Chimeric antigen receptor targeting CD33

Abstract

This invention provides a chimeric antigen receptor (CAR) for the treatment of CD33-related disorders, and a method for treating subjects with CD33-related disorders. [Solution] A chimeric antigen receptor is provided, comprising an extracellular component including a binding domain having a complementarity-determining region set of 1E6, 1H10, 1A9, 1D2, 1B9, 1H8, 2D3, or 2E3, an intracellular component including an effector domain, and a transmembrane domain linking the extracellular component to the intracellular component, wherein, in one embodiment, the binding domain includes a single-stranded variable fragment.

Description

[Technical Field]

[0001] This application claims priority to U.S. Provisional Patent Application No. 63 / 003,196, filed on 31 March 2020, the entirety of which is incorporated herein by reference.

[0002] Statement on federally supported research or development This invention was made with government support under CA234203 and CA245594 granted by the National Institutes of Health. The government has certain rights to this invention.

[0003] Areas of this disclosure This disclosure provides a novel set of chimeric antigen receptors (CARs) having binding domains derived from human CD33-binding antibodies, which may include optimized short spacer regions and intermediate spacer regions. This disclosure also provides a method for a cell expansion / activation process utilizing IL-2, IL-7, IL-15, and / or IL-21 to improve cell proliferation and cytolysis of the described CARs. [Background technology]

[0004] According to the World Health Organization, cancer is the second leading cause of death globally, accounting for an estimated 9.6 million deaths in 2018. Acute myeloid leukemia (AML) is a type of cancer caused by malignant tumors of clonal proliferative myeloblastic cells. In the United States, there are 20,000 new cases of AML annually, and 11,000 people die from AML each year (Siegel et al., 2021, CA Cancer J Clin. 71(1):7-33). While young patients with AML can achieve high complete remission rates of 60% to 80% with conventional chemotherapy (Dohner et al., 2017, Blood. 129(4):424-447), treatment outcomes for elderly patients aged 65 and over remain inadequate, with as many as 70% dying from the disease within one year of diagnosis (Meyers et al., Appl Health Economic Health Policy, 11:275-286, 2013). Unfortunately, due to the chemotherapy resistance of leukemia stem cells, relapse after conventional treatment is common (Eppert et al., 2011, Nat. Med. 17(9): pp. 1086-1093), and current treatment options for relapsed / refractory (R / R) AML are poor, resulting in a 12-month overall survival rate of less than 30%.

[0005] For many years, the treatment of choice for cancer has been surgery, chemotherapy, and / or radiation therapy. In recent years, many targeted therapies have emerged that specifically target cancer cells by identifying and utilizing specific molecular and / or immunophenotypic changes that are primarily found in cancer cells. For example, many cancer cells selectively express certain markers on their cell surface, and these markers have become targets for antibody-based therapies.

[0006] CD33 is a member of the sialic acid-binding immunoglobulin-like lectin (SIGLEC) protein family. It is a 67 kDa glycosylated transmembrane protein. CD33 (also known as SIGLEC-3) is a myeloid differentiation antigen found in at least some leukemia cells, and possibly in some cases in AML stem cells, in almost all patients with AML. Based on this widespread expression pattern, CD33 has been widely pursued as a therapeutic target in AML. Recent data from several randomized studies have demonstrated that gemtuzumab ozogamicin (GO), a CD33 antibody-drug conjugate, improves survival when added to chemotherapy in a defined subset of patients with newly diagnosed AML. This data demonstrates CD33 as the first (and so far only) target for immunotherapy in AML. In parallel with the development of new and more effective CD33-targeted therapies (e.g., antibody-drug conjugates, radioactive immunoconjugates, bispecific antibodies, chimeric antigen receptor [CAR]-modified T cells) to overcome the limitations pointed out by GO, there is growing interest in CD33 as a drug target for other malignant and non-malignant disorders. These efforts include targeting CD33 splice variants not recognized by GO, as well as targeting CD33+ tumor cells in other hematological malignancies, CD33+ myeloid-derived suppressor cells (MDSCs) in various diseases, and normal CD33+ microglia in Alzheimer's disease (Walter, Expert Opin Biol Ther. 2020, 20(9):955-958).

[0007] Full-length CD33 protein (CD33 FL CD33 is characterized by a membrane-distal V-set immunoglobulin (Ig)-like domain and a membrane-proximal C2-set Ig-like domain at the amino terminus of its extracellular portion (Figure 2). Shorter isoforms of CD33 exist. One variant of CD33 lacks exon 2, which encodes the V-set domain (CD33 ΔE2) is included. At least at the mRNA level, CD33 ΔE2 CD33 is widely expressed in bone marrow and peripheral blood myeloid cells of patients with AML. However, almost all commercially and clinically available CD33 antibodies currently recognize the immunodominant V-set Ig-like domain. This means that these antibodies do not recognize CD33 ΔE2 This means that shorter forms of CD33, such as those lacking the V-set domain, will not be recognized. ΔE2 Selective transcription and CD33 FL This can explain the observation made in one clinical trial of pediatric AML in that patients with a single nucleotide polymorphism in the CD33 gene, which results in reduced translation, did not benefit from the addition of GO (which also binds to the V set domain of CD33) to intensive chemotherapy.

[0008] Significant progress has been made in genetically engineering T cells of the immune system to target and kill undesirable cell types, such as cancer cells, beyond antibody-based therapies. Many of these T cells are genetically engineered to express chimeric antigen receptors (CARs). CARs are proteins containing several distinct subcomponents that enable genetically modified T cells to recognize and kill cancer cells. These subcomponents include at least an extracellular component and an intracellular component. The extracellular component includes a binding domain that specifically binds to markers selectively present on the surface of undesirable cells (e.g., CD33). Once the binding domain binds to such a marker, the intracellular component signals the T cell to destroy the bound cell. CARs further include a transmembrane domain that can link the extracellular component to the intracellular component.

[0009] Other minor components that can enhance the function of CARs may also be used. For example, spacer regions can provide additional structural flexibility to the CAR and often increase the ability of the binding domain to bind to the targeted cellular marker. The appropriate length of spacer regions within a particular CAR may depend on many factors, including how close or far the targeted marker is located from the undesirable cell membrane surface.

[0010] When performed ex vivo, T cell gene modification can involve numerous cell manipulation steps, and it has been observed that different manipulation conditions can affect the cancer cell-killing properties of the cells. Therefore, when designing CARs and genetically modifying cells to express them, many considerations must be taken into account, including the targeted cell markers, the presence and / or length of spacers, and the ex vivo manipulation procedure. [Prior art documents] [Non-patent literature]

[0011] [Non-Patent Document 1] Siegel et al., 2021, CA Cancer J Clin.71(1):7-33 [Non-Patent Document 2] Dohner et al., 2017. Blood. 129(4): 424-447. [Non-Patent Document 3] Meyers et al., Appl Health Economic Health Policy, 11: pp. 275-286, 2013. [Non-Patent Document 4] Eppert et al., 2011. Nat. Med. 17(9): 1086-1093. [Non-Patent Document 5] Walter, Expert Opin Biol Ther. 2020, 20(9):955-958. [Overview of the project] [Problems that the invention aims to solve]

[0012] (Summary of the Disclosure) The present disclosure provides chimeric antigen receptors (CARs) for the treatment of CD33-related disorders. The CARs include binding domains derived from a new series of anti-CD33 antibodies. In certain embodiments, the CARs include binding domains that bind CD33 regardless of the CD33 variant (CD33 FL or CD33 ΔE2 ) expressed by the patient. These CD33 binding domains are referred to as "pan" binders. In certain embodiments, the pan binder binds to the membrane-proximal C2-set Ig-like domain of CD33 (see Figure 2). In certain embodiments, these pan binders are derived from the antibodies: 1H10, 1A9, 1E6, 1D2 and 1B9 and can include single-chain variable fragments of these antibody binding domains. Additional newly developed CD33-targeting antibodies disclosed herein bind to the V-set domain of CD33. These antibodies include 1H8, 2D3 and 2E3, and provide additional CAR-based treatment options for patients expressing CD33 FL .

[0013] In certain embodiments, the present disclosure provides CARs having a short spacer region or an intermediate spacer region. In certain embodiments, the short spacer region includes the hinge region (12 amino acids) of IgG4. In certain embodiments, the intermediate spacer region includes the hinge region and CH3 domain of IgG4 (a total of 131 amino acids).

[0014] In certain embodiments, the present disclosure utilizes a combination of cytokines IL-7, IL-15 and IL-21 to provide expansion and activation of T cells genetically modified to express the CARs disclosed herein. In certain embodiments, the present disclosure utilizes a combination of cytokines including IL-2 to provide expansion and activation of T cells genetically modified to express the CARs disclosed herein.

[0015] The CARs disclosed herein may be used to treat acute myeloid leukemia (AML) and other CD33+ disorders. [Brief explanation of the drawing]

[0016] [Figure 1] This figure shows that CD33 is overexpressed in acute myeloid leukemia (AML) compared to hematopoietic stem cells, due to the expression of CD33 in AML and normal hematopoietic cells. Relative expression data matched from bloodspot.eu. HSPC: hematopoietic stem cells / progenitor cells; MPP: pluripotent progenitor cells. ****p<0.0001, ns are not significant by multiple t-tests using the Benjamini, Krieger, and Yekutieli two-step linear elevation procedure at Q=1%. [Figure 2] This figure shows CD33 (CD33ΔE2) with exon 2 deletion, resulting in the deletion of the full-length CD33 (CD33FL) and the V set domain. Antibodies bound to CD33FL only (anti-CD33FL), CD33ΔE2 only (anti-CD33ΔE2), or CD33FL and CD33ΔE2 (anti-CD33FL+ΔE2 or anti-CD33PAN) are shown. [Figure 3]This is a schematic diagram of an artificial CD33 molecule (CD33ΔE3-4) with deletions of exons 3 and 4 resulting in membrane-proximal rearrangement of the CD33FL and V set domains, or an insertion of two C2 set domains of CD22 ("CD33FL+CD22 2D") or an insertion of four C2 set domains of CD22 ("CD33FL+CD22 4D"). CD33ΔE3-4 was engineered using site-directed mutagenesis to splice out CD33 amino acids (aa) 140-232 of the human CD33FL extracellular domain (ECD). CD33FL+CD22 4D was generated using endogenous CD33 signal peptides (aa1-17), a 6-histidine tag, a 3x glycine linker, human CD33 ECD (aa18-259), a portion of human CD22 ECD including C2 type domains 3-6 (aa331-683), the CD33 transmembrane domain, and the CD33 intracellular domain (aa260-364). CD22 aa331-504 (C2 type domains 3 and 4) were removed from CD33FL+CD22 24 to generate CD33FL+CD22 2D. [Figure 4]This diagram illustrates the cloning strategy for generating CD33 CAR expression constructs. (i) Supernatants from antibody-secreting mouse hybridoma cells were screened for pan-CD33 binding activity, then for the lack of binding to CD33 null cell lines, and the best individual hybridomas were selected. Antibody isotyping was performed, RNA was extracted, and used for rapid amplification (RACE) amplification (Takara) of cDNA ends and subsequent isotype-specific polymerase chain reaction (PCR) based amplification. (ii) Cloning of antibody variable region sequences was performed using either pRACE (Takara) or TOPO (Invitrogen) standard cloning vectors. (iii) Plasmid DNA was purified from individual bacterial colonies and Sanger DNA sequencing was performed to obtain at least three individual identical cDNA sequences corresponding to each antibody variable region for both the heavy and light chains of the antibody. (iv) The antibody variable region cDNA sequences were translated into amino acid sequences using the ExPASy.org translation tool. (v) The amino acid sequences for each individual antibody variable region were submitted to the Integrated DNA Technologies (IDT, Coralville, IA) website for human codon optimization. (vi) The codon-optimized single-stranded variable region (scFv) nucleic acid sequences were then combined with nucleic acid sequences contained within the CAR lentiviral backbone-41BB-3z-T-CD19t, specifically the sequence within the EF1 promoter adjacent to the 5' end of the granulocyte-macrophage colony-stimulating factor receptor (GM-CSFR) signal peptide, which contains the GM-CSFR signal peptide, and then the linker sequence derived from the CAR lentiviral backbone-41BB-3z-T-CD19t, were added to the 5' of the antibody HC variable region, followed by the flexible Gly-Ser linker sequence, then the antibody LC variable region, and then the linker sequence derived from the CAR lentiviral backbone-41BB-3z-T-CD19t. (vii) The complete nucleotide sequences were then submitted to IDT for the synthesis of gBlocks. (viii) The gBlocks were cloned into TOPO and their nucleic acid sequences were confirmed by Sanger DNA sequencing.(ix) All gBlocks were generated to contain a universal priming sequence, CAR_universal_forward primer, and CAR_universal_reverse primer. (x) Next, using a suitable TOPO vector as a template, DNA polymerase was calibrated and PCR amplification was performed using the CAR_universal_forward primer and a reverse primer selected from two options: CAR_universal_rev_sh or CAR_universal_rev_int. In these primers, rev indicates reverse, sh indicates a short linker, and int indicates an intermediate-length linker within the CAR construct. (xi) The recipient plasmid (lentiviral skeleton with sh or int linker - 41BB-3z-T-CD19t) was digested with RsrII / NheI, the plasmid was cleaved, and the PCR product was gel-purified. (xii) Gibson assembly (NEB) was performed using the RsrII / NheI-digested recipient plasmid (lentiviral skeleton with sh or int linker - 41BB-3z-T-CD19t) and a suitable PCR amplicon. (xiii) Sanger DNA sequencing was used to verify the scFv and linker contained in each plasmid. (vii) Maxiprep and lentiviral packaging were then performed. [Figure 5A]This figure shows the enhanced antitumor effect of CD33 / CD3 BsAb on human myeloid leukemia cells by reducing the binding distance from the cell membrane. Human CD33+ myeloid leukemia cell lines ((5A)ML-1, (5B)HL-60, (5C)K562) with CRISPR / Cas9-mediated deletion of the endogenous CD33 locus were engineered to overexpress either CD33FL or CD33ΔE3-4 via lentiviral gene transfer. Relative expression of the target protein was assessed by flow cytometry with the V-set domain CD33 antibody, P67.6, and representative histograms are shown in the lower right panel. Cells were then treated with V-set domain targeting CD33 / CD3 BsAb at a concentration of 1000 pg / mL and healthy donor T cells enriched from unstimulated peripheral blood mononuclear cells collected from healthy adult volunteers with the shown effector:target (E:T) cell ratio (upper panel). Bone marrow cells were also treated with gemtuzumab ozogamicin (GO) at the indicated concentrations (lower left panel). Cytotoxicity was quantified by flow cytometry as the change in percentage of dead cells measured by 4',6-diamidino-2-phenylindole (DAPI) staining at 2 days (for BsAb) or 3 days (for GO). Anti-V set domain-directed CD33 / CD3 BsAb was constructed in scFv-scFv format using constructs referred herein as RC1 or A3, utilizing the sequence shown in Sequence ID No. 254 and described in U.S. Patent Application Publication US2016 / 0317657 A1. *p<0.05;**p<0.01;***p<0.001. [Figure 5B]This figure shows the enhanced antitumor effect of CD33 / CD3 BsAb on human myeloid leukemia cells by reducing the binding distance from the cell membrane. Human CD33+ myeloid leukemia cell lines ((5A)ML-1, (5B)HL-60, (5C)K562) with CRISPR / Cas9-mediated deletion of the endogenous CD33 locus were engineered to overexpress either CD33FL or CD33ΔE3-4 via lentiviral gene transfer. Relative expression of the target protein was assessed by flow cytometry with the V-set domain CD33 antibody, P67.6, and representative histograms are shown in the lower right panel. Cells were then treated with V-set domain targeting CD33 / CD3 BsAb at a concentration of 1000 pg / mL and healthy donor T cells enriched from unstimulated peripheral blood mononuclear cells collected from healthy adult volunteers with the shown effector:target (E:T) cell ratio (upper panel). Bone marrow cells were also treated with gemtuzumab ozogamicin (GO) at the indicated concentrations (lower left panel). Cytotoxicity was quantified by flow cytometry as the change in percentage of dead cells measured by 4',6-diamidino-2-phenylindole (DAPI) staining at 2 days (for BsAb) or 3 days (for GO). Anti-V set domain-directed CD33 / CD3 BsAb was constructed in scFv-scFv format using constructs referred herein as RC1 or A3, utilizing the sequence shown in Sequence ID No. 254 and described in U.S. Patent Application Publication US2016 / 0317657 A1. *p<0.05;**p<0.01;***p<0.001. [Figure 5C]This figure shows the enhanced antitumor effect of CD33 / CD3 BsAb on human myeloid leukemia cells by reducing the binding distance from the cell membrane. Human CD33+ myeloid leukemia cell lines ((5A)ML-1, (5B)HL-60, (5C)K562) with CRISPR / Cas9-mediated deletion of the endogenous CD33 locus were engineered to overexpress either CD33FL or CD33ΔE3-4 via lentiviral gene transfer. Relative expression of the target protein was assessed by flow cytometry with the V-set domain CD33 antibody, P67.6, and representative histograms are shown in the lower right panel. Cells were then treated with V-set domain targeting CD33 / CD3 BsAb at a concentration of 1000 pg / mL and healthy donor T cells enriched from unstimulated peripheral blood mononuclear cells collected from healthy adult volunteers with the shown effector:target (E:T) cell ratio (upper panel). Bone marrow cells were also treated with gemtuzumab ozogamicin (GO) at the indicated concentrations (lower left panel). Cytotoxicity was quantified by flow cytometry as the change in percentage of dead cells measured by 4',6-diamidino-2-phenylindole (DAPI) staining at 2 days (for BsAb) or 3 days (for GO). Anti-V set domain-directed CD33 / CD3 BsAb was constructed in scFv-scFv format using constructs referred herein as RC1 or A3, utilizing the sequence shown in Sequence ID No. 254 and described in U.S. Patent Application Publication US2016 / 0317657 A1. *p<0.05;**p<0.01;***p<0.001. [Figure 6]This figure illustrates the enhanced antitumor effect of CD33 / CD3 BsAb on human acute lymphoblastic leukemia cells engineered to express the CD33 protein by reducing the binding distance from the cell membrane. Human CD33neg acute lymphoblastic leukemia (ALL) cell line RS4;11 was engineered to overexpress either CD33FL or CD33ΔE3-4 via lentiviral gene transfer. Relative expression of the target proteins was assessed by flow cytometry using the V-set domain CD33 antibody, P67.6, with representative histograms shown in the lower panel. Cells were then treated with a concentration of 1000 pg / mL of V-set domain targeting CD33 / CD3 BsAb and healthy donor T cells enriched from unstimulated peripheral blood mononuclear cells collected from healthy adult volunteers with the indicated effector:target (E:T) cell ratio (upper panel). Cytotoxicity was quantified by flow cytometry at 2 days later as the change in the percentage of dead cells measured by 4',6-diamidino-2-phenylindole (DAPI) staining. Anti-V set domain-directed CD33 / CD3 BsAb was constructed in scFv-scFv format using constructs referred herein as RC1 or A3, which utilize the sequence shown in SEQ ID NO: 254 and described in U.S. Patent Application Publication US2016 / 0317657 A1. *p<0.05;**p<0.01;***p<0.001. [Figure 7]This figure shows the decreased antitumor effect of CD33 / CD3 BsAb with increased binding distance from the cell membrane. Human myeloid leukemia cell lines with CRISPR / Cas9-mediated deletion of the endogenous CD33 locus (ML-1 [upper panel], K562 [lower panel]) were engineered to overexpress CD33FL, CD33FL+CD22 2D, or CD33FL+CD22 4D via lentiviral gene transfer. Relative expression of CD33 constructs was assessed by flow cytometry using the V-set domain CD33 antibody, P67.6 (right panel). Cells were then treated with V-set domain targeting CD33 / CD3 BsAb at the indicated concentrations (pg / mL) and healthy donor T cells enriched from healthy donor unstimulated peripheral blood mononuclear cells with a 1:1 E:T cell ratio. Cytotoxicity was quantified by flow cytometry at 2 days later as the change in the percentage of dead cells measured by 4',6-diamidino-2-phenylindole (DAPI) staining. Anti-V set domain-directed CD33 / CD3 BsAb was constructed in scFv-scFv format using constructs referred herein as RC1 or A3, which utilize the sequence shown in SEQ ID NO: 254 and described in U.S. Patent Application Publication US2016 / 0317657 A1. *p<0.05;**p<0.01;***p<0.001. [Figure 8]This figure shows the enhanced antitumor effect of CD33 chimeric antigen receptor (CAR) T cells by reducing the binding distance from the cell membrane. Human myeloid leukemia cell line K562, with a CRISPR / Cas9-mediated deletion of the endogenous CD33 locus, was engineered to overexpress CD33FL or CD33ΔE3-4 via lentiviral gene transfer. Relative expression of the target protein, along with representative histograms, was evaluated by flow cytometry using the V-set domain CD33 antibody, P67.6. The efficacy of V-set domain-directed CAR T cells was assessed by chromium 51 release. To generate CAR T cells, healthy, donor-negative selected human CD8+ T cells were transduced with epHIV7 lentivirus encoding scFv from the CD33V set / CD3 BsAb described in Figures 3, 5A-5C and 6, ligated to an IgG4 CH3 domain spacer, CD28 transmembrane domain, CD3 zeta and 4-1BB intracellular signaling domain, and a truncated CD19 (tCD19) transduction marker. tCD19 CD8+ CAR-T cells were selected and expanded in IL-7 and IL-15 (10 ng / mL; Peprotech, Rocky Hill, NJ, USA) for 14 days each, with alternating medium and cytokines. The cytotoxicity of the CAR-T cells was evaluated after 4-hour incubation with a chromium-51 labeled target. [Figure 9] This figure shows different surface expression levels of CD33 in AML cell lines. CD33 expression, measured by Quantibrite-PE, was determined by staining human AML cell lines with PE conjugate p67.6 antibody. [Figure 10A]This is a diagram of the gene map of the chimeric antigen receptor (CAR) inserted into the lentiviral vector backbone. The VL and VH domains represent the variable light (VL) and variable heavy (VH) domains of the single-stranded variable fragment (scFv). The short (10A), intermediate (10B), and long (10C) domains differ by the insertion of hinge only (short); hinge and CH3 domain (intermediate); and the hinge, CH3, and CH2 domains (long) of human IgG4. All IgG4 domains contain mutant sequences that block binding to the human Fc receptor. This mutation involves substituting the first six amino acids of the CH2 domain of IgG4 (APEFLG, SEQ ID NO: 52) with the first five amino acids of IgG2 (APPVA, SEQ ID NO: 53). TM is the CD28 transmembrane domain. tCD19 represents truncated CD19 lacking intracellular signaling. CD3ζ and 4-1BB are the internal signaling domains of these proteins, and T2A represents the cleaved peptide. [Figure 10B] This is a diagram of the gene map of the chimeric antigen receptor (CAR) inserted into the lentiviral vector backbone. The VL and VH domains represent the variable light (VL) and variable heavy (VH) domains of the single-stranded variable fragment (scFv). The short (10A), intermediate (10B), and long (10C) domains differ by the insertion of hinge only (short); hinge and CH3 domain (intermediate); and the hinge, CH3, and CH2 domains (long) of human IgG4. All IgG4 domains contain mutant sequences that block binding to the human Fc receptor. This mutation involves substituting the first six amino acids of the CH2 domain of IgG4 (APEFLG, SEQ ID NO: 52) with the first five amino acids of IgG2 (APPVA, SEQ ID NO: 53). TM is the CD28 transmembrane domain. tCD19 represents truncated CD19 lacking intracellular signaling. CD3ζ and 4-1BB are the internal signaling domains of these proteins, and T2A represents the cleaved peptide. [Figure 10C]This is a diagram of the gene map of the chimeric antigen receptor (CAR) inserted into the lentiviral vector backbone. The VL and VH domains represent the variable light (VL) and variable heavy (VH) domains of the single-stranded variable fragment (scFv). The short (10A), intermediate (10B), and long (10C) domains differ by the insertion of hinge only (short); hinge and CH3 domain (intermediate); and the hinge, CH3, and CH2 domains (long) of human IgG4. All IgG4 domains contain mutant sequences that block binding to the human Fc receptor. This mutation involves substituting the first six amino acids of the CH2 domain of IgG4 (APEFLG, SEQ ID NO: 52) with the first five amino acids of IgG2 (APPVA, SEQ ID NO: 53). TM is the CD28 transmembrane domain. tCD19 represents truncated CD19 lacking intracellular signaling. CD3ζ and 4-1BB are the internal signaling domains of these proteins, and T2A represents the cleaved peptide. [Figure 11A] This figure shows the results of flow cytometry testing of (11A) human CD33PAN antibody clones (clones 1A9, 1H10, 1B9, 1E6, and 1D2) and (11B) human CD33V set antibody clones (clones 2E3, 2D3, and 1H8) against CD33+ parental ML-1 cells, ML-1 cells with CRISPR / Cas9-mediated deletion of CD33 ("CD33 KO"), and REH sublines engineered to express CD33FL or CD33ΔE2, as shown. A control without the primary antibody was included. [Figure 11B] This figure shows the results of flow cytometry testing of (11A) human CD33PAN antibody clones (clones 1A9, 1H10, 1B9, 1E6, and 1D2) and (11B) human CD33V set antibody clones (clones 2E3, 2D3, and 1H8) against CD33+ parental ML-1 cells, ML-1 cells with CRISPR / Cas9-mediated deletion of CD33 ("CD33 KO"), and REH sublines engineered to express CD33FL or CD33ΔE2, as shown. A control without the primary antibody was included. [Figure 12A](12A) This figure shows the reaction rate profile established from anti-CD33 antibody purified by surface plasmon resonance (SPR) technique from a Carterra instrument. SPR is an excellent method for estimating the reaction rate constant of the binding interaction, which can be fitted into a 1:1 Langmuir binding model to determine the on-rate (ka) and off-rate (kd). Both of these rate parameters allow for the calculation of the dissociation rate constant (kD), referred to as binding affinity. Antibody clones were captured as an array on protein A / G lones immobilized on an HC30M tip. The initial reaction rate experiments used full-length CD33 (CD33FL) antigen, starting at a concentration of 2 μM before 4-fold titration to 2 nM. After 10 HBSTE buffer blanks, six injections from low to high concentrations were subsequently flowed onto the array to evaluate the reaction rate of each clone printed on the array: 1 min baseline, 5 min association, and 10 min dissociation. Next, if any antigens remained bound to the array, preventing interaction with the second antigen of interest, the chip was regenerated in 0.85% phosphate pH 1.7 for a new reprint of the same array cloned onto a protein A / G loan, in which CD33ΔE2 was flowed onto the antibody array. Carterra reaction rate software was used to process the data to fit the raw data into reaction rate curves for each concentration of antigen injected onto the array. [Figure 12B-1](12B) This figure shows the SPR evaluation of purified ECD from CD33FL or CD33ΔE2 bound to captured 1H10 and 2D3. Experiments were performed at 25°C using a Biacore T100 instrument equipped with a Series S CM4 tip. 60 μg / mL of protein A / G in 10 mM sodium acetate, pH 4.0 was immobilized in two flow cells (1000 RU) using a standard amine coupling reaction. Capture reaction rate experiments were performed in 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% surfactant P20, and 0.1 mg / mL IgG-free bovine serum albumin buffer. 0.5 μg / mL anti-human CD33 antibody was injected at 10 μL / min for 30–40 seconds into a second flow cell of immobilized protein A / G to capture 40–58 RU of antibody for CD33FL binding experiments, or injected for 45–80 seconds to capture 75–95 RU of antibody for CD33ΔE2 binding experiments. Purified external domains for CD33FL and CD33ΔE2 were run as a concentration series at 50 μL / min on both the captured antibody and protein A / G alone (reference) surface. The CD33FL series was started at a high concentration of 160 nM at 2D3 and 1H10, while the CD33ΔE2 series was started at 40 nM at 1H10 and 300 nM at 2D3. CD33 was injected for 7 minutes and dissociated for 20 or 30 minutes for most pairs. Two consecutive 2-fold dilutions of the external domain concentration were performed in a series, randomized, and a buffer blank was included every four infusions. CM4 tips were refilled by injecting 0.85% H3PO4 at 50 μL / min for 30 seconds twice, and the antibody was recaptured before each CD33 infusion. Data were dual-referenced and analyzed in BiaEval 2.0.4 software using a 1:1 binding model with local Rmax. [Figure 12B-2](12B) This figure shows the SPR evaluation of purified ECD from CD33FL or CD33ΔE2 bound to captured 1H10 and 2D3. Experiments were performed at 25°C using a Biacore T100 instrument equipped with a Series S CM4 tip. 60 μg / mL of protein A / G in 10 mM sodium acetate, pH 4.0 was immobilized in two flow cells (1000 RU) using a standard amine coupling reaction. Capture reaction rate experiments were performed in 10 mM HEPES, pH 7.4, 150 mM NaCl, 3 mM EDTA, 0.05% surfactant P20, and 0.1 mg / mL IgG-free bovine serum albumin buffer. 0.5 μg / mL anti-human CD33 antibody was injected at 10 μL / min for 30–40 seconds into a second flow cell of immobilized protein A / G to capture 40–58 RU of antibody for CD33FL binding experiments, or injected for 45–80 seconds to capture 75–95 RU of antibody for CD33ΔE2 binding experiments. Purified external domains for CD33FL and CD33ΔE2 were run as a concentration series at 50 μL / min on both the captured antibody and protein A / G alone (reference) surface. The CD33FL series was started at a high concentration of 160 nM at 2D3 and 1H10, while the CD33ΔE2 series was started at 40 nM at 1H10 and 300 nM at 2D3. CD33 was injected for 7 minutes and dissociated for 20 or 30 minutes for most pairs. Two consecutive 2-fold dilutions of the external domain concentration were performed in a series, randomized, and a buffer blank was included every four infusions. CM4 tips were refilled by injecting 0.85% H3PO4 at 50 μL / min for 30 seconds twice, and the antibody was recaptured before each CD33 infusion. Data were dual-referenced and analyzed in BiaEval 2.0.4 software using a 1:1 binding model with local Rmax. [Figure 13] This figure shows the internalization of 1E6 and P67.6. AML cell lines were incubated with CD33 antibody at 37°C for the times indicated. A fluorescently labeled secondary antibody was then added to quantify the CD33 antibody remaining on the cell surface. Results are presented as the percentage of fluorescence signal present at time 0. [Figure 14]This figure shows the binding of recombinant fully human CD33V set antibody 1H8, which has a human IgG1 framework, to REH cells (human acute lymphoblastic leukemia cell line, endogenous CD33neg) that have been engineered to express human CD33FL. [Figure 15A] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15B]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15C] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15D]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15E] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15F]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15G] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15H]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15I] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15J]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15K] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15L]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15M] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15N]This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 15O] This figure shows CAR-T cells expressing humanized (1E6, 1A9, and 1H10) scFV directed towards CD33, which can be expanded in vitro with cytokines. Intermediate spacer CARs and CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv were sorted by fluorescence-activated cell sorting (FACS) 7 days after CD3 / CD28 bead stimulation and transduction. The cells were then expanded in IL-2 (50 ng / mL, 15A, 15D, 15G, 15J, and 15M); IL-7 and IL-15 (10 ng / mL each, 15B, 15E, 15H, 15K, and 15N); or IL-7, IL-15, and IL-21 (10 ng / mL each, 15C, 15F, 15I, 15L, and 15O). The culture medium and cytokines were refreshed every other day. Cells were evaluated every two days for cell number (15A-15F), cell growth (15G-15I), viability after propidium iodide exclusion (15J-15L), and diameter (15M-15O). [Figure 16A]This figure shows the in vitro antigen-specific lysis of 1E6, 1A9, and 1H10 CD33-oriented CAR-T cells with an intermediate spacer against multiple AML cell lines. KG1a and ML-1 cells, either expressing endogenous CD33 (Par) or lacking CD33 due to CRISPR-Cas9 gene deletion (KO), and labeled with chromium 51 (Cr51), were exposed for 4 hours to (16A) CD33-oriented CD8+ CAR-T cells enlarged in IL-2 (50 ng / mL), (16B) IL-7 and IL-15 (10 ng / mL each), or (16C) IL-7, IL-15, and IL-21 (10 ng / mL each) in various effector:target ratios. The supernatant was collected and analyzed for Cr51 concentration by scintillation. [Figure 16B] This figure shows the in vitro antigen-specific lysis of 1E6, 1A9, and 1H10 CD33-oriented CAR-T cells with an intermediate spacer against multiple AML cell lines. KG1a and ML-1 cells, either expressing endogenous CD33 (Par) or lacking CD33 due to CRISPR-Cas9 gene deletion (KO), and labeled with chromium 51 (Cr51), were exposed for 4 hours to (16A) CD33-oriented CD8+ CAR-T cells enlarged in IL-2 (50 ng / mL), (16B) IL-7 and IL-15 (10 ng / mL each), or (16C) IL-7, IL-15, and IL-21 (10 ng / mL each) in various effector:target ratios. The supernatant was collected and analyzed for Cr51 concentration by scintillation. [Figure 16C]This figure shows the in vitro antigen-specific lysis of 1E6, 1A9, and 1H10 CD33-oriented CAR-T cells with an intermediate spacer against multiple AML cell lines. KG1a and ML-1 cells, either expressing endogenous CD33 (Par) or lacking CD33 due to CRISPR-Cas9 gene deletion (KO), and labeled with chromium 51 (Cr51), were exposed for 4 hours to (16A) CD33-oriented CD8+ CAR-T cells enlarged in IL-2 (50 ng / mL), (16B) IL-7 and IL-15 (10 ng / mL each), or (16C) IL-7, IL-15, and IL-21 (10 ng / mL each) in various effector:target ratios. The supernatant was collected and analyzed for Cr51 concentration by scintillation. [Figure 17A] The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17B]The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17C] The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17D]The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17E] The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17F]The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17G] The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17H]The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 17I] The figure shows CD33-directed CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10 scFv being expanded in IL-2 (50 ng / mL, 17A, 17D, and 17G), IL-7 and 15 (10 ng / mL each, 17B, 17E, and 17H) or IL-7, IL-15, and IL-21 (10 ng / mL each, 17C, 17F, and 17I), and then exposed to endogenous CD33-rich irradiated AML cell lines (ML-1 or KG1a) (Par); CD33-deficient cell lines due to CRISPR-Cas9 gene deletion (KO); culture medium; or phorbol-12 myristate and ionomycin (PMA-Iono). Subsequently, the cell supernatant was collected after 24 hours and analyzed for levels of interferon-gamma (IFNγ, 17A-17C), interleukin-2 (IL-2, 17D-17F), or tumor necrosis factor alpha (TNFα, 17G-17I) by enzyme-linked immunosorbent assay (ELISA). [Figure 18A]This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 18B] This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 18C]This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 18D] This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 18E]This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 18F] This figure shows that CD33-directed CAR-T cells expanded in IL-7, IL-15, and IL-21 exhibit superior proliferation in vitro. CD8+ CAR-T cells expressing 1E6, 1A9, or 1H10-directed scFv and expanded in either IL-2 (50 ng / ml, 18A and 18D), IL-7 and 15 (10 ng / ml, 18B and 18E), or IL-7, 15, and 21 (10 ng / ml, 18C and 18F) were labeled with CFSE and exposed for 5 days to either ML-1 cells expressing endogenous CD33 (Par) or ML-1 cells lacking CD33 due to CRISPR-Cas9 gene deletion (KO) or culture medium alone. Cells were then analyzed for CFSE levels by flow cytometry. Summary of percentages of dividing cells (18D-18F), including undivided peaks isolated by proliferation modeling in FlowJo software. [Figure 19-1]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-2]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-3]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-4]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-5]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-6]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-7]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-8]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-9]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-10]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-11]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-12]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-13]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-14]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-15]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-16]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-17]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-18]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-19]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-20]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-21]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-22]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-23]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-24]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-25]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-26]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-27]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-28]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-29]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-30]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-31]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-32]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-33]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Figure 19-34]This figure shows sequences supporting this disclosure: 1H10 VH-VL scFv coding sequence (sequence number 2); 1A9 VH-VL scFv coding sequence (sequence number 3); 1E6 VH-VL scFv coding sequence (sequence number 4); 1D2 VH-VL scFv coding sequence (sequence number 5); 1B9 VH-VL scFv coding sequence (sequence number 6); 1H8 VH-VL scFv coding sequence (sequence number 7); 2D3 VH-VL scFv coding sequence (sequence number 8); 2E3 VH-VL scFv coding sequence (sequence number 9); signal peptide coding sequence (sequence number 188); G4Sx3 linker coding sequence (sequence number 189); IgK signal peptide (sequence number 158); 1H10 scFv VH-VL orientation (sequence number 190); 1H10 scFv VL-VH orientation (sequence number 191); 1A9 scFv VH-VL orientation (sequence number 192); 1A9 scFv VL-VH orientation (SEQ ID NO: 193); 1E6 scFv VH-VL orientation (SEQ ID NO: 194); 1E6 scFv VL-VH orientation (SEQ ID NO: 195); 2D3 scFv VH-VL orientation (SEQ ID NO: 196); 2D3 scFv VL-VH orientation (SEQ ID NO: 197); Human CD33 full-length DNA code (SEQ ID NO: 198); Human CD33 full-length protein (SEQ ID NO: 199); IgG4 hinge code sequence-A (SEQ ID NO: 10); IgG4 hinge code sequence-B (SEQ ID NO: 11); IgG4-int(DS) code sequence (SEQ ID NO: 12); IgG4-long code sequence (SEQ ID NO: 13); CD3ζ code sequence (SEQ ID NO: 14); CD3ζ protein-A (SEQ ID NO: 15); CD3ζ protein-B (SEQ ID NO: 16); 4-1BB signal transduction code sequence-A (SEQ ID NO: 17); 4- 1BB signaling pathway coding sequence-B (SEQ ID NO: 18); 4-1BB protein-A (SEQ ID NO: 19); 4-1BB protein-B (SEQ ID NO: 20); CD28TM coding sequence-A (SEQ ID NO: 21); CD28TM coding sequence-B (SEQ ID NO: 22); CD28TM coding sequence-C (SEQ ID NO: 23); CD28TM protein-A (SEQ ID NO: 24); CD28TM protein-B (SEQ ID NO: 25); tCD19 coding sequence (SEQ ID NO: 26); T2A coding sequence (SEQ ID NO: 27); Tosea signaling virus 2A (T2A) peptide (SEQ ID NO: 28);Porcine Tescovirus-1 2A(P2A) peptide (SEQ ID NO: 29); Equine Rhinitis A virus (ERAV) 2A(E2A) peptide (SEQ ID NO: 30); Foot-and-mouth disease virus 2A(F2A) peptide (SEQ ID NO: 31); EF1 promoter-A (SEQ ID NO: 32); EF1 promoter-B (SEQ ID NO: 33); Psi (SEQ ID NO: 34); RRE (SEQ ID NO: 35); Flap (SEQ ID NO: 36); GM-CSFR code sequence (SEQ ID NO: 37); WPRE (SEQ ID NO: 38); delU3 (SEQ ID NO: 39); R (SEQ ID NO: 40); U5 (SEQ ID NO: 41); AmpR (SEQ ID NO: 42); CoE1 origin of replication (SEQ ID NO: 43); SV40 (SEQ ID NO: 44); CMV (SEQ ID NO: 45); Glycosylation site; 1H10-intDS-41bb-3z-T-CD19t top strand (SEQ ID NO: 46); 1H10-sh-41bb-3z-T-CD19t top strand (Sequence) Number 47); 1A9-intDS-41bb-3z-T-CD19t top strand (sequence number 48); 1A9-sh-41bb-3z-T-CD19t top strand (sequence number 49); 1E6-intDS-41bb-3z-T-CD19t top strand (sequence number 50); 1E6-sh-41bb-3z-T-CD19t top strand (sequence number 51); 1H10-LvHv-intDS-4 1bb-3z-T-CD19t top strand (sequence number 200); 1A9-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 201); 1E6-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 202); 2D3-LvHv-intDS-41bb-3z-T-CD19t top strand (sequence number 203); CD33:CD22 4D protein (SEQ ID NO: 204); CD33:CD22 4D nucleotide (SEQ ID NO: 205); CD33:CD22 2D protein (SEQ ID NO: 206); CD33:CD22 2D nucleotide (SEQ ID NO: 207); CD33 V-set construct (exon 3 and 4 deletion) protein (SEQ ID NO: 208); CD33 V-set construct (exon 3 and 4 deletion) nucleotide (SEQ ID NO: 209); CD33 signal peptide (SEQ ID NO: 210); CD33 signal peptide coding sequence (SEQ ID NO: 211);6-histidine tag coding sequence (SEQ ID NO: 212); 3x glycine linker; 3x glycine linker coding sequence; CD33 ECD (SEQ ID NO: 213); CD33 ECD coding sequence (SEQ ID NO: 214); CD33 ECD with deletion of amino acids 140-232 (SEQ ID NO: 215); CD33 ECD with deletion of amino acid 140-232 coding sequence (SEQ ID NO: 216); CD33 transmembrane domain (SEQ ID NO: 217); CD33 transmembrane domain coding sequence (SEQ ID NO: 218); CD33 intracellular domain (SEQ ID NO: 219); CD33 intracellular domain coding sequence (SEQ ID NO: 220); CD22 containing CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6. Part of ECD (Sequence ID 221): CD22 domains defined as Ig-like C2 type 3, Ig-like C2 type 4, Ig-like C2 type 5, and Ig-like C2 type 6 coding sequences. Part of ECD (Sequence ID 222): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6. Part of ECD (Sequence ID 223): CD22 domains defined as Ig-like C2 type 5 and Ig-like C2 type 6 coding sequences. Part of ECD (SEQ ID NO: 224); 1H10, 1A9, 1E6 and / or 1B9 light chain signal peptide (SEQ ID NO: 225); 1D2 light chain signal peptide (SEQ ID NO: 226); 1H8 light chain signal peptide (SEQ ID NO: 227); 2D3 light chain signal peptide (SEQ ID NO: 228); 1H10 heavy chain signal peptide (SEQ ID NO: 229); 1A9 heavy chain signal peptide (SEQ ID NO: 230); 1E6 and / or 2E3 heavy chain signal peptide (SEQ ID NO: 231); 1D2 heavy chain signal peptide (SEQ ID NO: 232); 1B9 heavy chain signal peptide (SEQ ID NO: 233); 1H8 heavy chain signal peptide (SEQ ID NO: 234); 2D3 heavy chain signal peptide (SEQ ID NO: 235); My96_int_41BB_3z_TCD19 code sequence (SEQ ID NO: 236); My96 code sequence (SEQ ID NO: 237); V-set oriented CD33 / CD3 BsAb(RC1, Sequence ID 254); V-set oriented CD33 / CD3 without a leader array or His tag; BsAb(Sequence ID 255);Also included are the V-set oriented CD33 scFv protein sequence (SEQ ID NO: 256) and coding sequence (SEQ ID NO: 257). [Modes for carrying out the invention]

[0017] According to the World Health Organization, cancer is the second leading cause of death worldwide, accounting for an estimated 9.6 million deaths in 2018. Acute myeloid leukemia (AML) is a type of cancer that arises from malignant tumors of clonal proliferative myeloblastic cells. AML is also known as acute myeloid leukemia, acute myeloid leukemia, acute granulocytic leukemia, and acute nonlymphocytic leukemia.

[0018] In the United States, there are 20,000 new cases of AML annually (Kouchkovsky and Abdul-Hay, Blood Cancer J.6(7):e441, 2016), and 11,000 people die from AML each year (American Cancer Society, August 2018). In young patients with AML, conventional chemotherapy can achieve high complete remission rates of 60% to 80% (Dohner et al., 2017. Blood. 129(4):424-447), but the treatment outcomes for elderly patients aged 65 and over remain insufficient, with as many as 70% dying from the disease within one year of diagnosis (Meyers et al., Appl Health Economic Health Policy, 11:275-286, 2013). Unfortunately, due to the chemotherapy resistance of leukemia stem cells, relapse after conventional treatment is common (Eppert et al., 2011. Nat. Med. 17(9): pp. 1086-1093), and current treatment options for relapsed / refractory (R / R) AML are poor, resulting in a 12-month overall survival rate of less than 30%.

[0019] For many years, the treatment of choice for cancer has been surgery, chemotherapy, and / or radiation therapy. In recent years, many targeted therapies have emerged that specifically target cancer cells by identifying and exploiting specific molecular changes that are primarily found in these cells. For example, many cancer cells selectively express certain markers on their cell surface, and these markers have become targets for antibody-based therapies.

[0020] One of the keys to successful targeted therapy is the selection of target cancer cell markers. Ideal target markers are immunogenic, play a crucial role in proliferation and differentiation, are expressed only on the surface of all malignant cells and malignant stem cells, and the majority of patients should test positive for these markers (Cheever et al., 2009. Clin. Cancer Res. 15(17):5323-8337).

[0021] CD33 FL It is primarily presented on mature cells of the myeloid lineage and on mature cells, initially expressed on pluripotent myeloid progenitor cells. It is not found outside the hematopoietic system and is thought not to be expressed on pluripotent hematopoietic stem cells. Consistent with its role as a myeloid differentiation antigen, CD33 FL CD33 is widely expressed on malignant cells in patients with myeloid neoplasms; for example, in AML, it is found in at least a subset of AML blast cells in almost all cases, and may be found in leukemia stem cells in some cases. Due to this expression pattern, CD33 FLIt has been widely used as an antigen for targeted therapy of AML (Walter et al., Blood 119(26):pp. 6198-6208, 2012; Cowan et al., Front. Biosci. (Landmark Ed) 18:pp. 1311-1334, 2013; Laszlo et al., Blood Ref. 28(4):pp. 143-153, 2014; and Walter, Expert Opin Investig Drugs 27(4):pp. 339-348, 2018). While unconjugated monoclonal CD33 antibodies have proven clinically ineffective, several recent randomized trials using the CD33 antibody-drug conjugate (ADC) gemtuzumab ozogamicin (GO) have demonstrated improved survival rates in a subset of patients with AML, establishing the value of antibodies in this disease and establishing CD33 as the first, and so far, only therapeutic target for immunotherapy of AML. FL This was validated (Laszlo et al., Blood Rev.28(4):143-153, 2014; Godwin et al., Leukemia 31(9)31(9):1855-1868, 2017). This data validated CD33 as the first (and so far only) target for immunotherapy in AML. Interest in CD33 as a drug target for other malignant and non-malignant disorders is growing, in parallel with the development of new, more effective CD33-targeted therapies (e.g., antibody-drug conjugates, radioactive immunoconjugates, bispecific antibodies, chimeric antigen receptor [CAR]-modified T cells) to overcome the limitations pointed out by GO. These efforts include targeting CD33 splice variants not recognized by GO, as well as targeting CD33+ tumor cells in other hematological malignancies, CD33+ myeloid-derived suppressor cells (MDSCs) in various diseases, and normal CD33+ microglia in Alzheimer's disease (Walter, Expert Opin Biol Ther. 2020, 20(9):955-958).

[0022] However, some patients lack exon 2 and CD33 ΔE2 It expresses a truncated splice variant of CD33, which is referred to as CD33. ΔE2 It has been identified at the mRNA level in normal hematopoietic cells and leukemia cells. Regarding the latter, CD33 ΔE2 The mRNA was identified in 29 of the 29 AML patient samples tested, demonstrating universal expression in human AML. CD33 ΔE2 This contains the C2 set Ig-like domain of CD33 but does not contain the V set Ig-like domain (Figure 2). Further splice variants identified at the mRNA level include CD33 E7a and CD33 ΔE2 / E7a CD33 is included. E7a This utilizes selective exon 7 (E7a) to induce truncation of the intracellular domain of CD33. ΔE2 / E7a It lacks exon 2, and the intracellular domain of CD33 is also truncated.

[0023] However, currently, almost all commercially available diagnostic CD33 antibodies and clinically available CD33 antibody-based therapies recognize the immunodominant V-set Ig-like domain encoded by exon 2 (Figure 2). That is, CD33 lacking the V-set Ig-like domain. ΔE2 And other CD33 proteins are not recognized by almost all commercially and clinically available CD33 antibodies. This means that these antibodies do not recognize CD33 ΔE2 This means that shorter forms of CD33, such as those lacking the V-set domain, will not be recognized. ΔE2 Selective transcription and CD33 FL This can explain the observation made in one clinical trial in pediatric AML that patients with a single nucleotide polymorphism in the CD33 gene, which results in reduced translation, did not benefit from the addition of GO (which also binds to the V set domain of CD33) to intensive chemotherapy.

[0024] Antibodies that recognize and bind to the C2 set Ig-like domain of the CD33 protein, regardless of the presence or absence of the V set Ig-like domain (e.g., CD33 PAN CD33, also known as an antibody ΔE2 and CD33 FL Antibodies that bind to isoforms represent a significant advance in targeting all CD33 isoforms, resulting in broader therapeutic effects. These pan-binding antibodies also offer advantages because they bind closer to the cell membrane (Figure 2). For several therapeutic targets, the specificity of the targeted epitope has been shown to be crucial for antibody-based therapies, with proximal membrane epitopes yielding more potent antitumor effects than distal membrane epitopes, as demonstrated for CD20, CD22, CD25, and EpCAM. For example, see Cleary et al., J Immunol. 2017; 198(10): pp. 3999-4011; Lin, Pharmgenomics Pers Med. 2010; 3: pp. 51-59; Haso et al., Blood. 2013; 121(7): pp. 1165-1174; and Bluemel et al., Cancer Immunol Immunother. 2010; 59(8): pp. 1197-1209.

[0025] Significant progress has been made in genetically engineering T cells of the immune system to target and kill undesirable cell types, such as cancer cells, beyond antibody-based therapies. Many of these T cells are genetically engineered to express chimeric antigen receptors (CARs). CARs are proteins containing several distinct subcomponents that enable genetically modified T cells to recognize and kill cancer cells. These subcomponents include at least extracellular and intracellular components. The extracellular component includes a binding domain that specifically binds to a marker selectively present on the surface of undesirable cells (e.g., CD33). The binding domain is typically a single-stranded variable fragment (scFv) derived from a monoclonal antibody (mAb), but it can be based on other formats that include an antibody-like antigen-binding site.

[0026] When a binding domain binds to such a marker, intracellular components signal T cells to destroy the bound cell. The intracellular components provide such activation signals based on the inclusion of the effector domain. First-generation CARs utilized the cytoplasmic domain of CD3ζ as the effector domain. Second-generation CARs utilized the cytoplasmic domain of CD3ζ in combination with the differentiation cluster 28 (CD28) or 4-1BB (CD137) cytoplasmic domain, while third-generation CARs utilized the CD3ζ cytoplasmic domain in combination with the CD28 and 4-1BB cytoplasmic domains as the effector domain.

[0027] CAR further includes a transmembrane domain that can link extracellular components to intracellular components.

[0028] Other minor components that can enhance the function of CARs may also be used. For example, spacer regions can provide additional structural flexibility to the CAR and often increase the ability of the binding domain to bind to the targeted cellular marker. The appropriate length of spacer regions within a particular CAR may depend on many factors, including how close or far the targeted marker is located from the undesirable cell membrane surface.

[0029] When performed ex vivo, T cell gene modification can involve numerous cell manipulation steps, and it has been observed that different manipulation conditions can affect the cancer cell-killing properties of the cells. Therefore, when designing CARs and genetically modifying cells to express them, many considerations must be taken into account, including the targeted cell markers, the presence and / or length of spacers, and the ex vivo manipulation procedure.

[0030] This disclosure provides chimeric antigen receptors (CARs) for the treatment of CD33-related disorders such as AML. In certain embodiments, the CAR is a CD33 variant expressed by the patient (e.g., CD33 FL or CD33 ΔE2Regardless of the CD33 binding domain, the antibodies contain a binding domain that binds to CD33. These CD33 binding domains are referred to as "pan" conjugates. In certain embodiments, the pan conjugates bind to the membrane-proximal C2-set Ig-like domain of CD33. In certain embodiments, these pan conjugates are derived from newly developed antibodies: 1H10, 1A9, 1E6, 1D2, and 1B9, and may contain single-strand variable fragments of these antibodies. As described herein, many membrane-proximal bindings enhance the immunoeffector function of CARs for treating AML and other CD33+ disorders. Further newly developed CD33-targeting antibodies disclosed herein bind to the V-set domain of CD33 and include 1H8, 2D3, and 2E3. These antibodies bind to the CD33 FL This provides an additional CAR-based treatment option for patients who develop this condition.

[0031] The antibody binding domains used for treatment may be based on a combination of binding domains, depending on whether the subject expresses or lacks the V-set domain of CD33. For example, if the subject expresses the V-set domain, a combination therapy containing one or more binding domains from 1H10, 1A9, 1E6, 1D2, and 1B9 may be selected in combination with one or more from 1H8, 2D3, and 2E3.

[0032] In certain embodiments, the disclosure provides a CAR having a short spacer region or an intermediate spacer region. In certain embodiments, the short spacer region comprises the hinge region (12 amino acids) of IgG4. In certain embodiments, the intermediate spacer region comprises the hinge region and the CH3 domain (131 amino acids in total) of IgG4. The IgG4 domain used as a spacer region may include mutations that block binding to the human Fc receptor. In certain embodiments, these mutations include substituting the first six amino acids of the CH2 domain of IgG4 (APEFLG, SEQ ID NO: 52) with the first five amino acids of IgG2 (APPVA, SEQ ID NO: 53).

[0033] In certain embodiments, the disclosure provides expansion and activation of T cells genetically modified to express the CARs disclosed herein, utilizing a combination of cytokines IL-7, IL-15, and IL-21. In certain embodiments, the disclosure provides expansion and activation of T cells genetically modified to express the CARs disclosed herein, utilizing a combination of cytokines including IL-2. Expansion / activation with this combination of cytokines results in increased proliferation and antigen-specific cytolysis.

[0034] Herein, aspects of this disclosure are described in more substantive detail as follows: (i) immune cells; (ii) cell sampling and cell enrichment; (iii) genetic modification of cell populations to express chimeric antigen receptors (CARs); (iii-a) genetic engineering techniques; (iii-b) CAR subcomponents; (iii-bi) binding domains; (iii-b-ii) spacer regions; (iii-b-iii) transmembrane domains; (iii-b-iv) intracellular effector domains; (iii-bv) linkers; (iii-b-vi) regulatory functions including tag cassettes, transduction markers and / or suicide switches; (iv) cell activation culture conditions; (v) cell formulations manufactured ex vivo; (vi) methods of use; (vii) reference levels derived from control populations; (viii) exemplary embodiments; and (ix) the final paragraph. These headings are provided for compositional purposes only and do not limit the scope or interpretation of this disclosure.

[0035] (i) Immune cells. This disclosure describes cells that have been genetically modified to express CARs. Genetically modified cells may include T cells, B cells, natural killer (NK) cells, NK-T cells, monocytes / macrophages, lymphocytes, hematopoietic stem cells (HSCs), hematopoietic progenitor cells (HPCs), and / or mixtures of HSCs and HSCs (i.e., HSPCs). In certain embodiments, the genetically modified cells include T cells.

[0036] Several different subsets of T cells have been discovered, each with a different function. For example, most T cells possess a T cell receptor (TCR), which exists as a complex of several proteins. The actual T cell receptor is produced from independent T cell receptor alpha and beta (TCRα and TCRβ) genes and consists of two distinct peptide chains called the α-TCR chain and the β-TCR chain.

[0037] γδ T cells represent a small subset of T cells that possess different T cell receptors (TCRs) on their surface. In γδ T cells, the TCR consists of one γ chain and one δ chain. This group of T cells is less common than αβ T cells (2% of all T cells).

[0038] CD3 is expressed on all mature T cells. Activated T cells express 4-1BB (CD137), CD69, and CD25. CD5 and the transferrin receptor are also expressed on T cells.

[0039] T cells can be further classified into helper cells (CD4+ T cells) and cytotoxic T cells (CTLs, CD8+ T cells), including cytolytic T cells. T helper cells assist other leukocytes in immunological processes, including the maturation of B cells into plasma cells and the activation of cytotoxic T cells and macrophages. These cells are also known as CD4+ T cells because they express the CD4 protein on their surface. Helper T cells are activated when presented with peptide antigens by MHC class II molecules expressed on the surface of antigen-presenting cells (APCs). Once activated, they rapidly divide and secrete small proteins called cytokines that modulate or assist the active immune response.

[0040] Cytotoxic T cells destroy virus-infected cells and tumor cells and are also involved in graft rejection. These cells are also known as CD8+ T cells because they express the CD8 glycoprotein on their surface. These cells recognize their targets by binding to antigens associated with MHC class I, which are present on the surface of almost all cells in the body.

[0041] As used herein, “central memory” T cells (or “TCM”) refer to CTLs that have experienced an antigen and express CD62L or CCR7 and CD45RO on their surface, and do not express CD45RA or have reduced expression compared to naive cells. In certain embodiments, central memory cells are positive for CD62L, CCR7, CD25, CD127, CD45RO and CD95 expression, and have reduced CD45RA expression compared to naive cells.

[0042] As used herein, “effector memory” T cells (or “TEM”) refer to T cells that have experienced an antigen and which do not express or have reduced expression of CD62L on their surface compared to central memory cells, and do not express or have reduced expression of CD45RA compared to naive cells. In certain embodiments, effector memory cells are negative for CD62L and CCR7 expression and have variability in CD28 and CD45RA expression compared to naive or central memory cells. Effector T cells are positive for granzyme B and perforin compared to memory or naive T cells.

[0043] As used herein, “naive” T cells refer to T cells that have not experienced an antigen and, compared to central memory cells or effector memory cells, express CD62L and CD45RA but not CD45RO. In certain embodiments, naive CD8+ T lymphocytes are characterized by the expression of naive T cell phenotypic markers including CD62L, CCR7, CD28, CD127, and CD45RA.

[0044] Natural killer cells (NK cells, K cells, and killer cells) are activated in response to interferon or macrophage-derived cytokines. They function to suppress viral infections while antigen-specific cytotoxic T cells, which can eliminate infections through an adaptive immune response, are being generated. NK cells express CD8, CD16, and CD56, but do not express CD3.

[0045] NK cells include NK-T cells. NK-T cells are a specialized population of T cells that express a semi-invariant T cell receptor (TCR ab) and surface antigens that typically associate with natural killer cells. NK-T cells contribute to antimicrobial and antiviral immune responses and promote tumor-related immune surveillance or immunosuppression. Like natural killer cells, NK-T cells can also induce perforin-related, Fas-related, and TNF-related cytotoxicity. Activated NK-T cells can produce IFN-γ and IL-4. In certain embodiments, NK-T cells are CD3+ / CD56+.

[0046] Macrophages (and their precursor cells, monocytes) are present in all tissues of the body (in certain cases, as microglia, Kupffer cells, and osteoclasts) and engulf apoptotic cells, pathogens, and other non-self components. Monocytes / macrophages express CD11b, F4 / 80, CD68, CD11c, IL-4Rα, and / or CD163.

[0047] Immature dendritic cells (i.e., pre-activation) engulf antigens and other non-self components in the periphery, and then, in their activated form, migrate to the T cell region of lymphoid tissue to present antigens to T cells. Dendritic cells express CD1a, CD1b, CD1c, CD1d, CD21, CD35, CD39, CD40, CD86, CD101, CD148, CD209, and DEC-205.

[0048] Hematopoietic stem cells / progenitor cells, or HSPCs, refer to a combination of hematopoietic stem cells and hematopoietic progenitor cells.

[0049] Hematopoietic stem cells are undifferentiated hematopoietic cells that can self-replicate in vivo, proliferate essentially without limit in vitro, and differentiate into all other hematopoietic cell types.

[0050] Hematopoietic progenitor cells are cells derived from hematopoietic stem cells or fetal tissue that can further differentiate into mature cell types. In certain embodiments, hematopoietic progenitor cells are CD24 lo Lin - CD117 + Hematopoietic progenitor cells. HPCs can differentiate into (i) myeloid progenitor cells that ultimately give rise to monocytes and macrophages, neutrophils, basophils, eosinophils, erythrocytes, megakaryocytes / platelets or dendritic cells, or (ii) lymphoid progenitor cells that ultimately give rise to T cells, B cells and NK cells.

[0051] HSPCs may be positive for certain markers expressed at increased levels on HSPCs compared to other types of hematopoietic cells. For example, such markers include CD34, CD43, CD45RO, CD45RA, CD59, CD90, CD109, CD117, CD133, CD166, HLA DR, or combinations thereof. Conversely, HSPCs may be negative for expression markers compared to other types of hematopoietic cells. For example, such markers include Lin, CD38, or combinations thereof. Preferably, HSPCs are CD34 + It is a cell.

[0052] The statement that a cell or cell population is “positive” for a particular marker, or expresses a particular marker, refers to the detectable presence of the particular marker on or within the cell. When referring to a surface marker, the term may refer to the presence of surface expression detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting the antibody, wherein the stain is detectable by flow cytometry at a level substantially higher than the stain detected by performing the same procedure using an isotype-matched control under otherwise identical conditions, and / or at a level substantially similar to that for cells known to be positive for the marker, and / or at a level substantially higher than that for cells known to be negative for the marker.

[0053] The statement that a cell or cell population is “negative” for a particular marker, or lacks expression of the marker, means that there is substantially no detectable presence of the particular marker on or within the cell. When referring to a surface marker, this term may refer to the absence of surface expression detected by flow cytometry, for example, by staining with an antibody that specifically binds to the marker and detecting said antibody, wherein the staining is not detected by flow cytometry at a level substantially higher than the staining detected by the same procedure using an isotype-matched control under otherwise identical conditions, and / or at a level substantially lower than that for cells known to be positive for the marker, and / or at a level substantially similar to that for cells known to be negative for the marker.

[0054] Cells genetically modified in accordance with the teachings of this disclosure may be patient-derived (autologous) cells, or, where appropriate, allogeneic cells, and may be in vivo or ex vivo.

[0055] (ii) Collection and concentration of cell samples. Methods for collection and concentration are known to those skilled in the art. In some embodiments, the cells are derived from cell lines. In some embodiments, the cells are obtained from a heterologous source, such as mice, rats, non-human primates, or pigs. In certain embodiments, the cells are derived from humans.

[0056] In some embodiments, T cells are derived from or isolated from samples such as whole blood, peripheral blood mononuclear cells (PBMCs), leukocytes, bone marrow, thymus, tissue biopsy, tumors, leukemia, lymphoma, lymph nodes, intestinal lymphoid tissue, mucosa-associated lymphoid tissue, spleen, other lymphoid tissue, liver, lungs, stomach, intestines, colon, kidneys, pancreas, breast, bone, prostate, cervix, testes, ovaries, tonsils, or other organs and / or cells derived therefrom. In certain embodiments, cells from the circulating blood of the subject are obtained, for example, by apheresis or leukocyte apheresis. In certain embodiments, the sample contains lymphocytes including T cells, monocytes, granulocytes, B cells, other nucleated leukocytes, HSCs, HPCs, HSPCs, erythrocytes and / or platelets, and in some embodiments, it contains cells other than erythrocytes and platelets that require further processing.

[0057] In some embodiments, blood cells collected from the subject are washed, for example, to remove the plasma fraction and to place the cells in a suitable buffer or culture medium for subsequent processing steps. In certain embodiments, the cells are washed with phosphate-buffered saline (PBS). In some embodiments, the washing solution is deficient in calcium and / or magnesium and / or many or all divalent cations. Washing may be achieved using a semi-automatic "flow-through" centrifuge (e.g., Cobe 2991 cell processor, Baxter) according to the manufacturer's instructions. Tangential flow filtration (TFF) may also be performed. In certain embodiments, after washing, the cells may be resuspended in a variety of biocompatible buffers, such as Ca++ / Mg++-free PBS.

[0058] Isolation may comprise one or more of various cell preparation and separation steps, including separation based on one or more properties such as size, density, sensitivity or tolerance to a particular reagent and / or affinity, e.g., immunoaffinity to an antibody or other binding partner. In certain embodiments, isolation is performed sequentially and / or simultaneously in a single process stream using the same apparatus or equipment. In certain embodiments, isolation, culture, and / or operations of different populations are performed from the same starting composition or material, such as from the same sample.

[0059] In certain embodiments, a sample may be enriched for T cells by using density-based cell separation methods and related techniques. For example, leukocytes may be separated from other cell types in peripheral blood by lysing erythrocytes and centrifuging the sample using a Percoll or Ficoll gradient.

[0060] In certain embodiments, a bulk T cell population that is not enriched for a particular T cell type may be used. In certain embodiments, the selected T cell type may be enriched and / or isolated based on positive and / or negative selection based on a cell marker. In positive selection, cells bound to the cell marker are retained for further use. In negative selection, cells not bound to the cell marker by a capture agent such as an antibody are retained for further use. In some examples, both fractions may be retained for further use.

[0061] Isolation does not necessarily result in 100% enrichment or removal of a particular cell population or cells expressing a particular marker. For example, positive selection or enrichment of a particular type of cell refers to increasing the number or percentage of such cells, but does not necessarily result in the complete absence of cells that do not express the marker. Similarly, negative selection, removal, or depletion of a particular type of cell refers to decreasing the number or percentage of such cells, but does not necessarily result in the complete removal of all such cells.

[0062] In some cases, multiple rounds of isolation processes are performed, and fractions selected as positive or negative in one process are subjected to other isolation processes, such as subsequent positive or negative selection.

[0063] In some embodiments, antibodies or binding domains for cell markers bind to a solid support or matrix, such as magnetic or paramagnetic beads, to enable the separation of cells for positive and / or negative selection. For example, in some embodiments, cells and cell populations are separated or isolated using immunomagnetic (or affinity magnetic) separation techniques (as outlined in Methods in Molecular Medicine, vol. 58: Metastasis Research Protocols, Vol. 2: Cell Behavior In Vitro and In Vivo, pp. 17–25, edited by SABrooks and U.Schumacher (copyright) Humana Press Inc., Totowa, NJ). See also US4,452,773, US4,795,698, US5,200,084 and EP452342.

[0064] In some embodiments, affinity-based selection is performed by magnetically activated cell sorting (MACS) (Miltenyi Biotec, Auburn, CA). The MACS system enables high-purity selection of cells to which magnetized particles are attached. In certain embodiments, MACS operates in a mode in which non-target and target species are sequentially eluted after the application of an external magnetic field. That is, cells attached to magnetized particles are retained in place, while non-attached species are eluted. Next, after this initial elution step is complete, species that were trapped by the magnetic field and whose elution was prevented are released in some way so that they can be eluted and recovered. In certain embodiments, non-target cells are labeled and depleted from heterogeneous cell populations.

[0065] In some embodiments, the cell populations described herein are collected and concentrated (or depleted) by flow cytometry, in which cells stained for multiple cell surface markers are carried in a fluid flow. In some embodiments, the cell populations described herein are collected and concentrated (or depleted) by preparative scale (FACS) sorting. In certain embodiments, the cell populations described herein are collected and concentrated (or depleted) by the use of a microelectromechanical system (MEMS) chip combined with a FACS-based detection system (see, e.g., WO2010 / 033140, Cho et al. (2010) Lab Chip 10, pp. 1567-1573; and Godin et al. (2008) J Biophoton. 1(5): pp. 355-376). In either case, cells may be labeled with multiple markers, enabling highly purified isolation of clearly defined cell subsets.

[0066] Cell markers for different T cell subpopulations are described above. In certain embodiments, cells expressing a specific subpopulation of T cells, e.g., positive or high levels of one or more surface markers, e.g., CCR7, CD45RO, CD8, CD27, CD28, CD62L, CD127, CD4 and / or CD45RA T cells, are isolated by positive or negative selection techniques.

[0067] CD3+, CD28+ T cells can be positively selected and expanded using anti-CD3 / anti-CD28 conjugate magnetic beads (e.g., DYNABEADS® M-450 CD3 / CD28 T Cell Expander).

[0068] In certain embodiments, a CD8+ or CD4+ selection step is used to separate CD4+ helper and CD8+ cytotoxic T cells. Such CD8+ and CD4+ populations may be further sorted into subpopulations by positive or negative selection for markers expressed on or relatively highly expressed on one or more naive, memory, and / or effector T cell subpopulations.

[0069] In some embodiments, enrichment is performed for central memory T (TCM) cells. In certain embodiments, memory T cells are present in both CD8+ and CD62L subsets of peripheral blood lymphocytes. PBMCs may be enriched or depleted for the CD62L, CD8, and / or CD62L+CD8+ fractions, for example, by using anti-CD8 and anti-CD62L antibodies.

[0070] In some embodiments, enrichment of central memory T (TCM) cells is based on positive or high surface expression of CCR7, CD45RO, CD27, CD62L, CD28, CD3, and / or CD127; in some embodiments, it is based on negative selection of cells expressing or highly expressing CD45RA and / or granzyme B. In some embodiments, isolation of the enriched CD8+ population of TCM cells is performed by depletion of cells expressing CD4, CD14, and CD45RA, and positive selection or enrichment of cells expressing CCR7, CD45RO, and / or CD62L. In one embodiment, enrichment of central memory T (TCM) cells is performed starting with a negative fraction of cells selected based on CD4 expression, which is subjected to negative selection based on CD14 and CD45RA expression, and positive selection based on CD62L. Such selections are performed simultaneously in some embodiments and sequentially in any order in others. In some embodiments, the same CD4 expression-based selection step used in preparing a CD8+ cell population or subpopulation is also used to generate a CD4+ cell population or subpopulation, thereby selectively retaining both positive and negative fractions from CD4-based isolation after one or more further positive or negative selection steps.

[0071] In certain cases, PBMC samples or other leukocyte samples are subjected to CD4+ cell selection, retaining both negative and positive fractions. The negative fraction is then subjected to negative selection based on the expression of CD14 and CD45RA or RORl, and positive selection based on markers characteristic of central memory T cells such as CCR7, CD45RO and / or CD62L, with positive and negative selection performed in either order.

[0072] In certain embodiments, cell enrichment results in a bulk CD8+FACs-sorted cell population.

[0073] Other cell types can be enriched based on known marker profiles and techniques. For example, CD34+HSCs, HSPs, and HSPCs can be enriched using anti-CD34 antibodies directly or indirectly conjugated to magnetic particles connected to a magnetic cell separation device, such as the CliniMACS® Cell Separation System (Miltenyi Biotec, Bergisch Gladbach, Germany).

[0074] (iii) Genetic modification of a cell population to express a chimeric antigen receptor (CAR). The cell population is genetically modified to express a chimeric antigen receptor (CAR) as described herein.

[0075] (iii-a) Genetic engineering techniques. Any desired gene encoding a CAR disclosed herein can be introduced into cells by any method known in the art, including transfection, electroporation, microinjection, lipofection, calcium phosphate-mediated transfection, infection with a virus or bacteriophage vector containing a gene sequence, cell fusion, chromosome-mediated gene transfer, microcell-mediated gene transfer, spheroplast fusion, in vivo nanoparticle-mediated delivery, etc. Numerous techniques for introducing foreign genes into cells are known in the art (see, for example, Loeffler and Behr, 1993, Meth. Enzymol. 217, pp. 599-618; Cohen et al., 1993, Meth. Enzymol. 217: pp. 618-644; Cline, 1985, Pharmac. Ther. 29: pp. 69-92), and can be used as long as the necessary development and physiological functions of the recipient cells are not unnecessarily disrupted. This technology provides a stable introduction of genes into cells so that the genes are expressible by the cells, preferably heritable in certain cases, and expressible by the offspring of these cells.

[0076] The term “gene” refers to a nucleic acid sequence (used interchangeably with polynucleotides or nucleotide sequences) encoding a CAR containing the CD33-binding domain as described herein. This definition includes a variety of sequence polymorphisms, mutations, and / or sequence variants such as those which do not substantially affect the function of the encoded CAR. The term “gene” may also include regulatory regions such as promoters, enhancers, and termination regions, as well as coding sequences. The term may further include all introns and other DNA sequences spliced ​​from mRNA transcripts, along with variants arising from alternative splice sites. Gene sequences encoding molecules may be DNA or RNA that directs the expression of a chimeric molecule. These nucleic acid sequences may be DNA strand sequences transcribed into RNA or RNA sequences translated into proteins. Nucleic acid sequences include both full-length nucleic acid sequences and incomplete-length sequences derived from full-length proteins. Sequences may also include degenerate codons of native sequences(s) that may be introduced to provide codon selection in a particular cell type. As will be understood by those skilled in the art, portions of complete gene sequences are referred to throughout this disclosure.

[0077] Gene sequences encoding CARs are provided herein and can be readily prepared by synthetic or recombinant methods from related amino acid sequences and other descriptions provided herein. In embodiments, gene sequences encoding any of these sequences may also have one or more restriction enzyme sites at the 5' and / or 3' ends of the coding sequence to provide easy excision and substitution of the sequence-encoding gene sequence with another gene sequence encoding a different sequence. In embodiments, the sequence-encoding gene sequences may be codon-optimized for expression in mammalian cells.

[0078] "Code" refers to the characteristic of a particular sequence of nucleotides in a gene, such as cDNA or mRNA, to function as a template for the synthesis of other macromolecules, such as defined sequences of amino acids. Therefore, a gene codes for a protein if the transcription and translation of mRNA equivalent to a gene produces a protein in a cell or other biological system. A "protein-coding gene sequence" includes all nucleotide sequences that code for the same amino acid sequence(s) of substantially similar form and function, in degenerate form of each other.

[0079] Polynucleotide gene sequences encoding more than one portion of an expressed CAR can be operably ligated to each other and to related regulatory sequences. For example, a functional binding exists between a regulatory sequence and an exogenous nucleic acid sequence, which can result in the expression of the latter. In another example, if a first nucleic acid sequence is functionally related to a second nucleic acid sequence, the first nucleic acid sequence can be operably ligated to the second nucleic acid sequence. For example, if a promoter affects the transcription or expression of a coding sequence, the promoter is operably ligated to the coding sequence. Generally, operably ligated DNA sequences are adjacent and, if necessary or beneficial, ligate coding regions into the same reading frame.

[0080] In any of the embodiments described herein, the polynucleotide may include a polynucleotide encoding a self-cleaving polypeptide, which is located between the polynucleotide encoding the CAR construct and the polynucleotide encoding the transduction marker (e.g., tEGFR). Exemplary self-cleaving polypeptides include 2A peptides derived from porcine teschovirus-1 (P2A), Thosea asigna virus (T2A), equine rhinitis A virus (E2A), foot-and-mouth disease virus (F2A), or their variants (see Figure 19). Further exemplary nucleic acids and amino acid sequences of 2A peptides are described, for example, by Kim et al. (PLOS One 6:e18556 (2011)).

[0081] A "vector" is a nucleic acid molecule capable of transporting another nucleic acid. Vectors can be, for example, plasmids, cosmids, viruses, or phages. An "expression vector" is a vector that, when present in the appropriate environment, can direct the expression of proteins encoded by one or more genes carried by the vector.

[0082] "Lentivirus" refers to a genus of retroviruses that can infect both dividing and non-dividing cells. Some examples of lentiviruses include HIV (human immunodeficiency virus: including HIV type 1 and HIV type 2); equine infectious anemia virus; feline immunodeficiency virus (FIV); bovine immunodeficiency virus (BIV); and simian immunodeficiency virus (SIV).

[0083] A "retrovirus" is a virus that has an RNA genome. "Gamma retrovirus" refers to a genus of the family Retroviridae. Examples of gamma retroviruses include mouse stem cell virus, mouse leukemia virus, feline leukemia virus, feline sarcoma virus, and reticuloendotheliosis virus.

[0084] Retroviral vectors (see Miller et al., 1993, Meth. Enzymol. 217: pp. 581-599) may be used. In such embodiments, the gene to be expressed is cloned into a retroviral vector for delivery to cells. In certain embodiments, the retroviral vector includes all the cis-acting sequences necessary for packaging and incorporating the viral genome, namely (a) long terminal repeats (LTRs) or parts thereof at each end of the vector, (b) primer binding sites for the synthesis of minus and plus strand DNA, and (c) packaging signals necessary for the uptake of genomic RNA into virions. Further details on retroviral vectors can be found in Boesen et al., 1994, Biotherapy 6: pp. 291-302; Clowes et al., 1994, J. Clin. Invest. 93: pp. 644-651; Kiem et al., 1994, Blood 83: pp. 1467-1473; Salmons and Gunzberg, 1993, Human Gene Therapy 4: pp. 129-141; and Grossman and Wilson, 1993, Curr. Opin. in Genetics and Devel. 3: pp. 110-114. Adenoviruses, adeno-associated viruses (AAVs), and alphaviruses may also be used. See Kozarsky and Wilson, 1993, Current Opinion in Genetics and Development 3: pp. 499-503; Rosenfeld et al., 1991, Science 252: pp. 431-434; Rosenfeld et al., 1992, Cell 68: pp. 143-155; Mastrangeli et al., 1993, J. Clin. Invest. 91: pp. 225-234; Walsh et al., 1993, Proc. Soc. Exp. Bioi. Med. 204: pp. 289-300; and Lundstrom, 1999, J. Recept. Signal Transduct. Res. 19: pp. 673-686.Other methods of gene delivery include the use of mammalian artificial chromosomes (Vos, 1998, Curr. Op. Genet. Dev. 8: pp. 351-359); liposomes (Tarahovsky and Ivanitsky, 1998, Biochemistry (Mosc) 63: pp. 607-618); ribozymes (Branch and Klotman, 1998, Exp. Nephrol. 6: pp. 78-83); and triplex DNA (Chan and Glazer, 1997, J. Mol. Med. 75: pp. 267-282).

[0085] Numerous suitable viral vectors are available within the scope of this disclosure, including those identified for application in human gene therapy (see Pfeifer and Verma, 2001, Ann. Rev. Genomics Hum. Genet. 2: 177). Methods for packaging cells for transduction into mammalian host cells with viral particles containing CAR transgenes using retroviral and lentiviral vectors are described, for example, US8, 119, 772; Walchli et al., 2011, PLoS One 6: 327930; Zhao et al., 2005, J. Immunol. 174: 4415; Engels et al., 2003, Hum. Gene Ther. 14: 1155; Frecha et al., 2010, Mol. Ther. 18: 1748; and Verhoeyen et al., 2009, Methods Mol. Biol. 506: 97. Retrovirus and lentivirus vector constructs and expression systems are also commercially available.

[0086] Targeted genetic engineering approaches may also be available. The CRISPR (clustered, regularly arranged, short palindromic sequence repeats) / Cas (CRISPR-related protein) nuclease system is an engineered nuclease system used for genetic engineering based on bacterial systems. Information on the CRISPR-Cas system and its components can be found, for example, in US8697359, US8771945, US8795965, US8865406, US8871445, US8889356, US8889418, US8895308, US8906616, US8932814, US8945839, US8993233 and US8999641, as well as to these and related applications; as well as WO2014 / 018423, WO2014 / 093595, WO2014 / 093622, WO2014 / 093635, WO2014 / 093655, WO2014 / 093661, WO2014 / 093694, WO2014 / 093701, WO2014 / 093709, WO2014 / 093712, WO2014 / 093718, WO2014 / 145599, WO2014 / 204723, WO2014 / 204724, WO2014 / 204725, WO2014 / 204726, WO 2014 / 204727, WO2014 / 204728, WO2014 / 204729, WO2015 / 065964, WO2015 / 089351, WO2015 / 089354 This is described in WO2015 / 089364, WO2015 / 089419, WO2015 / 089427, WO2015 / 089462, WO2015 / 089465, WO2015 / 089473 and WO2015 / 089486, WO2016205711, WO2017 / 106657, WO2017 / 127807 and related applications.

[0087] Certain embodiments utilize zinc finger nucleases (ZFNs) as gene editing agents. ZFNs are a class of site-specific nucleases engineered to bind to and cleave DNA at specific locations. ZFNs are used to introduce double-strand breaks (DSBs) at specific sites within a DNA sequence, which allows ZFNs to target unique sequences within the genome in various different cells. For further information regarding ZFNs and useful ZFNs within the scope of the teachings of this disclosure, see, for example, US6,534,261;US6,607,882;US6,746,838;US6,794,136;US6,824,978;6,866,997;US6,933,113;6,979,539;US7,013,219;US7,030,215;US7,220,719;US7,241,573;US7,241,574;US7,585,849;US7,595,376;US6,903,185;US6,479,626;US2003 / 0232410 and US2009 / 0203140 and Gaj et al., Nat Methods, 2012, 9(8):805-807; Ramirez et al., Nucle Acids Res, 2012, 40(12):5560-558; Kim et al., Genome Res, 2012, 22(7):1327-133; Urnov et al., Nature Reviews Genetics, 2010, 11:636-646; Miller et al., Nature biotechnology 25, 778-785 (2007); Bibikova et al., Science 300, 764 (2003); Bibikova et al., Genetics 161, 1169-1175 (2002); Wolfe et al., Annual review of biophysics and biomolecular structure 29, 183-212 (2000); Kim et al., Proceedings of the National Academy of Sciences of See The United States of America 93, pp. 1156–1160 (1996); and Miller et al., The EMBO journal 4, pp. 1609–1614 (1985).

[0088] In certain embodiments, activator-like effector nucleases (TALENs) may be used as gene editing agents. A TALEN refers to a fusion protein comprising an activator-like effector (TALE) DNA-binding protein and a DNA-cleavage domain. TALENs are used to edit genes and genomes by inducing two double-strand breaks (DSBs) within the DNA, which induce intracellular repair mechanisms. Generally, the two TALENs must bind to each side of the target DNA site, with the DNA-cleavage domains flanking each other, so that they dimerize and induce the DSBs. For further information on TALEN, see US8,440,431;US8,440,432;US8,450,471;US8,586,363; and US8,697,853; as well as Joung and Sander, Nat Rev Mol Cell Biol, 2013, 14(l):49-55; Beurdeley et al., Nat Commun, 2013, 4:1762; Scharenberg et al., Curr Gene Ther, 2013, 13(4):291-303; Gaj et al., Nat Methods, 2012, 9(8):805-807; Miller et al., Nature biotechnology 29, 143-148 (2011); Christian et al., Genetics 186, 757-761 (2010); Boch et al., Science See also Moscou and Bogdanove, Science 326, p. 1509–1512 (2009); and Science 326, p. 1501 (2009).

[0089] In certain embodiments, MegaTAL can be used as a gene editing agent. MegaTAL has an sc-rare cleavage nuclease structure in which TALE is fused with the DNA cleavage domain of a meganuclease. Meganucleases, also known as homing endonucleases, are single peptide chains that possess both DNA recognition and nuclease functions within the same domain. In contrast to TALENs, megaTAL requires only the delivery of a single peptide chain for functional activity.

[0090] Nanoparticles that result in selective in vivo gene modification of targeted cell types are described and may be used within the scope of the teachings of this disclosure. In certain embodiments, the nanoparticles may be those described in WO2014153114, WO2017181110 and WO201822672.

[0091] (iii-b) CAR subcomponents. As previously described, CAR molecules contain several distinct subcomponents that enable genetically modified cells to recognize and kill undesirable cells, such as cancer cells. The subcomponents include at least an extracellular component and an intracellular component. The extracellular component includes a binding domain that specifically binds to markers selectively present on the surface of undesirable cells. Once the binding domain binds to such a marker, the intracellular component activates the cell to destroy the bound cell. CAR further includes a transmembrane domain that links the extracellular component to the intracellular component, and other subcomponents that can enhance the function of CAR. For example, by including a spacer region and / or one or more linker sequences, CAR can have further conformational flexibility, which can often increase the ability of the binding domain to bind to targeted cellular markers.

[0092] (iii-bi) Binding Domain. This disclosure provides a novel binding domain for use in CAR based on an antibody that binds to CD33. The antibody is produced from two genes, a heavy chain gene and a light chain gene. Generally, the antibody contains two identical copies of the heavy chain and two identical copies of the light chain. Within the variable heavy chain and variable light chain, segments referred to as complementarity-determining regions (CDRs) determine epitope binding. Each heavy chain has three CDRs (i.e., CDRH1, CDRH2, and CDRH3), and each light chain has three CDRs (i.e., CDRL1, CDRL2, and CDRL3). The CDR regions are adjacent to framework residues (FRs). The precise amino acid sequence boundaries of a given CDR or FR can be easily determined using one of several well-known schemes, including: Kabat et al. (1991) "Sequences of Proteins of Immunological Interest", 5th edition, Public Health Service, National Institutes of Health, Bethesda, MD. (Kabat numbering scheme); Al-Lazikani et al. (1997) J Mol Biol 273: pp. 927-948 (Chothia numbering scheme); North et al. (2011) J Mol Biol 406(2): pp. 228-256 (North numbering scheme); Maccallum et al. (1996) J Mol Biol 262: pp. 732-745 (Contact numbering scheme); Martin et al. (1989) Proc. Natl. Acad. Sci., 86: pp. 9268-9272 (AbM numbering scheme); Lefranc This includes those described in MP et al. (2003) Dev Comp Immunol 27(1):55-77 (IMGT numbering scheme) and Honegger and Pluckthun (2001) J Mol Biol 309(3):657-670 ("Aho" numbering scheme). The boundary between a given CDR or FR may vary depending on the scheme used for identification. For example, the Kabat scheme is based on structural alignment, while the Chothia scheme is based on structural information.The numbering for both the Kabat and Chothia schemes is based on the most common antibody region sequence lengths, with insertions corresponding to insertion letters, e.g., "30a," and deletions appearing in some antibodies. The two schemes result in different numbering because they place certain insertions and deletions ("indels") in different positions. The Contact scheme is based on the analysis of complex crystal structures and is similar in many ways to the Chothia numbering scheme. In certain embodiments, the antibody CDR sequences disclosed herein follow Kabat numbering. CDR residues can be identified using software programs such as ABodyBuilder.

[0093] The CD33-binding domain for use in CAR is derived from antibody 1H10, 1A9, 1E6, 1D2, 1B9, 1H8, 2D3, or 2E3. In certain embodiments, the antibody comprises the following CDR set. A CDR set refers to three light chain CDRs and three heavy chain CDRs that together produce binding to CD33.

[0094] [Table 1] JPEG2026108769000002.jpg218150 JPEG2026108769000003.jpg17150

[0095] [Table 2] JPEG2026108769000005.jpg194150

[0096] [Table 3] JPEG2026108769000007.jpg158150

[0097] [Table 4] JPEG2026108769000009.jpg210150 JPEG2026108769000010.jpg128150

[0098] [Table 5] JPEG2026108769000012.jpg212150 JPEG2026108769000013.jpg91150

[0099] Specific embodiments include scFv derived from CDR, VL, or VH of 1H10, 1A9, 1E6, 1D2, 1B9, 1H8, 2D3, or 2E3 for use in CAR. Examples of such scFv are provided in Figure 19. The scFv may be formed in VH-VL orientation or VL-VH orientation. The scFv for use in CAR may also be formulated from the variable chains of these antibodies.

[0100] In a particular embodiment, the 1H10 antibody has the following sequence:

[0101] [ka] Variable light chains and sequences including:

[0102] [ka] It includes variable heavy chains.

[0103] In a particular embodiment, the 1A9 antibody has the following sequence:

[0104] [ka] Variable light chains and sequences including:

[0105] [ka] It includes variable heavy chains.

[0106] In a particular embodiment, the 1E6 antibody has the following sequence:

[0107] [ka] Variable light chains and sequences including:

[0108] [ka] It includes variable heavy chains.

[0109] In a particular embodiment, the 1D2 antibody has the following sequence:

[0110] [ka] Variable light chains and sequences including:

[0111] [ka] It includes variable heavy chains.

[0112] In a particular embodiment, the 1B9 antibody has the following sequence:

[0113] [ka] Variable light chains and sequences including:

[0114] [ka] It includes variable heavy chains.

[0115] In a particular embodiment, CD33 Vセット The antibody contains 1H8. In certain embodiments, the 1H8 antibody has the following sequence:

[0116] [ka] Variable light chains and sequences including:

[0117] [ka] It includes variable heavy chains.

[0118] In a particular embodiment, CD33 Vセット The antibody contains 2D3. In certain embodiments, the 2D3 antibody has the sequence:

[0119] [ka] Variable light chains and sequences including:

[0120] [ka] It includes variable heavy chains.

[0121] In a particular embodiment, CD33 Vセット The antibody contains 2E3. In certain embodiments, the 2E3 antibody has the sequence:

[0122] [ka] Variable light chains and sequences including:

[0123] [ka] It includes variable heavy chains.

[0124] In some cases, further scFvs based on binding domains for use in CARs, as described herein, may be prepared according to methods known in the art (see, for example, Bird et al., (1988) Science 242: pp. 423-426 and Huston et al., (1988) Proc. Natl. Acad. Sci. USA 85: pp. 5879-5883). ScFv molecules may be produced by linking and joining the VH and VL regions of an antibody using a flexible polypeptide linker. When a short polypeptide linker (e.g., 5-10 amino acids) is used, intrachain folding is inhibited. Interchain folding is also required for the two variable regions to join together to form a functional epitope binding site. For examples of linker orientation and size, see, for example, Hollinger et al., 1993, Proc Natl Acad.Sci.USA90:6444-6448, US2005 / 0100543, US2005 / 0175606, US2007 / 0014794, and WO2006 / 020258 and WO2007 / 024715. More specifically, the linker sequences used to connect the VL and VH of scFv are generally 5-35 amino acid lengths. In certain embodiments, the VL-VH linker contains 5-35, 10-30, or 15-25 amino acids. Varying the linker length can maintain or increase activity and may result in superior efficacy in activity testing. scFv is commonly used as a CAR binding domain.

[0125] Other binding fragments such as Fv, Fab, Fab', and F(ab')2 may also be used within the CARs disclosed herein. Further examples of antibody-based binding domain formats for use in CARs include scFv-based grababody and soluble VH domain antibodies. These antibodies use only the heavy chain variable region to form the binding region. See, for example, Jespers et al., Nat. Biotechnol. 22:1161, 2004; Cortez-Retamozo et al., Cancer Res. 64:2853, 2004; Baral et al., Nature Med. 12:580, 2006; and Barthelemy et al., J. Biol. Chem. 283:3639, 2008.

[0126] A functional variant comprises the addition or substitution of one or more residues that does not substantially affect the physiological effects of the protein. A functional fragment comprises one or more deletions or truncations that do not substantially affect the physiological effects of the protein. The absence of substantial effect can be confirmed by observing experimentally equivalent results in activation or binding studies. Functional variants and functional fragments of intracellular domains (e.g., intracellular signaling domains), when in the activated state of this disclosure, transmit activation or inhibitory signals equivalent to those of the wild-type reference. Functional variants and functional fragments of binding domains bind to their alloantigens or ligands at levels equivalent to those of the wild-type reference.

[0127] In certain embodiments, the VL region in the binding domain of this disclosure is derived from or based on the VL of the antibody disclosed herein and contains one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) insertions, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) deletions, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (e.g., conservative amino acid substitutions) or combinations thereof, compared to the VL of the antibody disclosed herein. Insertions, deletions, or substitutions may be located anywhere in the VL region, including the amino-terminus, carboxyl-terminus, or both ends of this region, provided that each CDR contains no changes, or contains one, two, or three or fewer changes, and that the binding domain containing the modified VL region is still capable of specifically binding to its target with the same affinity as the wild-type binding domain.

[0128] In certain embodiments, the binding domain VH region of the Disclosure may be derived from or obtained based on the VH of an antibody disclosed herein and may contain one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) insertions, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) deletions, one or more (e.g., 2, 3, 4, 5, 6, 7, 8, 9, 10) amino acid substitutions (e.g., conserved or non-conserved amino acid substitutions) or combinations thereof, compared to the VH of an antibody disclosed herein. The insertions, deletions, or substitutions may be located at any site in the VH region, including the amino-terminus, carboxyl-terminus, or both ends of this region, provided that each CDR contains no changes, or one, two, or three or fewer changes, and the binding domain containing the modified VH region can still specifically bind to its target with the same affinity as the wild-type binding domain.

[0129] In certain embodiments, the binding domain includes or is a sequence that is at least 90%, at least 91%, at least 92%, at least 93%, at least 94%, at least 95%, at least 96%, at least 97%, at least 98%, at least 99%, at least 99.5%, or 100% identical to the light chain variable region (VL) or the heavy chain variable region (VH), or the amino acid sequence of both, and each CDR does not include any changes from the antibodies disclosed herein or their fragments or derivatives that bind specifically to CD33, or includes one, two, or three or fewer changes.

[0130] (iii-b-ii) The spacer region is used to create appropriate distance and / or flexibility from other CAR subcomponents. As shown, in certain embodiments, the length of the spacer region is customized to bind to and mediate disruption of CD33-expressing cells. In certain embodiments, the length of the spacer region may be selected based on the location of the cell marker epitope, the affinity of the binding domain to the epitope, and / or the ability of the CD33 targeting agent to mediate cell disruption after CD33 binding.

[0131] The spacer region typically includes those containing 10-250 amino acids, 10-200 amino acids, 10-150 amino acids, 10-100 amino acids, 10-50 amino acids, or 10-25 amino acids.

[0132] In certain embodiments, the spacer regions are 5 amino acids, 8 amino acids, 10 amino acids, 12 amino acids, 14 amino acids, 20 amino acids, 21 amino acids, 26 amino acids, 27 amino acids, 45 amino acids, or 50 amino acids. These lengths are considered short spacer regions.

[0133] In certain embodiments, the spacer regions are 100 amino acids, 110 amino acids, 120 amino acids, 125 amino acids, 128 amino acids, 131 amino acids, 135 amino acids, 140 amino acids, 150 amino acids, 160 amino acids, or 170 amino acids. These lengths are considered intermediate spacer regions.

[0134] An exemplary spacer region includes all or part of an immunoglobulin hinge region. The immunoglobulin hinge region may be a wild-type immunoglobulin hinge region or a modified wild-type immunoglobulin hinge region. In certain embodiments, the immunoglobulin hinge region is a human immunoglobulin hinge region. As used herein, “wild-type immunoglobulin hinge region” refers to naturally occurring upper hinge amino acid sequences and middle hinge amino acid sequences that interpose between the CH1 and CH2 domains (for IgG, IgA, and IgD) to connect them, or between the CH1 and CH3 domains (for IgE and IgM) found in the heavy chain of an antibody to connect them.

[0135] The immunoglobulin hinge region may be an IgG, IgA, IgD, IgE, or IgM hinge region. The IgG hinge region may be an IgG1, IgG2, IgG3, or IgG4 hinge region. Sequences from IgG1, IgG2, IgG3, IgG4, or IgD may be used alone or in combination with all or part of a CH2 region, all or part of a CH3 region, or all or part of a CH2 region and all or part of a CH3 region.

[0136] In certain embodiments, the spacer is a short spacer including an IgG4 hinge region. In certain embodiments, the short spacer is coded by either Seq...

Claims

1. According to North, IMGT, Kabat, or Chothia, extracellular components containing a binding domain having a set of complementarity-determining regions (CDRs) for antibodies 1E6, 1H10, 1A9, 1D2, 1B9, 1H8, 2D3, or 2E3, Intracellular components including effector domains, and Transmembrane domains that link extracellular components to intracellular components A chimeric antigen receptor (CAR) that includes this receptor.

2. The CAR according to claim 1, wherein the binding domain includes a single-stranded variable fragment (scFv).

3. scFv is, The 1H10 scFv code sequence shown in sequence number 2, The 1A9 scFv code sequence shown in sequence number 3, The 1E6 scFv code sequence shown in sequence number 4, The 1D2 scFv code sequence shown in sequence number 5, The 1B9 scFv code sequence shown in sequence number 6, The 1H8 scFv code sequence shown in sequence number 7, The 2D3 scFv code sequence shown in sequence number 8, or The 2E3 scFv code sequence shown in sequence number 9 The CAR according to claim 2, which is coded by

4. scFv is, 1H10 scFv in VH-VL orientation as shown in Sequence ID No. 190, 1H10 scFv in VL-VH orientation as shown in Sequence ID No. 191, 1A9 scFv in VH-VL orientation as shown in Sequence ID No. 192, 1A9 scFv in VL-VH orientation as shown in Sequence ID No. 193, 1E6 scFv in VH-VL orientation shown in Sequence ID No. 194, 1E6 scFv in VL-VH orientation shown in Sequence ID No. 195, 2D3 scFv in VH-VL orientation as shown in Sequence ID No. 196, or 2D3 scFv in VL-VH orientation as shown in Sequence ID No. 197 The CAR according to claim 2.

5. The CAR according to claim 1, wherein the extracellular component further comprises a spacer region.

6. The CAR according to claim 5, wherein the spacer region is less than 135 amino acids or less than 16 amino acids.

7. The CAR according to claim 5, wherein the spacer region consists of less than 131 amino acids and comprises the hinge region and CH3 domain of IgG4.

8. The CAR according to claim 5, wherein the spacer region consists of less than 12 amino acids and comprises an IgG4 hinge region.

9. The CAR according to claim 7, wherein IgG4 is human IgG4.

10. The CAR according to claim 5, wherein the spacer region is coded by the sequence shown in sequence number 10, sequence number 11, or sequence number 12.

11. The CAR according to claim 1, wherein the effector domain includes all or part of the signaling domain of CD3ζ, all or part of the signaling domain of 4-1BB, all or part of the signaling domain of CD28, all or part of the signaling domains of CD3ζ and 4-1BB, all or part of the signaling domains of CD3ζ and CD28, or all or part of the signaling domains of CD3ζ, 4-1BB and CD28.

12. The CAR according to claim 1, wherein the effector domain comprises all or part of the signal transduction domains of CD3ζ and 4-1BB.

13. The CAR according to claim 11, wherein the CD3ζ signaling domain is encoded by the CD3ζ coding sequence shown in Sequence ID No.

14.

14. The CAR according to claim 11, wherein CD3ζ has the sequence shown in sequence number 15 or sequence number 16.

15. The CAR according to claim 11, wherein the 4-1BB signaling domain is encoded by the sequence shown in SEQ ID NO: 17 or SEQ ID NO:

18.

16. The CAR according to claim 11, wherein 4-1BB has the sequence shown in sequence number 19 or sequence number 20.

17. The CAR according to claim 1, wherein the transmembrane domain includes a CD28 transmembrane domain.

18. The CAR according to claim 17, wherein the CD28 transmembrane domain is encoded by the sequence shown in SEQ ID NO: 21, SEQ ID NO: 22, or SEQ ID NO:

23.

19. The CAR according to claim 17, wherein the CD28 transmembrane domain has the sequence shown in SEQ ID NO: 24 or SEQ ID NO:

25.

20. The CAR according to claim 1, further comprising a control function selected from a tag cassette, a transduction marker, and / or a suicide switch.

21. A gene construct encoding CAR as described in claim 1.

22. The gene construct according to claim 21, comprising the sequence shown in SEQ ID NO: 46, SEQ ID NO: 47, SEQ ID NO: 48, SEQ ID NO: 49, SEQ ID NO: 50, SEQ ID NO: 51, SEQ ID NO: 200, SEQ ID NO: 201, SEQ ID NO: 202, or SEQ ID NO:

203.

23. Nanoparticles encapsulating the gene construct according to claim 21 or 22.

24. Cells genetically modified to express the CAR described in claim 1.

25. The cell according to claim 24, wherein the target is an autologous cell or an allogeneic cell.

26. The cell according to claim 24, which is in vivo or ex vivo.

27. The cell according to claim 24, which is a T cell, a B cell, a natural killer (NK) cell, an NK-T cell, a monocyte / macrophage, a hematopoietic stem cell (HSC), or a hematopoietic progenitor cell (HPC).

28. The cell according to claim 24, which is a T cell selected from CD3+ T cells, CD4+ T cells, CD8+ T cells, central memory T cells, effector memory T cells and / or naive T cells.

29. The cell according to claim 24, which is a CD8+ T cell.

30. The cells according to claim 24, incubated in a cell culture medium containing IL-2, IL-7, IL-15 and / or IL-21.

31. The cells according to claim 30, incubated in a cell medium containing IL-2.

32. The cell according to claim 31, wherein the cell culture medium contains 10 to 100 ng / mL of IL-2.

33. The cell according to claim 32, wherein the cell culture medium contains 50 ng / mL of IL-2.

34. The cells according to claim 30, incubated in a cell medium containing IL-7 and IL-15.

35. The cell according to claim 34, wherein the cell culture medium contains 5 to 15 ng / mL of IL-7 and 5 to 15 ng / mL of IL-15.

36. The cell according to claim 35, wherein the cell culture medium contains 10 ng / mL of IL-7 and 10 ng / mL of IL-15.

37. The cell according to claim 30, wherein the cell culture medium contains IL-7, IL-15, and IL-21.

38. The cell according to claim 37, wherein the cell culture medium contains 5 to 15 ng / mL of IL-7, 5 to 15 ng / mL of IL-15, and 5 to 15 ng / mL of IL-21.

39. The cell according to claim 38, wherein the cell culture medium contains 10 ng / mL of IL-7, 10 ng / mL of IL-15, and 10 ng / mL of IL-21.

40. A population of cells according to claim 24, formulated for administration to a target.

41. A method for treating a subject having a CD33-related disorder, comprising administering a therapeutically effective amount to a population of nanoparticles according to claim 23 or cells according to claim 40, thereby treating the subject having a CD33-related disorder.

42. The method according to claim 41, wherein the group of cells includes autologous cells or allogeneic cells.

43. The method according to claim 41, wherein the CD33-expressing cells include acute myeloid leukemia (AML) cells.

44. The method according to claim 41, wherein the CD33-related disorder includes acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CML), mast cell leukemia, myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), or megakaryocytic leukemia.

45. To determine whether the subject expresses or lacks the V-set domain of CD33, and If the subject expresses the V-set domain of CD33, select a combination therapy comprising a composition containing one or more binding domains from among 1H10, 1A9, 1E6, 1D2, and 1B9, and one or more binding domains from among 1H8, 2D3, and 2E3. The method according to claim 41, further comprising:

46. To determine whether the subject expresses or lacks the V-set domain of CD33, and If the subject does not express the V-set domain of CD33, select a therapy that includes a composition containing one or more binding domains from among 1H10, 1A9, 1E6, 1D2, and 1B9. The method according to claim 41, further comprising:

47. A method for activating an immune response to CD33-expressing cells in a subject requiring activation of an immune response to CD33-expressing cells, comprising administering a therapeutically effective amount of nanoparticles according to claim 23 or a population of cells according to claim 40 to a subject to activate an immune response to CD33-expressing cells in the subject requiring activation.

48. The method according to claim 47, wherein the cell population includes autologous cells or allogeneic cells.

49. The method according to claim 47, wherein the CD33-expressing cells include acute myeloid leukemia (AML) cells.

50. The method according to claim 47, wherein the CD33-expressing cells include acute lymphoblastic leukemia (ALL), chronic myeloid leukemia (CML), chronic myelomonocytic leukemia (CML), mast cell leukemia, myelodysplastic syndrome (MDS), B-cell acute lymphoblastic leukemia (B-ALL), T-cell acute lymphoblastic leukemia (T-ALL), or megakaryocytic leukemia.

51. To determine whether the subject expresses or lacks the V-set domain of CD33, and The method according to claim 47, further comprising selecting a combination therapy comprising a composition containing one or more binding domains from among 1H10, 1A9, 1E6, 1D2, and 1B9 and one or more binding domains from among 1H8, 2D3, and 2E3, if the subject expresses the V set domain of CD33.

52. To determine whether the subject expresses or lacks the V-set domain of CD33, and The method according to claim 47, further comprising selecting a therapy comprising a composition containing one or more binding domains from among 1H10, 1A9, 1E6, 1D2, and 1B9 if the subject does not express the V set domain of CD33.

53. A kit comprising a nucleotide sequence encoding a CAR containing one or more binding domains from among 1H10, 1A9, 1E6, 1D2, and 1B9, and a nucleotide sequence encoding a CAR containing one or more binding domains from among 1H8, 2D3, and 2E3.

Images