequence of the long chain of bispecific antibody HELAB03 used for mechanism validation is SEQ ID NO: 24 and the amino acid sequence of the short chain is SEQ ID NO: 25.

[0090] Trispecific antibody HELAB04 has anti-HEL antibody as basic structure, anti-CD16a single chain antibody is connected at N-terminus or C-terminus of antibody heavy chain, and SIRPaD1 protein is connected at N-terminus or C-terminus of antibody light chain. In one embodiment, long chain structure of trispecific antibody HELAB04 is: anti-HEL antibody heavy chain-linker-anti-CD16a single chain antibody, and short chain structure is: anti-HEL antibody light chain-linker-SIRPaD1 protein. In one specific embodiment, amino acid sequence of long chain of trispecific antibody HELAB04 used as mechanism verification is SEQ ID NO: 26, and amino acid sequence of short chain is SEQ ID NO: 27. In one alternative embodiment, short chain structure of trispecific antibody HELAB04 is replaced by: SIRPaD1 protein-linker-anti-HEL antibody light chain, preferably, amino acid sequence of replaced short chain is SEQ ID NO: 85.

[0091] Trispecific antibody HELAB05 has anti-HEL antibody as basic structure, IL-15 and anti-CD16a nanobody are connected at N-terminus or C-terminus of antibody heavy chain, and SIRPaD1 protein is connected at N-terminus or C-terminus of antibody light chain. In one embodiment, long chain structure of trispecific antibody HELAB05 is: anti-HEL antibody heavy chain-linker-human IL-15-anti-CD16a nanobody, and short chain structure is: anti-HEL antibody light chain-linker-SIRPaD1 protein. In one specific embodiment, amino acid sequence of long chain of trispecific antibody HELAB05 used as mechanism verification is SEQ ID NO: 28, and amino acid sequence of short chain is SEQ ID NO: 29. In one alternative embodiment, short chain structure of bispecific antibody HELAB05 is replaced by: SIRPaD1 protein-linker-anti-HEL antibody light chain, preferably, amino acid sequence of replaced short chain is SEQ ID NO: 85.

[0092] III. Complete immune antibody

[0093] Connecting the variable region of an antibody of adaptive immunity or the variable region of a tumor-associated antigen antibody to the innate immune super connector (ISE) described herein can result in a complete immune antibody (CIA). As shown in Table 1, in some embodiments, the variable region of an antibody of adaptive immunity or the variable region of a tumor-associated antigen antibody is connected to ISE-A to result in a complete immune antibody ISE-A-I or ISE-A-II. For example, see Figure 2 CIA configuration of A1 / A2. In some embodiments, the variable region of an antibody of adaptive immunity or the variable region of a tumor-associated antigen antibody is connected to ISE-B to result in a complete immune antibody ISE-B-I or ISE-B-II. For example, see Figure 1CIA configuration of B1 / B2. In some embodiments, the post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-C to obtain a complete immune antibody ISE-C-I or ISE-C-II configuration. For example, see Figure 2 CIA configuration of C1 / C2. Similarly, the post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-D to obtain a complete immune antibody ISE-D-I or ISE-D-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-E to obtain a complete immune antibody ISE-E-I or ISE-E-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-F to obtain a complete immune antibody ISE-F-I or ISE-F-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-G to obtain a complete immune antibody ISE-G-I or ISE-G-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-H to obtain a complete immune antibody ISE-H-I or ISE-H-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-I to obtain a complete immune antibody ISE-I-I or ISE-I-II. The post-immune antibody variable region or the tumor-associated antigen antibody variable region is connected on the basis of ISE-J to obtain a complete immune antibody ISE-J-I or ISE-J-II. In the above embodiments, the post-immune antibody includes but is not limited to antibodies of PD-1, CTLA-4, B7-H4, B7-H3, 4-1BB, LAG-3, TIGIT, TIM-3, LILRB2, CD24, CD40 or CD40L, etc. In the above embodiments, the tumor-associated antigen antibody includes but is not limited to antibodies of PD-L1, Her2, VEGF, EGFR, CD38, 5T4, DLL3, CD30, Claudin18.2, Claudin6, CD19, MSLN, Trop-2, CD20, Nectin-4, BCMA, MUC17, MUC16 or c-Met, etc. The skilled person can select a specific post-immune antibody or tumor-associated antigen antibody as needed to prepare a complete immune antibody specific to a specific target, so as to achieve specific targeted killing of different tumors.

[0094] In another aspect, the present application takes PD-L1, PD-1, CD38, CD20, Claudin18.2, Trop-2, CTLA-4, BCMA, Nectin-4, EGFR, VEGF and Her2 as examples, respectively, to prepare complete immune antibodies that specifically bind to tumor-specific antigens, CD16a and CD47.

[0095] In one embodiment, there is provided a fully immune antibody TriAtezo01 with anti-PD-L1 antibody as backbone, which connects anti-CD16a single chain antibody through linker at N- or C-terminus of heavy chain of anti-PD-L1 antibody, and connects SIRPaD1 protein through linker at N- or C-terminus of light chain of anti-PD-L1 antibody. Preferably, the long chain structure of the fully immune antibody TriAtezo01 is: anti-PD-L1 antibody heavy chain-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-L1 antibody light chain-linker-SIRPaD1 protein. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo01 comprises or is SEQ ID NO: 68, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 69.

[0096] In one embodiment, there is provided a fully immune antibody TriAtezo02 with anti-PD-L1 antibody as backbone, which connects human IL-15 and anti-CD16a nanobody through linker at N- and / or C-terminus of heavy chain of anti-PD-L1 antibody, and connects SIRPaD1 protein through linker at N- or C-terminus of light chain of anti-PD-L1 antibody. Preferably, the long chain structure of the fully immune antibody is: anti-PD-L1 antibody heavy chain-linker-human IL-15-linker-anti-CD16a nanobody, and the short chain structure is: anti-PD-L1 antibody light chain-linker-SIRPaD1 protein. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo02 comprises or is SEQ ID NO: 70, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 69.

[0097] In one embodiment, there is provided a fully immune antibody TriAtezo03 with anti-PD-L1 antibody as backbone, which connects human IL-15Rα, human IL-15 and anti-CD16a nanobody through a linker at N-terminus and / or C-terminus of the heavy chain of the anti-PD-L1 antibody, and connects SIRPαD1 protein through a linker at N-terminus or C-terminus of the light chain of the anti-PD-L1 antibody. Preferably, the long chain structure of the fully immune antibody is: anti-PD-L1 antibody heavy chain-linker-human IL-15Rα-linker-human IL-15-linker-anti-CD16a nanobody, and the short chain structure is: anti-PD-L1 antibody light chain-linker-SIRPαD1 protein. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo03 comprises or is SEQ ID NO: 71, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 69.

[0098] In one embodiment, there is provided a fully immune antibody TriAtezo04 with anti-PD-L1 antibody as backbone, which connects anti-CD16a single chain antibody through a linker at N-terminus or C-terminus of the heavy chain of the anti-PD-L1 antibody, and connects SIRPαD1 protein through a linker at N-terminus or C-terminus of the light chain of the anti-PD-L1 antibody. Preferably, the long chain structure of the fully immune antibody is: anti-PD-L1 antibody heavy chain-linker-anti-CD16a single chain antibody, and the short chain structure is: SIRPαD1 protein-linker-anti-PD-L1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo04 comprises or is SEQ ID NO: 72, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 73.

[0099] In one embodiment, there is provided a fully immune antibody TriAtezo05 with anti-PD-L1 antibody as backbone, which connects anti-CD16a single chain antibody and SIRPαD1 protein through a linker at N-terminus and / or C-terminus of the heavy chain of the anti-PD-L1 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPαD1 protein-linker-anti-PD-L1 antibody heavy chain-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-L1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo05 comprises or is SEQ ID NO: 74, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 75.

[0100] In an embodiment, there is provided a fully immune antibody TriAtezo06 with anti-PD-L1 antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-PD-L1 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-PD-L1 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-L1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo06 comprises or is SEQ ID NO: 76, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 75.

[0101] In an embodiment, there is provided a fully immune antibody TriAtezo07 with anti-PD-L1 antibody as backbone, which connects SIRPaD1 protein, human IL-15Ra, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-PD-L1 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-PD-L1 antibody heavy chain-linker-human IL-15Ra-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-L1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo07 comprises or is SEQ ID NO: 77, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 75.

[0102] In an embodiment, there is provided a fully immune antibody TriAtezo12 with anti-PD-L1 antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-PD-L1 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-PD-L1 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-L1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAtezo12 comprises or is SEQ ID NO: 78, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 75.

[0103] In one embodiment, there is provided a fully immune antibody TriCetu03 with anti-EGFR antibody as backbone, which connects SIRPaD1 protein and anti-CD16a single chain antibody through a linker at N-terminus and / or C-terminus of anti-EGFR antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-EGFR antibody heavy chain-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-EGFR antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriCetu03 comprises or is SEQ ID NO: 79, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 80.

[0104] In one embodiment, there is provided a fully immune antibody TriCetu04 with anti-EGFR antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through a linker at N-terminus and / or C-terminus of anti-EGFR antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-EGFR antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-EGFR antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriCetu04 comprises or is SEQ ID NO: 81, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 80.

[0105] In one embodiment, there is provided a fully immune antibody TriCetu12 with anti-EGFR antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through a linker at N-terminus and / or C-terminus of anti-EGFR antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-EGFR antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-EGFR antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriCetu12 comprises or is SEQ ID NO: 120, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 80.

[0106] In one embodiment, there is provided a fully immune antibody TriAB04 with anti-Her2 antibody as backbone, which connects anti-CD16a single chain antibody through linker at N- or C-terminus of anti-Her2 antibody heavy chain, and connects SIRPaD1 protein through linker at N- or C-terminus of anti-Her2 antibody light chain. Preferably, the long chain structure of the fully immune antibody is: anti-Her2 antibody heavy chain-linker-anti-CD16a single chain antibody, and the short chain structure is: SIRPaD1 protein-linker-anti-Her2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriAB04 comprises or is SEQ ID NO: 82, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 83. In an alternative embodiment, the fully immune antibody TriAB04 replaces the short chain structure with: anti-Her2 antibody light chain-linker-SIRPaD1 protein, and preferably the amino acid sequence of the replaced short chain is SEQ ID NO: 84.

[0107] In one embodiment, there is provided a fully immune antibody TriTrastu06 with anti-Her2 antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N- and / or C-terminus of anti-Her2 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Her2 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Her2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriTrastu06 comprises or is SEQ ID NO: 110, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 111.

[0108] In one embodiment, there is provided a fully immune antibody TriTrastu12 with anti-Her2 antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N- and / or C-terminus of anti-Her2 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Her2 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Her2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriTrastu12 comprises or is SEQ ID NO: 112, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 111.

[0109] In one embodiment, there is provided a TriKey06 fully immune antibody with anti-PD-1 antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-PD-1 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-PD-1 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the TriKey06 fully immune antibody comprises or is SEQ ID NO: 86, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 87.

[0110] In one embodiment, there is provided a TriKey12 fully immune antibody with anti-PD-1 antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-PD-1 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-PD-1 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-PD-1 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the TriKey12 fully immune antibody comprises or is SEQ ID NO: 88, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 87.

[0111] In one embodiment, there is provided a TriDaratu06 fully immune antibody with anti-CD38 antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-CD38 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CD38 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CD38 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the TriDaratu06 fully immune antibody comprises or is SEQ ID NO: 89, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 90.

[0112] In one embodiment, there is provided a fully immune antibody TriDaratu12 with an anti-CD38 antibody as a backbone, which is connected with a linker to a SIRPaD1 protein, a human IL-2 and an anti-CD16a single chain antibody at the N-terminus and / or C-terminus of the heavy chain of the anti-CD38 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CD38 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CD38 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriDaratu12 comprises or is SEQ ID NO: 91, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 90.

[0113] In one embodiment, there is provided a fully immune antibody TriRituxi06 with an anti-CD20 antibody as a backbone, which is connected with a linker to a SIRPaD1 protein, a human IL-15 and an anti-CD16a single chain antibody at the N-terminus and / or C-terminus of the heavy chain of the anti-CD20 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CD20 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CD20 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriRituxi06 comprises or is SEQ ID NO: 92, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 93.

[0114] In one embodiment, there is provided a fully immune antibody TriRituxi12 with an anti-CD20 antibody as a backbone, which is connected with a linker to a SIRPaD1 protein, a human IL-2 and an anti-CD16a single chain antibody at the N-terminus and / or C-terminus of the heavy chain of the anti-CD20 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CD20 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CD20 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriRituxi12 comprises or is SEQ ID NO: 94, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 93.

[0115] In an embodiment, there is provided a fully immune antibody TriZolbetu06 with anti-Claudin 18.2 antibody as a backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-Claudin 18.2 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Claudin 18.2 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Claudin 18.2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriZolbetu06 comprises or is SEQ ID NO: 95, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 96.

[0116] In an embodiment, there is provided a fully immune antibody TriZolbetu12 with anti-Claudin 18.2 antibody as a backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-Claudin 18.2 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Claudin 18.2 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Claudin 18.2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriZolbetu12 comprises or is SEQ ID NO: 97, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 96.

[0117] In an embodiment, there is provided a fully immune antibody TriSacitu06 with anti-Trop-2 antibody as a backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-Trop-2 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Trop-2 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Trop-2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriSacitu06 comprises or is SEQ ID NO: 98, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 99.

[0118] In an embodiment, there is provided a fully immune antibody TriSacitu12 with an anti-Trop-2 antibody as a backbone, which is connected with a SIRPaD1 protein, a human IL-2 and an anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-Trop-2 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Trop-2 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Trop-2 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriSacitu12 comprises or is SEQ ID NO: 100, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 99.

[0119] In an embodiment, there is provided a fully immune antibody TriIpilimu06 with an anti-CTLA-4 antibody as a backbone, which is connected with a SIRPaD1 protein, a human IL-15 and an anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-CTLA-4 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CTLA-4 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CTLA-4 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriIpilimu06 comprises or is SEQ ID NO: 101, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 102.

[0120] In an embodiment, there is provided a fully immune antibody TriIpilimu12 with an anti-CTLA-4 antibody as a backbone, which is connected with a SIRPaD1 protein, a human IL-2 and an anti-CD16a single chain antibody through a linker at the N-terminus and / or C-terminus of the heavy chain of the anti-CTLA-4 antibody. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-CTLA-4 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-CTLA-4 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriIpilimu12 comprises or is SEQ ID NO: 103, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 102.

[0121] In one embodiment, there is provided a fully immune antibody TriBelanta06 with anti-BCMA antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-BCMA antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-BCMA antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-BCMA antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriBelanta06 comprises or is SEQ ID NO: 104, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 105.

[0122] In one embodiment, there is provided a fully immune antibody TriBelanta12 with anti-BCMA antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-BCMA antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-BCMA antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-BCMA antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriBelanta12 comprises or is SEQ ID NO: 106, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 105.

[0123] In one embodiment, there is provided a fully immune antibody TriEnfortu06 with anti-Nectin-4 antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N terminal and / or C terminal of anti-Nectin-4 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Nectin-4 antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Nectin-4 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriEnfortu06 comprises or is SEQ ID NO: 107, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 108.

[0124] In one embodiment, there is provided a fully immune antibody TriEnfortu 12 with anti-Nectin-4 antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N-terminus and / or C-terminus of anti-Nectin-4 antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-Nectin-4 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-Nectin-4 antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriEnfortu 12 comprises or is SEQ ID NO: 109, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 108.

[0125] In one embodiment, there is provided a fully immune antibody TriBeva 06 with anti-VEGF antibody as backbone, which connects SIRPaD1 protein, human IL-15 and anti-CD16a single chain antibody through linker at N-terminus and / or C-terminus of anti-VEGF antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-VEGF antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-VEGF antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriBeva 06 comprises or is SEQ ID NO: 113, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 114.

[0126] In one embodiment, there is provided a fully immune antibody TriBeva 12 with anti-VEGF antibody as backbone, which connects SIRPaD1 protein, human IL-2 and anti-CD16a single chain antibody through linker at N-terminus and / or C-terminus of anti-VEGF antibody heavy chain. Preferably, the long chain structure of the fully immune antibody is: SIRPaD1 protein-linker-anti-VEGF antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody, and the short chain structure is: anti-VEGF antibody light chain. In a particular embodiment, the amino acid sequence of the long chain of the fully immune antibody TriBeva 12 comprises or is SEQ ID NO: 115, and the amino acid sequence of the short chain comprises or is SEQ ID NO: 114.

[0127] In any of the embodiments described herein, the heavy chain variable region of the anti-CD16a single chain antibody comprises a CDR1 set forth in SEQ ID NO: 1, a CDR2 set forth in SEQ ID NO: 2, and a CDR3 set forth in SEQ ID NO: 3; and the light chain variable region comprises a CDR1 set forth in SEQ ID NO: 4, a CDR2 set forth in SEQ ID NO: 5, and a CDR3 set forth in SEQ ID NO: 6.

[0128] In any of the embodiments described herein, the heavy chain variable region of the anti-CD16a single chain antibody comprises a CDR1 set forth in SEQ ID NO: 1, a CDR2 set forth in SEQ ID NO: 2, and a CDR3 set forth in SEQ ID NO: 3; and the light chain variable region comprises a CDR1 set forth in SEQ ID NO: 4, a CDR2 set forth in SEQ ID NO: 5, and a CDR3 set forth in SEQ ID NO: 6.

[0129] In any of the embodiments described herein, the anti-CD16a single chain antibody comprises an amino acid sequence set forth in SEQ ID NO: 9.

[0130] In any of the embodiments described herein, the anti-CD16a nanobody comprises a CDR1 set forth in SEQ ID NO: 10, a CDR2 set forth in SEQ ID NO: 11, and a CDR3 set forth in SEQ ID NO: 12.

[0131] In any of the embodiments described herein, the anti-CD16a nanobody comprises an amino acid sequence set forth in SEQ ID NO: 13.

[0132] In any of the embodiments described herein, the SIRPaDl protein comprises an amino acid sequence set forth in SEQ ID NO: 14.

[0133] In any of the embodiments described herein, the human IL-15 protein fragment comprises an amino acid sequence set forth in SEQ ID NO: 15 or SEQ ID NO: 16.

[0134] In any of the embodiments described herein, the human IL-15Ra protein fragment comprises an amino acid sequence set forth in SEQ ID NO: 17.

[0135] In any of the embodiments described herein, the CpG ODN comprises a sequence set forth in SEQ ID NO: 117.

[0136] In any of the embodiments described herein, the IL-2 mutant comprises an amino acid sequence set forth in SEQ ID NO: 118.

[0137] In any of the embodiments described herein, the IFNa mutant comprises an amino acid sequence set forth in SEQ ID NO: 119.

[0138] In any of the embodiments described herein, the macrophage activating agent, the NK cell linking and activating agent, the PRR agonist, and the immune cytokine can be linked to the base antibody structure with a suitable linker, which can be a flexible linker or a flexible peptide linker, which is preferably a polypeptide of at least 5 amino acids in length. In some embodiments, the peptide linker is PSGQAGAAASESLFVSNHAY (SEQ ID NO: 19); in other embodiments, the peptide linker is (GxS)n or (GxS)nGm, where G = glycine, S = serine, and (x = 3, n = 3, 4, 5 or 6 and m = 0, 1, 2 or 3) or (x = 4 and n = 2, 3, 4 or 5 and m = 0, 1, 2 or 3), preferably x = 4 and n = 2 or 3, more preferably x = 4 and n = 3. In some preferred embodiments, the amino acid sequence of the peptide linker is (G4S)3 or (G4S)4, such as GGGGSGGGGSGGGGS (SEQ ID NO: 18).

[0139] Exemplary embodiments:

[0140] 1. An innate immune super connector (ISE) for recruiting or activating innate immune cells, comprising two or more of the following components: (a) an anti-CD16a antibody, (b) a SIRPaDl protein, (c) an immune cytokine, (d) a pattern recognition receptor agonist.

[0141] 2. A complete immune antibody (CIA) for recruiting or activating innate and adaptive immune cells to kill tumor cells, comprising (a) an ISE or fragment thereof according to embodiment 1, (b) an adaptive immune antibody or a tumor associated antigen antibody.

[0142] 3. The innate immune super connector according to embodiment 1, wherein a traditional antibody is used as a base structure, the variable region of the antibody is removed or there is no targeting antibody in the human body, an anti-CD16a antibody or a SIRPaDl protein is connected to the N-terminus or C-terminus of the heavy chain constant region of the antibody through a linker, and a SIRPaDl protein or an anti-CD16a antibody is connected to the N-terminus or C-terminus of the light chain constant region of the antibody through a linker.

[0143] 4. The innate immune super connector according to embodiment 1, wherein a traditional antibody is used as a base structure, the variable region of the antibody is removed or there is no targeting antibody in the human body, one or more of a human immune cytokine, a human immune cytokine receptor, a SIRPaDl protein and an anti-CD16a antibody are connected to the N-terminus and / or C-terminus of the heavy chain constant region and / or the light chain constant region of the antibody through a linker.

[0144] 5. The innate immunity super connector according to any one of the preceding embodiments, wherein a pattern recognition receptor agonist, including but not limited to a Toll-like receptor (TLRs) agonist, a NOD-like receptor (NLRs) agonist, a RIG-I-like receptor (RLRs) agonist, a C-type lectin receptor (CLRs) agonist, an absent in melanoma 2 (AIM2)-like receptor (ALRs) agonist, is connected at the N-terminus or C-terminus of the ISE via a linker.

[0145] 6. The innate immunity super connector according to any one of the preceding embodiments, wherein human IL-2 or IFNa or its mutants are connected at the N-terminus or C-terminus of the ISE via a linker.

[0146] 7. The fully immune antibody according to embodiment 2, wherein the ISE is used as the basic structure, and an antibody variable region of adaptive immunity or an antibody variable region of tumor associated antigen is added at the N-terminus of the ISE.

[0147] 8. The fully immune antibody according to embodiment 7, wherein an antibody of adaptive immunity or an antibody of tumor associated antigen is used as the basic skeleton, and one or more of human immune cytokine, human immune cytokine receptor, SIRPaD1 protein and anti-CD16a antibody are connected at the N-terminus and / or C-terminus of the antibody heavy chain and / or antibody light chain via a linker.

[0148] 9. The fully immune antibody according to any one of the preceding embodiments, wherein the tumor associated antigen is, for example but not limited to, PD-L1, Her2, VEGF, EGFR, CD38, Claudin 18.2, Claudin 6, CD19, MSLN, Trop-2, CD20, Nectin-4, c-Met, MUC16, MUC17, 5T4, BCMA, DLL3 or CD30.

[0149] 10. The fully immune antibody according to any one of the preceding embodiments, wherein the antibody of adaptive immunity is an antibody targeting, for example but not limited to, PD-1, CTLA-4, B7-H4, B7-H3, 4-1BB, LAG-3, TIGIT, TIM-3, LILRB2, CD24, CD40 or CD40L.

[0150] 11. The innate immunity super connector or the fully immune antibody according to any one of the preceding embodiments, wherein the human immune cytokine is, for example but not limited to, IL-15, IL-18, IL-2, IL-7, IL-10 or IL-12.

[0151] 12. The innate immunity super connector of embodiment 11, wherein the anti-CD16a antibody, human IL-15 or IL-15Ra, or SIRPaDl protein is connected to the N- or C-terminus of the heavy chain constant region of a traditional antibody, or a human antibody without targeting antibody in the human body, through a linker.

[0152] 13. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the anti-CD16a antibody is an anti-CD16a single chain antibody or an anti-CD16a nanobody.

[0153] 14. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the heavy chain variable region of the anti-CD16a single chain antibody comprises a CDR1 of SEQ ID NO: 1, a CDR2 of SEQ ID NO: 2, and a CDR3 of SEQ ID NO: 3; and the light chain variable region comprises a CDR1 of SEQ ID NO: 4, a CDR2 of SEQ ID NO: 5, and a CDR3 of SEQ ID NO: 6.

[0154] 15. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the heavy chain variable region of the anti-CD16a single chain antibody comprises an amino acid sequence of SEQ ID NO: 7, and the light chain variable region comprises an amino acid sequence of SEQ ID NO: 8.

[0155] 16. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the anti-CD16a single chain antibody comprises an amino acid sequence of SEQ ID NO: 9.

[0156] 17. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the anti-CD16a nanobody comprises a CDR1 of SEQ ID NO: 10, a CDR2 of SEQ ID NO: 11, and a CDR3 of SEQ ID NO: 12.

[0157] 18. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the anti-CD16a nanobody comprises an amino acid sequence of SEQ ID NO: 13.

[0158] 19. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the SIRPaDl protein comprises the amino acid sequence set forth in SEQ ID NO: 14.

[0159] 20. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the human IL-15 protein fragment comprises the amino acid sequence set forth in SEQ ID NO: 15 or SEQ ID NO: 16, and the human IL-15Ra protein fragment comprises the amino acid sequence set forth in SEQ ID NO: 17.

[0160] 21. The innate immunity super connector of embodiment 5, wherein the pattern recognition receptor agonist is a CpG ODN, preferably the CpG ODN comprises the sequence set forth in SEQ ID NO: 117.

[0161] 22. The innate immunity super connector of embodiment 6, wherein the IL-2 mutant comprises the amino acid sequence set forth in SEQ ID NO: 118, and / or the IFNa mutant comprises the amino acid sequence set forth in SEQ ID NO: 119.

[0162] 23. The innate immunity super connector of any of the preceding embodiments, wherein the basic framework is free of targeting antibodies in human body, such as anti-hen egg lysozyme (HEL) antibody, anti-pathogenic microorganism antibody, such as anti-virus, anti-bacteria antibody, and the like.

[0163] 24. The innate immunity super connector of embodiment 23, wherein the anti-HEL antibody is the basic framework, and one or more of the human immune cytokine, the human immune cytokine receptor, the SIRPaDl protein, and the anti-CD16a antibody are connected to the N-terminus or C-terminus of the heavy chain of the antibody and / or the N-terminus or C-terminus of the light chain of the antibody via a linker.

[0164] 25. The innate immunity superconnector of embodiment 24, wherein the amino acid sequence of the long chain comprises SEQ ID NO: 20, the amino acid sequence of the short chain comprises SEQ ID NO: 21, or the amino acid sequence of the long chain comprises SEQ ID NO: 22, the amino acid sequence of the short chain comprises SEQ ID NO: 23, or the amino acid sequence of the long chain comprises SEQ ID NO: 24, the amino acid sequence of the short chain comprises SEQ ID NO: 25, or the amino acid sequence of the long chain comprises SEQ ID NO: 26, the amino acid sequence of the short chain comprises SEQ ID NO: 27, or the amino acid sequence of the long chain comprises SEQ ID NO: 28, the amino acid sequence of the short chain comprises SEQ ID NO: 29.

[0165] 26. The innate immunity superconnector according to any one of the preceding embodiments, characterized in that it is in the configuration of ISE-A, ISE-B, ISE-C, ISE-D, ISE-E, ISE-F, ISE-G, ISE-H, ISE-I or ISE-J, preferably the amino acid sequence of the long chain of the innate immunity connector ISE-A comprises SEQ ID NO: 30 or SEQ ID NO: 32 and the amino acid sequence of the short chain comprises SEQ ID NO: 31 or SEQ ID NO: 33, the amino acid sequence of the long chain of the innate immunity connector ISE-B comprises SEQ ID NO: 34 or SEQ ID NO: 36 and the amino acid sequence of the short chain comprises SEQ ID NO: 35 or SEQ ID NO: 37, the amino acid sequence of the long chain of the innate immunity connector ISE-C comprises SEQ ID NO: 38 or SEQ ID NO: 40 and the amino acid sequence of the short chain comprises SEQ ID NO: 39 or SEQ ID NO: 41, the amino acid sequence of the long chain of the innate immunity connector ISE-D comprises SEQ ID NO: 42 or SEQ ID NO: 44 and the amino acid sequence of the short chain comprises SEQ ID NO: 43 or SEQ ID NO: 45, the amino acid sequence of the long chain of the innate immunity connector ISE-E comprises SEQ ID NO: 46 or SEQ ID NO: 48 and the amino acid sequence of the short chain comprises SEQ ID NO: 47 or SEQ ID NO: 49, the amino acid sequence of the long chain of the innate immunity connector ISE-F comprises SEQ ID NO: 50 or SEQ ID NO: 52 and the amino acid sequence of the short chain comprises SEQ ID NO: 51 or SEQ ID NO: 53, the amino acid sequence of the long chain of the innate immunity connector ISE-G comprises SEQ ID NO: 54 or SEQ ID NO: 56 and the amino acid sequence of the short chain comprises SEQ ID NO: 55 or SEQ ID NO: 57, the amino acid sequence of the long chain of the innate immunity connector ISE-H comprises SEQ ID NO: 58 and the amino acid sequence of the short chain comprises SEQ ID NO: 59 or SEQ ID NO: 60, the amino acid sequence of the long chain of the innate immunity connector ISE-I comprises SEQ ID NO: 61 or SEQ ID NO: 63 and the amino acid sequence of the short chain comprises SEQ ID NO: 62 or SEQ ID NO: 64, the amino acid sequence of the long chain of the innate immunity connector ISE-J comprises SEQ ID NO: 65 and the amino acid sequence of the short chain comprises SEQ ID NO: 66 or SEQ ID NO: 67.

[0166] 27. The fully immune antibody according to Embodiment 7, characterized in that the amino acid sequence of the long chain is a heavy chain variable region of a tumor-associated antigen antibody or an antibody against an antigen acquired after birth introduced on the basis of the amino acid sequence of the ISE, and the amino acid sequence of the short chain is a light chain variable region of a tumor-associated antigen antibody or an antibody against an antigen acquired after birth introduced on the basis of the amino acid sequence of the ISE.

[0167] 28. The fully immunized antibody according to embodiment 27, characterized in that the fully immunized antibody is the fully immunized antibody TriAtezo01, TriAtezo02, TriAtezo03, TriAtezo04, TriAtezo05, TriAtezo06, TriAtezo07, TriAtezo12, TriCetu03, TriCetu04 and TriCetu12, the amino acid sequence of the long chain of which comprises SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 81 and SEQ ID NO: 120, respectively.The short chain amino acid sequences comprise SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 80, SEQ ID NO: 80, and SEQ ID NO: 80, respectively, and / or the fully immune antibodies are fully immune antibodies TriKey06, TriKeyl 2, TriDaratu06, TriDaratu 12, TriRituxi06, TriRituxi 12, TriZolbetu06, TriZolbetu 12, TriSacitu06, TriSacitu 12, TriTrastu06, TriTrastu 12, TriBeva06, TriBeva 12, TriEnfortu06, TriEnfortu 12, TriBelanta06, TriBelanta 12, TriIpilimu06, and TriIpilimu 12, whose long chain amino acid sequences comprise SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 101, and SEQ ID NO: 103, respectively; the short chain amino acid sequences comprise SEQ ID NO: 87, SEQ ID NO: 87, SEQ ID NO: 90, SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 93, SEQ ID NO: 96, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 99, SEQ ID NO: 111, SEQ ID NO: 111, SEQ ID NO: 114, SEQ ID NO: 114, SEQ ID NO: 108, SEQ ID NO: 108, SEQ ID NO: 105, SEQ ID NO: 105, SEQ ID NO: 102, and SEQ ID NO: 102, respectively.

[0168] 29. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the Fc fragment of the antibody incorporates L234A and L235A mutations, or further incorporates M252Y, S254T, and T256E mutations in the Fc fragment.

[0169] 30. The innate immunity super connector or fully immune antibody of any of the preceding embodiments, wherein the linker comprises, but is not limited to, PSGQAGAAASESLFVSNHAY or (G4S) n wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

[0170] 31. A nucleic acid molecule encoding the innate immunity super connector or fully immune antibody of any of the preceding embodiments.

[0171] 32. A vector comprising the nucleic acid molecule of embodiment 31.

[0172] 33. A cell comprising the nucleic acid molecule of embodiment 31 or the vector of embodiment 32.

[0173] 34. A method of making the innate immunity super connector or fully immune antibody of any of the preceding embodiments, the method comprising the steps of:

[0174] (1) amplifying the two chains of ISE or CIA, respectively, using PCR technology; introducing mutations L234A, L235A, M252Y, S254T, and T256E in the Fc fragment;

[0175] (2) cloning the DNA fragments obtained in step (1) into pcDNA series vectors or other vectors for mammalian cell expression systems to obtain recombinant vectors;

[0176] (3) transfecting the recombinant vector plasmids obtained in step (2) into mammalian cells for expression of the recombinant proteins, and obtaining the purified target proteins after affinity chromatography.

[0177] 35. Use of the innate immunity super connector or fully immune antibody of any of the preceding embodiments in the manufacture of a medicament for treating solid tumors and hematological tumors in a subject.

[0178] 36. The use according to embodiment 35, wherein the solid and hematological tumors include, but are not limited to, respiratory tract tumors, digestive tract tumors, reproductive system tumors, nervous system tumors, urinary system tumors, skin tumors, head and neck tumors, hematological tumors, preferably, the solid and hematological tumors include, but are not limited to, nasopharyngeal carcinoma, laryngeal carcinoma, lung cancer, esophageal cancer, gastric cancer, colorectal cancer, prostate cancer, ovarian cancer, testicular cancer, brain tumor, glioma, renal cancer, bladder cancer, ureter cancer, epithelial tumor, melanoma, ocular tumor, nasal tumor, ear tumor, oral tumor, leukemia, multiple myeloma, lymphoma, breast cancer, epidermal cancer.

[0179] The sequences involved in the present application are summarized as follows:

[0180] SEQ ID NO: 1 (anti-CD16a single-chain antibody heavy chain CDR1 amino acid sequence)

[0181] GYTFTSYY

[0182] SEQ ID NO: 2 (anti-CD16a single-chain antibody heavy chain CDR2 amino acid sequence)

[0183] INPSGGST

[0184] SEQ ID NO: 3 (anti-CD16a single-chain antibody heavy chain CDR3 amino acid sequence)

[0185] ARGSAYYYDFADY

[0186] SEQ ID NO: 4 (anti-CD16a single-chain antibody light chain CDR1 amino acid sequence)

[0187] NIGSKN

[0188] SEQ ID NO: 5 (anti-CD16a single-chain antibody light chain CDR2 amino acid sequence)

[0189] QDN

[0190] SEQ ID NO: 6 (anti-CD16a single-chain antibody light chain CDR3 amino acid sequence)

[0191] QVWDNYSVL

[0192] SEQ ID NO: 7 (anti-CD16a single-chain antibody heavy chain variable region amino acid sequence)

[0193] EVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSS

[0194] SEQ ID NO: 8 (anti-CD16a single-chain antibody light chain variable region amino acid sequence)

[0195] QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0196] SEQ ID NO: 9 (anti-CD16a single-chain antibody amino acid sequence)

[0197] EVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0198] SEQ ID NO: 10 (anti-CD16a nanobody CDR1 amino acid sequence)

[0199] GLTFSSYNMG

[0200] SEQ ID NO: 11 (anti-CD16a nanobody CDR2 amino acid sequence)

[0201] SITWSGRDTFYADSVKG

[0202] SEQ ID NO: 12 (anti-CD16a nanobody CDR3 amino acid sequence)

[0203] NPWPVAAPRSGTY

[0204] SEQ ID NO: 13 (anti-CD16a Nanobody amino acid sequence)

[0205] QVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0206] SEQ ID NO: 14 (SIRPaDl protein amino acid sequence)

[0207] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0208] SEQ ID NO: 15 (human IL-15 protein fragment amino acid sequence)

[0209] NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS

[0210] SEQ ID NO: 16 (human IL-15 protein fragment amino acid sequence)

[0211] GIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS

[0212] SEQ ID NO: 17 (human IL-15Ra protein fragment amino acid sequence)

[0213] MSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTA

[0214] SEQ ID NO: 18 (flexible linker amino acid sequence)

[0215] GGGGSGGGGSGGGGS

[0216] SEQ ID NO: 19 (flexible linker amino acid sequence)

[0217] PSGQAGAAASESLFVSNHAY

[0218] SEQ ID NO: 20 (HELAB01 long chain amino acid sequence, anti-HEL antibody heavy chain)

[0219] EVQLQQSGAELMKPGASVKISCKASGYTFSDYWIEWVKQRPGHGLEWIGEILPGSGSTNYHERFKGKATFTADTSSSTAYMQLNSLTSEDSGVYYCLHGNYDFDGWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0220] SEQ ID NO: 21 (HELAB01 short chain amino acid sequence, anti-HEL antibody light chain)

[0221] DIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0222] SEQ ID NO:22 (HELAB02 long-chain amino acid sequence, anti-HEL antibody heavy chain)

[0223] EVQLQQSGAELMKPGASVKISCKASGYTFSDYWIEWVKQRPGHGLEWIGEILPGSGSTNYHERFKGKATFTADTSSSTAYMQLNSLTSEDSGVYYCLHGNYDFDGWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0224] SEQ ID NO:23 (HELAB02 short-chain amino acid sequence, anti-HEL antibody light chain - linker - SIRPαD1 protein)

[0225] DIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0226] SEQ ID NO:24 (HELAB03 long-chain amino acid sequence, anti-HEL antibody heavy chain-linker-anti-CD16a single-chain antibody)

[0227] EVQLQQSGAELMKPGASVKISCKASGYTFSDYWIEWVKQRPGHGLEWIGEILPGSGSTNYHERFKGKATFTADTSSSTAYMQLNSLTSEDSGVYYCLHGNYDFDGWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0228] SEQ ID NO:25 (HELAB03 short-chain amino acid sequence, anti-HEL antibody light chain)

[0229] DIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0230] SEQ ID NO: 26 (HELAB04 long chain amino acid sequence, anti-HEL antibody heavy chain - linker - anti-CD 16a single chain antibody)

[0231] EVQLQQSGAELMKPGASVKISCKASGYTFSDYWIEWVKQRPGHGLEWIGEILPGSGSTNYHERFKGKATFTADTSSSTAYMQLNSLTSEDSGVYYCLHGNYDFDGWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0232] SEQ ID NO:27 (Short chain amino acid sequence of HELAB04, light chain of anti-HEL antibody-linker-SIRPαD1 protein)

[0233] DIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0234] SEQ ID NO: 28 (HELAB05 long chain amino acid sequence, anti-HEL antibody heavy chain - linker - human IL-15 - anti-CD 16a nanobody)

[0235] EVQLQQSGAELMKPGASVKISCKASGYTFSDYWIEWVKQRPGHGLEWIGEILPGSGSTNYHERFKGKATFTADTSSSTAYMQLNSLTSEDSGVYYCLHGNYDFDGWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0236] SEQ ID NO:29 (Short chain amino acid sequence of HELAB05, light chain of anti-HEL antibody-linker-SIRPαD1 protein)

[0237] DIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0238] SEQ ID NO: 30 (ISE-A1 long chain amino acid sequence, SIRPaD1 protein - linker - antibody heavy chain constant region - linker - anti-CD16a single chain antibody)

[0239] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0240] SEQ ID NO:31 (ISE-A1 short chain amino acid sequence, antibody kappa chain constant region)

[0241] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0242] SEQ ID NO:32 (ISE-A2 long chain amino acid sequence, antibody heavy chain constant region - linker - anti-CD16a single chain antibody)

[0243] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0244] SEQ ID NO:33 (ISE-A2 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0245] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0246] SEQ ID NO: 34 (ISE-B1 long chain amino acid sequence, SIRPαD1 protein-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a nanobody)

[0247] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGV HTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYT LPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLII LANNSLSSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0248] SEQ ID NO: 35 (ISE-B1 short chain amino acid sequence, antibody kappa chain constant region)

[0249] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0250] SEQ ID NO: 36 (ISE-B2 long chain amino acid sequence, antibody heavy chain constant region - linker - human IL-15 - linker - anti-CD16a nanobody)

[0251] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0252] SEQ ID NO: 37 (ISE-B2 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0253] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0254] SEQ ID NO: 38 (ISE-C1 long chain amino acid sequence, SIRPaD1 protein - linker - antibody heavy chain constant region - linker - human IL-15Ra - linker - human IL-15 - linker - anti-CD16a nanobody)

[0255] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTASGGSGGGGSGGGSGGGGSLQGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGESLTLSCVVAGSIFSFAMSWYRQAPGKERELVARIGSDDRVTYADSVKGRFTISRDNIKRTAGLQMNSLKPEDTAVYYCNAQTDLRDWTVREYWGQGTQVTVSS

[0256] SEQ ID NO: 39 (ISE-C1 short chain amino acid sequence, antibody kappa chain constant region)

[0257] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0258] SEQ ID NO: 40 (ISE-C2 long chain amino acid sequence, antibody heavy chain constant region - linker - human IL-15Ra - linker - human IL-15 - linker - anti-CD 16a nanobody)

[0259] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTASGGSGGGGSGGGSGGGGSLQGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGESLTLSCVVAGSIFSFAMSWYRQAPGKERELVARIGSDDRVTYADSVKGRFTISRDNIKRTAGLQMNSLKPEDTAVYYCNAQTDLRDWTVREYWGQGTQVTVSS

[0260] SEQ ID NO:41 (ISE-C2 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0261] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0262] SEQ ID NO:42 (ISE-D1 long chain amino acid sequence, SIRPaD1 protein - linker - antibody heavy chain constant region - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0263] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0264] SEQ ID NO: 43 (ISE-D1 short chain amino acid sequence, antibody kappa chain constant region)

[0265] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0266] SEQ ID NO:44 (ISE-D2 long chain amino acid sequence, antibody heavy chain constant region - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0267] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0268] SEQ ID NO:45 (ISE-D2 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0269] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0270] SEQ ID NO: 46 (ISE-E1 long chain amino acid sequence, CpG ODN-linker- antibody heavy chain constant region-linker-anti-CD16a single chain antibody)

[0271] gggggACGATCGTCggggggGGGGGTEPLASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0272] SEQ ID NO:47 (ISE-E1 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0273] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0274] SEQ ID NO:48 (ISE-E2 long chain amino acid sequence, CpG ODN - linker - antibody heavy chain constant region - linker - anti-CD16a single chain antibody)

[0275] gggggACGATCGTCggggggGGGGGTEPLASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0276] SEQ ID NO:49 (ISE-E2 short-chain amino acid sequence, SIRPaD1 protein-linker-antibody kappa chain constant region)

[0277] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0278] SEQ ID NO: 50 (ISE-F1 long chain amino acid sequence, antibody heavy chain constant region - linker - anti-CD16a single chain antibody)

[0279] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0280] SEQ ID NO: 51 (ISE-F1 short chain amino acid sequence, CpG ODN - linker - antibody kappa chain constant region - linker - SIRPaD1 protein)

[0281] gggggACGATCGTCggggggGGGGGTEPLRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0282] SEQ ID NO: 52 (ISE-F2 long chain amino acid sequence, antibody heavy chain constant region - linker - anti-CD16a single chain antibody)

[0283] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0284] SEQ ID NO: 53 (ISE-F2 short chain amino acid sequence, SIRPaD1 protein - linker - antibody kappa chain constant region - linker - CpG ODN)

[0285] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECPETGGGGGggggggACGATCGTCggggg

[0286] SEQ ID NO: 54 (ISE-G1 long chain amino acid sequence, antibody heavy chain constant region - linker - anti-CD16a single chain antibody - linker - CpG ODN)

[0287] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVLPETGGGGGggggggACGATCGTCggggg

[0288] SEQ ID NO: 55 (ISE-G1 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0289] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0290] SEQ ID NO: 56 (ISE-G2 long chain amino acid sequence, antibody heavy chain constant region - linker - anti-CD16a single chain antibody - linker - CpG ODN)

[0291] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVLPETGGGGGggggggACGATCGTCggggg

[0292] SEQ ID NO: 57 (ISE-G2 short chain amino acid sequence, SIRPaD1 protein - linker - antibody kappa chain constant region)

[0293] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0294] SEQ ID NO: 58 (ISE-H1 long chain amino acid sequence, human IL-2 mutant-linker- antibody heavy chain constant region-linker-anti-CD16a single chain antibody)

[0295] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0296] SEQ ID NO: 59 (ISE-H1 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0297] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGG GGEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0298] SEQ ID NO: 60 (ISE-H2 short chain amino acid sequence, SIRPαD1 protein-linker-antibody kappa chain constant region)

[0299] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGG GGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0300] SEQ ID NO: 61 (ISE-I1 long chain amino acid sequence, human IL-2 mutant-linker-antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a nanobody)

[0301] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0302] SEQ ID NO: 62 (ISE-I1 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0303] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSGGGGEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0304] SEQ ID NO: 63 (ISE-I2 long chain amino acid sequence, human IL-2 mutant-linker- antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a nanobody)

[0305] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0306] SEQ ID NO: 64 (ISE-I2 short chain amino acid sequence, SIRPaD1 protein - linker - antibody kappa chain constant region)

[0307] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0308] SEQ ID NO: 65 (ISE-J1 long chain amino acid sequence, human IL-2 mutant-linker- antibody heavy chain constant region-linker-human IL-15-linker-anti-CD16a single chain antibody)

[0309] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVESGGGLVQPGESLTLSCVVAGSIFSFAMSWYRQAPGKERELVARIGSDDRVTYADSVKGRFTISRDNIKRTAGLQMNSLKPEDTAVYYCNAQTDLRDWTVREYWGQGTQVTVSSGGGGSGGGGSGGGGSMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTASGGSGGGGSGGGSGGGGSLQGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTS

[0310] SEQ ID NO: 66 (ISE-J1 short chain amino acid sequence, antibody kappa chain constant region - linker - SIRPaD1 protein)

[0311] RTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSGGGGEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK SEQ ID NO: 67 (ISE-J2 short chain amino acid sequence, SIRPaD1 protein-linker- antibody kappa chain constant region) EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK GGGGSGGGGSGGGGSRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0312] SEQ ID NO: 68 (TriAtezo01 long chain amino acid sequence, anti-PD-L1 antibody heavy chain-linker-anti-CD16a single chain antibody)EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSN KALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSG GSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0313] SEQ ID NO: 69 (TriAtezo01 short chain amino acid sequence, anti-PD-L1 antibody light chain-linker-SIRPαD1 protein)

[0314] DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQ PDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSD LTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0315] SEQ ID NO: 70 (TriAtezo02 long chain amino acid sequence, anti-PD-Ll antibody heavy chain - linker - human IL-15 - linker - anti-CD 16a nanobody)

[0316] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSQVQLVESGGGLVQPGGSLRLSCAASGLTFSSYNMGWFRQAPGQGLEAVASITWSGRDTFYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYYCAANPWPVAAPRSGTYWGQGTLVTVSS

[0317] SEQ ID NO:71 (TriAtezo03 long chain amino acid sequence, anti-PD-L1 antibody heavy chain - linker - human IL-15Ra - linker - human IL-15 - linker - anti-CD16a nanobody)

[0318] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSMSVEHADIWVKSYSLYSRERYICNSGFKRKAGTSSLTECVLNKATNVAHWTTPSLKCIRDPALVHQRPAPPSTVTTASGGSGGGGSGGGSGGGGSLQGIHVFILGCFSAGLPKTEANWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGESLTLSCVVAGSIFSFAMSWYRQAPGKERELVARIGSDDRVTYADSVKGRFTISRDNIKRTAGLQMNSLKPEDTAVYYCNAQTDLRDWTVREYWGQGTQVTVSS

[0319] SEQ ID NO:72 (TriAtezo04 long chain amino acid sequence, anti-PD-L1 antibody heavy chain - linker - anti-CD16a single chain antibody)

[0320] EVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0321] SEQ ID NO:73 (TriAtezo04 short chain amino acid sequence, SIRPaDl protein - linker - anti-PD-Ll antibody light chain)

[0322] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0323] SEQ ID NO: 74 (TriAtezo05 long chain amino acid sequence, SIRPaD1 protein - linker - anti-PD-Ll antibody heavy chain - linker - anti-CD16a single chain antibody)

[0324] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0325] SEQ ID NO:75 (TriAtezo05 short chain amino acid sequence, anti-PD-L1 antibody light chain)

[0326] DIQMTQSPSSLSASVGDRVTITCRASQDVSTAVAWYQQKPGKAPKLLIYSASFLYSGVPS RFSGSGSGTDFTLTISSLQPEDFATYYCQQYLYHPATFGQGTKVEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0327] SEQ ID NO: 76 (TriAtezo06 long chain amino acid sequence, SIRPaDl protein - linker - anti-PD-Ll antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0328] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0329] SEQ ID NO: 77 (TriAtezo07 long chain amino acid sequence, SIRPaD1 protein - linker - anti-PD-Ll antibody heavy chain - linker - human IL-15Ra - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0330]

[0331] SEQ ID NO: 78 (TriAtezo12 long chain amino acid sequence, SIRPaD1 protein - linker - anti-PD-Ll antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0332] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSDSWIHWVRQAPGKGLEWVAWISPYGGSTYYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCARRHWPGGFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0333] SEQ ID NO: 79 (TriCetu03 long chain amino acid sequence, SIRPaD1 protein - linker - anti-EGFR antibody heavy chain - linker - anti-CD16a single chain antibody)

[0334] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSD LT KRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGG GS QVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPF TSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSV FPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVP S S SLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYIT REPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNG KEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVE WESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSL SPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQG LEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFA DYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSK NVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDN YS VLFGGGTKLTVL

[0335] SEQ ID NO: 80 (TriCetu03 short chain amino acid sequence, anti-EGFR antibody light chain)

[0336] DILLTQSPVILSVSPGERVSFSCRASQSIGTNIHWYQQRTNGSPRLLIKYASESISGIPSRFSGSGSGTDFTLSINSVESEDIADYYCQQNNNWPTTFGAGTKLELKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0337] SEQ ID NO: 81 (TriCetu04 long chain amino acid sequence, SIRPaDl protein - linker - anti-EGFR antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0338] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0339] SEQ ID NO: 82 (TriAB04 long chain amino acid sequence, anti-Her2 antibody heavy chain - linker - CD16a single chain antibody)

[0340] EVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0341] SEQ ID NO: 83 (TriAB04 short chain amino acid sequence, SIRPaD1 protein - linker - anti-Her2 antibody light chain)

[0342] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSDIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0343] SEQ ID NO: 84 (TriAB04 replacement short chain amino acid sequence, anti-Her2 antibody light chain - linker - SIRPaD1 protein)

[0344] DIQMTQSPSSLSASVGDRVTITCRASQDVNTAVAWYQQKPGKAPKLLIYSASFLYSGVPSRFSGSRSGTDFTLTISSLQPEDFATYYCQQHYTTPPTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGECGGGGSGGGGSGGGGSEEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAK

[0345] SEQ ID NO: 85 (HELAB02, HELAB04, and HELAB05 replacement short chain amino acid sequence, SIRPaD1 protein - linker - anti-HEL antibody light chain)

[0346] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSDIVLTQSPAIMSASPGEKVTMTCSASSSVNYMYWYQQKSGTSPKRWIYDTSKLASGVPVRFSGSGSGTSYSLTISSMETEDAATYYCQQWGRNPTFGGGTKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0347] SEQ ID NO: 86 (TriKey06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-PD-1 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0348] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGS NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0349] SEQ ID NO: 87 (TriKey06 short chain amino acid sequence, anti-PD-1 antibody light chain)

[0350] EIVLTQSPAT LSLSPGERAT LSCRASKGVS TSGYSYLHWY QQKPGQAPRL LIYLASYLESGVPARFSGSGSGTDFTLTISSLEPEDFAVYYCQHSRDLPLTFGGGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0351] SEQ ID NO: 88 (TriKey12 long chain amino acid sequence, SIRPaD1 protein-linker-anti-PD-1 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0352] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLVQSGVEVKKPGASVKVSCKASGYTFTNYYMYWVRQAPGQGLEWMGGINPSNGGTNFNEKFKNRVTLTTDSSTTTAYMELKSLQFDDTAVYYCARRDYRFDMGFDYWGQGTTVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0353] SEQ ID NO: 89 (TriDaratu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-CD38 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0354] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAVSGFTFNSFM SWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0355] SEQ ID NO: 90 (TriDaratu06 short chain amino acid sequence, anti-CD38 antibody light chain)

[0356] EIVLTQSPAT LSLSPGERAT LSCRASQSVS SYLAWYQQKP GQAPRLLIYD ASNRATGIPAR FSGSGSGTDFTLTISSLEPEDFAVYYCQQRSNWPPTFGQGTKVEIKRTVAAPSVFIFPPS DEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTL TLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0357] SEQ ID NO: 91 (TriDaratu12 long chain amino acid sequence, SIRPaDl protein - linker - anti-CD38 antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0358] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLLESGGGLVQPGGSLRLSCAVSGFTFN SFAMSWVRQAPGKGLEWVSAISGSGGGTYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAVYFCAKDKILWFGEPVFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0359] SEQ ID NO: 92 (TriRituxi06 long chain amino acid sequence, SIRPaDl protein - linker - anti-CD20 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0360] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQT PGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARST YYGGDWYFNW GAGTTVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0361] SEQ ID NO: 93 (TriRituxi06 short chain amino acid sequence, anti-CD20 antibody light chain)

[0362] QIVLSQSPAILSA SPGEKVTMTCRASSSVSYIHWFQQKPG SSPKPWIYATS N LASGVPVRFSGSGSGTSY SLTISRVEAE DAATYYCQQW TSNPPTFGGG TKLEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0363] SEQ ID NO: 94 (TriRituxi12 long chain amino acid sequence, SIRPaD1 protein-linker-anti-CD20 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0364] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSQVQLQQPGAELVKPGASVKMSCKASGYTFTSYNMHWVKQT PGRGLEWIGAIYPGNGDTSYNQKFKGKATLTADKSSSTAYMQLSSLTSEDSAVYYCARST YYGGDWYFNW GAGTTVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0365] SEQ ID NO: 95 (TriZolbetu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-Claudin 18.2 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0366] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGS NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0367] SEQ ID NO: 96 (TriZolbetu06 short chain amino acid sequence, anti-Claudin 18.2 antibody light chain)

[0368] DIVMTQSPSSLTVTAGEKVTMSCKSSQSLLNSGNQKNYLTWYQQKPGQPPKLLIYWAS TRESGVPDRFTGSGSGTDFTLTISSVQAEDLAVYYCQNDYSYPFTFGSGTKLEIKRTVA APSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDS KDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0369] SEQ ID NO: 97 (TriZolbetu12 long chain amino acid sequence, SIRPαD1 protein - linker - anti-Claudin 18.2 antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0370] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLQQPGAELVRPGASVKLSCKASGYTFTSYWINWVKQRPGQGLEWIGNIYPSDSYTNYNQKFKDKATLTVDKSSSTAYMQLSSPTSEDSAVYYCTRSWRGNSFDYWGQGTTLTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0371] SEQ ID NO: 98 (TriSacitu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-Trop-2 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0372] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSQVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0373] SEQ ID NO: 99 (TriSacitu06 short chain amino acid sequence, anti-Trop-2 antibody light chain)

[0374] DIQLTQSPSS LSASVGDRVS ITCKASQDVS IAVAWYQQKP GKAPKLLIYS ASYRYTGVPDRFSGSGSGTD FTLTISSLQP EDFAVYYCQQ HYITPLTFGA GTKVEIKRTV AAPSVFIFPP SDEQLKSGTASVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLTLSKADYEKHK VYACEVTHQGLSSPVTKSFN RGEC

[0375] SEQ ID NO: 100 (TriSacitu12 long chain amino acid sequence, SIRPaDl protein - linker - anti-Trop-2 antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0376] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLQQSGSELKKPGASVKVSCKASGYTFTNYGMNWVKQAPGQGLKWMGWINTYTGEPTYTDDFKGRFAFSLDTSVSTAYLQISSLKADDTAVYFCARGGFGSSYWYFDVWGQGSLVTVSSASTK GPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0377] SEQ ID NO: 101 (TriIpilimu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-CTLA-4 antibody heavy chain - linker - human IL-15 - linker - anti-CD 16a single chain antibody)

[0378] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTV SDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGG GGSGGGGS QVQLVESGGGVVQPGRSLRL SCAASGFTFS SYTMHWVRQA PGKGLEWVTF ISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGS NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0379] SEQ ID NO: 102 (TriIpilimu06 short chain amino acid sequence, anti-CTLA-4 antibody light chain)

[0380] EIVLTQSPGT LSLSPGERAT LSCRASQSVG SSYLAWYQQK PGQAPRLLIY GAFSRATGIP DRFSGSGSGT DFTLTISRLE PEDFAVYYCQ QYGSSPWTFG QGTKVEIKRT VAAPSVFIFP PSDEQLKSGTASVVCLLNNF YPREAKVQWK VDNALQSGNS QESVTEQDSK DSTYSLSSTLTLSKADYEKH KVYACEVTHQGLSSPVTKSF NRGEC

[0381] SEQ ID NO: 103 (TriIpilimu12 long chain amino acid sequence, SIRPaD1 protein-linker-anti-CTLA-4 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0382] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTV SDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGG GGSGGGGS QVQLVESGGGVVQPGRSLRL SCAASGFTFS SYTMHWVRQA PGKGLEWVTF ISYDGNNKYYADSVKGRFTISRDNSKNTLYLQMNSLRAEDTAIYYCARTGWLGPFDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0383] SEQ ID NO: 104 (TriBelanta06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-BCMA antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0384] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGS NWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0385] SEQ ID NO: 105 (TriBelanta06 short chain amino acid sequence, anti-BCMA antibody light chain)

[0386] DIQMTQSPSS LSASVGDRVT ITCSASQDIS NYLNWYQQKP GKAPKLLIYY TSNLHSGVPSRFSGSGSGTD FTLTISSLQP EDFATYYCQQ YRKLPWTFGQ GTKLEIKRTV AAPSVFIFPP SDEQLKSGTASVVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLLSSTLTLSKADYEKHK VYACEVTHQGLSSPVTKSFN RGEC

[0387] SEQ ID NO: 106 (TriBelanta12 long chain amino acid sequence, SIRPαD1 protein-linker-anti-BCMA antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0388] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGS QVQLVQSGAEVKKPGSSVKVSCKASGGTFSNYWMHWVRQAPGQGLEWMGATYRGHSDTYYNQKFKGRVTITADKSTSTAYMELSSLRSEDTAVYYCARGAIYDGYDVLDNWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGS QPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0389] SEQ ID NO: 107 (TriEnfortu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-Nectin-4 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0390] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFS SYNMNWVRQAPGKGLEWVSYISSSSSTIYYADSVKGRFTISRDNAKNSLSLQMNSLRDEDTAVYYCARAYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0391] SEQ ID NO: 108 (TriEnfortu06 short chain amino acid sequence, anti-Nectin-4 antibody light chain)

[0392] DIQMTQSPSS VSASVGDRVT ITCRASQGIS GWLAWYQQKP GKAPKFLIYA ASTLQSGVPS RFSGSGSGTD FTLTISSLQP EDFATYYCQQ ANSFPPTFGG GTKVEIKRTV AAPSVFIFPP SDEQLKSGTAS VVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT LS KADYEKHK VYACEVTHQG LSSPVTKSFN RGEC

[0393] SEQ ID NO: 109 (TriEnfortu12 long chain amino acid sequence, SIRPαD1 protein-linker-anti-Nectin-4 antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0394] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFTFSNYNMNWVRQAPGKGLEWVSYISSSSSTIYYADSVKGRFTISRDNAKNSLSLQMNSLRDEDTAVYYCARAYYYGMDVWGQGTTVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLMVTVREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRWSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTV

[0395] SEQ ID NO: 110 (TriTrastu06 long chain amino acid sequence, SIRPaD1 protein - linker - anti-Her2 antibody heavy chain - linker - human IL-15 - linker - anti-CD16a single chain antibody)

[0396] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0397] SEQ ID NO: 111 (TriTrastu06 short chain amino acid sequence, anti-Her2 antibody light chain)

[0398] DIQMTQSPSS LSASVGDRVT ITCRASQDVN TAVAWYQQKP GKAPKLLIYS ASFLYSGVPS RFSGSRSGTD FTLTISSLQP EDFATYYCQQ HYTTPPTFGQ GTKVEIKRTV AAPSVFIFPP SDEQLKSGTAS VVCLLNNFY PREAKVQWKV DNALQSGNSQ ESVTEQDSKD STYSLSSTLT LS KADYEKHK VYACEVTHQG LSSPVTKSFN RGEC

[0399] SEQ ID NO: 112 (TriTrastu12 long chain amino acid sequence, SIRPaD1 protein - linker - anti-Her2 antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0400] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGFNIKDTYIHWVRQAPGKGLEWVARIYPTNGYTRYADSVKGRFTISADTSKNTAYLQMNSLRAEDTAVYYCSRWGGDGFYAMDYWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0401] SEQ ID NO: 113 (TriBeva06 long chain amino acid sequence, SIRPaD1 protein-linker-anti-VEGF antibody heavy chain-linker-human IL-15-linker-anti-CD16a single chain antibody)

[0402] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSNWVNVISDLKKIEDLIQSMHIDATLYTESDVHPSCKVTAMKCFLLELQVISLESGDASIHDTVENLIILANNSLSSNGNVTESGCKECEELEEKNIKEFLQSFVHIVQMFINTSGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0403] SEQ ID NO: 114 (TriBeva06 short chain amino acid sequence, anti-VEGF antibody light chain)

[0404] DIQMTQSPSSLSASVGDRVTITCSASQDISNYLNWYQQKPGKAPKVLIYFTSSLHSGVPSRFSGSGSGTDFTLTISSLQPEDFATYYCQQYSTVPWTFGQGTKVEIKRTVAAPSVFIFPPSDEQLKSGTASVVCLLNNFYPREAKVQWKVDNALQSGNSQESVTEQDSKDSTYSLSSTLTLSKADYEKHKVYACEVTHQGLSSPVTKSFNRGEC

[0405] SEQ ID NO: 115 (TriBeva12 long chain amino acid sequence, SIRPaD1 protein - linker - anti-VEGF antibody heavy chain - linker - human IL-2 - linker - anti-CD16a single chain antibody)

[0406] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSGGGGSEVQLVESGGGLVQPGGSLRLSCAASGYTFTNYGMNWVRQAPGKGLEWVGWINTYTGEPTYAADFKRRFTFSLDTSKSTAYLQMNSLRAEDTAVYYCAKYPHYYGSSHWYFDVWGQGTLVTVSSASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGGGGSGGGGSGGGGSAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0407] SEQ ID NO: 116 (ISE-Negative Control long chain amino acid sequence, antibody heavy chain constant region)

[0408] ASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPGK

[0409] SEQ ID NO: 117 (CpG ODN sequence)

[0410] gggggACGATCGTCgggggg

[0411] SEQ ID NO: 118 (IL-2 mutant amino acid sequence)

[0412] APTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLT

[0413] SEQ ID NO: 119 (IFN alpha mutant amino acid sequence)

[0414] CDLPQTHSLGSRRTLMLLAQMRRISLFSCLKDRHDFGFPQEEFGNQFQKAETIPVLHEMIQQIFNLFSTKDSSAAWDETLLDKFYTELYQQLNDLEACVIQGVGVTETPLMNEDSILAVRKYFQRITLYLKEKKYSPCAWEVVAAEIMRSFSLSTNLQESLRSKE

[0415] SEQ ID NO: 120 (TriCetu12 long chain amino acid sequence, SIRPaD1 protein-linker-anti-EGFR antibody heavy chain-linker-human IL-2-linker-anti-CD16a single chain antibody)

[0416] EEELQVIQPDKSVLVAAGETATLRCTATSLIPVGPIQWFRGAGPGRELIYNQKEGHFPRVTTVSDLTKRNNMDFSIRIGAITPADAGTYYCVKFRKGSPDDVEFKSGAGTELSVRAKGGGGSGGGGSGGGGSQVQLKQSGPGLVQPSQSLSITCTVSGFSLTNYGVHWVRQSPGKGLEWLGVIWSGGNTDYNTPFTSRLSINKDNSKSQVFFKMNSLQSNDTAIYYCARALTYYDYEFAYWGQGTLVTVSAASTKGPSVFPLAPSSKSTSGGTAALGCLVKDYFPEPVTVSWNSGALTSGVHTFPAVLQSSGLYSLSSVVTVPSSSLGTQTYICNVNHKPSNTKVDKKVEPKSCDKTHTCPPCPAPELLGGPSVFLFPPKPKDTLYITREPEVTCVVVDVSHEDPEVKFNWYVDGVEVHNAKTKPREEQYNSTYRVVSVLTVLHQDWLNGKEYKCKVSNKALPAPIEKTISKAKGQPREPQVYTLPPSREEMTKNQVSLTCLVKGFYPSDIAVEWESNGQPENNYKTTPPVLDSDGSFFLYSKLTVDKSRWQQGNVFSCSVMHEALHNHYTQKSLSLSPPSGQAGAAASESLFVSNHAYAPTSSSTKKTQLQLEHLLLDLQMILNGINNYKNPKLTAMLTFKFYMPKKATELKHLQCLEEELKPLEEVLNLAQSKNFHFDPRDVVSNINVFVLELKGSETTFMCEYADETATIVEFLNRWITFCQSIISTLTGGGGSGGGGSGGGGSEVQLVQSGAEVKKPGESLKVSCKASGYTFTSYYMHWVRQAPGQGLEWMGIINPSGGSTSYAQKFQGRVTMTRDTSTSTVYMELSSLRSEDTAVYYCARGSAYYYDFADYWGQGTLVTVSSGGGGSGGGGSGGGGSGGGGSQPVLTQPSSVSVAPGQTATISCGGHNIGSKNVHWYQQRPGQSPVLVIYQDNKRPSGIPERFSGSNSGNTATLTISGTQAMDEADYYCQVWDNYSVLFGGGTKLTVL

[0417] It is hereby declared that the above technical route, features and sequences of the present application, and the technical features specifically described hereinafter (such as the examples) can be combined with each other within the scope of the present application to form new or preferred technical solutions. Due to the limited space, they will not be repeated here. BRIEF DESCRIPTION OF DRAWINGS

[0418] Figure 3 Structure diagram of the innate immune super connector of the present application;

[0419] Figure 4 Structure diagram of the fully immune antibody of the present application;

[0420] Figure 5 Structure diagram of the HEL antibody of the present application;

[0421] Figure 6 SDS-PAGE electropherogram of the anti-HEL antibody innate immune super connector variant of the present application;

[0422] Figure 7 SDS-PAGE electropherogram of the variable region-free innate immune super connector variant and the fully immune antibody variant of the present application;

[0423] Figure 8 Binding of the fully immune antibody with PD-L1 protein based on the anti-PD-L1 antibody framework of the present application;

[0424] Figure 9 Binding of the fully immune antibody with CD47 protein based on the anti-PD-L1 antibody framework of the present application;

[0425] Figure 10 Binding of the fully immune antibody with CD16a protein based on the anti-PD-L1 antibody framework of the present application;

[0426] Figure 11 Binding of the fully immune antibody with CD16b protein based on the anti-PD-L1 antibody framework of the present application;

[0427] Figure 12 Binding of the fully immune antibody with FcRn protein based on the anti-PD-L1 antibody framework of the present application;

[0428] Figure 13 ADCC activity of the variable region-free innate immune super connector of the present application;

[0429] Figure 14 ADCC activity of the human non-targeting innate immune super connector of the present application for mechanism verification;

[0430] Figure 15ADCC activity of the fully humanized antibody of the application with anti-PD-L1 antibody as the framework on tumor cells expressing PD-L1 (HCC-1954);

[0431] Figure 16 Proliferation effect of the fully humanized antibody TriAtezo06 of the application with anti-PD-L1 antibody as the framework on mouse T lymphocyte CTLL-2;

[0432] Figure 17 ADCC activity of the fully humanized antibody of the application with anti-EGFR antibody as the framework on tumor cells expressing EGFR (A. human epidermal carcinoma cell 431, B. human breast cancer cell MDA-MB-468);

[0433] Figure 18 ADCC activity of the fully humanized antibody of the application with anti-Claudin18.2 antibody as the framework on tumor cells NCI-N87 expressing Claudin18.2 gastric cancer;

[0434] Figure 19 ADCC activity of the fully humanized antibody of the application with anti-CTLA-4 antibody as the framework on human breast cancer cells HCC-1954;

[0435] Figure 20 ADCC activity of the fully humanized antibody of the application with anti-BCMA antibody as the framework on tumor cells expressing BCMA (human lymphoma cell Raji);

[0436] Figure 21 ADCC activity of the fully humanized antibody of the application with anti-CD20 antibody as the framework on tumor cells expressing CD20 (human lymphoma cell Raji);

[0437] Figure 22 ADCC activity of the fully humanized antibody of the application with anti-CD38 antibody as the framework on tumor cells expressing CD38 (human lymphoma cell Raji);

[0438] Figure 23 ADCC activity of the fully humanized antibody of the application with anti-Trop-2 antibody as the framework on tumor cells expressing Trop-2 (human gastric cancer cell NCI-N87);

[0439] Figure 24 ADCC activity of the fully humanized antibody of the application with anti-Nectin-4 antibody as the framework on tumor cells expressing Nectin-4 (human gastric cancer cell NCI-N87);

[0440] Figure 25ADCC activity of the fully humanized antibody with anti-PD-1 antibody as the framework on human gastric cancer cell NCI-N87 of the present application;

[0441] Figure 26 ADCC activity of the fully humanized antibody with anti-Her2 antibody as the framework on Her2 tumor cells (human gastric cancer cell NCI-N87) of the present application;

[0442] Figure 27 Pharmacodynamic study of the fully humanized antibody with anti-Her2 antibody as the framework in Ex Vivo human pleural effusion model of the present application;

[0443] Figure 1 Pharmacodynamic study of the fully humanized antibody with anti-Her2 antibody as the framework in Ex Vivo human peritoneal effusion model of the present application;

[0444] Figure 2 Organ toxicity study of the fully humanized antibody with anti-PD-L1 antibody as the framework in mice of the present application. Example

[0445] The present application is further illustrated by the following drawings and specific examples, which should be understood as merely illustrative of the present application and not limiting the scope of the present application.

[0446] Example 1 Structure design of innate immune super connector and fully humanized antibody

[0447] In the structure of innate immune super connector, in addition to CD16a antibody targeting the surface of NK cells and SIRPa protein blocking the "don't eat me" signal of macrophages, immune cytokines and pattern recognition receptor agonists are also introduced. The structure of ISE is shown in Figure 3 The structure of fully humanized antibody is based on innate immune super connector and connects with adaptive immune antibody or tumor antigen antibody. The structure of fully humanized antibody is shown in Figure 4

[0448] Example 2 Antibody structure and expression of innate immune super connector without targeting in human body

[0449] ​In one aspect, in order to prove the possibility of the innate immune super connector concept, the present application takes the innate immune super connector as the skeleton, adds the antibody without targeting in the human body, the variable region of the anti-hen egg lysozyme antibody (anti-hen egg lysozyme HEL antibody) to the innate immune super connector, and prepares five antibodies of the main structure of the innate immune super connector: HE LAB01, HE LAB02, HE LAB03, HE LAB04 and HE LAB05, the purpose is to show the superposition function of different parts of ISE and the function of cytokines. They are anti-HEL specific antibody, anti-HEL and SIRPa (anti-CD47) bispecific fusion antibody, anti-HEL and anti-CD16a bispecific antibody, anti-HEL, SIRPa (anti-CD47) and CD16a trispecific fusion antibody, and anti-HEL, SIRPa (anti-CD47) and CD16a tetraspecific fusion antibody with the introduction of human IL-15. The antibody structure is shown in Figure 5 SDS-PAGE electrophoretogram (reduced and non-reduced) is shown in Figure 5 In this embodiment, the amino acid sequences of the long chain of HE LAB01, HE LAB02, HE LAB03, HE LAB04 and HE LAB05 are SEQ ID NO: 20, SEQ ID NO: 22, SEQ ID NO: 24, SEQ ID NO: 26 and SEQ ID NO: 28 respectively, and the amino acid sequences of the short chain are SEQ ID NO: 21, SEQ ID NO: 23, SEQ ID NO: 25, SEQ ID NO: 27, SEQ ID NO: 29 respectively.

[0450] Example 3 Expression and purification of innate immune super connector and complete immune antibody

[0451] The gene is amplified by conventional molecular biology technology PCR, and the two chain genes are connected to the pcDNA3.4 vector by homologous recombination technology. After the positive clone after sequencing is extracted, the plasmid is transfected into Expi293F cells, which are cultured at 37°C / 5% CO2 / 125 rpm shaker for 7 days, and after the culture is completed, the supernatant is collected and purified by protein A affinity chromatography to obtain the purified innate immune super connector and complete immune antibody, and the antibody concentration is determined by UV280 combined with the theoretical extinction coefficient. The results of SDS-PAGE electrophoretogram are shown in Figure 5In this example, the amino acid sequences of the long chains of the innate immunity superconnector ISE-Negative Control, ISE-A2, and ISE-D2 are SEQ ID NO: 116, SEQ ID NO: 32, and SEQ ID NO: 44, respectively; and the amino acid sequences of the short chains are SEQ ID NO: 31, SEQ ID NO: 33, and SEQ ID NO: 45, respectively Figure 6 A-B reduction and non-reducing SDS-PAGE). The amino acid sequences of the long chains of the complete immunity antibodies TriAtezo01, TriAtezo02, TriAtezo03, TriAtezo04, TriAtezo05, TriAtezo06, TriAtezo07, TriAtezo12, TriCetu03, TriCetu04, and TriCetu12 are SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 81, and SEQ ID NO: 120, respectively; and the amino acid sequences of the short chains are SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 80, SEQ ID NO: 80, and SEQ ID NO: 80, respectively Figure 7C-D non-reducing SDS-PAGE). The long chain amino acid sequences of the fully immune antibodies TriKey06, TriKey12, TriDaratu06, TriDaratu12, TriRituxi06, TriRituxi12, TriZolbetu06, TriZolbetu12, TriSacitu06, TriSacitu12, TriTrastu06, TriTrastu12, TriBeva06, TriBeva12, TriEnfortu06, TriEnfortu12, TriBelanta06, TriBelanta12, TriIpilimu06 and TriIpilimu12 are SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 101 and SEQ ID NO: 103, respectively; the short chain amino acid sequences are SEQ ID NO: 87, SEQ ID NO: 87, SEQ ID NO: 90, SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 93, SEQ ID NO: 96, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 99, SEQ ID NO: 111, SEQ ID NO: 111, SEQ ID NO: 114, SEQ ID NO: 114, SEQ ID NO: 108, SEQ ID NO: 108, SEQ ID NO: 105, SEQ ID NO: 105, SEQ ID NO: 102 and SEQ ID NO: 102, respectively; the innate immune super connectors with different targets of the fully immune antibodies were successfully expressed and the purity of SDS-PAGE after one-step affinity chromatography reached about 80%.

[0452] Example 4 ELISA detection of the binding of fully immune antibodies with anti-PD-L1 antibody as the skeleton to PD-L1 protein

[0453] Coat 2 ug / ml PD-L1 protein 100ul / well at 4°C overnight; block with 3% skim milk powder at 37°C for 1h; add different concentrations (10000 ng / ml, 2500 ng / ml, 625 ng / ml, 156.25 ng / ml, 39.06 ng / ml, 9.77 ng / ml, 2.44 ng / ml, 0) of anti-PD-L1 complete immune antibodies and other control samples each 100ul, incubate at 37°C for 1h; wash 3 times with PBST after incubation at 37°C for 1h; then add 100ul of goat anti-human IgG-HRP diluted 1:8000, incubate at 37°C for 1h, then wash 6 times, add 100ul TMB to each well for color development for 5min, then add 50ul stop solution, read OD450 on the enzyme marker. The results are shown in Figure 8 As shown in the figure, the binding ability of the complete immune antibody to PD-L1 protein is at the same level as that of the monoclonal antibody.

[0454] Example 5 ELISA detection of the binding of complete immune antibodies with CD47 protein as the framework of anti-PD-L1 antibodies

[0455] Coat different concentrations of 2 ug / ml CD47 protein, 100ul / well at 4°C overnight; block with 3% skim milk powder at 37°C for 1h, then wash 3 times with PBST; add different concentrations (10000 ng / ml, 2500 ng / ml, 625 ng / ml, 156.25 ng / ml, 39.06 ng / ml, 9.77 ng / ml, 2.44 ng / ml, 0) of anti-PDL-1 complete immune antibodies and other control samples each 100ul, incubate at 37°C for 1h, then wash 3 times with PBST; then add 100ul of goat anti-human IgG-HRP diluted 1:8000, incubate at 37°C for 1h, then wash 6 times, add 100ul TMB to each well for color development for 5min, then add 50ul stop solution, read OD450 on the enzyme marker. The results are shown in Figure 8 As shown in the figure, the binding ability of the complete immune antibody to CD47 protein is at the same level as that of SIRPa fusion molecule (TTI-622). The position of SIRPa D1 protein in the complete immune antibody with other targets as the framework is the same as that of the complete immune antibody, so the binding to CD47 protein is also at the same level. This will not be repeated here.

[0456] Example 6 ELISA detection of the binding of complete immune antibodies with CD16a protein as the framework of anti-PD-L1 antibodies

[0457] CD16a amino acid at position 158 has two types, V (valine) strong affinity, F (phenylalanine) weak affinity. Coated with 20 ug / ml CD16a protein, 100ul / well at 4°C overnight; 3% skimmed milk powder was blocked at 37°C for 1h; different concentrations (10000ng / ml, 2500ng / ml, 625ng / ml, 156.25ng / ml, 39.06ng / ml, 9.77ng / ml, 2.44ng / ml, 0) of anti-PDL-1 complete immune antibody and other control samples were added to each well, 100ul, 37°C for 1h; after incubation at 37°C for 1h, PBST was washed for 3 times; then 100ul of 1:8000 diluted goat anti-human IgG-HRP was added, incubated at 37°C for 1h, then washed for 6 times, 100ul TMB was added to each well for color development for 5min, then 50ul stop solution was added, and OD450 was read on the enzyme marker. The results are shown in Figure 8 , Figure 9 A is the binding of the molecule to CD16a-V protein, Figure 10 B is the binding of the molecule to CD16a-F protein. The complete immune antibody has a strong binding level with CD16a protein. The position of the anti-CD16a single-chain antibody in the complete immune antibody with other target skeletons is the same as that of the complete immune antibody, so the binding with CD16a protein is also at the same level, which is not described here.

[0458] Example 7 ELISA detection of the binding of complete immune antibody with anti-PD-L1 antibody as the skeleton to CD16b protein

[0459] Coated with 20 ug / ml CD16b protein, 100ul / well at 4°C overnight; 3% skimmed milk powder was blocked at 37°C for 1h; different concentrations (10000ng / ml, 2500ng / ml, 625ng / ml, 156.25ng / ml, 39.06ng / ml, 9.77ng / ml, 2.44ng / ml, 0) of anti-PDL-1 complete immune antibody and other control samples were added to each well, 100ul, 37°C for 1h; after incubation at 37°C for 1h, PBST was washed for 3 times; then 100ul of 1:8000 diluted goat anti-human IgG-HRP was added, incubated at 37°C for 1h, then washed for 6 times, 100ul TMB was added to each well for color development for 5min, then 50ul stop solution was added, and OD450 was read on the enzyme marker. The results are shown in Figure 11 As shown in the results, the complete immune antibody does not bind to CD16b protein, indicating that the CD16a antibody in the complete immune antibody has strong specificity.

[0460] Example 8 ELISA detection of the binding of complete immune antibody TriAtezo06 to FcRn protein

[0461] Coat 2 ug / ml FcRn protein 100ul / well at 4°C overnight; block with 3% skim milk powder at 37°C for 1h; add different concentrations (10000ng / ml, 2500ng / ml, 625ng / ml, 156.25ng / ml, 39.06ng / ml, 9.77ng / ml, 2.44ng / ml, 0) TriAtezo06 and other control samples 100ul per well, incubate at 37°C for 1h; wash 3 times with PBST after incubation at 37°C for 1h; then add goat anti-human IgG-HRP diluted 1:8000 100ul, incubate at 37°C for 1h, wash 6 times, add 100ul TMB for color development per well, add 50ul stop solution, and read OD450 on the enzyme label instrument. The results are shown in Figure 1 Compared with Atezolizumab, the fully immune antibody binds strongly with FcRn protein, indicating that the fully immune antibody has a longer half-life than Atezolizumab in vivo.

[0462] Example 9 ADCC activity study of innate immune super connector without variable region

[0463] Culture and expand human breast cancer cells HCC-1954 to the required amount, plate the cells into a 96-well plate at 4000 / well, adhere overnight, resuspend fresh human PBMC in 1640 culture medium, add the antibody to the 96-well plate according to the effector to target ratio of 10:1, and the drug concentration gradient is 500nM, 50nM, 5nM, 0.5nM, 0.05nM, 0.005nM and 0.0005nM respectively, continue to culture for 5 days, wash the plate 3 times with PBS, detect the number of living cells by CCK-8 reagent, and calculate the IC 50 . The results are shown in Figure 12 Without the presence of ISE components (SIRPa D1 protein, CD16a antibody and cytokine IL-15), ISE-Negative Control (as shown in Figure 13 ) has no ADCC activity; in the presence of SIRPa D1 protein and CD16a antibody at the same time, ISE-A2 only shows weak ADCC activity at high concentration; in the presence of all ISE components (SIRPa D1 protein, CD16a antibody and cytokine IL-15), ISE-D2 shows very strong ADCC activity, and the activity is significantly better than that of the cytokine fusion molecule. It proves the superiority of the ISE structure.

[0464] Example 10 Innate immune super connector antibody ADCC activity verified by human target-free antibody mechanism

[0465] HCC-1954 cells were expanded in culture to the desired amount, plated into 96-well plates at 4000 / well, and allowed to adhere overnight. Fresh human PBMCs were resuspended in 1640 medium, and the antibodies were added to the 96-well plates according to an effector-to-target ratio of 10:1. The drug concentration gradient was 500 nM, 50 nM, 5 nM, 0.5 nM, 0.05 nM, 0.005 nM, and 0.0005 nM, respectively. After 5 days of continued culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using a CCK-8 reagent. The IC 50 Results are shown in Figure 13 As can be seen from the figure, the HEL antibody (HELAB01) had no ADCC activity in the absence of ISE components (SIRPa D1 protein, CD16a antibody, and cytokine IL-15). HE LAB02 had some ADCC activity in the presence of only SIRPa D1 protein. HE LAB03 did not exhibit obvious ADCC activity in the presence of only CD16a antibody, indicating that the activity of macrophages in the donor PBMCs was strong, and the activity of NK cells was weak. The killing effect of HE LAB04 was stronger than that of HE LAB02 and HE LAB03 in the presence of both SIRPa D1 protein and CD16a antibody, indicating that the recruitment of multiple immune effector cells had a stronger killing effect than the recruitment of only a single immune effector cell. The killing effect of HE LAB05 was further enhanced compared to HE LAB04 in the presence of all ISE components, demonstrating that the introduced IL-15 had a proliferative effect on immune cells.

[0466] Example 11 ADCC activity of a complete immune antibody with an anti-PD-L1 antibody as a skeleton

[0467] PD-1 / PD-L1 ADCC activity model in vitro: HCC-1954 cells were expanded in culture to the desired amount, plated into 96-well plates at 4000 / well, and allowed to adhere overnight. Human PBMCs were incubated with SEB in vitro for 7 days, and when the PD1 expression was greater than or equal to 16% or more, the PBMCs were resuspended in 1640 medium. The antibodies were added to the 96-well plates according to an effector-to-target ratio of 10:1. The drug concentration gradient was 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continued culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using a CCK-8 reagent. As Figure 14As shown in -A, Atezolizumab itself has certain ADCC activity, and after being combined with TTI-622 (Fc-SIRPa fusion molecule), the ADCC activity of human breast cancer cells HCC-1954 (PD-L1 positive) is improved to a certain extent; TriAtezo01 (with Atezolizumab as the basic skeleton, SIRPa connected at the C-terminus of the light chain, and anti-CD16a single-chain antibody connected at the C-terminus of the heavy chain) has significantly better ADCC activity than Atezolizumab; TriAtezo02 (with Atezolizumab as the basic skeleton, SIRPa connected at the C-terminus of the light chain, and cytokine IL-15 and anti-CD16a nanoantibody connected at the C-terminus of the heavy chain) has more significant ADCC activity than TriAtezo01. Figure 15 As shown in -B, atezolizumab itself has certain ADCC activity. The ADCC activity of TriAtezo05 (based on atezolizumab, with SIRPa linked to the N-terminus of the heavy chain and an anti-CD16a single-chain antibody linked to the C-terminus of the heavy chain) is significantly better than that of atezolizumab. The ADCC activity of TriAtezo06 (based on atezolizumab, with SIRPa linked to the N-terminus of the heavy chain and an anti-CD16a single-chain antibody linked to the C-terminus of the heavy chain) is significantly better than that of atezolizumab. The ADCC activity of TriAtezo07 (based on atezolizumab, with SIRPα linked to the N-terminus of the heavy chain and the C-terminus of the heavy chain linked to the cytokine IL-15 and its receptor IL-15R and anti-CD16a single-chain antibody) was superior to that of TriAtezo05. The ADCC activity of TriAtezo12 (based on atezolizumab, with SIRPα linked to the N-terminus of the heavy chain and the C-terminus of the heavy chain linked to the cytokine IL-2 and anti-CD16a single-chain antibody) was even more significantly improved. These results indicate that the ISE components of fully immune antibodies (SIRPα protein, anti-CD16a antibody, cytokine IL-15 or IL-2) can significantly enhance the ADCC activity of anti-PD-L1 antibodies.

[0468] Example 12 Study on cytokine activity in the fully immune antibody TriAtezo06 with anti-PD-L1 antibody as the backbone

[0469] Mouse T lymphocytes CTLL-2 were cultured and expanded, 5E5 cells / well were plated in 96-well plates, and the drug concentrations were 100nM, 10nM, 1nM, 0.1nM, 0.01nM, 0.001nM and 0.0001nM respectively. The culture was continued for 72h, and the cell viability was detected by CCK-8 according to the instructions. The results are shown in the figure. Figure 15As shown, the proliferation activity of TriAtezo06 on mouse T lymphocyte CTLL-2 was significantly weaker than wild type IL-15 and Fc-IL-15. It is illustrated that the structural design of the fully immunized antibody affects the activity of the cytokine IL-15, which can significantly reduce the toxic side effects of the molecule in clinical application.

[0470] Example 13 Study on ADCC activity of fully immunized antibody with anti-EGFR antibody as the skeleton

[0471] 13.1 Study on ADCC activity against human epidermal carcinoma cell A431

[0472] The expanded cells were cultured to the required amount, plated into 96-well plates at 1500 / well, and adhered overnight. Fresh human PBMC was resuspended in 1640 culture medium, and the antibodies were added to the 96-well plate according to the effector-to-target ratio of 10:1. The drug concentration gradient was 500nM, 5nM, 0.05nM, 0.0005nM, 0.000005nM, 0.00000005nM, and 0.0000000005nM, respectively. After 5 days of continuous culture, the plate was washed with PBS for 3 times, and the number of living cells was detected by CCK-8 reagent. The IC 50 was calculated according to the pharmacodynamic measurement. The results are shown in Figure 16 A. In the ADCC activity experiment against tumor cell line A431 with high expression of EGFR, Cetuximab showed a certain ADCC activity. The ADCC activity of Fc-IL-15 and Fc-IL-2 was only better than that of Cetuximab at the highest concentration, but TriCetu03 (with Cetuximab as the basic skeleton, connecting SIRPa at the N terminus of the heavy chain, and connecting anti-CD16a single-chain antibody at the C terminus of the heavy chain) showed ADCC activity better than Cetuximab. The ADCC activity of TriCetu04 (with Cetuximab as the basic skeleton, connecting SIRPa at the N terminus of the heavy chain, and connecting cytokine IL-15 and anti-CD16a single-chain antibody at the C terminus of the heavy chain) and TriCetu12 (with Cetuximab as the basic skeleton, connecting SIRPa at the N terminus of the heavy chain, and connecting cytokine IL-2 and anti-CD16a single-chain antibody at the C terminus of the heavy chain) was improved more significantly.

[0473] 13.2 Study on ADCC activity against human breast cancer cell MDA-MB-468

[0474] Culture the expanded cells to the required amount, plate the cells into 96-well plates at 4000 / well, adhere overnight, resuspend fresh human PBMC in 1640 culture medium, add the antibodies to the 96-well plates according to the effector-to-target ratio of 10:1, the drug concentration gradient is 500nM, 5nM, 0.05nM, 0.0005nM, 0.000005nM, 0.00000005nM and 0.0000000005nM respectively, continue to culture for 5 days, then wash the plates 3 times with PBS, detect the number of living cells using CCK-8 reagent, and calculate the IC 50 . The results are shown in Figure 17 In the ADCC activity experiment against EGFR-expressing tumor cell line MDA-MB-468, Cetuximab showed a certain ADCC activity, and the ADCC activity of Fc-IL-15 and Fc-IL-2 was better than that of Cetuximab only at the highest concentration, but TriCetu03 (with Cetuximab as the basic framework, SIRPa connected at the N-terminus of the heavy chain, and anti-CD16a single-chain antibody connected at the C-terminus of the heavy chain) showed ADCC activity better than Cetuximab; the ADCC activity of TriCetu04 (with Cetuximab as the basic framework, SIRPa connected at the N-terminus of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminus of the heavy chain) and TriCetu12 (with Cetuximab as the basic framework, SIRPa connected at the N-terminus of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminus of the heavy chain) was improved more significantly.

[0475] The ADCC activity of the above two EGFR-expressing tumor cell lines showed that the ISE components of the fully immune antibody (SIRPaD1 protein, anti-CD16a antibody, cytokine IL-15 or IL-2) can significantly improve the ADCC activity of the anti-EGFR antibody.

[0476] Example 14 ADCC activity study of fully immune antibody with anti-Claudin18.2 antibody as the framework

[0477] Culture the expanded human gastric cancer cells NCI-N87 to the required amount, plate the cells into 96-well plates at 4000 / well, adhere overnight, resuspend fresh human PBMC in 1640 culture medium, add the antibodies to the 96-well plates according to the effector-to-target ratio of 10:1, the drug concentration gradient is 500nM, 5nM, 0.05nM, 0.0005nM, 0.000005nM, 0.00000005nM and 0.0000000005nM respectively, continue to culture for 5 days, then wash the plates 3 times with PBS, detect the number of living cells using CCK-8 reagent. The results are shown in Figure 18The complete immune antibodies TriZolbetu06 (with Zolbetuximab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriZolbetu12 (with Zolbetuximab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the Claudin 18.2 tumor-related antigen showed good ADCC activity against human gastric cancer cells NCI-N87 (Claudin 18.2 positive).

[0478] Example 15 ADCC activity study of complete immune antibodies with anti-CTLA-4 antibodies as the framework

[0479] The human breast cancer cells HCC-1954 were cultured and expanded to the required amount, and the cells were plated into 96-well plates at 4000 / well, adhered overnight, and fresh human PBMCs were resuspended in 1640 culture medium. The antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1, and the drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using the CCK-8 reagent. The results are shown in Figure 19 The complete immune antibodies TriIpilimu06 (with Ipilimumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriIpilimu12 (with Ipilimumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the CTLA-4 acquired immune checkpoint showed good ADCC activity against human breast cancer cells HCC-1954.

[0480] Example 16 ADCC activity study of complete immune antibodies with anti-BCMA antibodies as the framework

[0481] The human Burkitt's lymphoma cells Raji were cultured and expanded to the required amount, and the cells were plated into 96-well plates at 10000 / well, adhered overnight, and fresh human PBMCs were resuspended in 1640 culture medium. The antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1, and the drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using the CCK-8 reagent. The results are shown inFigure 20 The complete immune antibodies TriBelanta06 (taking Belantamab as the basic framework, connecting SIRPa at the N-terminal of the heavy chain, and connecting the cytokine IL-15 and the anti-CD16a single-chain antibody at the C-terminal of the heavy chain) and TriBelabta12 (taking Belantamab as the basic framework, connecting SIRPa at the N-terminal of the heavy chain, and connecting the cytokine IL-2 and the anti-CD16a single-chain antibody at the C-terminal of the heavy chain) targeting the BCMA tumor-associated antigen showed good ADCC activity against human lymphoma cells Raji (BCMA positive).

[0482] Example 17 ADCC activity study of complete immune antibodies taking anti-CD20 antibodies as the framework

[0483] The human Burkitt's lymphoma cells Raji were cultured and expanded to the required amount, and the cells were plated into 96-well plates at 10,000 / well, and adhered overnight. Fresh human PBMCs were resuspended in 1640 culture medium, and the antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1. The drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of living cells was detected by using the CCK-8 reagent. The results are shown in Figure 21 The complete immune antibodies TriRituxi06 (taking Rituximab as the basic framework, connecting SIRPa at the N-terminal of the heavy chain, and connecting the cytokine IL-15 and the anti-CD16a single-chain antibody at the C-terminal of the heavy chain) and TriRituxi12 (taking Rituximab as the basic framework, connecting SIRPa at the N-terminal of the heavy chain, and connecting the cytokine IL-2 and the anti-CD16a single-chain antibody at the C-terminal of the heavy chain) targeting the CD20 tumor-associated antigen showed good ADCC activity against human lymphoma cells Raji (CD20 positive).

[0484] Example 18 ADCC activity study of complete immune antibodies taking anti-CD38 antibodies as the framework

[0485] The human Burkitt's lymphoma cells Raji were cultured and expanded to the required amount, and the cells were plated into 96-well plates at 10,000 / well, and adhered overnight. Fresh human PBMCs were resuspended in 1640 culture medium, and the antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1. The drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of living cells was detected by using the CCK-8 reagent. The results are shown in Figure 22The complete immune antibodies TriDaratu06 (with Daratumumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriDaratu12 (with Daratumumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the CD38 tumor-associated antigen showed good ADCC activity against human lymphoma cells Raji (CD38 positive).

[0486] Example 19 ADCC activity study of complete immune antibodies with anti-Trop-2 antibodies as the framework

[0487] The human gastric cancer cells NCI-N87 were cultured and expanded to the required amount, and then plated into 96-well plates at 4000 / well, and adhered overnight. Fresh human PBMCs were resuspended in 1640 culture medium, and the antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1. The drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using the CCK-8 reagent. The results are shown in Figure 23 The complete immune antibodies TriSacitu06 (with Sacituzumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriSacitu12 (with Sacituzumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the Trop-2 tumor-associated antigen showed strong ADCC activity against human gastric cancer cells NCI-N87 (Trop-2 positive).

[0488] Example 20 ADCC activity study of complete immune antibodies with anti-Nectin-4 antibodies as the framework

[0489] The human gastric cancer cells NCI-N87 were cultured and expanded to the required amount, and then plated into 96-well plates at 4000 / well, and adhered overnight. Fresh human PBMCs were resuspended in 1640 culture medium, and the antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1. The drug concentration gradients were 500 nM, 5 nM, 0.05 nM, 0.0005 nM, 0.000005 nM, 0.00000005 nM, and 0.0000000005 nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using the CCK-8 reagent. The results are shown in Figure 24The full immune antibodies TriEnfortu06 (with Enfortumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriEnfortu12 (with Enfortumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the Nectin-4 tumor-related antigen showed good ADCC activity against human gastric cancer cells NCI-N87 (Nectin-4 positive).

[0490] Example 21 ADCC activity study of full immune antibodies with anti-PD-1 antibody as the framework

[0491] Human gastric cancer cells NCI-N87 were cultured and expanded to the required amount, and plated into 96-well plates at 4000 / well, and adhered overnight. Human PBMCs were incubated in vitro with SEB for 7 days, and when the PD1 expression was greater than or equal to 16% or more, the PBMCs were resuspended in 1640 culture medium, and the antibodies were added to the 96-well plates according to the effector-to-target ratio of 10:1, with the drug concentration gradient being 500nM, 5nM, 0.05nM, 0.0005nM, 0.000005nM, 0.00000005nM, and 0.0000000005nM, respectively. After 5 days of continuous culture, the plates were washed 3 times with PBS, and the number of viable cells was detected using the CCK-8 reagent. The results are shown in Figure 25 The full immune antibodies TriKey06 (with Pembrolizumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-15 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) and TriKey12 (with Pembrolizumab as the basic framework, SIRPa connected at the N-terminal of the heavy chain, and the cytokine IL-2 and anti-CD16a single-chain antibody connected at the C-terminal of the heavy chain) targeting the PD-1 acquired immunity showed good ADCC activity against human gastric cancer cells NCI-N87.

[0492] Example 22 ADCC activity study of full immune antibodies with anti-Her2 antibody as the framework

[0493] The human gastric cancer cell NCI-N87 was expanded to the required amount, and plated into a 96-well plate at 4000 / well, and adhered overnight. Fresh human PBMC was resuspended in 1640 culture medium, and the antibody was added to the 96-well plate according to the effector-to-target ratio of 10:1. The drug concentration gradient was 500nM, 5nM, 0.05nM, 0.0005nM, 0.000005nM, 0.00000005nM, and 0.0000000005nM, respectively. After 5 days of continuous culture, the plate was washed 3 times with PBS, and the number of viable cells was detected by CCK-8 reagent. The IC 50 was calculated according to the pharmacodynamic measurement. The results are shown in Figure 25 . The fully immune antibodies TriTrastu06 (using Trastuzumab as the basic framework, connecting SIRPa at the N-terminus of the heavy chain, and connecting the cytokine IL-15 and the anti-CD16a single-chain antibody at the C-terminus of the heavy chain) and TriTrastu12 (using Trastuzumab as the basic framework, connecting SIRPa at the N-terminus of the heavy chain, and connecting the cytokine IL-2 and the anti-CD16a single-chain antibody at the C-terminus of the heavy chain) targeting the Her2 tumor-associated antigen showed good ADCC activity against human gastric cancer cells NCI-N87 (Her2 positive).

[0494] Example 23 Pharmacodynamic study of anti-Her2-based fully immune antibodies in ex vivo organoids

[0495] 23.1 Her2 2 + Breast cancer patient pleural effusion sample

[0496] The Her2 2 + breast cancer pleural effusion sample was collected, and the cells in the sample, including tumor cells and immune cells, were separated. The candidate and control drugs were co-incubated with the tumor cell immune cell complex in vitro. After 4-5 days of incubation, the culture supernatant was collected to detect tumor cell granzyme B and IFN-γ, and the tumor cells and immune cells were analyzed. The amino acid sequence of the long chain of the anti-Her2 fully immune antibody TriAB04 (using Trastuzumab as the basic framework, connecting SIRPa at the C-terminus of the light chain, and connecting the anti-CD16a single-chain antibody at the C-terminus of the heavy chain) used in this example was SEQ ID NO: 82, and the amino acid sequence of the short chain was SEQ ID NO: 83. The results are shown in Figure 25 . The sample immune cell phenotype Figure 25 (A): NK cells 6.5%; macrophages 2.11% (M1 0.06%, M2 0.33%); CD3 + T cells 5.47%; CD4 + T cells 2.93%; CD8 +T cells 1.66%. Anti-Her2 complete immune antibody increased the killing effect of Her2 positive tumor cells by 27% compared with the baseline ( Figure 25 -B), anti-Her2 complete immune antibody, anti-PD-1 antibody or Herceptin combined with TTI-622 (Fc-SIRPa fusion molecule) had a weak ability to induce IFN-γ secretion, while combined with anti-PD-1 antibody, it could significantly enhance IFN-γ secretion ( Figure 26 -D). Combination with anti-PD-1 antibodies also significantly increased granzyme B levels ( Figure 26 Since the combination of TTI-622 and Herceptin did not show significant tumor-killing effects, while the anti-Her2 fully immune antibody showed significant anti-tumor activity, combined with the immune cell analysis results, which showed 6.5% NK cells (within the normal range of peripheral blood levels) and 2.11% macrophages (significantly lower than normal peripheral blood levels), it can be inferred that the anti-CD16a single-chain antibody portion of the fully immune antibody played a key role in this tumor-killing effect.

[0497] 23.2 Ascites samples from patients with Her2-negative ovarian cancer

[0498] Ascites samples from Her2-negative ovarian cancer were collected and cells in the samples, including tumor cells and immune cells, were isolated. Candidate and control drugs were co-incubated with tumor cell and immune cell complexes in vitro. After 4-5 days of incubation, the culture supernatant was collected to detect tumor cell granzyme B and IFN-γ, and tumor cells (EpCAM+) and immune cells were analyzed. The results are shown in Figure 26 The immune cell phenotype of this sample ( Figure 26 -A): NK cells 0.5%; macrophages 10.74% (M1 0.41%, M2 0.05%); CD3 + T cells 5.98%; CD4 + T cells 4.36%; CD8 + T cells were 0.97%. The results showed that compared with the non-administered group, the anti-PD-1 antibody had no significant tumor cell killing function, while the anti-Her2 complete immune antibody group or the Herceptin and TTI-622 combination group had significantly improved tumor cell killing ability (about 2.5 times), while the combination of anti-Her2 complete immune antibody and anti-PD-1 antibody did not further improve the efficacy ( Figure 26 -B). Anti-Her2 complete immune antibody or anti-PD-1 antibody cannot increase granzyme B ( Figure 27 -C) or IFN-γ( Figure 27The anti-Her2 fully immune antibody combined with the anti-PD-1 antibody can significantly increase the secretion of granzyme B or IFN-g. Since the NK cell proportion of this patient is 0.5% (significantly lower than the normal level of peripheral blood), the macrophage proportion is 10.74% (within the normal level of peripheral blood), and the combination of Herceptin and TTI-622 shows significant tumor killing function, it can be inferred that the SIRPa part in the fully immune antibody (blocking the "don't eat me" signal to activate macrophages) plays a key role in tumor killing.

[0499] Example 24 Organ toxicity study of fully immune antibody with anti-PD-L1 antibody as a skeleton in vivo in mice

[0500] The 7-week-old Balb / c mice meeting the test conditions were grouped, a total of 9 groups, 3 animals in each group, intravenous injection was performed on Day 0 and Day 4, and daily observation was performed on the clinical state of the animals, including mental state, activity, diet, and water, etc. until the animals were euthanized. On Day 7, the mice were euthanized with carbon dioxide, and the liver, spleen, and kidney were weighed. The results are shown in Table 1 Figure 27 . Spleen weight: The fully immune antibody TriAtezo06 (containing cytokine IL-15) at high dose (16.99 mg / kg) and low dose (8.49 mg / kg) did not significantly change the spleen weight of mice after two administrations, while the Fc-IL-15 administration group significantly increased the spleen weight of mice; the fully immune antibody TriAtezo12 (containing cytokine IL-2) slightly increased the spleen weight after two administrations, while the Fc-IL-2 significantly increased the spleen weight after two administrations Figure 27 -A). Liver weight: The fully immune antibody TriAtezo06 (containing cytokine IL-15) at high dose (16.99 mg / kg) and low dose (8.49 mg / kg) and the fully immune antibody TriAtezo12 (containing cytokine IL-2) at high dose (17.33 mg / kg) and low dose (8.66 mg / kg) did not significantly change the liver weight of mice after two administrations, and the Fc-IL-15 administration group did not significantly change the liver weight of mice; while the high dose group (5.63 mg / kg) of Fc-IL-2 significantly increased the liver weight after two administrations https: / / www.gilead.com / news-and-press / company-statements / gilead--B). Kidney weight: The kidney weights of mice in the high-dose (16.99 mg / kg) and low-dose (8.49 mg / kg) complete immune antibody TriAtezo06 (containing the cytokine IL-15) and the high-dose (17.33 mg / kg) and low-dose (8.66 mg / kg) complete immune antibody TriAtezo12 (containing the cytokine IL-2) did not change significantly after two administrations, and the kidney weights of mice in the Fc-IL-15 administration group also did not change significantly; while the kidney weights of mice in the high-dose (5.63 mg / kg) and low-dose (2.82 mg / kg) Fc-IL-2 groups increased significantly after two administrations statement-on-the-discontinuation-of-magrolimab-study-in-aml-with-tp53- -C). These organ toxicity results show that the complete immune antibodies TriAtezo06 and TriAtezo12 both exhibit good safety compared with wild-type cytokines.

[0501] It should be noted that the above only illustrates the technical idea of the present application and cannot limit the protection scope of the present application. For those skilled in the art, without departing from the principles of the present application, a number of improvements and refinements can be made, which fall within the scope of protection of the claims of the present application.

[0502] References:

[0503] [1] Danyang Li, Minghua Wu. Pattern recognition receptors in health and diseases. Signal Transduction and Targeted Therapy. 6: 291 (2021)

[0504] [2] Liu YJ. IPC: professional type 1 interferon-producing cells and plasmacytoid dendritic cell precursors. Annu Rev Immunol. 2005; 23: 275-306

[0505] [3] A. Ribas, I. Mehmi, T. Medina, C. Lao et al. Phase lb / ll study of the combination of SD-101 and pembrolizumab in patients with advanced melanoma who had progressive disease on or after prior anti-PD-1 therapy. Annals of Oncology. Volume 29, Supplement 8, October 2018.

[0506] [4] Yoji Murata, Yasuyuki Saito, Takenori Kotani, Takashi Matozaki. Blockade of CD47 or SIRPa: a new cancer immunotherapy. Expert Opinion on Therapeutic Targets.

[0507] [5] mutations https: / / www.affimed.com / affimed-receives-fda-fast-track- designation-for-afm13-in-combination-with-allonk-for-the-treatment-of-

[0508] [6] Cai Zhang, Yuan Hu, Chongdeng Shi. Targeting Natural Killer Cells for Tumor Immunotherapy. Frontiers in Immunology. Article 60 Volume 11 February 2020.

[0509] [7] patients-with-relapsed-or-refractory-hodgkin-lymphoma / ​ ​

[0510] [8] Yawu Jing, Zhenya Ni, Jianming Wu, LeeAnn Higgins et al. Identification of an ADAM17 cleavage region in human CD16 (FcyRIII) and the engineering of a non-cleavable version of the receptors in NK cells.

Claims

1. An innate immune super connector (ISE) comprising Recruit or activate innate immune cells, the innate immune super connector comprises two or more than two components: (a) anti-CD16a antibody, (b) SIRPαD1 protein, (c) immune cytokine, (d) pattern recognition receptor agonist.

2. A complete immune antibody (CIA) characterized in that Recruit or activate innate and adaptive immune cells to kill tumor cells, the complete immune antibody comprises (a) the ISE or its fragment of claim 1, (b) an adaptive immune antibody or a tumor associated antigen antibody.

3. The innate immunity superconnector of claim 1, wherein, Take the traditional antibody as the basic structure, remove the variable region of the antibody or use the antibody without targeting in the human body, connect the anti-CD16a antibody or the SIRPαD1 protein through the linker at the N terminal or C terminal of the antibody heavy chain constant region, and connect the SIRPαD1 protein or the anti-CD16a antibody through the linker at the N terminal or C terminal of the antibody light chain constant region.

4. The innate immunity superconnector of claim 1, wherein, Take the traditional antibody as the basic structure, remove the variable region of the antibody or use the antibody without targeting in the human body, connect one or more of human immune cytokine, human immune cytokine receptor, SIRPαD1 protein and anti-CD16a antibody through the linker at the N terminal and / or C terminal of the antibody heavy chain constant region and / or the antibody light chain constant region.

5. The innate immunity superconnector of any one of the preceding claims, wherein, Connect the pattern recognition receptor agonist at the N terminal or C terminal of the ISE through the linker, which includes but is not limited to Toll-like receptor (TLRs) agonist, NOD-like receptor (NLRs) agonist, RIG-I-like receptor (RLRs) agonist, C-type lectin receptor (CLRs) agonist, melanoma deficiency factor 2 (AIM2) like receptor (ALRs) agonist.

6. The innate immunity superconnector of any one of the preceding claims, wherein, Connect human IL-2 or IFNα or its mutant at the N terminal or C terminal of the ISE through the linker.

7. The fully human antibody of claim 2, wherein Take the ISE as the basic structure, add the adaptive immune antibody variable region or the tumor associated antigen antibody variable region at the N terminal of the ISE.

8. The fully human antibody of claim 7, wherein, Take the adaptive immune antibody or the tumor associated antigen antibody as the basic skeleton, connect one or more of human immune cytokine, human immune cytokine receptor, SIRPαD1 protein and anti-CD16a antibody through the linker at the N terminal and / or C terminal of the antibody heavy chain and / or the antibody light chain.

9. The fully humanized antibody according to any one of the preceding claims, characterized in that, The tumor associated antigen is for example but not limited to PD-L1, Her2, VEGF, EGFR, CD38, Claudin18.2, Claudin6, CD19, MSLN, Trop-2, CD20, Nectin-4, c-Met, MUC16, MUC17, 5T4, BCMA, DLL3 or CD30.

10. The fully human antibody of any one of the preceding claims, wherein, The adaptive immune antibody is the antibody targeting for example but not limited to the following target points: PD-1, CTLA-4, B7-H4, B7-H3, 4-1BB, LAG-3, TIGIT, TIM-3, LILRB2, CD24, CD40 or CD40L.

11. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The human immune cytokine is for example but not limited to IL-15, IL-18, IL-2, IL-7, IL-10 or IL-12.

12. The innate immunity superconnector of claim 11, wherein, The anti-CD16a antibody is an anti-CD16a single-chain antibody or an anti-CD16a nanobody.

13. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The heavy chain variable region of the anti-CD16a single-chain antibody comprises CDR1 shown in SEQ ID NO: 1, CDR2 shown in SEQ ID NO: 2 and CDR3 shown in SEQ ID NO: 3; and the light chain variable region comprises CDR1 shown in SEQ ID NO: 4, CDR2 shown in SEQ ID NO: 5 and CDR3 shown in SEQ ID NO:

6.

14. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The heavy chain variable region of the anti-CD16a single-chain antibody comprises the amino acid sequence shown in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:

8.

15. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The anti-CD16a single-chain antibody comprises the amino acid sequence shown in SEQ ID NO:

9.

16. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The anti-CD16a nanobody comprises CDR1 shown in SEQ ID NO: 10, CDR2 shown in SEQ ID NO: 11 and CDR3 shown in SEQ ID NO:

12.

17. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The anti-CD16a nanobody comprises the amino acid sequence shown in SEQ ID NO:

13.

18. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The SIRPαD1 protein comprises the amino acid sequence shown in SEQ ID NO:

14.

19. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The human IL-15 protein fragment comprises the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 16, and the human IL-15Rα protein fragment comprises the amino acid sequence shown in SEQ ID NO:

17.

20. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The pattern recognition receptor agonist is CpG ODN, preferably, the CpG ODN comprises the sequence shown in SEQ ID NO:

117.

21. The innate immunity superconnector of claim 5, wherein, The IL-2 mutant comprises the amino acid sequence shown in SEQ ID NO: 118, and / or the IFNα mutant comprises the amino acid sequence shown in SEQ ID NO:

119.

22. The innate immunity superconnector of claim 6, wherein, The basic framework is a non-targeting antibody in human body, such as an anti-hen egg lysozyme (anti-HEL) antibody, an anti-pathogenic microorganism antibody such as an anti-virus, bacterial antibody, etc. The anti-CD16a antibody is an anti-CD16a single-chain antibody or an anti-CD16a nanobody.

23. The innate immunity superconnector of any one of the preceding claims, wherein, The heavy chain variable region of the anti-CD16a single-chain antibody comprises CDR1 shown in SEQ ID NO: 1, CDR2 shown in SEQ ID NO: 2 and CDR3 shown in SEQ ID NO: 3; and the light chain variable region comprises CDR1 shown in SEQ ID NO: 4, CDR2 shown in SEQ ID NO: 5 and CDR3 shown in SEQ ID NO:

6.

24. The innate immunity superconnector of claim 23, wherein, The heavy chain variable region of the anti-CD16a single-chain antibody comprises the amino acid sequence shown in SEQ ID NO: 7, and the light chain variable region comprises the amino acid sequence shown in SEQ ID NO:

8. The anti-CD16a single-chain antibody comprises the amino acid sequence shown in SEQ ID NO:

9. The anti-CD16a nanobody comprises CDR1 shown in SEQ ID NO: 10, CDR2 shown in SEQ ID NO: 11 and CDR3 shown in SEQ ID NO:

12. The anti-CD16a nanobody comprises the amino acid sequence shown in SEQ ID NO:

13. The SIRPαD1 protein comprises the amino acid sequence shown in SEQ ID NO:

14. The human IL-15 protein fragment comprises the amino acid sequence shown in SEQ ID NO: 15 or SEQ ID NO: 16, and the human IL-15Rα protein fragment comprises the amino acid sequence shown in SEQ ID NO:

17. The pattern recognition receptor agonist is CpG ODN, preferably, the CpG ODN comprises the sequence shown in SEQ ID NO:

117. The IL-2 mutant comprises the amino acid sequence shown in SEQ ID NO: 118, and / or the IFNα mutant comprises the amino acid sequence shown in SEQ ID NO:

119. The basic framework is a non-targeting antibody in human body, such as an anti-hen egg lysozyme (anti-HEL) antibody, an anti-pathogenic microorganism antibody such as an anti-virus, bacterial antibody, etc. The anti-CD16a antibody is an anti-CD16a single-chain antibody or an anti-CD16a nanobody.

25. The innate immunity superconnector of claim 24, wherein, The long chain amino acid sequence comprises SEQ ID NO: 20, the short chain amino acid sequence comprises SEQ ID NO: 21, or the long chain amino acid sequence comprises SEQ ID NO: NO:22, the short chain amino acid sequence comprises SEQ ID NO:23, or the long chain amino acid sequence comprises SEQ ID NO:24, the short chain amino acid sequence comprises SEQ ID NO:25, or the long chain amino acid sequence comprises SEQ ID NO:26, the short chain amino acid sequence comprises SEQ ID NO:27, or the long chain amino acid sequence comprises SEQ ID NO:28, the amino acid sequence of the short chain comprises SEQ ID NO:

29.

26. The innate immunity superconnector of any one of the preceding claims, wherein, It is ISE-A, ISE-B, ISE-C, ISE-D, ISE-E, ISE-F, ISE-G, ISE-H, ISE-I or ISE-J configuration, preferably, the amino acid sequence of the long chain of the innate immunity connector ISE-A comprises SEQ ID NO: 30 or SEQ ID NO: 32, the amino acid sequence of the short chain comprises SEQ ID NO: 31 or SEQ ID NO: 33, the amino acid sequence of the long chain of the innate immunity connector ISE-B comprises SEQ ID NO: 34 or SEQ ID NO: 36, the amino acid sequence of the short chain comprises SEQ ID NO: 35 or SEQ ID NO: 37, the amino acid sequence of the long chain of the innate immunity connector ISE-C comprises SEQ ID NO: 38 or SEQ ID NO:40, short chain amino acid sequence comprising SEQ ID NO:39 or SEQ ID NO:41, innate immunity connector The long chain amino acid sequence of ISE-D comprises SEQ ID NO: 42 or SEQ ID NO: 44, the short chain amino acid sequence comprises SEQ ID NO: 43 or SEQ ID NO: 45, and the long chain amino acid sequence of the innate immunity connector ISE-E comprises SEQ ID NO: 46 or SEQ ID NO: 48, the short chain amino acid sequence comprises SEQ ID NO: 47 or SEQ ID NO:49, the amino acid sequence of the long chain of the innate immunity connector ISE-F comprises SEQ ID NO:50 or SEQ ID NO:52, the short chain amino acid sequence comprises SEQ ID NO:51 or SEQ ID NO:53, the long chain amino acid sequence of the innate immunity connector ISE-G comprises SEQ ID NO:54 or SEQ ID NO:56, the short chain amino acid sequence comprises SEQ ID NO:55 or SEQ ID NO:57, the long chain amino acid sequence of the innate immunity connector ISE-H comprises SEQ ID NO:58 or SEQ ID NO:

59. NO: 58, the amino acid sequence of the long chain of the innate immunity connector ISE-I comprises SEQ ID NO: 61 or SEQ ID NO: 63, the amino acid sequence of the short chain comprises SEQ ID NO: 62 or SEQ ID NO: 64, the amino acid sequence of the long chain of the innate immunity connector ISE-J comprises SEQ ID NO: 65, the amino acid sequence of the short chain comprises SEQ ID NO: 66 or SEQ ID NO:

67.

27. The fully immunized antibody of claim 7, wherein The amino acid sequence of the long chain is introduced with the heavy chain variable region of the tumor associated antigen antibody or the postnatal immune antibody on the basis of the amino acid sequence of the ISE, and the amino acid sequence of the short chain is introduced with the light chain variable region of the tumor associated antigen antibody or the postnatal immune antibody on the basis of the amino acid sequence of the ISE.

28. The fully human antibody of claim 27, wherein, TriAtezo01, TriAtezo02, TriAtezo03, TriAtezo04, TriAtezo05, TriAtezo06, TriAtezo07, TriAtezo12, TriCetu03, TriCetu04 and TriCetu12, the long chain of which comprises the amino acid sequence of SEQ ID NO: 68, SEQ ID NO: 70, SEQ ID NO: 71, SEQ ID NO: 72, SEQ ID NO: 74, SEQ ID NO: 76, SEQ ID NO: 77, SEQ ID NO: 78, SEQ ID NO: 79, SEQ ID NO: 81 and SEQ ID NO: 120, respectively, the short chain of which comprises the amino acid sequence of SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 69, SEQ ID NO: 73, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 75, SEQ ID NO: 80, SEQ ID NO: 80 and SEQ ID NO: 80, respectively, and / or the fully immune antibody is TriKey06, TriKey12, TriDaratu06, TriDaratu12, TriRituxi06, TriRituxi12, TriZolbetu06, TriZolbetu12, TriSacitu06, TriSacitu12, TriTrastu06, TriTrastu12, TriBeva06, TriBeva12, TriEnfortu06, TriEnfortu12, TriBelanta06, TriBelanta12, TriIpilimu06 and TriIpilimu12, the long chain of which comprises the amino acid sequence of SEQ ID NO: 86, SEQ ID NO: 88, SEQ ID NO: 89, SEQ ID NO: 91, SEQ ID NO: 92, SEQ ID NO: 94, SEQ ID NO: 95, SEQ ID NO: 97, SEQ ID NO: 98, SEQ ID NO: 100, SEQ ID NO: 110, SEQ ID NO: 112, SEQ ID NO: 113, SEQ ID NO: 115, SEQ ID NO: 107, SEQ ID NO: 109, SEQ ID NO: 104, SEQ ID NO: 106, SEQ ID NO: 101 and SEQ ID NO: 103, respectively.The short chain amino acid sequences comprise SEQ ID NO: 87, SEQ ID NO: 87, SEQ ID NO: 90, SEQ ID NO: 90, SEQ ID NO: 93, SEQ ID NO: 93, SEQ ID NO: 96, SEQ ID NO: 96, SEQ ID NO: 99, SEQ ID NO: 99, SEQ ID NO: 111, SEQ ID NO: 111, SEQ ID NO: 114, SEQ ID NO: 114, SEQ ID NO: 108, SEQ ID NO: 108, SEQ ID NO: 105, SEQ ID NO: 105, SEQ ID NO: 102, and SEQ ID NO: 102, respectively.

29. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The Fc fragment of the antibody is introduced with the L234A and L235A mutations, or further introduced with the M252Y, S254T and T256E mutations in the Fc fragment.

30. The innate immunity superconnector or fully immune antibody of any one of the preceding claims, wherein, The linker includes, but is not limited to, PSGQAGAAASESLFVSNHAY or (G4S) n wherein n is 1, 2, 3, 4, 5, 6, 7, 8, 9, or 10.

31. A nucleic acid molecule encoding the innate immunity super connector or the complete immune antibody of any one of the preceding claims.

32. A vector comprising the nucleic acid molecule of claim 31.

33. A cell comprising the nucleic acid molecule of claim 31 or the vector of claim 32.

34. A method for preparing the innate immunity super connector or the complete immune antibody of any one of the preceding claims, the method comprising the following steps: (1) amplifying the two chains of the ISE or the CIA respectively by using the PCR technology; introducing the mutations L234A, L235A, M252Y, S254T and T256E in the Fc fragment; (2) cloning the DNA fragments obtained in step (1) into the pcDNA series of vectors or other vectors for mammalian cell expression system to obtain a recombinant vector; (3) transfecting the recombinant vector plasmid obtained in step (2) into mammalian cells for expression of the recombinant protein, and obtaining the purified target protein after affinity chromatography.

35. Use of the innate immunity super connector or the complete immune antibody of any one of the preceding claims in the preparation of a medicament for treating solid tumors and hematological tumors in a subject.

36. The use according to claim 35, wherein the solid tumors and hematological tumors include but are not limited to respiratory tract tumors, digestive tract tumors, reproductive system tumors, nervous system tumors, urinary system tumors, skin tumors, head and neck tumors, blood tumors, preferably, the solid tumors and hematological tumors include but are not limited to nasopharyngeal carcinoma, laryngeal carcinoma, lung cancer, esophageal cancer, gastric cancer, colorectal cancer, prostate cancer, ovarian cancer, testicular cancer, brain tumor, glioma, renal cancer, bladder cancer, ureter cancer, epithelial tumor, melanoma, ocular tumor, nasal tumor, ear tumor, oral tumor, leukemia, multiple myeloma, lymphoma, breast cancer, epidermal cancer. ​

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