Intravenous solution stabilizer

Abstract

The present disclosure provides a method of administering a low dose soluble protein to a subject in need thereof, wherein the method comprises (a) adding an intravenous solution stabilizer ("IVSS") to an isotonic aqueous solution in a container to form a pretreated container, (b) adding an amount from a protein stock solution to the pretreated container of step (a) to form a low dose protein solution, and (c) administering the low dose protein solution of step (b) to the subject. Optionally, the dose of the soluble protein to be administered to the subject is an ultra-low dose protein, wherein the low dose protein solution undergoes a further dilution step. In particular, the method of administering an ultra-low dose soluble protein comprises: (a) adding an IVSS to an isotonic aqueous solution in a first container to form a first pretreated container, (b) adding an amount from the protein stock solution to the first pretreated container to form a low dose protein solution; (c) adding an IVSS to an isotonic aqueous solution in a second container to form a second pretreated container; and (d) adding an amount of the low dose protein solution in step (b) to the second pretreated container of step (c) to form an ultra-low dose protein solution, and (e) administering the low dose protein solution of step (b) to an individual in need thereof.

Description

Atty. Docket No. 0282-0010W01INTRAVENOUS SOLUTION STABILIZER FIELD OF THE INVENTION

[0001] The present disclosure provides a method of administering a low dose soluble protein to a subject in need thereof, wherein the method comprises: (a) adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a container to form a pretreated container, wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate, about 0.5% (w / v) polysorbate 80, water, and has a pH of about 5.8 to about 6.2, (b) adding an amount from a protein stock solution to the pretreated container of step (a) to form a low dose protein solution, and (c) administering the low dose protein solution of step (b) to the subject.

[0002] In some embodiments, the dose of the soluble protein to be administered to a subject in need thereof is an ultra-low dose. In such cases, the low dose protein solution undergoes a dilution step. In particular, the method of administering an ultra-low dose soluble protein comprises: (a) adding an IVSS to an isotonic aqueous solution in a first container to form a first pretreated container, (b) adding an amount from the protein stock solution to the first pretreated container to form the low dose protein solution; (c) adding an IVSS to an isotonic aqueous solution in a second container to form a second pretreated container; (d) adding a desired amount of the low dose protein solution of step (b) to the second pretreated container of step (c) to form an ultra-low dose protein solution; and (e) administering the ultra-low dose protein solution of step (d) to the subject.

[0003] The disclosure further provides a method of treatment, IVSS formulations, compositions comprising IVSS, packages comprising IVSS, and kits comprising IVSS.BACKGROUND

[0004] Protein-based therapeutics have been increasingly utilized over the past few decades for the effective treatment of numerous types of diseases, including infectious diseases, autoimmune disorders, genetic disorders, cancer, and others. However, protecting such therapeutic drugs from effects such as degradation and denaturation that occurs during processing, storage and delivery remain significant challenges. Such protein-based drugs include antibody-based drugs, Fc fusion proteins, recombinant interferons and interleukins, and the like.Atty. Docket No. 0282-0010W01

[0005] New treatments for cancer and other diseases are emerging that utilize immune mobilizing T-cell receptor proteins against diseases. By identify and / or optimizing human T-cell receptor (“TCR”) proteins that bind to target peptide MHC complexes and fusing or combining them with an effector arm that engages T cells, soluble TCRs can overcome the limitations of the natural immune system to provide new therapeutic approaches. These new treatments provide critical treatment options for patients. Recently, a bispecific TCR protein tebentafusp was approved for treating adults with uveal melanoma. Additional TCR bispecific proteins such as the bispecific T cell engager brenetafusp (PRAME X CD3) (See e.g., U. S. Patent No. 11 / 718,657, the disclosure of which is herein incorporated by reference in its entirety), the first soluble T-cell receptor (TCR) bispecific protein targeting PRAME, are also being developed.

[0006] The therapeutically effective doses of TCR proteins used as therapeutic agents are typically very low. In some instances, these low or ultra-low dose ranges make administration of TCR proteins (e.g., intravenously) prone to adsorption to the surface of storage or delivery containers, such as intravenous solution bags. The adsorption of TCR proteins may result in inconsistent dosing and product waste. Currently, human serum albumin (“HSA”) is used in some intravenous stabilizing solutions (“IVSS”) for some intravenous delivery systems to protect against surface adsorption. However, the use of HSA may result in complexity for dose preparation and administration of a TCR protein. In addition, there are ethical concerns regarding handling and disposal of formulations that contain HSA.SUMMARY OF THE INVENTION

[0007] In some aspects, the present disclosure is directed to a method of administering a low dose soluble protein to a subject in need thereof, the method comprising: (a) adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a container to form a pretreated container, wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate; about 0.5% (w / v) polysorbate 80; and water, wherein the IVSS has a pH of about 5.8 to about 6.2, (b) adding an amount from a protein stock solution to the pretreated container of step (a) to form a low dose protein solution; and (c) administering the low dose protein solution of step (b) to the subject.

[0008] In some aspects, the present disclosure is directed to a method of administering an ultra-low dose soluble protein to a subject in need thereof, the method comprising: (a) addingAtty. Docket No. 0282-0010W01an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a first container to form a first pretreated container, wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate; about 0.5% (w / v) polysorbate 80; and water, wherein the IVSS has a pH of about 5.8 to about 6.2, (b) adding an amount from a protein stock solution to the first pretreated container of step (a) to form a low dose protein solution; (c) adding an IVSS to an isotonic aqueous solution in a second container to form a second pretreated container; (d) adding an amount of the low dose protein solution of step (b) to the second pretreated container of step (c) to form an ultra-low dose protein solution; and (e) administering the ultra-low dose protein solution of step (d) to the subject.

[0009] In some aspects, the protein stock solution comprises about 50 pg / mL to about 500 pg / mL of the protein. In some aspects, the low dose protein solution comprises 20 pg to about 800 pg of the protein. In some aspects, the ultra-low dose protein solution comprises about 0.2 pg to about 20 pg of the protein. In some aspects, the amount of the protein stock solution added to the pretreated container is at least 100 pL. In some aspects, the amount of the low dose protein solution added to the second pretreated container is at least 100 pL [OOlOJIn some aspects, the IVSS comprises 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate, and 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate.

[0011] In some aspects, the low dose or the ultra-low dose soluble protein is a soluble TCR protein. In some aspects, the soluble TCR protein is a heterodimeric TCR protein. In some aspects, the soluble TCR protein is a bispecific fusion protein. In some aspects, the soluble TCR protein is a bispecific T-cell engager, such as a gplOO peptide-HLA-directed CD3 T-cell engager.

[0012] In some aspects, the soluble TCR protein comprises an alpha chain complementarity determining region (CDR) sequence according to SEQ ID NOs: 6-8 and a beta chain CDR sequence according to SEQ ID NOs: 9- 11. In some aspects, the soluble TCR protein is tebentafusp. In some aspects, the tebentafusp is added to the pretreated container from a solution at a concentration of about 50 pg / mL to about 500 pg / mL. In some aspects, the tebentafusp is administered to the subject at a dose amount of about 20 pg to about 800 pg. In some aspects, the tebentafusp is administered to the subject at a dose amount of about 0.2 pg to about 800 pg.Atty. Docket No. 0282-0010W01

[0013] In some aspects, the soluble TCR protein targets preferentially expressed antigen of melanoma (PRAME). In some aspects, the soluble TCR protein comprises an alpha chain CDR sequence according to SEQ ID NOs: 31-33, SEQ ID NOs: 59-61, or SEQ ID NOs: 65-67; and a beta chain CDR sequence according to SEQ ID NOs: 39-41; SEQ ID NOs: 62-64; or SEQ ID NOs: 68-70. In some aspects, the soluble TCR protein is brenetafusp, IMC-P115C, or IMC-T119C. In some aspects, the soluble TCR protein is brenetafusp. In some aspects, the brenetafusp is added to the pretreated container from a solution at a concentration of about 50 pg / mL to about 500 pg / mL. In some aspects, the brenetafusp is administered to the subject at a dose amount of about 20 pg to about 800 pg. In some aspects, the brenetafusp is administered to the subject at a dose amount of about 0.2 pg to about 800 pg.

[0014] In some aspects, the soluble TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator. In some aspects, the soluble TCR protein comprises an alpha chain CDR sequence according to SEQ ID NOs: 71-73, and a beta chain CDR sequence according to SEQ ID NOs: 74-76. In some aspects, the PIWIL1 modulator is IMC-R117C. In some aspects, the IMC-R117C is added to the pretreated container from a solution at a concentration of about 50 pg / mL to about 500 pg / mL. In some aspects, the IMC-R117C is administered to the subject at a dose amount of about 20 pg to about 800 pg. In some aspects, the IMC-R117C is administered to the subject at a dose amount of about 0.2 µg to about 800 µg.

[0015] In some aspects, the method in the present disclosure comprises adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a container to form a pretreated container. In some aspects, the isotonic aqueous solution comprises one or more of normal saline, Ringer’s lactate solution, dextrose, or a combination thereof. In some aspects, the IVSS comprises about 25 mM sodium citrate.

[0016] In some aspects, the low dose protein is provided in a liquid. In some aspects, the protein stock solution is prepared from a concentrated solution. In some aspects, the protein stock solution is prepared from a lyophilized TCR protein. In some aspects, the lyophilized TCR protein is reconstituted prior to adding to the pretreated container.

[0017] In some aspects, the present disclosure is directed to a pretreated container suitable for administering a low dose or an ultra-low dose protein, wherein the pretreated container comprises a composition comprising: an isotonic aqueous base; about 0.2 mM to about 0.3Atty. Docket No. 0282-0010W01mM sodium citrate; and about 0.005% (w / v) polysorbate 80, wherein the composition has a pH of about 5.8 to about 6.2. In some aspects, the composition further comprises the low dose or the ultra-low dose protein, wherein the low dose or the ultra-low dose protein is a soluble TCR protein. In some aspects, the soluble TCR protein is tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, or IMC-R117C.

[0018] In some aspects, the composition comprises about 20 pg to about 800 pg of tebentafusp. In some aspects, the composition comprises about 0.2 pg to about 20 pg of tebentafusp. In some aspects, the composition comprises about 20 pg to about 800 pg of brenetafusp. In some aspects, the composition comprises about 0.2 pg to about 20 pg of brenetafusp. In some aspects, the composition comprises about 20 pg to about 800 pg of IMC-R117C. In some aspects, the composition comprises about 0.2 pg to about 20 pg of IMC-R117C.

[0019] In some aspects, the present disclosure is directed to an intravenous solution stabilizer (“IVSS”) for pretreating a container, the IVSS comprising: a. 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate; b. 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate; and c. 5 mg / mL PS 80, wherein the IVSS has a pH of about 5.8 to about 6.2.

[0020] In some aspects, the present disclosure is directed to a package comprising an intravenous solution stabilizer (“IVSS”), wherein the package comprises a container comprising the IVSS comprising: a.0.7 mg / mL to 0.8 mg / mL citric acid monohydrate; b. 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate; and c. 5 mg / mL PS80, wherein the IVSS has a pH of about 5.8 to about 6.2. In some aspects, the package does not transmit visible light or UV light. In some aspects, the package comprises a layer that blocks greater than 90% of UV light. In some aspects, the package further comprises instructions for preparing a low dose or an ultra-low dose protein solution in the container, wherein the instructions comprise: (i) adding the IVSS to the container, wherein the container comprises an isotonic aqueous solution in a container to form a pretreated container; (ii) adding a protein solution to the pretreated container to form a low dose protein solution, and, optionally, (iii) adding the low dose protein solution to a second pretreated container to form an ultra-low dose protein solution. In some aspects, the IVSS comprises the low dose or the ultra-low dose protein. In some aspects, the low dose or the ultra-low dose protein is a soluble TCR protein. In some aspects, the soluble TCR protein is tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, or IMC-R117C.Atty. Docket No. 0282-0010W01

[0021] In some aspects, the present disclosure is directed to a kit comprising: (a) a lyophilized T-cell receptor (“TCR”) protein; and (b) an intravenous solution stabilizer (“IVSS”), wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate; about 0.5% (w / v) polysorbate 80; and water, wherein the IVSS has a pH of about 6.0; and (c) a container. In some aspects, the IVSS comprises about 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate. In some aspects, the IVSS comprises 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate. In some aspects, the lyophilized TCR protein is a bispecific T-cell engager, such as a gplOO peptide-HLA-directed CD3 T-cell engager. In some aspects, the bispecific T-cell engager is tebentafusp. In some aspects, the container is pre-treated with the IVSS. In some aspects, the tebentafusp is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pre-treated containers. In some aspects, the lyophilized TCR protein is a PRAME targeting TCR protein.

[0022] In some aspects, the PRAME targeting TCR protein is selected from the group consisting of brenetafusp, IMC-P115C, and IMC-T119C. In some aspects, the PRAME targeting TCR protein is brenetafusp. In some aspects, the brenetafusp is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pretreated containers. In some aspects, the lyophilized TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator. In some aspects, the PIWIL1 modulator is IMC-R117C. In some aspects, the IMC-R117C is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pre-treated containers. In some aspects, the kit further comprises a medical grade plastic bag, wherein the IVSS is disposed within the medical grade bag. In some aspects, the medical grade bag comprises a layer that does not transmit visible light or UV light. In some aspects, the kit comprises a first medical grade bag and a second medical grade bag, wherein the IVSS is disposed in the first medical grade bag and the lyophilized TCR protein is disposed in the second medical grade bag.

[0023] In some aspects, the present disclosure is directed to a method of treating a disease or condition in a subject in need thereof comprising administering to the subject a soluble protein administered at a low dose or ultra-low dose, i.e., a low dose protein or a ultra-low dose protein, wherein the low dose protein or the ultra-low dose protein is prepared with the use of an intravenous solution stabilizer (“IVSS”), wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate; about 0.5% (w / v) polysorbate 80; and water, wherein the IVSS has a pH of about 5.8 to about 6.2. In some aspects, the low dose protein or the ultraAtty. Docket No. 0282-0010W01low dose protein is a soluble TCR protein. In some aspects, the soluble TCR protein is a heterodimeric TCR protein. In some aspects, the soluble TCR protein is a bispecific fusion protein. In some aspects, the soluble TCR protein is a bispecific CD3 T-cell engager. In some aspects, the soluble TCR protein targets gplOO. In some aspects, the soluble TCR protein is tebentafusp. In some aspects, the soluble TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator. In some aspects, the soluble TCR protein is IMC-R117C. In some aspects, the soluble TCR protein targets preferentially expressed antigen of melanoma (PRAME). In some aspects, the soluble TCR protein is selected from the group consisting of brenetafusp, IMC-P115C, and IMC-T119C. In some aspects, the disease is a PRAME-positive cancer.

[0024] In some aspects, the present disclosure is directed to a composition comprising: (a) an intravenous solution stabilizer (“IVSS”), wherein the IVSS comprises about 20 mM to about 30 mM sodium citrate; about 0.5% (w / v) polysorbate 80; and water, wherein the IVSS has a pH of about 6.0; and (b) a low dose or an ultra-low dose soluble protein.BRIEF DESCRIPTION OF THE DRAWINGS

[0025] The following drawings form part of the present specification and are included to further demonstrate exemplary embodiments of certain aspects of the present disclosure.

[0026] FIG. 1A is a reverse phase high performance liquid chromatography graph showing the visible light overlay for Formulation 1 IVSS. As demonstrated by the poor overlay, polysorbate 80 was susceptible to chemical degradation in the IVSS Fl composition when exposed to visible light stress.

[0027] FIG. 1B is a reverse phase high performance liquid chromatography graph showing the ultraviolet light stress overlay for Formulation 1 IVSS. As demonstrated by the poor overlay, polysorbate 80 was susceptible to chemical degradation in the IVSS Fl composition when exposed to visible light stress. UV = ultraviolet.

[0028] FIG. 2 is a reverse phase high performance liquid chromatography (HPLC) graph showing the overlay for Formulation 2 IVSS stored for 12 months at 5 °C (gray line) versus 25 °C (blue line). At 12 months, Formulation 2 IVSS F2 shows signs of oxidation at 25 °C.

[0029] FIG. 3 is a reverse phase high performance liquid chromatography graph showing the overlay for an exemplary IVSS stored for 12 months at 5 °C (gray line) versus 25 °C (blueAtty. Docket No. 0282-0010W01line). The IVSS Fl composition showed good chemical stability of polysorbate 80 even up to 12 months.DETAILED DESCRIPTION

[0030] Unless otherwise defined herein, scientific and technical terms used in the present disclosure shall have meanings that are commonly understood by one of ordinary skill in the art. Further, unless otherwise required by context, singular terms shall include pluralities, and plural terms shall include the singular.

[0031] The articles “a” and “an” are used herein to refer to one or to more than one (i.e., to at least one) of the grammatical object of the article. By way of example, “an element” means one element or more than one element.

[0032] The use of the term “or” in the claims is used to mean “and / or” unless explicitly indicated to refer only to alternatives or the alternatives are mutually exclusive, although the disclosure supports a definition that refers to only alternatives and “and / or.”

[0033] As used herein, the terms “comprising” (and any variant or form of comprising, such as “comprise” and “comprises”), “having” (and any variant or form of having, such as “have” and “has”), “including” (and any variant or form of including, such as “includes” and “include”) or “containing” (and any variant or form of containing, such as “contains” and “contain”) are inclusive or open-ended and do not exclude additional, unrecited, elements or method steps.

[0034] The use of the term “for example” and its corresponding abbreviation “e.g.” means that the specific terms recited are representative examples and embodiments of the disclosure that are not intended to be limited to the specific examples referenced or cited unless explicitly stated otherwise.

[0035] As used herein, “about” can mean plus or minus 10% of the provided value. Where ranges are provided, they are inclusive of the boundary values. “About” can additionally or alternately mean either within 10% of the stated value, or within 5% of the stated value, or in some cases within 2.5% of the stated value; or “about” can mean rounded to the nearest significant digit.Atty. Docket No. 0282-0010W01

[0036] As used herein, “between” is a range inclusive of the ends of the range. For example, a number between x and y explicitly includes the numbers x and y and any numbers that fall within x and y.

[0037] In some embodiments, the methods, containers, packages and kits described herein can be used in the administration of a TCR protein to a subject. The term “administration” or “administering” refers to routes of introducing a compound or composition provided herein to a subject to perform its intended function. An example of a route of administration that can be used includes, but is not limited to, e.g., intravenous infusion (e.g., intravenous cannula (IVC) or central venous access device (CVAD)), intramuscular, subcutaneous, oral, sublingual, intranasal, transdermal administration, or any combination thereof. In some embodiments, the term “administration” or “administering” refers to intravenous infusion (e.g., intravenous cannula (IVC) or central venous access device (CVAD)). An example of a route of administration that can be used includes, but is not limited to, intravenous infusion.

[0038] The term “subject” means any subject, particularly a mammalian subject, in need of treatment. In some embodiments, the term “subject” refers to a human subject. In some embodiments, the term “subject” refers to an adult human subject. In some embodiments, the term “subject” refers to a male human subject. In some embodiments, the term “subject” refers to a female human subject. In some embodiments, the term “subject” refers to administration to a subject in need thereof, i.e., a subject having non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma and / or a subject having a PRAME-positive cancer. As used herein, a “subject in need thereof’ can refer to the subject for whom it is desirable to treat, e.g., a subject being diagnosed with non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma, and / or a PRAME-positive cancer as described herein. In some embodiments, the term “subject in need thereof’ can refer to a subject having one or more symptoms associated with non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma, and / or a PRAME-positive cancer, e.g., a subject having the following symptoms: coughing that gets worse or does not go away, chest pain, shortness of breath, wheezing, coughing up blood, abnormal vaginal discharge or bleeding after menopause, bloating or swollen feeling in the stomach, feeling full very soon after starting to eat, new urinary frequency, new constipation or other changes in bowel movements, discomfort or pain in the pelvic area, abdomen or lower back, fatigue, unexplained weight loss, poor or blurred vision in one eye, loss of peripheral vision,Atty. Docket No. 0282-0010W01brown or dark patches on the white of the eye, etc. In some embodiments, the term “subject in need thereof’ can refer to a subject at high risk for suffering from non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma, and / or a PRAME-positive cancer suitable to treatment with the soluble TCR proteins described herein.

[0039] In some embodiments, the subject is HLA-A*02 positive. The term “HLA-A*02 positive” refers to a subject having TCRs that bind to the SLLQHLIGL-HLA-A*02 complex. In some embodiments, the subject is HLA-A* 02:01 positive. The term “HLA-A*02:01 positive refers to a subject that expresses the HLA-A*02 genotype with the HLA-A*02:01 allele. In some embodiments, the subject is HLA*A0201 positive. The term “HLA*A0201 positive” refers to a subject having TCRs that bind to the YLEPGPVTA-HLA-A*0201 complex.

[0040] The term “treating” refers to administering an active agent with the purpose to cure, heal, alleviate, relieve, alter, remedy, ameliorate, improve, or affect a condition (e.g., a disease), the symptoms of the condition, or to prevent or delay the onset of the symptoms, complications, biochemical indicia of a disease, or otherwise arrest or inhibit further development of the cancer in a statistically significant manner, e.g., a non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma, and / or a subject having a PRAME-positive cancer in a statistically significant manner.

[0041] In some embodiments, an “effective amount” corresponds to a therapeutically effective amount. A therapeutically effective amount means an amount that is effective in therapy, or an amount sufficient to provide a therapeutic effect. In some embodiments, an amount that is effective in therapy is an amount which produces a biological activity and will depend, among other things, on the individual. In some embodiments, the typical daily dose of the active substance (e.g., a therapeutic agent) varies and will depend on various factors such as the individual requirements of the subjects, the mode of administration and disease.

[0042] In some embodiments, provided herein is a method of administering a low dose protein to a subject in need thereof, the method comprising: (a) adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a container to form a pretreated container, (b) adding an amount from a protein stock solution to the pretreated container of step (a) to form a low dose protein solution; and (c) administering the low dose protein solution of step (b) to a subject in need thereof.Atty. Docket No. 0282-0010W01

[0043] In some embodiments, the low dose protein is administered to a subject in a dose amount less than 1000 pg less than 800 pg, less than 700 pg, less than 600 pg, less than 500 pg, less than 450 pg, less than 400 pg, less than 350 pg, less than 300 pg, less than 250 pg, or less than 200 pg. In some embodiments, the low dose protein is administered to a subject at 1000 pg to 20 pg, 800 pg to 20 pg, 700 pg to 20 pg, 600 pg to 20 pg, 500 pg to 20 pg, 400 pg to 20 pg, 300 pg to 20 pg, or 200 pg to 20 pg. In some embodiments, the low dose protein is a soluble protein such as T-cell receptor (“TCR”) proteins. In some embodiments, the disclosure provides a method of pretreating a container for administration of drug products or drug product candidates comprising: (a) providing a soluble protein, e.g., a soluble TCR protein, (b) providing an intravenous solution stabilizer (“IVSS”), (c) adding the IVSS to the container, which in some embodiments can already contain an isotonic aqueous solution such that the IVSS and isotonic aqueous solution together forms a pretreated container, and (d) adding the soluble protein, e.g., a soluble TCR protein, to the pretreated container to form a low dose protein solution. While not intending to be bound by theory, in some embodiments, the IVSS can prevent adsorption of the low dose protein, e.g., a soluble TCR protein, onto the surface of the container thereby increasing efficiency of delivery of the low dose protein, e.g., a soluble TCR protein, even in highly diluted concentrations and / or very small dose amounts. In some embodiments, the IVSS can prevent adsorption of the TCR protein onto the surface of the container thereby increasing consistency in delivering the expected dosing amount.

[0044] In embodiments, 0.5 mL of IVSS is added to a container, e.g., an IV bag(s). In some embodiments, 1 mL of IVSS is added to each container, e.g., an IV bag(s). In some embodiments, the amount to be administered to the subject is 100 mL of solution comprising the relevant dose of the low dose protein. In specific embodiments wherein the dose of the soluble protein to be administered is 20 pg or higher, the present disclosure is directed to a one-step process dilution comprising:• adding IVSS to a 100 mL bag;• adding a desired amount from a stock solution of the soluble protein to 100 mL bag that has been pretreated with the IVSS; and• administering the entire 100 mL of solution containing the soluble protein to the subject.Atty. Docket No. 0282-0010W01

[0045] In embodiments, the one-step dilution process is used for administration of a soluble protein in the dose range for approved products likely to encompass ranges of 20 pg to 800 pg. In some embodiments, within this range, approved products are likely to be within 20 pg to 300 pg. Exemplary amounts of stock solution to add for exemplary doses in range of 20 pg to 800 pg are presented in Table 1:Table 1. Low dose protein administration by one-step processDose to be Volume of Stock Solution (100 pg / 0.5 mL = 200 pg / mL = administered 0.2 mg / mL) to add to 100 mL aqueous saline solution20 pg 0.1 mL30 pg 0.15 mL68 pg 0.34 mL40 pg 0.2 mL140 pg 0.7 mL200 pg 1.0 mL240 pg 1.2 mL280 pg 1.4 mL300 pg 1.5 mL400 pg 2.0 mL500 pg 2.5 mL600 pg 3.0 mL700 pg 3.5 mL800 pg 4.0 mLAtty. Docket No. 0282-0010W01

[0046] In some embodiments, provided herein is a method of administering an ultra-low dose soluble protein to a subject in need thereof, the method comprising: (a) adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a first container to form a first pretreated container (b) adding an amount from a protein stock solution to the first pretreated container of step (a) to form a low dose protein solution; (c) adding an IVSS to an isotonic aqueous solution in a second container to form a second pretreated container; (d) adding an amount of the low dose protein solution of step (b) to the second pretreated container of step (c) to form an ultra-low dose protein solution; and (e) administering the ultra-low dose protein solution of step (d) to the subject.

[0047] In some embodiments, the ultra-low dose protein is administered to a subject in a dose amount less than 25 pg, less than 20 pg, less than 15 pg, less than 10 pg, less than 5 pg, less than 1 pg, less than 0.8 pg, less than 0.6 pg, less than 0.5 pg, less than 0.4 pg, or less than 0.3 pg. In some embodiments, the ultra-low dose protein is administered to a subject at 30 pg to 0.2 pg, 25 pg to 0.2 pg, 20 pg to 0.2 pg, 600 pg to 0.2 pg, 500 pg to 0.2 pg, 400 pg to 0.2 pg, 300 pg to 0.2 pg, or 200 pg to 0.2 pg. In some embodiments, the ultra-low dose protein is a soluble protein such as T-cell receptor (“TCR”) proteins. In some embodiments, the disclosure provides a method of pretreating a container for administration of drug products or drug product candidates comprising: (a) providing a soluble protein, e.g., a soluble TCR protein, (b) providing an intravenous solution stabilizer (“IVSS”), (c) adding the IVSS to a first container, which in some embodiments can already contain an isotonic aqueous solution such that the IVSS and isotonic aqueous solution together forms a first pretreated container, (d) adding the soluble protein, e.g., a soluble TCR protein, to the first pretreated container to form a low dose protein solution, (e) adding the IVSS to a second container, which in some embodiments can already contain an isotonic aqueous solution such that the IVSS and isotonic aqueous solution together forms a second pretreated container; and (f) adding a desired amount from the low dose solution of the first pretreated container to the second pretreated container to form an ultra-low dose protein solution. While not intending to be bound by theory, in some embodiments, the IVSS can prevent adsorption of the ultra-low dose protein, e.g., a soluble TCR protein, onto the surface of the container thereby increasing efficiency of delivery of the ultra-low dose protein, e.g., a soluble TCR protein, even in highly diluted concentrations and / or very small dose amounts. In some embodiments, the IVSS can prevent adsorption of the TCR protein onto the surface of the container thereby increasing consistency in delivering the expected dosing amount.Atty. Docket No. 0282-0010W01

[0048] In embodiments, 0.5 mL of IVSS is added to a container, e.g., an IV bag(s). In some embodiments, 1 mL of IVSS is added to each IV bag(s). In some embodiments, the amount to be administered to the subject is 100 mL of solution comprising the relevant dose of the ultralow dose protein. In specific embodiments wherein the dose of the soluble protein to be administered is 20 pg or lower, the present disclosure is directed to a two-step dilution process comprising:• adding IVSS to a 50 mL bag (first container);• adding a desired amount from a stock solution of the soluble protein to a first container that has been pretreated with IVSS;• adding a desired amount from the first container to a 100 mL bag (second container) that has been pretreated with IVSS; and• administering the entire 100 mL of solution containing the soluble protein to the subject.

[0049] In embodiments, the two-step dilution process is used for administration of a soluble protein in the dose range for approved products likely to encompass ranges of 0.2 pg to 20 pg. In some embodiments, within this range, approved products are likely to be within 0.2 pg to 8 pg. Exemplary amounts of stock solution to add for exemplary doses less than 20 pg are presented in Table 2:Table 2: Ultra-low dose protein administration by two-step processDose Amount of Stock Solution to add Amount of 50 mL solution to add to 50 mL container (first to 100 mL container (second container) container)0.2 µg 0.1 mL 0.5 mL0.4 pg 0.1 mL 1.0 mL1 µg 0.2 mL 1.25 mL2 µg 0.4 mL 1.25 mL4 µg 1 mL 1.0 mL5 µg 1 mL 1.25 mLAtty. Docket No. 0282-0010W016 pg 1 mL 1.5 mL10 µg 1 mL 2.5 mL12 pg 1.5 mL 2.0 mL16 pg 2.0 mL 2.0 mL

[0050] In embodiments, the amount of stock solution to add to the 50 mL container (first container) is in the range of about 0.1 mL to 2.5 mL, preferably 2.0 mL or less, 1.5 mL or less, 1.0 mL or less, 0.8 mL or less, 0.6 mL or less, 0.5 mL or less, 0.4 mL or less, 0.3 mL or less, 0.2 mL or less, 0.1 mL or less.

[0051] In embodiments, the amount of 50 mL solution (from the first container) to add to the 100 mL solution (second container) is in the range of about 0 to 5 mL, preferably 3.0 mL or less, 2.5 mL or less, 2.0 mL or less, 1.5 mL or less, 1.25 mL or less, 1.0 mL or less, 0.5 mL or less.

[0052] To administer a final low dose as required, one of ordinary skill in the art would be able to calculate the amount of a stock solution that would need to be added using a one step or two step process in order to administer the required low or ultra-low dose. The dilution could be achieved in more than one way that would be apparent to one of ordinary skill in the art. For example, 10 pg could be delivered by putting in 1 mL of a 0.2 mg / .mL stock solution to 50 mL, adding 2.5 mL of that 50 mL solution to the 100 mL bag, and then administering the 100 mL intravenously. In another example, 10 pg could be delivered by putting 2.0 mL of a 0.2 mg / mL stock solution into 50 mL, adding 1.25 mL of that 50 mL solution to the 100 mL bag, and then administering the 100 mL intravenously. The amount that is added to each bag or container can be adjusted depending on factors including: (1) the volume of the solution being added to a container relative to the volume of the solution in the container (to ensure that whatever volume added does not significantly impact the dilution, and thus the amount of therapeutic, being calculated), (2) the volume capacity of the container, and (3) the minimum amount that can be extracted from the stock solution. In embodiments, the amount of a stock solution that would need to be added using a one step or two step process is at least 100 pL.Atty. Docket No. 0282-0010W01

[0053] In embodiments, the soluble protein in the stock solution is at a concentration of about 50 pg / mL to about 1000 pg / mL, about 100 pg / mL to about 800 pg / mL, about 150 pg / mL to about 500 pg / mL. In embodiments, the soluble protein in the stock solution is at a concentration of about 100 pg / mL, about 200, pg / mL, about 300 pg / mL, about 400 pg / mL, about 500 pg / mL, about 600 pg / mL, about 700 pg / mL, or about 800 pg / mL.

[0054] Suitable containers to which the IVSS is added to form a pretreated container can include, for example, bottles, vials, syringes, intravenous solution bag, etc. In some embodiments, the container is an intravenous solution bag, such as a medical grade bag. The containers can be made from a variety of materials, such as glass or plastic. The container holds or contains a composition that comprises the IVSS. In some embodiments, the container comprises an isotonic aqueous solution, to which the IVSS is then added. In some embodiments, the container is of a size to accommodate the volume of both the isotonic aqueous solution and the IVSS.

[0055] In some embodiments, the IVSS comprises a buffering agent and a surfactant. In some embodiments, the buffering agent is sodium citrate. In some embodiments, the IVSS comprises about 20 mM to about 30 mM, e.g., about 21 mM to about 29 mM, about 22 mM to about 28 mM, about 23 mM to about 27 mM or about 24 mM to about 26 mM sodium citrate. In some embodiments, the IVSS comprises about 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, 25 mM, 26 mM, 27 mM, 28 mM, 29 mM, or 30 mM sodium citrate. In some embodiments, the IVSS comprises about 25 mM sodium citrate.

[0056] In some embodiments, the term sodium citrate refers to one or more acid salts of citric acid, including monosodium citrate, disodium citrate, and / or trisodium citrate. In some embodiments, the sodium citrate is provided as a combination of acid salts of citric acid. In some embodiments, the sodium citrate is formed with citric acid, such as, but not limited to citric acid monohydrate.

[0057] In some embodiments, the IVSS includes citric acid monohydrate in an amount of about 0.7 mg / mL to about 0.8 mg / mL, about 0.72 mg / mL to about 0.78 mg / mL, or about 0.74 mg / mL to about 0.76 mg / mL (e.g., 0.7 mg / mL, 0.71 mg / mL, 0.72 mg / mL, 0.73 mg / mL, 0.74 mg / mL, 0.75 mg / mL, 0.76 mg / mL, 0.77 mg / mL, 0.78 mg / mL, 0.79 mg / mL, 0.8 mg / mL,). In some embodiments, the IVSS includes trisodium citrate dihydrate in an amount of about 6.2 mg / mL to about 6.3 mg / mL, 6.2 mg / mL to 6.28 mg / mL, 6.2 mg / mL to 6.26 mg / mL, 6.2Atty. Docket No. 0282-0010W01mg / mL to 6.24 mg / mL, 6.2 mg / mL to 6.22 mg / mL, 6.22 mg / mL to 6.30 mg / mL, 6.22 mg / mL to 6.28 mg / mL, 6.22 mg / mL to 6.26 mg / mL, 6.22 mg / mL to 6.24 mg / mL, 6.24 mg / mL to 6.3 mg / mL, 6.24 mg / mL to 6.28 mg / mL, or 6.24 mg / mL to 6.26 mg / mL (e.g., 6.2 mg / mL, 6.21 mg / mL, 6.22 mg / mL, 6.23 mg / mL, 6.24 mg / mL, 6.25 mg / mL, 6.26 mg / mL, 6.27 mg / mL, 6.28 mg / mL, 6.29 mg / mL, 6.30 mg / mL).

[0058] In some embodiments, the buffering agent in the IVSS is sodium succinate. In some embodiments, the IVSS comprises about 15 mM to about 25 mM or about 18 mM to about 22 mM, e.g., 15 mM, 16 mM, 17 mM, 18 mM, 19 mM, 20 mM, 21 mM, 22 mM, 23 mM, 24 mM, or 25 mM sodium succinate. In some embodiments, the IVSS comprises about 20 mM sodium succinate. In some embodiments, the IVSS comprises about 0.5% weight to volume (“w / v”) polysorbate 80. In some embodiments, the IVSS comprises sodium succinate at about 15 mM to about 25 mM, about 0.5% w / v polysorbate 80, and water, and has a pH from about 6.4 to about 6.6.

[0059] In some embodiments, the IVSS comprises about 0.07 mg / mL to about 0.08 mg / mL succinic acid. In some embodiments, the IVSS comprises about 3.1 mg / mL to about 3.2 mg / mL disodium succinate (e.g., 3.1 mg / mL, 3.12 mg / mL, 3.14 mg / mL, 3.16 mg / mL, 3.18 mg / mL, 3.2 mg / mL, 3.1 mg / mL to 3.18 mg / mL, 3.1 mg / mL to 3.16 mg / mL, 3.1 mg / mL to 3.14 mg / mL, 3.1 mg / mL to 3.12 mg / mL, 3.12 mg / mL to about 3.2 mg / mL, 3.12 mg / mL to 3.18 mg / mL, 3.12 mg / mL to 3.16 mg / mL, or 3.12 mg / mL to 3.14 mg / mL). In some embodiments, the IVSS comprises about 0.07 mg / mL to about 0.08 mg / mL succinic acid and about 3 mg / mL to about 4 mg / mL disodium succinate.

[0060] In some embodiments, the surfactant is a nonionic surfactant, such as polysorbate 80 (also referred to herein as “PS80”). In some embodiments, the IVSS comprises sodium citrate, polysorbate 80, and water, and has a pH of about 5.8 to about 6.2, about 5.9 to about 6.1, or about 6.0, e.g., 5.8, 5.85, 5.9, 5.95, 6.0, 6.05, 6.1, 6.15, or 6.2. In some embodiments, the IVSS has a pH of about 6.0 (e.g., a pH of 6.0). In some embodiments, the IVSS comprises about 0.4% to about 0.6%, e.g., 0.5%, weight to volume (“w / v”) polysorbate 80. In some embodiments, the IVSS comprises sodium citrate at about 20 mM to about 30 mM, about 0.5% w / v polysorbate 80, and water, and has a pH from about 5.8 to about 6.0.

[0061] In some embodiments, the IVSS comprises 0.7 mg / mL to about 0.8 mg / mL, trisodium citrate dihydrate in an amount between 6.2 mg / mL and about 6.3 mg / mL, about 0.5% w / v polysorbate 80 and sufficient water to bring the IVSS up to the desired volume. In someAtty. Docket No. 0282-0010W01embodiments, the IVSS comprises sodium succinate, polysorbate 80 and water and has a pH of about 6.4 to about 6.6, e.g., 6.4, 6.45, 6.5, 6.55, or 6.6. In some embodiments, the IVSS has a pH of about 6.5.

[0062] In some embodiments, the disclosure provides a container, wherein the container comprises an isotonic aqueous solution, and the IVSS is added to the isotonic aqueous solution to form a pretreated container. Suitable isotonic aqueous solutions as described herein are solutions having substantially the same or similar osmotic pressure or solute concentration as compared to bodily fluids and include, but are not limited to, normal saline, Ringer’s lactate solution, dextrose, or a combination thereof. For instance, in some embodiments, the isotonic aqueous solution is 0.9% normal saline.

[0063] The IVSS described herein may be used to pretreat containers, such as medical grade bags (intravenous solution bags), to protect against container surface adsorption of therapeutic agents, such as low dose or ultra-low dose proteins including soluble TCR proteins, to be administered to patients in need thereof. In some embodiments, this pretreatment is especially suitable for some therapeutic agents, such as a soluble TCR protein, which are administered to subjects in small amounts, in a very diluted form. As such, the IVSS described herein enable higher recovery of the soluble TCR proteins and, therefore, more efficient and consistent delivery of the soluble TCR proteins to the subject for the treatment or prevention of, for example, ocular melanoma, endometrial tumors, and the like.

[0064] In some embodiments, the TCR protein is a soluble TCR protein (i.e. having no transmembrane of cytoplasmic domains). In some embodiments, the soluble TCR protein can recognize intracellular epitopes in addition to extracellular epitopes. In some embodiments, the soluble TCR protein is added to container that is pretreated with the IVSS. The soluble TCR protein may be added to the container using manual injection, pouring, injecting, or any other art standard technique. In some embodiments, the IVSS pretreatment prevents or significantly reduces the adsorption of the soluble TCR protein onto the surface of the container as compared to an unpretreated container. Therefore, in some embodiments, the IVSS improves soluble TCR protein recovery, and therefore can improve the efficiency and consistency of delivery of the soluble TCR protein.

[0065] In some embodiments, the soluble TCR protein of the disclosure can be a bispecific TCR protein. The term “bispecific TCR protein” refers herein to a therapeutic agent thatAtty. Docket No. 0282-0010W01target and bind to two antigens or two epitopes. In some embodiments, the soluble TCR protein comprises both a TCR alpha variable domain and a TCR beta variable domain. In some embodiments, the TCR protein is a heterodimeric TCR protein comprising an alphabeta heterodimer. In some embodiments, the bispecific TCR protein comprises the sequences of any one of the TCR proteins presented in Table 3. In some embodiments, the TCR protein comprises any one of the CDR sequences in Table 4. In some embodiments, the bispecific TCR protein targets CD3 and PReferentially Expressed antigen of MElanoma (“PRAME”). In some embodiments, the bispecific TCR protein targets CD3 and gpIOO. In some embodiments the bispecific TCR protein targets piwi like RNA-mediated gene silencing 1 (“PIWIL1”). In some embodiments, the bispecific TCR protein that targets gpIOO is tebentafusp (also referred to as tebentafusp-tebn or KIMMTRAK®). In some embodiments, the bispecific TCR protein that targets PRAME is brenetafusp (also referred to as IMC-F106C), IMC-P115C, or IMC-T119C. In some embodiments, the bispecific TCR protein that targets PIWIL1 is IMC-R117C. In some embodiments, pretreating the container with the IVSS of the disclosure will protect against surface adsorption of the soluble TCR protein and enable efficient and consistent dosing of the soluble TCR protein, e.g., tebentafusp or brenetafusp.Table 3. TCR proteins.TCR protein Target Protein Alpha chain Beta Chain Beta chain- (variable + (variable + antiCD3 fusion constant constant (anti-CD3 (underlined)) (underlined) +linkers italicized and underlined) Tebentafusp GpIOO AQQGEEDPQA DGGITQSPKY AIOMTOSPSSL gPplOO X LSIQEGENAT LFRKEGQNVT SASVGDRVTIT CD3 / HLA-A2 MNCSYKTSIN LSCEQNLNHD CRASQDIRNYL NLQWYRQNS AMYWYRQDP NWYOOKPGKA GRGLVHLILIR GQGLRLIYYS PKLLIYYTSRLE SNEREKHSGR WAQGDFQKG SGVPSRFSGSG LRVTLDTSKK DIAEGYSVSR SGTDYTLTISSL SSSLLITASRA EKKESFPLTVT QPEDFATYYC ADTASYFCAT SAQKNPTAFY OOGNTLPWTF DGSTPMQFGK LCASSWGAPY GOGTKVEIKG GTRLSVIANIQ EQYFGPGTRL GGGSGGGGSG KPDPAVYQLR TVTEDLKNVF GGGSGGGGSG DSKSSDKSVC PPEVAVFEPSE GGSEVOLVESG LFTDFDSQTN AEISHTQKAT GGLVQPGGSL VSQSKDSDVY LVCLATGFYP RLSCAASGYSFITDKCVLDMR DHVELSWWV TGYTMNWVRQ-Atty. Docket No. 0282-0010W01SMDFKSNSAV NGKEVHSGVC APGKGLEWVA AWSNKSDFAC *TDPQPLKEQP LINPYKGVSTY ANAFNNSIIPE ALNDSRYALS NQKFKDRFTIS DT (SEQ ID SRLRVSATFW VDKSKNTAYLQ NO: 1) QDPRNHFRCQ MNSLRAEDTAV VQFYGLSEND YYCARSGYYGD EWTQDRAKP SDWYFDVWGQ VTOIVSAEAW GTLVTVSSGGG GRAD GSDGGITQSPK(SEQ ID NO: 3) YLFRKEGQNV TLSCEQNLNH DAMYWYRQD PGQGLRLIYY SWAQGDFQK GDIAEGYSVS REKKESFPLT VTSAQKNPTA FYLCASSWGA PYEQYFGPGT RLTVTEDLKN VFPPEVAVFEP SEAEISHTQKA TLVCLATGFY PDHVELSWW VNGKEVHSG VCTDPQPLKE QPALNDSRYA LSSRLRVSATF WQDPRNHFR CQVQFYGLSE NDEWTQDRA KPVTQIVSAE AWGRAD(SEQ ID NO: 4) Brenetafusp PRAME GDAKTTQPNS DGGITQSPKY AIOMTOSPSSL (IMC-F106C) MESNEEEPVH LFRKEGQNVT SASVGDRVTIT PRAMEX LPCNHSTISGT LSCEQNLNHD CRASQDIRNYL CD3 / HLA-A2 DYIHWYRQLP AMYWYRQDP NWYQQKPGKA SQGPEYVIHG GQGLRLIYYS PKLLIYYTSRLE LTSNVNNRM QIMGDEQKGD SGVPSRFSGSG ASLAIAEDRK IAEGYSVSRE SGTDYTLTISSL SSTLILHRATL KKESFPLTVTS QPEDFATYYC RDAAVYYCIL AQKNPTAFYL OOGNTLPWTF ILGHSRLGNYI CASSWWTGG GQGTKVEIKG ATFGKGTKLS ASPIRFGPGTR GGGSGGGGSG VIPNIQNPDPA LTVTEDLKNV GGGSGGGGSG VYOLRDSKSS FPPEVAVFEPS GGSEVQLVESG DKSVCLFTDF EAEISHTQKA GGLVQPGGSL DSQTNVSQSK TLVCLATGFY RLSCAASGYSFDSDVYITDKC PDHVELSWW TGYTMNWVRQ.Atty. Docket No. 0282-0010W01VLDMRSMDF VNGKEVHSG APGKGLEWVA KSNSAVAWS VCTDPQPLKE LINPYKGVSTY NKSDFACANA QPALNDSRYA NQKFKDRFTIS FNNSIIPEDT LSSRLRVSATF VDKSKNTAYLQ(SEQ ID NO: WQDPRNHFR MNSLRAEDTAV 28) CQVQFYGLSE YYCARSGYYGD NDEWTQDRASDWYFDVWGQ KPVTQIVSAE GTLVTVSSGGG AWGRAD GSDGGITQSPK(SEQ ID NO: YLFRKEGQNV 38) TLSCEQNLNH DAMYWYRQD PGQGLRLIYY SQIMGDEQKG DIAEGYSVSR EKKESFPLTVT SAQKNPTAFY LCASSWWTG GASPIRFGPGT RLTVTEDLKN VFPPEVAVFEP SEAEISHTQKA TLVCLATGFY PDHVELSWW VNGKEVHSG VCTDPQPLKE QPALNDSRYA LSSRLRVSATF WQDPRNHFR CQVQFYGLSE NDEWTQDRA KPVTQIVSAE AWGRAD(SEQ ID NO: 29)IMC-P115C PRAME GDAKTTQPNS DGGITQSPKY AIQMTQSPSSL PRAMEX MESNEEEPVH LFRKEGQNVT SASVGDRVTIT CD3 / HLA-A2- LPCQHSTISGT LSCEQNLNHD CRASQDIRNYL HLE DYIHWYRQLP AMYWYRQDP NWYQQKPGKA SQGPEYVIHG GQGLRLIYYS PKLLIYYTSRLE LTSNVNNRM QIMGDEQKGD SGVPSRFSGSG ASLAIAEDRK IAEGYSVSRE SGTDYTLTISSL SSTLILHRATL KKESFPLTVTS QPEDFATYYC RDAAVYYCIL AQKNPTAFYL QQGNTLPWTF ILGHSRLGNYI CASSWWTGG GQGTKVEIKG ATFGKGTKLS ASPIRFGPGTR GGGSGGGGSG VIPNIQNPDPA LTVTEDLKNV GGGSGGGGSG VYQLRDSKSS FPPEVAVFEPS GGSEVQLVESGDKSVCLFTDF EAEISHTQKA GGLVQPGGSLAtty. Docket No. 0282-0010W01DSQTQVSQSK TLVCLATGFY RLSCAASGYSF DSDVYITDKC PDHVELSWW TGYTMNWVRO VLDMRSMDF VNGKEVHSG APGKGLEWVA KSNSAVAWS VCTDPQPLKE LINPYKGVSTY QKSDFACANA QPALQDSRYA NQKFKDRFTIS FQNSIIPEDT LSSRLRVSATF VDKSKNTAYLQ(SEQ ID NO: WQDPRNHFR MNSLRAEDTAV 49) CQVQFYGLSE YYCARSGYYGD NDEWTQDRA SDWYFDVWGQ KPVTQIVSAE GTLVTVSSGGG AWGRAD GSDGGITQSPK(SEQ ID NO: YLFRKEGQNV 50) TLSCEQNLNH DAMYWYRQD PGQGLRLIYY SQIMGDEQKG DIAEGYSVSR EKKESFPLTVT SAQKNPTAFY LCASSWWTG GASPIRFGPGT RLTVTEDLKN VFPPEVAVFEP SEAEISHTQKA TLVCLATGFY PDHVELSWW VNGKEVHSG VCTDPQPLKE QPALQDSRYA LSSRLRVSATF WQDPRNHFR CQVQFYGLSE NDEWTQDRA KPVTQIVSAE AWGRAD(SEQ ID NO: 51)IMC-T119C PRAME AQSVTQLDSH DSGVTQTPKH AIQMTQSPSSL PRAMEX VSVSEGTPVL LIKATGQRVT SASVGDRVTIT CD3 / HLA-A24 LRCNYSSSYSP LRCSPRSGDY CRASQDIRNYL SLFWYVQHPN SVYWYQQSL NWYQQKPGKA KGLQLLLKYI DQGPQFLFQY PKLLIYYTSRLE GNVTLVKGIQ YNAEERAKG SGVPSRFSGSG GFEAEFKKSE NIPERFSAQQF SGTDYTLTISSL TSFHLTKPSA PDLHSELNLSS QPEDFATYYC HMSDAAEYFC LELGDSALYIC QQGNTLPWTF WGAPHHND ASSIWSIGGAS GQGTKVEIKG KIIFGKGTRLH SGNLSFGEGS GGGSGGGGSGILPNIQNPDPA RLTVLEDMKN GGGSGGGGSGAtty. Docket No. 0282-0010W01VYQLRDSKSS VFPPEVAVFEP GGSEVQLVESG DKSVCLFTDF SEAEISHTQKA GGLVQPGGSL DSQTNVSQSK TLVCLATGFY RLSCAASGYSF DSDVYITDKC PDHVELSWW TGYAMNWVRO VLDMRSMDF VNGKEVHSG APGKGLEWVA KSNSAVAWS VCTDPQPLKE LINPYKGVSTY NKSDFACANA QPALNDSRYA NQKFKDRFTF FNNSIIPEDT LSSRLRVSATF SVDKSKNTAYL(SEQ ID NO: WQDPRNHFR QMNSLRAEDT 52) CQVQFYGLSE AVYYCARSGYY NDEWTQDRA GDSDWYFDVW KPVTQIVSAE GQGTLVTVSSG AWGRAD GGG. SDSGVTO (SEQ ID NO: TPKHLIKATG 53) QRVTLRCSPR SGDYSVYWY QQSLDQGPQF LFQYYNAEER AKGNIPERFSA QQFPDLHSEL NLSSLELGDS ALYICASSIWS IGGASSGNLSF GEGSRLTVLE DMKNVFPPEV AVFEPSEAEIS HTQKATLVCL ATGFYPDHVE LSWWVNGKE VHSGVCTDPQ PLKEQPALND SRYALSSRLR VSATFWQDPR NHFRCQVQFY GLSENDEWTQ DRAKPVTQIV SAEAWGRAD(SEQ ID NO: 55)IMC-R117C PIWIL AAKTTQPISM EAGVAQSPRY AIQMTQSPSSL PIWILX DSYEGQEVNI KIIEKGQSVAF SASVGDRVTIT CD3 / HLA-A2 PCSHNYIAAN WCNPISGHGT CRASQDIRNYL DFITWYQQFP LYWYQQILGQ NWYOOKPGKA SQGPRFFIQGY GPKLLIQFHEE PKLLIYYTSRLE KTNVQNEVAS GVVDDSQLPK SGVPSRFSGSG LFISADRKSST DRFSAERLKG SGTDYTLTISSLLSLPRVSLSDT VDSTLKIQPA QPEDFATYYC.Atty. Docket No. 0282-0010W01AVYYCLAWG KLEDSAVYLC QQGNTLPWTF GTDLLPFGTG ASSVDWVGD GQGTKVEIKG TRLQVFPNIQ GERQYFGPGT GGGSGGGGSG NPDPAVYQLR RLLVLEDLKN GGGSGGGGSG DSKSSDKSVC VFPPEVAVFEP GGSEVQLVESG LFTDFDSQTN SEAEISHTQKA GGLVQPGGSL VSQSKDSDVY TLVCLATGFY RLSCAASGYSF ITDKCVLDMR PDHVELSWW TGYAMNWVRQ SMDFKSNSAV VNGKEVHSG APGKGLEWVA AWSNKSDFAC VCTDPQPLKE LINPYKGVSTY ANAFNNSIIPE QPALNDSRYA NQKFKDRFTF DT (SEQ ID LSSRLRVSATF SVDKSKNTAYL NO: 56) WQDPRNHFR QMNSLRAEDT CQVQFYGLSE AVYYCARSGYY NDEWTQDRA GDSDWYFDVW KPVTQIVSAE GQGTLVTVSSG AWGRAD GGGSEAGNAQ(SEQ ID NO: SPRYKIIEKGQ 57) SVAFWCNPIS GHGTLYWYQ QILGQGPKLLI QFHEEGVVDD SQLP LNHDA KDRFSAERLK GVDSTLKIQP AKLEDSAVYL CASSVDWVG DGERQYFGPG TRLLVLEDLK NVFPPEVAVF EPSEAEISHTQ KATLVCLATG FYPDHVELSW WVNGKEVHS GVCTDPQPLK EQPALNDSRY ALSSRLRVSA TFWQDPRNHF RCQVQFYGLS ENDEWTQDR AKPVTQIVSA EAWGRAD(SEQ ID NO:58)Atty. Docket No. 0282-0010W01Table 4. CDR sequences of TCR proteins.TCR Alpha Alpha Alpha Beta Beta Beta protein chain chain chain chain chain chain CDR1 CDR2 CDR3 CDR1 CDR2 CDR3Tebentafusp TSINN IRS (SEQ ATDGST LNHDA SWAQGD TSINN (SEQ ID ID NO: 7) PMQ (SEQ ID (SEQ ID (SEQ ID NO: 6) (SEQ ID NO: 9) NO: 10) NO: 11)NO: 8)Brenetafusp TISGT GLTSN CILILGH LNHDA SQIMGD CASSWW DY (SEQ ID SRLGNYI (SEQ ID E (SEQ ID TGGASPI (IMC- (SEQ ID NO: 32) ATF (SEQ NO: 39) NO: 40) RF (SEQ F106C) NO: 31) ID NO: ID NO:33) 41)IMC-P115C TISGT GLTSN CILILGH LNHDA SQIMGD CASSWW DY (SEQ ID SRLGNYI (SEQ ID E (SEQ ID TGGASPI (SEQ ID NO: 60) ATF (SEQ NO: 62) NO: 63) RF (SEQ NO: 59) ID NO: ID NO:61) 64) IMC-T119C SSYSPS YIGNVTL VVGAPH YYNAEE SGDYS ASSIWSI (SEQ ID V (SEQ ID HNDKIIF (SEQ ID (SEQ ID GGASSG NO: 65) NO: 66) (SEQ ID NO: 68) NO: 69) NLS (SEQ NO: 67) ID NO:70) IMC-R117C YIAAN QGYKTN LAWGGT SGHGT FHEEGV ASSVDW DF (SEQ ID DLLP (SEQ ID (SEQ ID VGDGER (SEQ ID NO: 72) (SEQ ID NO: 74) NO: 75) QY (SEQ NO: 71) NO: 73) ID NO:76)

[0066] In some embodiments, the soluble TCR protein comprises a multi-domain molecule. In some embodiments, the soluble TCR protein comprises a T cell engaging immune effector domain. In some embodiments, the soluble TCR protein comprises a peptide-major histocompatibility (pMHC) binding domain. In some embodiments, the soluble TCR protein comprises a half-life extending domain. In some embodiments, the soluble TCR protein comprises aPD-1 binding domain. In some embodiments, the soluble TCR protein comprises a CDl-a binding domain. In some embodiments, the soluble TCR protein comprises a PRAME complex domain, such as, but not limited to a PRAME-HLA complex bindingAtty. Docket No. 0282-0010W01domain, a PRAME-HLA-A2 complex binding domain, a PRAME-HLA-A24 complex binding domain, or a combination thereof. In some embodiments, the soluble TCR protein comprises a PIWIL-HLA complex binding domain. In some embodiments, the soluble TCR protein comprises a PIWIL-HLA-A2 complex binding domain. In some embodiments, the soluble TCR protein comprises agp-100 binding domain.

[0067] As noted above, the soluble TCR protein can be a bispecific TCR protein. One arm engages T cells via a CD3 receptor while the other binds to a tumor cell specifically via a peptide HLA complex where the peptide is from a tumor specific molecule. In some embodiments, the tebentafusp. In some embodiments, the soluble TCR protein targets CD3 and gplOO. In specific embodiments, the soluble TCR protein is tebentafusp.

[0068] Tebentafusp is an immune mobilizing monoclonal T-cell receptors against cancer (“ImmTAC”). ImmTACs detect tumor cells that present a specific p-HLA, recruiting T cells to lyse the targeted cells. Tebentafusp is directed to a gplOO peptide united to HLA-A*0201 that prolongs overall survival in subjects. Tebentafusp targets tumor cells that express a peptide of gplOO presented by HLA*A0201, creating an immune synapse that kills targeted tumor cells. The specific p-HLA complex detected by tebentafusp is YLEPGPVTA-HLA-A*02:01 (SEQ ID NO: 54), which is an immunogenic peptide derived from gplOO.(Martinez-Perez et al. “Gp-100 as a Novel Therapeutic Target in Uveal Melanoma,” Cancers (Basel) 13(23): 5968 (2021)). In some embodiments, tebentafusp is used to treat or prevent unresectable or metastatic uveal melanoma. In some embodiments, tebentafusp is used to treat or prevent adjuvant uveal (ocular) melanoma or advanced cutaneous melanoma. In particular embodiments, tebentafusp is administered to the subject intravenously.

[0069] Tebentafusp is composed of an alpha chain and a beta chain (i.e., tebentafusp is a heterodimeric TCR protein). The alpha chain comprises a TCR alpha chain variable region domain and a constant region domain. The beta chain comprises a TCR beta chain variable region domain and a constant region domain where the TCR beta chain is linked to a singlechain variable fragment (“scFv”) anti-CD3 antibody at its N-terminus. The two chains are covalently bonded via a disulfide bond between the cysteine at position 157 on the alpha chain (al57) amino acid sequence of SEQ ID NO: 1 and the cysteine in position 427 on the anti-CD3 scFv beta chain (P427) of SEQ ID NO:2 (indicated with asterisks, *, in the amino acid sequences below).Atty. Docket No. 0282-0010W01

[0070] The amino acid sequence of tebentafusp is provided below, and tebentafusp is further described in International Patent Publication No. WO 2011 / 001152 and U. S. Patent Nos. 8,519,100 and 9,068,178, each of which is hereby incorporated by reference in its entirety. The alpha chain is composed of 195 amino acid residues and the beta chain is composed of 500 amino acid residues. The linker sequence between the two variable domains of the scFv antibody (24 amino acids) and the linker sequence between the scFv and TCR P chain variable region (5 amino acids) are included below. The initiator methionine residues are post-translationally removed from both alpha and beta chains.

[0071] Amino acid sequence of tebentafusp:

[0072] Alpha chain:AQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKH SGRLRVTLDTSKKSSSLLITASRAADTASYFCATDGSTPMQFGKGTRLSVIANIQKPD PAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKC*VLDMRSMDFKSNSA VAWSNKSDFACANAFNNSIIPEDT (SEQ ID NO: 1)

[0073] Wild type gplOO-specific TCR alpha chain with KI 13 substituted forN113:SQQGEEDPQALSIQEGENATMNCSYKTSINNLQWYRQNSGRGLVHLILIRSNEREKH SGRLRVTLDTSKKSSSLLITASRAADTASYFCATDGSTPMQFGKGTRLSVIANIQKPD PAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKTVLDMRSMDFKSNSA VAWSNKSDFACANAFNNSIIPEDTFFPSPESS (SEQ ID NO: 2)

[0074] Wild-type gplOO-specific TCR beta chain:DGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSWAQGDF QKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSWGAPYEQYFGPGTRLTVT EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGV STDPQPLKEQPALNDSRYALSSRLRVSATFWQDPRNHFRCQVQFYGLSENDEWTQD RAKPVTQIVSAEAWGRAD (SEQ ID NO: 3)

[0075] Anti-CD3 scFv Beta chain:AIQMTQSPSSLSASVGDRVTITCRASQDIRNYLNWYQQKPGKAPKLLIYYTSRLESGV PSRFSGSGSGTDYTLTISSLQPEDFATYYCOQGNTLPWTFGOGTKVEIKGGGGSGGG GSGGGGSGGGGSGGGSEVQLVESGGLVQPGGSLRLSCAASGYSFTGYTMNWVRO APGKGLEWVALINPYKGVSTYNQKFKDRFTISVDKSKNTAYLQMNSLRAEDTAVYY CARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSDGGITQSPKYLFRKEGQNVTLSCAtty. Docket No. 0282-0010W01EQNLNHDAMYWYRQDPGQGLRLIYYSWAQGDFQKGDIAEGYSVSREKKESFPLTV TSAQKNPTAFYLCASSWGAPYEQYFGPGTRLTVTEDLKNVFPPEVAVFEPSEAEISHT QKATLVCLATGFYPDHVELSWWVNGKEVHSGVC*TDPQPLKEQPALNDSRYALSSR LRVSATFWQDPRNHFRCQVQFYGLSENDEWTQDRAKPVTQIVSAEAWGRAD (SEQ ID NO: 4)

[0076] Beta chain:DGGITQSPKYLFRKEGQNVTLSCEQNLNHDAMYWYRQDPGQGLRLIYYSWAQGDF QKGDIAEGYSVSREKKESFPLTVTSAQKNPTAFYLCASSWGAPYEQYFGPGTRLTVT EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVC*TDPQPLKEQPALNDSRYALSSRLRVSATFWQDPRNHFRCQVQFYGLSENDEWTQ DRAKPVTQIVSAEAWGRAD (SEQ ID NO: 5)

[0077] Complementarity determining region (“CDR”) sequences of tebentafusp:Alpha Alpha Alpha chain Beta chain Beta chain Beta chain CDR3 chain chain CDR3 CDR1 CDR2 (SEQ ID NO: 11) CDR1 CDR2 (SEQ ID NO: 8) (SEQ ID (SEQ ID(SEQ ID (SEQ ID NO: 9) NO: 10)NO: 6) NO: 7)TSINN IRS ATDGSTPMQ LNHDA SWAQGD ASSWGAPYEQY

[0078] Framework region sequences of tebentafusp:Alpha chain Alpha Alpha chain FR3 Alpha chain FR4 (SEQ ID NO: 15) FR1 chain (SEQ ID NO: 14)(SEQ ID FR2NO: 12) (SEQID NO:13)QQGEEDP LQWY NEREKHSGRL GKGTRLSVIANIQKPDPAVYQLRDSKSS QALSIQE RQNS RVTLDTSKKSS DKSVCLFTDFDSQTNVSQSKDSDVYITD GENATM GRGL SLLITASRAAD KC*VLDMRSMDFKSNSAVAWSNKSDFANCSYK VHLIL TASYFC CANAFNNSIIPEDTBeta Beta Beta chain Beta chain FR4 (SEQ ID NO: 19)chain chain FR3 (SEQ IDFR1 FR2 NO: 18)(SEQ ID (SEQNO: 16) IDAtty. Docket No. 0282-0010W01NO:17)DGGITQ MYW FQKGDIAE FGPGTRLTVTEDLKNVFPPEVAVFEPSEAEISHT SPKYLF YRQ GYSVSREK QKATLVCLATGFYPDHVELSWWVNGKEVHSG RKEGQ DPGQ KESFPLTV VC*TDPQPLKEQPALNDSRYALSSRLRVSATFW NVTLSC GERE TSAQKNPT QDPRNHFRCQVQFYGLSENDEWTQDRAKPVTQEQN IYY AFYLC IVSAEAWGRAD

[0079] Additional linker sequences are as follows: GGGSG (SEQ ID NO: 20), GGSGG (SEQ ID NO: 21), GSGGG (SEQ ID NO: 22), GSGGGP (SEQ ID NO: 23), GGEPS (SEQ ID NO: 24), GGEGGGP (SEQ ID NO: 25), and GGEGGGSEGGGS (SEQ ID NO: 26).

[0080] In some embodiments, the TCR protein for use in the methods herein comprises: a TCR alpha chain amino acid sequence of SEQ ID NO: 1 or a TCR alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 1, anda TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 5 or a TCR beta chain-anti-CD3 amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 5,wherein the TCR alpha chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 6, 7 and 8 respectively and the TCR beta chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 9, 10 and 11, respectively.

[0081] In some embodiments, tebentafusp is added to the pretreated container, wherein the tebentafusp in the pretreated container is about 0.2 pg / mL to about 10 pg / mL. e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the tebentafusp is added to the pretreated container, wherein the tebentafusp in the pretreated container is about 0.2 pg / mL to about 20 pg / mL, e.g., 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16Atty. Docket No. 0282-0010W01pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0082] In some embodiments, tebentafusp is added to the pretreated container from a tebentafusp stock solution, wherein the tebentafusp stock solution concentration is about 50 pg / mL to about 500 pg / mL, e.g., 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, 85 pg / mL, 90 pg / mL, 95 pg / mL, 100 pg / mL, 105 pg / mL, 110 pg / mL, 115 pg / mL, 120 pg / mL, 125 pg / mL, 130 pg / mL, 135 pg / mL, 140 pg / mL, 145 pg / mL, 150 pg / mL, 155 pg / mL, 160 pg / mL, 165 pg / mL, 170 pg / mL, 175 pg / mL, 180 pg / mL, 185 pg / mL, 190 pg / mL, 195 pg / mL, 200 pg / mL, 205 pg / mL, 210 pg / mL, 215 pg / mL, 220, pg / mL 225 pg / mL, 230 pg / mL, 235 pg / mL, 240 pg / mL, 245 pg / mL, 250 pg / mL, 255 pg / mL, 260 pg / mL, 265 pg / mL, 270 pg / mL, 275 pg / mL, 280 pg / mL, 285 pg / mL, 290 pg / mL, 295 pg / mL, 300 pg / mL, 305 pg / mL, 310 pg / mL, 315 pg / mL, 320 pg / mL, 325 pg / mL, 330 pg / mL, 335 pg / mL, 340 pg / mL, 345 pg / mL, 350 pg / mL, 355 pg / mL, 360 pg / mL, 365 pg / mL, 370 pg / mL, 375 pg / mL, 380 pg / mL, 390 pg / mL, 400 pg / mL, 405 pg / mL, 410 pg / mL, 415 pg / mL, 420 pg / mL, 425 pg / mL, 430 pg / mL, 435 pg / mL, 440 pg / mL, 445 pg / mL, 450 pg / mL, 455 pg / mL, 460 pg / mL, 465 pg / mL, 470 pg / mL, 475 pg / mL, 480 pg / mL, 485 pg / mL, 490 pg / mL, 495 pg / mL, or 500 pg / mL. In embodiments, the tebentafusp in the stock solution is at a concentration of about 50 pg / mL to about 1000 pg / mL, about 100 pg / mL to about 800 pg / mL, about 150 pg / mL to about 500 pg / mL. In embodiments, the tebentafusp in the stock solution is at a concentration of about 100 pg / mL, about 200, pg / mL, about 300 pg / mL, about 400 pg / mL, about 500 pg / mL, about 600 pg / mL, about 700 pg / mL, or about 800 pg / mL.

[0083] In embodiments, tebentafusp is administered to the subject in a dosing regimen. In embodiments, the tebentafusp is administered weekly, biweekly, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks. In embodiments, the dosing regimen comprises a step-up dose sequence, wherein the subsequent doses administered are increased (“stepped up”). An example of a tebentafusp dosing regimen using a stock solution of 200 pg / mL is illustrated in the table below. The required amounts from the stock solution to add to 100 mL of 0.9% sodiumAtty. Docket No. 0282-0010W01chloride solution in an IV bag pretreated with IVSS and administer the desired low dose of tebentafusp is presented in Table 5. It is to be understood that a person of ordinary skill in the art would add the necessary amount of stock solution to be able to deliver the prescribed doses.Table 5. Step up dose sequence for tebentafusp.Day Dose Amount of stock solution Day 1 20 pg 0.1 mbDay 8 30 pg 0.15 mbDay 15 and thereafter 68 pg 0.34 mb

[0084] In some embodiments, tebentafusp is added to a first pretreated container, wherein the tebentafusp in the first pretreated container is about 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the tebentafusp is added to the first pretreated container, wherein the tebentafusp in the first pretreated container is about 0.2 pg / mL to about 20 pg / mL, e.g., 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0085] In further embodiments, tebentafusp from the first pretreated container is added to a second pretreated container, wherein the tebentafusp in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, e.g., 0.002 pg / mL, 0.003 pg / mL, 0.004 pg / mL,Atty. Docket No. 0282-0010W010.005 pg / mL, 0.006 pg / mL. 0.007 pg / mL, 0.008 pg / mL, 0.009 pg / mL, 0.01 pg / mL, 0.02 pg / mL, 0.03 pg / mL, 0.04 pg / mL, 0.05 pg / mL, 0.06 pg / mL, 0.07 pg / mL, 0.08 pg / mL, 0.09 pg / mL, or 0.1 pg / mL. In some embodiments, the tebentafusp is added to the second pretreated container, wherein the tebentafusp in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, 0.002 pg / mL to 0.18 pg / mL, 0.002 pg / mL to 0.16, 0.002 pg / mLto 0.14, 0.002 pg / mL to 0.12 pg / mL, 0.002 pg / mL to 0.1 pg / mL, 0.002 pg / mL to 0.08 pg / mL, 0.002 pg / mL to 0.06 pg / mL, 0.002 pg / mL to 0.04 pg / mL, 0.004 pg / mL to 0.2 pg / mL, 0.004 pg / mL to 0.18 pg / mL, 0.004 pg / mLto 0.16 pg / mL, 0.004 pg / mLto 0.14 pg / mL, 0.004 pg / mL to 0.12 pg / mL, 0.004 pg / mLto 0.1 pg / mL, 0.004 pg / mL to 0.08 pg / mL, 0.004 pg / mL to 0.06 pg / mL, 0.006 pg / mL to 0.02 pg / mL, 0.006 pg / mL to 0.018 pg / mL, 0.006 pg / mL to 0.016 pg / mL, 0.006 pg / mLto 0.014 pg / mL, 0.006 pg / mLto 0.012 pg / mL, 0.006 pg / mL to 0.010 pg / mL, 0.006 pg / mL to 0.008 pg / mL.

[0086] In some embodiments, the soluble TCR protein targets PRAME. PRAME is a cancertestis antigen that is frequently highly expressed in a range of solid and hematologic malignancies including melanoma, ovarian carcinoma, uterine carcinoma, small-cell and nonsmall cell lung cancer, triple-negative breast cancer, and several rare tumor types. In some embodiments, PRAME targeting soluble TCR proteins suitable for use herein are brenetafusp (IMC-F106C), IMC-P115C, and IMC-T119C. In some embodiments, the PRAME targeting soluble TCR proteins are used to treat or prevent cutaneous melanoma, ovarian tumors, advanced endometrial cancer, or multiple solid tumors.

[0087] In some embodiments, the soluble TCR protein is brenetafusp. Brenetafusp is a T cell redirecting bispecific therapeutic agent comprising a soluble affinity enhanced TCR protein that binds to the SLLQHLIGL peptide-HLA-A*02 complex, fused to an anti-CD3 scFv. The targeting end of brenetafusp (the soluble TCR) binds to a peptide fragment of the PRAME antigen presented by HLA-A*02 on the surface of cancer cells. HLA molecules are polymorphic; approximately 47% of Caucasian individuals in the US and European countries express the HLA-A*02 genotype with the HLA-A*02:01 allele detected in more than 95% of HLA-A* 02-positive individuals. The effector end of brenetafusp (anti-CD3 scFv) can bind to CD3 on any T cell, redirecting the T cell to produce effector cytokines and / or kill the cell presenting the target. In addition, brenetafusp-mediated tumor lysis may prime an endogenous anti-tumor immune response. ImmTAC® molecules such as brenetafusp are highly potent molecules, with redirection of T-cell activity observed against tumor cell linesAtty. Docket No. 0282-0010W01presenting as few as 10 to 50 target peptide: HLA complexes. Brenetafusp has been shown to selectively redirect T cell activity in the presence of HLA-A* 02:01 -positive / PRAME-positive cell lines, leading to T cell activation and killing of PRAME-positive cancer cells, at concentrations as low as 1 pM to 10 pM. As described above, the HLA-A* 02 restricted peptide SLLQHLIGL (SEQ ID NO:27) is derived from the germline cancer antigen PRAME. Brenetafusp has a TCR alpha chain amino acid sequence of SEQ ID NO: 28 and a TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 29.

[0088] The sequences referred to herein are as follows:

[0089] SEQ ID NO: 27 HLA-A*02 restricted peptide: SLLQHLIGL

[0090] SEQ ID NO: 30 Amino acid sequence of the TCR alpha chain variable domain of brenetafusp. CDRs (CDR1, CDR2 and CDR3) are underlined and are designated SEQ ID NO: 31, 32, and 33 respectively, framework regions (FR1, FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 34, 35, 36, and 37, respectively. Mutations with respect to native alpha chain are in bold.10091]GDAKTTOPNSMESNEEEPVHLPCNHSTISGYDYIHWYROLPSOGPEYVIHGLYSNV NNRMASLAIAEDRKSSTLILHRATLRDAAVYYCIULGHSRLGNYIAAYGKGTKLSVIP

[0092] SEQ ID NO: 38 Amino acid sequence of the TCR beta chain domain of the brenetafusp. CDRs (CDR1, CDR2 and CDR3) are underlined and are designated SEQ ID NO: 39, 40, 41, respectively, framework regions (FR1, FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 42, 43, 44, and 45, respectively. Mutations with respect to native beta chain are in bold. The constant region is double underlined.

[0093] DGGITQSPKYLFRKEGONVTLSCEONiAAGD MYWYRODPGQGLR LTTSQIMGD EOKGDIAEGYSVSREKKESFPLTVTSAOKNPTAFYLCASSWWYGGASPIRFGPGTRLTVT EDLKNVFPPEVAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGV CTDPOPLKEQPALNDSRYALSSRLRVSATFWQDPRNHFRCQVQFYGLSENDEWTOD RAKPVTQIVSAEAWGRAD

[0094] SEQ ID NO: 28 Amino acid sequence of the TCR alpha chain of the brenetafusp. CDRs (CDR1, CDR2 and CDR3) are underlined and are designated SEQ ID NO: 31, 32, and 33, respectively, framework regions (FR1, FR2, FR3 and FR4) are in italics and areAtty. Docket No. 0282-0010W01designated SEQ ID NO: 34, 35, 36, and 37, respectively. The constant region is shown in bold and is designated SEQ ID NO: 48. Within the constant region, the nonnative cysteine residue is double underlined (at position 48 of constant region).10095]GDAKTTOPNSMESNEEEPVHLPCNHSTISGYDYIHWYROLPSOGPEYVIHGLYSNV NNRAUSLAIAEDRKSSTLILHRATLRDAAVYYClULGHSRLGNYlATFGKGTKLSVIPmO NPDPAVYQLRDSKSSDKSVCLFTDFDSQTNVSQSKDSDVYITDKCVLDMRSMDF KSNSAVAWSNKSDFACANAFNNSIIPEDT

[0096] SEQ ID NO: 29 Amino acid sequence of the TCR beta chain-anti-CD3 of the brenetafusp. Anti-CD3 scFv (amino acids 1-253) is shown in bold and underlined and is designated SEQ ID NO: 46. The linker (GGGGS) appears immediately after the scFv, is shown in paler text and is designated SEQ ID NO: 47. CDRs (CDR1, CDR2 and CDR3) are underlined and are designated SEQ ID NO: 39, 40, and 41, respectively, framework regions (FR1, FR2, FR3 and FR4) are in italics and are designated SEQ ID NO: 42, 43, 44 and 45, respectively. Constant region is shown in bold (no underline) and is designated SEQ ID NO: 46. Within the constant region, the nonnative cysteine residue is double underlined (at position 57 of constant region). Additional non-native amino acids at position 75 and position 89 of the constant region are also double underlined.10097] AIQMTOSPSSLSASVGDRVTITCRASODIRNYLNWYQQKPGKAPKLLIYYT SRLESGVPSRFSGSGSGTDYTLTISSLQPEDFATYYCOOGNTLPWTFGOGTKVEI KGGGGSGGGGSGGGGSGGGGSGGGSEVQLVESGGGLVQPGGSLRLSCAASGY SFTGYTMNWVROAPGKGLEWVALINPYKGVSTYNOKFKDRFTISVDKSKNTAY LQMNSLRAEDTAVYYCARSGYYGDSDWYFDVWGQGTLVTVSSGGGGSDGGW SPKYLFRKEGONVTLSCEONLNHDAMYWYRODPGOGLRLIYYSQIMGDEOKGDIAEGYS VSREKKESFPLTVTSAOKNPTAFYLCASSWWYGGASPIRFGPGTRLTVTEDLKNVFPPE VAVFEPSEAEISHTQKATLVCLATGFYPDHVELSWWVNGKEVHSGVCTDPQPL KEQPALNDSRYALSSRLRVSATFWQDPRNHFRCQVQFYGLSENDEWTQDRAKP VTQIVSAEAWGRAD

[0098] CDR sequences of the brenetafusp:Atty. Docket No. 0282-0010W01Alpha Alpha chain Alpha chain Beta chain Beta chain Beta chain chain CDR2 CDR3 CDR1 CDR2 CDR3 CDR1TISGTDY GLTSN CILILGHSR LNHDA SQIMGDE CASSWWT(SEQ ID (SEQ ID LGNYIATF (SEQ ID (SEQ ID GGASPIRF NO:31) NO: 32) (SEQ ID NO: 39) NO: 40) (SEQ IDNO: 33) NO: 41)

[0099] Framework region sequences of brenetafusp:Alpha chain FR1 Alpha chain Alpha chain FR3 Alpha chain FR4FR2GDAKTTQPNSMES IHWYRQLP VNNRMASLAIA GKGTKLSVIP NEEEPVHLPCNHS SQGPEYVIH EDRKSSTLILHR (SEQ ID NO: 37) (SEQ ID NO:34) (SEQ ID NO: ATLRDAAVYY35) (SEQ ID NO: 36)Beta chain FR1 Beta chain FR2 Beta chain FR3 Beta chain FR4 DGGITQSPKYLFRK MYWYRQDP QKGDIAEGYSV GPGTRLTVT EGQNVTLSCEQN GQGLRLIYY SREKKESFPLTV (SEQ ID NO: 45) (SEQ ID NO:42) (SEQ ID NO: TSAQKNPTAFYL43) (SEQ ID NO: 44)

[0100] Additional linker sequences:

[0101] GGGSG (SEQ ID NO: 20), GGSGG (SEQ ID NO: 21), GSGGG (SEQ ID NO: 22), GSGGGP (SEQ ID NO: 23), GGEPS (SEQ ID NO: 24), GGEGGGP (SEQ ID NO: 25), and GGEGGGSEGGGS (SEQ ID NO: 26).

[0102] In some embodiments, the TCR protein for use in the methods herein comprises:a TCR alpha chain amino acid sequence of SEQ ID NO: 28 or a TCR alpha chain amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 28, anda TCR beta chain-anti-CD3 amino acid sequence of SEQ ID NO: 29 or a TCR beta chain-anti-CD3 amino acid sequence that has at least 90%, at least 95% or 100% identity to the amino acid sequence of SEQ ID NO: 29,wherein the TCR alpha chain variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 31, 32 and 33 respectively and the TCR beta chainAtty. Docket No. 0282-0010W01variable domain comprises CDRs 1, 2 and 3 having the amino acid sequences of SEQ ID NOs: 39, 40 and 41, respectively.

[0103] In some embodiments, the brenetafusp is added to the pretreated container, wherein the brenetafusp in the pretreated container is about is about 0.2 pg / mL to about 10 pg / mL, e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the brenetafusp is added to the pretreated container, wherein the brenetafusp in the pretreated container is about 0.2 pg / mL to about 20 pg / mL, e.g., 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0104] In some embodiments, brenetafusp is added to the pretreated container from a brenetafusp stock solution, wherein the brenetafusp stock solution concentration is about 50 pg / mL to about 500 pg / mL, e.g., 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, 85 pg / mL, 90 pg / mL, 95 pg / mL, 100 pg / mL, 105 pg / mL, 110 pg / mL, 115 pg / mL, 120 pg / mL, 125 pg / mL, 130 pg / mL, 135 pg / mL, 140 pg / mL, 145 pg / mL, 150 pg / mL, 155 pg / mL, 160 pg / mL, 165 pg / mL, 170 pg / mL, 175 pg / mL, 180 pg / mL, 185 pg / mL, 190 pg / mL, 195 pg / mL, 200 pg / mL, 205 pg / mL, 210 pg / mL, 215 pg / mL, 220, pg / mL 225 pg / mL, 230 pg / mL, 235 pg / mL, 240 pg / mL, 245 pg / mL, 250 pg / mL, 255 pg / mL, 260 pg / mL, 265 pg / mL, 270 pg / mL, 275 pg / mL, 280 pg / mL, 285 pg / mL, 290 pg / mL, 295 pg / mL, 300 pg / mL, 305 pg / mL, 310 pg / mL, 315 pg / mL, 320 pg / mL, 325 pg / mL, 330 pg / mL, 335 pg / mL, 340 pg / mL, 345 pg / mL, 350 pg / mL, 355 pg / mL, 360 pg / mL, 365 pg / mL, 370 pg / mL, 375 pg / mL, 380 pg / mL, 390 pg / mL, 400 pg / mL, 405 pg / mL, 410 pg / mL, 415 pg / mL, 420 pg / mL, 425 pg / mL, 430 pg / mL, 435 pg / mL, 440 pg / mL, 445 pg / mL, 450 pg / mL, 455 pg / mL, 460 pg / mL, 465 pg / mL, 470 pg / mL, 475Atty. Docket No. 0282-0010W01pg / mL, 480 pg / mL, 485 pg / mL, 490 pg / mL, 495 pg / mL, or 500 pg / mL. In embodiments, the brenetafusp in the stock solution is at a concentration of about 50 pg / mL to about 1000 pg / mL, about 100 pg / mL to about 800 pg / mL, about 150 pg / mL to about 500 pg / mL. In embodiments, the brenetafusp in the stock solution is at a concentration of about 100 pg / mL, about 200, pg / mL, about 300 pg / mL, about 400 pg / mL, about 500 pg / mL, about 600 pg / mL, about 700 pg / mL, or about 800 pg / mL.

[0105] In embodiments, brenetafusp is administered to the subject in a dosing regimen. In embodiments, the brenetafusp is administered weekly, biweekly, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks. In embodiments, the dosing regimen comprises a step-up dose sequence, wherein the subsequent doses administered are increased (“stepped up”). An example of a brenetafusp dosing regimen is illustrated in Table 6. It is to be understood that a person of ordinary skill in the art would add the necessary amount of stock solution to be able to deliver the prescribed doses.Table 6 Step-up dosing sequence for brenetafusp.Day DoseDay 1 20 pg or 3 pgDay 8 40 pg or 10 pgDay 15 and thereafter 160 pg or 40 pg

[0106] In some embodiments, brenetafusp is added to a first pretreated container, wherein the brenetafusp in the first pretreated container is about 0.2 pg / mL to about 10 pg / mL, e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the brenetafusp is added to the first pretreated container, wherein the brenetafusp in the first pretreated container is about 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL,Atty. Docket No. 0282-0010W010.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mLto 1.8 pg / mL, 0.8 pg / mLto 1.6 pg / mL, 0.8 pg / mLto 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mLto 1.6 pg / mL, 1.0 pg / mLto 1.4 pg / mL, 1.0 pg / mLto 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0107] In further embodiments, brenetafusp from the first pretreated container is added to a second pretreated container, wherein the brenetafusp in the second pretreated container is about 0.002 pg / mLto about 0.2 pg / mL, e.g., 0.002 pg / mL, 0.003 pg / mL, 0.004 pg / mL, 0.005 pg / mL, 0.006 pg / mL, 0.007 pg / mL, 0.008 pg / mL, 0.009 pg / mL, 0.01 pg / mL, 0.02 pg / mL, 0.03 pg / mL, 0.04 pg / mL, 0.05 pg / mL, 0.06 pg / mL, 0.07 pg / mL, 0.08 pg / mL, 0.09 pg / mL, or 0.1 pg / mL. In some embodiments, the brenetafusp is added to the second pretreated container, wherein the brenetafusp in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, 0.002 pg / mL to 0.18 pg / mL, 0.002 pg / mL to 0.16, 0.002 pg / mLto 0.14, 0.002 pg / mLto 0.12 pg / mL, 0.002 pg / mLto 0.1 pg / mL, 0.002 pg / mLto 0.08 pg / mL, 0.002 pg / mL to 0.06 pg / mL, 0.002 pg / mL to 0.04 pg / mL, 0.004 pg / mL to 0.2 pg / mL, 0.004 pg / mLto 0.18 pg / mL, 0.004 pg / mLto 0.16 pg / mL, 0.004 pg / mLto 0.14 pg / mL, 0.004 pg / mLto 0.12 pg / mL, 0.004 pg / mLto 0.1 pg / mL, 0.004 pg / mLto 0.08 pg / mL, 0.004 pg / mL to 0.06 pg / mL, 0.006 pg / mL to 0.02 pg / mL, 0.006 pg / mL to 0.018 pg / mL, 0.006 pg / mLto 0.016 pg / mL, 0.006 pg / mLto 0.014 pg / mL, 0.006 pg / mLto 0.012 pg / mL, 0.006 pg / mL to 0.010 pg / mL, 0.006 pg / mL to 0.008 pg / mL.

[0108] In some embodiments, the soluble TCR protein is IMC-T119C. IMC-T119C is a T cell redirecting bispecific therapeutic agent comprising a soluble affinity enhanced TCR protein that binds to the PYLGQMINL (SEQ ID NO: 78) complex, fused to an anti-CD3 scFv. The peptide PYLGQMINL (SEQ ID NO: 78) corresponds to amino acids 254-262 of the full length PRAME protein (UniProt id: P78395) and is presented on the cell surface in complex with HLA-A24 (i.e., “HLA-A*24:02”, also referred to as “HLA-A*24”). This peptide-HLA complex provides a useful target for TCR-based immunotherapeutic intervention.

[0109] In some embodiments, the IMC-T119C is added to the pretreated container, wherein the IMC-T119C in the pretreated container is about is about 0.2 pg / mL to about 10 pg / mL,Atty. Docket No. 0282-0010W01e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the IMC-T119C is added to the pretreated container, wherein the IMC-T119C in the pretreated container is about 0.2 pg / mL to about 20 pg / mL, e.g., 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0110] In some embodiments, IMC-T119C is added to the pretreated container from a IMC-T119C stock solution, wherein the IMC-T119C stock solution concentration is about 50 pg / mL to about 500 pg / mL, e.g., 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, 85 pg / mL, 90 pg / mL, 95 pg / mL, 100 pg / mL, 105 pg / mL, 110 pg / mL, 115 pg / mL, 120 pg / mL, 125 pg / mL, 130 pg / mL, 135 pg / mL, 140 pg / mL, 145 pg / mL, 150 pg / mL, 155 pg / mL, 160 pg / mL, 165 pg / mL, 170 pg / mL, 175 pg / mL, 180 pg / mL, 185 pg / mL, 190 pg / mL, 195 pg / mL, 200 pg / mL, 205 pg / mL, 210 pg / mL, 215 pg / mL, 220, pg / mL 225 pg / mL, 230 pg / mL, 235 pg / mL, 240 pg / mL, 245 pg / mL, 250 pg / mL, 255 pg / mL, 260 pg / mL, 265 pg / mL, 270 pg / mL, 275 pg / mL, 280 pg / mL, 285 pg / mL, 290 pg / mL, 295 pg / mL, 300 pg / mL, 305 pg / mL, 310 pg / mL, 315 pg / mL, 320 pg / mL, 325 pg / mL, 330 pg / mL, 335 pg / mL, 340 pg / mL, 345 pg / mL, 350 pg / mL, 355 pg / mL, 360 pg / mL, 365 pg / mL, 370 pg / mL, 375 pg / mL, 380 pg / mL, 390 pg / mL, 400 pg / mL, 405 pg / mL, 410 pg / mL, 415 pg / mL, 420 pg / mL, 425 pg / mL, 430 pg / mL, 435 pg / mL, 440 pg / mL, 445 pg / mL, 450 pg / mL, 455 pg / mL, 460 pg / mL, 465 pg / mL, 470 pg / mL, 475 pg / mL, 480 pg / mL, 485 pg / mL, 490 pg / mL, 495 pg / mL, or 500 pg / mL. In embodiments, the IMC-T119C in the stock solution is at a concentration of about 50 pg / mL to about 1000 pg / mL, about 100 pg / mL to about 800 pg / mL, about 150 pg / mL to about 500 pg / mL. In embodiments, the IMC-T119C in the stock solution is at a concentration of about 100 pg / mL,Atty. Docket No. 0282-0010W01about 200, pg / mL, about 300 pg / mL, about 400 pg / mL, about 500 pg / mL, about 600 pg / mL, about 700 pg / mL, or about 800 pg / mL.

[0111] In embodiments, IMC-T119C is administered to the subject in a dosing regimen. In embodiments, the IMC-T119C is administered weekly, biweekly, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks. In embodiments, the dosing regimen comprises a step-up dose sequence, wherein the subsequent doses administered are increased (“stepped up”). One of ordinary skill in the art would be able to calculate the amount of stock solution to add in view of the needed and / or prescribed doses.

[0112] In some embodiments, IMC-T119C is added to a first pretreated container, wherein the IMC-T119C in the first pretreated container is about 0.2 pg / mL to about 10 pg / mL, e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the IMC-T119C is added to the first pretreated container, wherein the IMC-T119C in the first pretreated container is about 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0113] In further embodiments, IMC-T119C from the first pretreated container is added to a second pretreated container, wherein the IMC-T119C in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, e.g., 0.002 pg / mL, 0.003 pg / mL, 0.004 pg / mL, 0.005 pg / mL, 0.006 pg / mL, 0.007 pg / mL, 0.008 pg / mL, 0.009 pg / mL, 0.01 pg / mL, 0.02 pg / mL, 0.03 pg / mL, 0.04 pg / mL, 0.05 pg / mL, 0.06 pg / mL, 0.07 pg / mL, 0.08 pg / mL, 0.09 pg / mL, or 0.1 pg / mL. In some embodiments, the IMC-T119C is added to the secondAtty. Docket No. 0282-0010W01pretreated container, wherein the IMC-T119C in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, 0.002 pg / mL to 0.18 pg / mL, 0.002 pg / mL to 0.16, 0.002 pg / mLto 0.14, 0.002 pg / mL to 0.12 pg / mL, 0.002 pg / mL to 0.1 pg / mL, 0.002 pg / mL to 0.08 pg / mL, 0.002 pg / mL to 0.06 pg / mL, 0.002 pg / mL to 0.04 pg / mL, 0.004 pg / mL to 0.2 pg / mL, 0.004 pg / mLto 0.18 pg / mL, 0.004 pg / mLto 0.16 pg / mL, 0.004 pg / mLto 0.14 pg / mL, 0.004 pg / mLto 0.12 pg / mL, 0.004 pg / mLto 0.1 pg / mL, 0.004 pg / mLto 0.08 pg / mL, 0.004 pg / mL to 0.06 pg / mL, 0.006 pg / mL to 0.02 pg / mL, 0.006 pg / mL to 0.018 pg / mL, 0.006 pg / mLto 0.016 pg / mL, 0.006 pg / mLto 0.014 pg / mL, 0.006 pg / mLto 0.012 pg / mL, 0.006 pg / mL to 0.010 pg / mL, 0.006 pg / mL to 0.008 pg / mL.

[0114] In some embodiments, the soluble TCR protein is a piwi like RNA-mediated silencing 1 (“PIWIL1”) modulator. In some embodiments, the PIWIL1 modulator is IMC-R117C. In some embodiments, IMC-R117C is used to treat or prevent colorectal or gastrointestinal cancers. In some embodiments the IMC-R117C comprises a TCR alpha chain variable domain and a TCR beta chain variable domain, which associate together to form a TCR binding site which is capable of binding to the SLSNRLYYL (SEQ ID NO: 77)-HLA-A*02 complex.

[0115] In some embodiments, IMC-R117C is added to the pretreated container, wherein the IMC-R117C in the pretreated container is about is about 0.2 pg / mL to about 10 pg / mL, e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9 pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the IMC-R117C is added to the pretreated container, wherein the IMC-R117C in the pretreated container is about 0.2 pg / mL to about 20 pg / mL, e.g., 0.2 pg / mLto 20, 0.2 pg / mLto 18, 0.2 pg / mLto 16, 0.2 pg / mLto 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mLto 14 pg / mL, 0.4 pg / mLto 12 pg / mL, 0.4 pg / mLto 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mLto 1.4 pg / mL, 0.8 pg / mLto 1.2 pg / mL, 0.8 pg / mLto 1.0 pg / mL, 1.0 pg / mLto 2.0 pg / mL, 1.0 pg / mLto 1.8 pg / mL, 1.0 pg / mLto 1.6 pg / mL, 1.0 pg / mLto 1.4Atty. Docket No. 0282-0010W01pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0116] In some embodiments, IMC-R117C is added to the pretreated container from a IMC-R117C stock solution, wherein the IMC-R117C stock solution concentration is about 50 pg / mL to about 500 pg / mL, e.g., 50 pg / mL, 55 pg / mL, 60 pg / mL, 65 pg / mL, 70 pg / mL, 75 pg / mL, 80 pg / mL, 85 pg / mL, 90 pg / mL, 95 pg / mL, 100 pg / mL, 105 pg / mL, 110 pg / mL, 115 pg / mL, 120 pg / mL, 125 pg / mL, 130 pg / mL, 135 pg / mL, 140 pg / mL, 145 pg / mL, 150 pg / mL, 155 pg / mL, 160 pg / mL, 165 pg / mL, 170 pg / mL, 175 pg / mL, 180 pg / mL, 185 pg / mL, 190 pg / mL, 195 pg / mL, 200 pg / mL, 205 pg / mL, 210 pg / mL, 215 pg / mL, 220, pg / mL 225 pg / mL, 230 pg / mL, 235 pg / mL, 240 pg / mL, 245 pg / mL, 250 pg / mL, 255 pg / mL, 260 pg / mL, 265 pg / mL, 270 pg / mL, 275 pg / mL, 280 pg / mL, 285 pg / mL, 290 pg / mL, 295 pg / mL, 300 pg / mL, 305 pg / mL, 310 pg / mL, 315 pg / mL, 320 pg / mL, 325 pg / mL, 330 pg / mL, 335 pg / mL, 340 pg / mL, 345 pg / mL, 350 pg / mL, 355 pg / mL, 360 pg / mL, 365 pg / mL, 370 pg / mL, 375 pg / mL, 380 pg / mL, 390 pg / mL, 400 pg / mL, 405 pg / mL, 410 pg / mL, 415 pg / mL, 420 pg / mL, 425 pg / mL, 430 pg / mL, 435 pg / mL, 440 pg / mL, 445 pg / mL, 450 pg / mL, 455 pg / mL, 460 pg / mL, 465 pg / mL, 470 pg / mL, 475 pg / mL, 480 pg / mL, 485 pg / mL, 490 pg / mL, 495 pg / mL, or 500 pg / mL. In embodiments, the IMC-R117C in the stock solution is at a concentration of about 50 pg / mL to about 1000 pg / mL, about 100 pg / mL to about 800 pg / mL, about 150 pg / mL to about 500 pg / mL. In embodiments, the IMC-R117C in the stock solution is at a concentration of about 100 pg / mL, about 200, pg / mL, about 300 pg / mL, about 400 pg / mL, about 500 pg / mL, about 600 pg / mL, about 700 pg / mL, or about 800 pg / mL.

[0117] In embodiments, IMC-R117C is administered to the subject in a dosing regimen. In embodiments, the IMC-R117C is administered weekly, biweekly, every 3 weeks, every 4 weeks, every 5 weeks, every 6 weeks, every 7 weeks, every 8 weeks, every 9 weeks, or every 10 weeks. In embodiments, the dosing regimen comprises a step-up dose sequence, wherein the subsequent doses administered are increased (“stepped up”). One of ordinary skill in the art would be able to calculate the amount of stock solution to add in view of the needed and / or prescribed doses.

[0118] In some embodiments IMC-R117C is added to a first pretreated container, wherein the IMC-R117C in the first pretreated container is about 0.2 pg / mL to about 10 pg / mL, e.g., 0.2 pg / mL, 0.3 pg / mL, 0.4 pg / mL, 0.5 pg / mL, 0.6 pg / mL, 0.7 pg / mL, 0.8 pg / mL, 0.9Atty. Docket No. 0282-0010W01pg / mL, 1 pg / mL, 2 pg / mL, 3 pg / mL, 4 pg / mL, 5 pg / mL, 6 pg / mL, 7 pg / mL, 8 pg / mL, 9 pg / mL, or 10 pg / mL. In some embodiments, the IMC-R117C is added to the first pretreated container, wherein the IMC-R117C in the first pretreated container is about 0.2 pg / mL to 20, 0.2 pg / mL to 18, 0.2 pg / mL to 16, 0.2 pg / mL to 14, 0.2 pg / mL to 12 pg / mL, 0.2 pg / mL to 10 pg / mL, 0.2 pg / mL to 8 pg / mL, 0.2 pg / mL to 6 pg / mL, 0.2 pg / mL to 4 pg / mL, 0.4 pg / mL to 20 pg / mL, 0.4 pg / mL to 18 pg / mL, 0.4 pg / mL to 16 pg / mL, 0.4 pg / mL to 14 pg / mL, 0.4 pg / mL to 12 pg / mL, 0.4 pg / mL to 10 pg / mL, 0.4 pg / mL to 8 pg / mL, 0.4 pg / mL to 6 pg / mL, 0.6 pg / mL to 20 pg / mL, 0.6 pg / mL to 18 pg / mL, 0.6 pg / mL to 16 pg / mL, 0.6 pg / mL to 14 pg / mL, 0.6 pg / mL to 12 pg / mL, 0.6 pg / mL to 10 pg / mL, 0.6 pg / mL to 8 pg / mL, 0.8 pg / mL to 20 pg / mL, 0.8 pg / mL to 1.8 pg / mL, 0.8 pg / mL to 1.6 pg / mL, 0.8 pg / mL to 1.4 pg / mL, 0.8 pg / mL to 1.2 pg / mL, 0.8 pg / mL to 1.0 pg / mL, 1.0 pg / mL to 2.0 pg / mL, 1.0 pg / mL to 1.8 pg / mL, 1.0 pg / mL to 1.6 pg / mL, 1.0 pg / mL to 1.4 pg / mL, 1.0 pg / mL to 1.2 pg / mL, 1.2 pg / mL to 2.0 pg / mL, 1.2 pg / mL to 1.8 pg / mL, 1.2 pg / mL to 1.6 pg / mL, or 1.2 pg / mL to 1.4 pg / mL.

[0119] In further embodiments, IMC-R117C from the first pretreated container is added to a second pretreated container, wherein the IMC-R117C in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, e.g., 0.002 pg / mL, 0.003 pg / mL, 0.004 pg / mL, 0.005 pg / mL, 0.006 pg / mL, 0.007 pg / mL, 0.008 pg / mL, 0.009 pg / mL, 0.01 pg / mL, 0.02 pg / mL, 0.03 pg / mL, 0.04 pg / mL, 0.05 pg / mL, 0.06 pg / mL, 0.07 pg / mL, 0.08 pg / mL, 0.09 pg / mL, or 0.1 pg / mL. In some embodiments, the IMC-R117C is added to the second pretreated container, wherein the IMC-R117C in the second pretreated container is about 0.002 pg / mL to about 0.2 pg / mL, 0.002 pg / mL to 0.18 pg / mL, 0.002 pg / mL to 0.16, 0.002 pg / mL to 0.14, 0.002 pg / mL to 0.12 pg / mL, 0.002 pg / mL to 0.1 pg / mL, 0.002 pg / mL to 0.08 pg / mL, 0.002 pg / mL to 0.06 pg / mL, 0.002 pg / mL to 0.04 pg / mL, 0.004 pg / mL to 0.2 pg / mL, 0.004 pg / mL to 0.18 pg / mL, 0.004 pg / mL to 0.16 pg / mL, 0.004 pg / mL to 0.14 pg / mL, 0.004 pg / mL to 0.12 pg / mL, 0.004 pg / mL to 0.1 pg / mL, 0.004 pg / mL to 0.08 pg / mL, 0.004 pg / mL to 0.06 pg / mL, 0.006 pg / mL to 0.02 pg / mL, 0.006 pg / mL to 0.018 pg / mL, 0.006 pg / mL to 0.016 pg / mL, 0.006 pg / mL to 0.014 pg / mL, 0.006 pg / mL to 0.012 pg / mL, 0.006 pg / mL to 0.010 pg / mL, 0.006 pg / mL to 0.008 pg / mL.

[0120] “Identity” as known in the art is the relationship between two or more polypeptide sequences or two or more polynucleotide sequences, as determined by comparing the sequences. In the art, identity also means the degree of sequence relatedness betweenAtty. Docket No. 0282-0010W01polypeptide or polynucleotide sequences, as the case can be, as determined by the match between strings of such sequences. While there exist a number of methods to measure identity between two polypeptide or two polynucleotide sequences, methods commonly employed to determine identity are codified in computer programs. Preferred computer programs to determine identity between two sequences include, but are not limited to, GCG program package (Devereux, et al., " A comprehensive set of sequence analysis programs for the VAX," Nucleic Acids Research, 12, 387-95 (1984)), BLASTP, BLASTN, and FASTA (Atschul et al., “Basic local alignment search tool,” J. Molec. Biol. 215(3), 403-10 (1990)).

[0121] One skilled in the art can use a program such as the CLUSTAL program to compare amino acid sequences. This program compares amino acid sequences and finds the optimal alignment by inserting spaces in either sequence as appropriate. It is possible to calculate amino acid identity or similarity (identity plus conservation of amino acid type) for an optimal alignment. A program like BLASTx will align the longest stretch of similar sequences and assign a value to the fit. It is thus possible to obtain a comparison where several regions of similarity are found, each having a different score. Both types of identity analysis are contemplated in the present disclosure.

[0122] The percent identity of two amino acid sequences or of two nucleic acid sequences is determined by aligning the sequences for optimal comparison purposes (e.g., gaps can be introduced in the first sequence for best alignment with the sequence) and comparing the amino acid residues or nucleotides at corresponding positions. The “best alignment” is an alignment of two sequences which results in the highest percent identity. The percent identity is determined by the number of identical amino acid residues or nucleotides in the sequences being compared (i.e., % identity = number of identical positions / total number of positions x 100).

[0123] The determination of percent identity between two sequences can be accomplished using a mathematical algorithm known to those of skill in the art. An example of a mathematical algorithm for comparing two sequences is the algorithm of Karlin and Altschul (1990) Proc. Natl. Acad. Sci. USA 87:2264-2268, modified as in Karlin and Altschul (1993) Proc. Natl. Acad. Sci. USA 90:5873-5877. The NBLAST and XBLAST programs of Altschul, et al. (1990) J. Mol. Biol. 215:403-410 have incorporated such an algorithm.BLAST nucleotide searches can be performed with the NBLAST program, score = 100, wordlength = 12 to obtain nucleotide sequences homologous to nucleic acid molecules.Atty. Docket No. 0282-0010W01BLAST protein searches can be performed with the XBLAST program, score = 50, wordlength = 3 to obtain amino acid sequences homologous to protein molecules for use in the disclosure. To obtain gapped alignments for comparison purposes, Gapped BLAST can be utilized as described in Altschul et al. (1997) Nucleic Acids Res. 25:3389-3402.Alternatively, PSI-Blast can be used to perform an iterated search which detects distant relationships between molecules (Id.). When utilizing BLAST, Gapped BLAST, and PSI-Blast programs, the default parameters of the respective programs (e.g., XBLAST and NBLAST) can be used. See http: / / www.ncbi.nlm.nih.gov. Another example of a mathematical algorithm utilized for the comparison of sequences is the algorithm of Myers and Miller, CABIOS (1989). The ALIGN program (version 2.0) which is part of the CGC sequence alignment software package has incorporated such an algorithm. Other algorithms for sequence analysis known in the art include ADVANCE and ADAM as described in Torellis and Robotti (1994) Comput. Appl. Biosci., 10:3-5; and FASTA described in Pearson and Lipman (1988) Proc. Natl. Acad. Sci. 85:2444-8. Within FASTA, ktup is a control option that sets the sensitivity and speed of the search.

[0124] Mutations, including conservation and tolerated substitutions, insertions, and deletions, can be introduced into the sequences provided using any appropriate method including, but not limited to, those based on polymerase chain reaction (PCR), restriction enzyme-based cloning, or ligation independent cloning (LIC) procedures. These methods are detailed in many of the standard molecular biology texts. For further details regarding polymerase chain reaction (PCR) and restriction enzyme-based cloning, see Sambrook & Russell, (2001) Molecular Cloning - A Laboratory Manual (3rdEd.) CSHL Press. Further information on ligation independent cloning (LIC) procedures can be found in Rashtchian, (1995) Curr Opin Biotechnol 6(1): 30-6. The TCR sequences provided by the disclosure can be obtained from solid state synthesis, or any other appropriate method known in the art.

[0125] In some embodiments, the TCR protein is provided in a liquid, e.g., the TCR protein is in a liquid solution or suspension before being added to the pretreated container. In some embodiments, the TCR protein is provided in a liquid, wherein the liquid comprises a salt solution, e.g., a sodium chloride solution, such as, but not limited to 0.9% sodium chloride solution.

[0126] In embodiments, the TCR protein provided herein is in dry form or in the form of a dry powder before being added to the pretreated container. In embodiments, the soluble TCRAtty. Docket No. 0282-0010W01protein provided herein is a lyophilized powder. Lyophilization is a water removal process typically used to preserve perishable materials, to extend shelflife or make the material more convenient for transport. In some embodiments, the soluble TCR protein is a lyophilized T-cell receptor protein. In some embodiments, the lyophilized T-cell receptor protein is reconstituted in a liquid, e.g., a buffered liquid, an isotonic liquid, and / or a saline solution, prior to being added to the pretreated container.

[0127] In some embodiments, a pretreated container suitable for administering a soluble TCR protein is provided. In some embodiments, the isotonic aqueous solution and IVSS are combined in a container to form a pretreated container (e.g., an intravenous solution bag). In such embodiments, the IVSS can be diluted by the isotonic aqueous solution, for example, 1:2 to 1:1,000, e.g., 1:2, 1:5, 1:10, 1:15, 1:25, 1:50, 1:100, 1:200, 1:300, 1:400, 1:500, 1:600, 1:700, 1:800, 1:900, or 1: 1,000. For instance, in one exemplary embodiment, the combined isotonic aqueous solution and IVSS result in a 1: 100 dilution of the IVSS composition. In some embodiments, the pretreated container comprises an isotonic aqueous solution, about 0.2 mM to about 0.3 mM sodium citrate, and about 0.005% w / v polysorbate 80, wherein the composition has a pH of about 5.8 to about 6.2. The isotonic aqueous solution can include, but is not limited to, normal saline, Ringer’s lactate solution, dextrose, or a combination thereof.

[0128] In some embodiments, one or more of the soluble TCR proteins discussed herein is added to the pretreated container. In some embodiments, the composition comprises an isotonic aqueous solution, about 0.2 mM to about 0.3 mM sodium citrate, and about 0.005% w / v polysorbate 80, and a soluble TCR protein.

[0129] In some embodiments, the container contains an isotonic aqueous solution, and the IVSS is added to the isotonic aqueous solution to form a pretreated container. Suitable isotonic aqueous solutions for use in intravenous solution bags and other medical containers are solutions having substantially the same or similar osmotic pressure or solute concentration as compared to bodily fluids and include, but are not limited to, normal saline, Ringer’s lactate solution, dextrose, or a combination thereof. For instance, in some embodiments, the isotonic aqueous solution is 0.9% normal saline. In some embodiments the soluble TCR protein is tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, and / or IMC-R117C.Atty. Docket No. 0282-0010W01

[0130] In some embodiments, a package is provided herein that includes an IVSS. For instance, the package may include a container, such as an intravenous solution bag (primary bag and, optionally, a secondary bag). In some embodiments, the package may include a container for delivery of a medicament (e.g., an intravenous solution bag), a container for storage of the IVSS (e.g., vial, tube, bottle, etc.), and a container for an isotonic aqueous solution (e.g., vial, tube, bottle, etc.), wherein the IVSS and the isotonic aqueous solution are then added to the container for delivery when ready to use, wherein the combining of the IVSS and isotonic aqueous solution in the container forms the pretreated container. In some embodiments, an isotonic aqueous isotonic solution is disposed within the container and the IVSS is stored in a separate container (e.g., vial, tube, bottle, etc.) and must be added to the container for pretreatment. In some embodiments, the container includes an injection port, and extender, and the like. In some embodiments, the IVSS provided in the package as described herein includes citric acid monohydrate, trisodium citrate dihydrate, and polysorbate 80. In some embodiments, the IVSS comprises 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate, 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate, 5 mg / mL polysorbate 80, and has a pH of about 5.8 to about 6.2.

[0131] In some embodiments, the package comprises a container comprising the IVSS. In such embodiments, the IVSS may include 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate, 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate, 5 mg / mL Polysorbate 80 and has a pH of about 5.8 to about 6.2. In some embodiments, the package does not transmit visible light or ultraviolet light.

[0132] Ultraviolet light can cause oxidation, hydrolysis, and instability of a solution in a container, such as the IVSS. In some embodiments, the package and / or container may be made of or comprising a layer that blocks ultraviolet light, such as, but not limited to ultraviolet blocking polyethylene. In some embodiments, the container comprises light-resistant glass or plastic. In some embodiments, the container and / or package is coated with ultraviolet blocking material. In some embodiments, a dark amber cover is disposed over the outer surface of the container and / or package. For instance, the package may be covered in ultraviolet light-blocking material to prevent ultraviolet light damage during storage. In some embodiments, the package includes one or more layers that block greater than 90% of the ultraviolet light, e.g., 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, 99.5%, 99.7% or more ultraviolet light. In some embodiments, the package includes aAtty. Docket No. 0282-0010W01layer that blocks greater than 95% of the ultraviolet light. In some embodiments, the container itself is made of or comprises a layer that blocks greater than 90% of ultraviolet light. In some embodiments, the container itself is made of or comprises a layer that blocks greater than 95% of ultraviolet light.

[0133] In some embodiments, the package further includes instructions for combining the IVSS with a soluble TCR protein. In such embodiments, the instructions include (i) adding the IVSS to the container, which contains an isotonic aqueous solution that, when combined with the IVSS, forms a pretreated container, and (ii) adding the soluble TCR protein to the pretreated container. In some embodiments, the soluble TCR protein is one or more of tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, and IMC-R117C.

[0134] In some aspects, the disclosure herein provides a kit comprising: a lyophilized TCR protein and an IVSS, wherein the IVSS contains about 20 mM to about 30 mM sodium citrate, about 0.5% w / v polysorbate 80, water, and has a pH of about 6.0. In some embodiments, the IVSS includes about 25 mM sodium citrate. In some embodiments, the kit includes a first container (e.g., a bottle or vial) comprising the lyophilized TCR protein and a second container (e.g., an intravenous solution bag) comprising the IVSS.

[0135] In some embodiments, the kit includes instructions for reconstituting the lyophilized TCR protein and adding the reconstituted TCR protein to the container comprising the IVSS.

[0136] In some embodiments, the IVSS in the kit comprises about 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate. In some embodiments, the IVSS in the kit comprises 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate.

[0137] In some embodiments, the lyophilized TCR protein in the kit is a tebentafusp in an amount less than about 0.2 pg / mL or greater than about 0.2 pg / mL to about 20 pg / mL when reconstituted and combined with the IVSS.

[0138] In some embodiments, the lyophilized TCR protein in the kit is a PRAME targeting TCR. In some embodiments, the PRAME targeting TCR comprises brenetafusp, IMC-P115C, or IMC-T119C. In some embodiments, the PRAME targeting TCR is brenetafusp.

[0139] In some embodiments, the lyophilized TCR protein in the kit is a PIWIL1 modulator. In some embodiments, the PIWIL1 modulator is IMC-R117C.Atty. Docket No. 0282-0010W01

[0140] In some embodiments, the kit includes a medical grade bag. In some embodiments, the medical grade bag comprises a layer that does not transmit visible light or ultraviolet light. In some embodiments, the kit comprises a first medical grade bag and a second medical grade bag, wherein the IVSS is disposed in the first medical grade bag and the lyophilized TCR protein is disposed in the second medical grade bag. In some embodiments, the kit includes instructions that comprise the steps of (i) reconstituting the lyophilized TCR; and (ii) adding the reconstituted TCR to the first medical grade bag containing the IVSS.

[0141] The disclosure provides for a method of administering to a subject in need thereof a therapeutic composition comprising one or more of the soluble TCR proteins discussed herein. The method can include pretreating a container with the IVSS of the present disclosure and then adding the therapeutic agent, such as one of the TCR proteins described above, to the pretreated container. The soluble TCR protein can then be administered to the subject intravenously. In some embodiments, the subject is a human subject. In some embodiments, the subject has been diagnosed with an autoimmune disease, human immunodeficiency virus or acquired immune deficiency syndrome, cancer (e.g., non-small or small cell lung cancer, ocular melanoma, advanced endometrial, ovarian carcinoma, and / or a PRAME-positive cancer).

[0142] In some embodiments, the low dose or ultra-low dose protein, e.g., a soluble TCR protein, is administered using a “two container system” to dose patients with very low concentrations of the protein.

[0143] In some embodiments, the two-container system uses two containers of fluids containing the IVSS. The low-concentration protein is diluted first into the first container, e.g., a first bag, to form a first diluted low-concentration protein. A defined amount is taken from the first diluted low-concentration protein and placed in the second container, e.g., a second bag, to form a second diluted low-concentration protein. The second diluted low-concentration protein can then be administered to the subject. Serial dilution of the low-concentration protein into containers which both contain the IVSS can result in greater efficiency and can provide enhanced predictability in the amount administered.

[0144] In some embodiments, the two-container system uses two containers of fluids containing the same amount of the IVSS but different concentrations of the TCR protein. In some embodiments, the two bags of fluid contain IVSS, but only one bag contains the soluble TCR protein. By adjusting the rates of each bag, clinicians can immediately provide aAtty. Docket No. 0282-0010W01customized infusion rate while keeping the fluid delivery constant. In specific embodiments, a 2.5 mL fill is chosen in the present disclosure to match intended commercial fill volumes and to ensure a single vial of IVSS could be used to prepare drug product for patient infusion using the two-bag dilution method while maintaining a suitable dead volume.

[0145] As noted above, the IVSS of the disclosure can protect against adsorption of the soluble TCR proteins onto a container surface. In some embodiments, the IVSS as provided herein can exhibit superior stability even after exposed to forced agitation relative to other known pretreatment solutions. In some embodiments the IVSS of the disclosure are free from visible particles after being subjected to one or more stresses, including, but not limited to freeze-thaw cycling and / or agitation. The term “practically free from visible particles” means that the number of particles that were observed met acceptable industry limits. This term is readily understood by one having ordinary skill in the art according to the Investigational Medicinal Product Dossier that is submitted for regulatory purposes.

[0146] In some embodiments, the IVSS remained free from visible particles or practically free from visible particles when subjected to agitation of at least about 60 rpm for at least about 4 hours. In some embodiments, IVSS remained free from visible particles or practically free from visible particles when subjected to agitation of at least about 100 rpm for at least about 4 hours. In some embodiments, IVSS remained free from visible particles or practically free from visible particles when subjected to agitation of at least about 300 rpm for at least about 4 hours. In some embodiments, IVSS remained free from visible particles or practically free from visible particles when subjected to agitation of at least about 500 rpm for at least about 4 hours. In some embodiments, IVSS remained free from visible particles or practically free from visible particles when subjected to agitation of at least about 600 rpm for at least about 4 hours.

[0147] In some embodiments, the IVSS remained free from visible particles or practically free from visible particles when subjected to at least two (2) freeze-thaw cycles. In some embodiments, the IVSS remained free from visible particles or practically free from visible particles when subjected to at least three (3) freeze-thaw cycles. In some embodiments, the IVSS remained free from visible particles or practically free from visible particles when subjected to at least four (4) freeze-thaw cycles. In some embodiments, the IVSS remained free from visible particles or practically free from visible particles when subjected to at least five (5) freeze-thaw cycles.Atty. Docket No. 0282-0010W01

[0148] In some embodiments, the IVSS are chemically and physically stable after storage at 5 °C for at least 1 month, or at least 2 months, or at least 3 months, or at least 4 months, or at least 5 months, or at least 6 months, or at least 7 months, or at least 8 months, or at least 9 months, or at least 10 months, or at least 11 months, or at least 12 months. In some embodiments, the IVSS are chemically and physically stable after storage at 25 °C for at least 1 month, or at least 2 months, or at least 3 months, or at least 4 months, or at least 5 months, or at least 6 months, or at least 7 months, or at least 8 months, or at least 9 months, or at least 10 months, or at least 11 months, or at least 12 months.

[0149] In some embodiments, the IVSS of the disclosure remained chemically stable after storage. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 1 month at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 2 months at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 3 months at 5 °C or colder, or after at least 6 months at 5 °C or colder, or after at least 7 months at 5 °C or colder or after at least 8 months at 5 °C or colder, or after at least 9 months at 5 °C or colder, or after at least 10 months at 5 °C or colder, or after at least 11 months at 5 °C or colder, or after at least 12 months at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.2 points after storage for at least 1 month at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.2 points after storage for at least 2 months at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.2 points after storage for at least 3 months at 5 °C or colder, or after at least 6 months at 5 °C or colder, or after at least 7 months at 5 °C or colder or after at least 8 months at 5 °C or colder, or after at least 9 months at 5 °C or colder, or after at least 10 months at 5 °C or colder, or after at least 11 months at 5 °C or colder, or after at least 12 months at 5 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 1 month at 25 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 2 months at 25 °C or colder. In some embodiments, the IVSS exhibited a change in pH of less than 0.5 points after storage for at least 3 months at 25 °C or colder, or after at least 6 months at 25 °C or colder, or after at least 7 months at 25 °C or colder or after at least 8 months at 25 °C or colder, or after at least 9 months at 25 °C or colder, or after at least 10 months at 25 °C or colder, or after at least 11 months at 25 °C or colder, or after at least 12 months at 25 °C or colder.Atty. Docket No. 0282-0010W01

[0150] In some embodiments, the IVSS exhibited less than 50 particles (≥10 microns) / mL after storage for at least 1 month at 5 °C or colder as measured by, e.g., light obscuration. In some embodiments, the IVSS exhibit less than 40 particles (≥10 microns) / mL after storage for at least 1 month at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 2 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 40 particles (≥10 microns) / mL after storage for at least 2 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 3 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 40 particles (≥10 microns) / mL after storage for at least 3 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 4 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 40 particles (≥10 microns) / mL after storage for at least 4 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 5 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 40 particles (≥10 microns) / mL after storage for at least 5 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 6 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 9 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 50 particles (≥10 microns) / mL after storage for at least 12 months at 5 °C or colder. In some embodiments, the IVSS exhibit less than 30 particles (≥25 microns) / mL after storage for at least 1 month at 5 °C or colder, or at least 2 months at 5 °C or colder, or at least 3 months at 5 °C or colder, or at least 6 months at 5 °C or colder, or at least 9 months at 5 °C or colder, or at least 12 months at 5 °C or colder.

[0151] As mentioned above, a container pretreated with the IVSS of the disclosure has increased protection against surface adsorption of therapeutic compositions comprising soluble TCR proteins. In some embodiments, at least 80% by weight of the soluble TCR protein is recovered from a container pretreated with the IVSS of the disclosure. In some embodiments, the percent recovery of the soluble TCR protein is at least 85%, e.g., 86%, 87%, 88%, 89%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 97.5%, 98%, 98.5%, 99%, or more by weight. In some embodiments, the recovery of the soluble TCR protein from a container pretreated with the IVSS provided herein is at least 90%, or at least 95% by weight.Atty. Docket No. 0282-0010W01

[0152] Various modifications of the disclosure in addition to those shown and described herein will become apparent to those skilled in the art from the foregoing description and fall within the scope of the appended claims. Preferred features of each aspect of the disclosure are as for each of the other aspects mutatis mutandis. The documents referred to herein are incorporated by reference to the fullest extent permitted by law.EXAMPLES

[0153] The present disclosure has been described with respect to representative examples that are to be considered illustrative embodiments that do not limit the scope of the disclosure which is defined solely by the claims. All references to publications, including scientific publications, treatises, textbooks, patent applications and issued patents are hereby incorporated by reference for all purposes.Example 1. Stability of IVSS following stress-testing

[0154] To determine if the IVSS of the disclosure remained stable and intact after exposure to forced agitation, the non-limiting, exemplary formulations described in Table 7 were stress-tested. The stress-test conditions were designed to mimic common stresses to which IVSS were likely to be exposed during manufacture, transport, and administration to subjects. The degree to which the composition characteristics changed due to stress application was assessed. The formulation of each of the IVSS tested is summarized in Table 7. The 2.5 mL fill was chosen to match intended commercial fill volumes and to ensure a single vial of IVSS could be used to prepare drug product for patient infusion using a two-bag dilution method while maintaining a suitable dead volume. Soluble bispecific T cell engager molecules are potent molecules that can require a wide range of dosing and two bag dilution steps are occasionally needed to dose patients with low masses of drug, <2 pg. This study tested commercial fill volumes, the effect of terminal sterilization was investigated and a limited IUS study was also run on the stressed samples.Table 7. IVSSBuffer Surfactant pH Formulation 1 (Fl): 0.7565 mg / mL citric 5 mg / mL 6.025 mM sodium acid monohydrate polysorbate 80citrate, 0.5% (w / v)tween 80 pH 6.0Atty. Docket No. 0282-0010W016.2937 mg / mLtrisodium citratedihydrateFormulation 2 (F2) 0.0791 mg / mL 5 mg / mL 6.520 mM sodium succinic acid polysorbate 80succinate, 0.5%(w / v) tween 80 pH 3.1324 mg / mL6.5 disodium succinate

[0155] For each stress test, 2.3 mL of each of the above-described IVSS was prepared in a 2R glass vial. The stress-test study consisted of five (5) stress tests applied to separate samples of each composition. The stresses included gentle agitation, vigorous agitation, freeze-thaw cycling, visible light stress, and ultra-violet (UV) light stress. For comparison, both dark control samples (no visible exposure and no UV light exposure) and unstressed control samples were prepared. The brief description of each stress test is summarized in Table 8.Table 8. Stress-Test DescriptionsStress Type Descriptionvigorous agitation The 2R glass vial was shaken on an agitator for four (4) hours at 600 rpm at room temperature (15 °C to 25 °C).gentle agitation The 2R glass vial was shaken on an agitator for four (4) hours at 60 rpm at room temperature (15 °C to 25 °C).freeze -thaw The 2R glass vial was subjected to five (5) freeze-thaw cycles.For each cycle, the sample was left at -80 °C, or less, for at least two (2) hours and then defrosted at room temperature (15 °C to 25 °C) for at least two (2) hours.visible light stress The sample was subjected to greater than 1,200,000 lux hours in a temperature-controlled environment. In particular, the sample was subjected to visible light for 144 hours at 25 °C and 60% relative humidity. The sample was placed in a Memmert ICH 110L stability chamber filled with three (3) DL (daylight, Sylvania) bulbs. The sample vial was left unlabeled to ensure maximum and even light exposure.UV light stress The sample was placed in a Memmert ICH 110L stability chamber filled with two (2) UV (ultraviolet Blacklight BL368 Toughcout, Sylvania) bulbs. The sample vial was left unlabeled to ensure maximum and even UV light exposure.dark control The sample was placed in a Memmert ICH 110L stability chamber, but wrapped in foil to protect the sample from visible light or UV light.unstressed control The 2R glass vial was stored at 2 °C to 8 °C during testing of thetest samples to act as a control sample.Atty. Docket No. 0282-0010W01

[0156] Two separate preparations of each of the three IVSS summarized in Table 7 were prepared. Duplicate vials of each IVSS undergoing testing were prepared and stressed at the same time. The stresses applied to each batch are summarized in Table 9.Table 9. Summary of IVSS and the stresses applied| Stress Applied | Preparation 1 Preparation 2 | control - no stress | Yes Yes| freeze-thaw | Yes No| gentle agitation | Yes | No| vigorous agitation | Yes | No ^visible light stress | No| visible light dark control | No| UV light stress | No| UV light dark control | No

[0157] The results of the light stress for Formulation 1 under visible light and UV light are found in FIG. 1A and FIG. 1B respectively.Visual Analysis

[0158] During testing, a manual, visible inspection was performed on each vial to determine whether visible particles were present in the vial. Vials where no visible particles were detected were determined to be “free from visible particles.” Vials where no more than the specified number of visible particles as deemed acceptable in the industry for IVSS were detected were determined to be “practically free from visible particles.” Finally, the descriptor “visible particles observed” was used for a vial when the number of visible particles that were detected exceeded industry acceptable limits. These descriptors are artstandard according to the Investigational Medicinal Product Dossier submitted for regulatory purposes. In addition to particles, the color and clarity for each vial was described. The results of the visual analysis are summarized in Table 10.Table 10. Visual analysis following agitative / temperature stress testing.Composition Stress Color Clarity Particle AnalysisAtty. Docket No. 0282-0010W01Fl control colorless clear free from visible particles F2 control colorless clear free from visible particles Fl vial 1 freeze-thaw colorless clear practically free from visible particlesFl vial 2 freeze-thaw colorless clear free from visible particles F2 vial 1 freeze-thaw colorless clear free from visible particles F2 vial 2 freeze-thaw colorless clear practically free from visible particlesFl vial 1 gentle agitation colorless clear free from visible particles Fl vial 2 gentle agitation colorless clear free from visible particles F2 vial 1 gentle agitation colorless clear free from visible particles F2 vial 2 gentle agitation colorless clear visible particles observed Fl vial 1 vigorous agitation colorless clear free from visible particles Fl vial 2 vigorous agitation colorless clear free from visible particles F2 vial 1 vigorous agitation colorless clear free from visible particles F2 vial 2 vigorous agitation colorless clear practically free from visible particles

[0159] As shown above, all of the vials for the F 1 composition were either free from visible particles or practically free from visible particles.. Moreover, the results in Table 10 demonstrate that both the Fl and F2 compositions met the criteria for each of the agitation / temperature stress tests (with the exception of one vial of the F2 composition in which visible particles were observed after gentle agitation, but this may have been caused by a dirty vial given that that (i) the other vial was free from visible particles and (ii) this composition passed the vigorous agitation test). These results demonstrate the chemical stability of the IVSS in response to agitation / temperature stress.

[0160] IVSS of the disclosure were also subjected to visible and UV light stress. As summarized in Table 8, test samples were placed in a test chamber and subjected to visible light or UV light in a temperature-controlled environment. Samples that were wrapped in foil were used as dark controls. The results are summarized in Table 11.Table 11. Visual Analysis following light stress testing.Composition Stress Color Clarity Particle AnalysisFl vial 1 control colorless clear Visible particles observedAtty. Docket No. 0282-0010W01Fl vial 2 control colorless clear Visible particles observed F2 vial 1 control colorless clear Visible particles observed F2 vial 2 control colorless clear Free from visible particles Fl vial 1 UV light colorless clear Visible particles observed Fl vial 2 UV light colorless clear Visible particles observed F2 vial 1 UV light colorless clear Visible particles observed F2 vial 2 UV light colorless clear Visible particles observed Fl vial 1 UV dark control colorless clear Visible particles observed Fl vial 2 UV dark control colorless clear Visible particles observed F2 vial 1 UV dark control colorless clear Visible particles observed F2 vial 2 UV dark control colorless clear Visible particles observed Fl vial 1 visible light colorless clear Visible particles observed Fl vial 2 visible light colorless clear Visible particles observed F2 vial 1 visible light colorless clear Visible particles observed F2 vial 2 visible light colorless clear Visible particles observed Fl vial 1 visible light dark colorless clear Practically free from control visible particlesFl vial 2 visible light dark colorless clear Visible particles observed controlF2 vial 1 visible light dark colorless clear Visible particles observed controlF2 vial 2 visible light dark colorless clear Practically free from control visible particles

[0161] The results of the light stress study revealed that the compositions exhibit minor chemical instability when exposed to both UV and visible light. A reverse phase high pressure liquid chromatography with charged aerosol detection (RP-HPLC-CAD) was used to quantify the amount of polysorbate 80 present in each of the IVSS and measure the chemical composition after the application of the light stress (both visible and UV) to determine if the minor chemical instability of the IVSS following light stress was attributed to the polysorbate 80. Deviations in chemical profiles between control and stressed samples were assumed to be due to polysorbate 80 degradation and visually assessed. Degradation was indicated by extra peaks, changes to the peak shape, changes to the peak area, and / or changes to peak retention time when the test sample profile was overlayed on the control sample profile. Overlays examined for the control compositions and the agitation / temperature stress conditionsAtty. Docket No. 0282-0010W01compared to controls showed no measurable effect on polysorbate chemical stability (data not shown). On the other hand, RP-HPLC-CAD did not reveal that light stress had any impact on the chemical stability of polysorbate 80 in the F2 composition. In any case, this minor instability can be easily mitigated by manufacturing in dark environments and storage in containers, such as cardboard, that block exposure to light.

[0162] Osmolality and pH Analysis

[0163] Osmolality analysis was performed on the IVSS to measure the concentration of particles dissolved in the composition following exposure to agitation / temperature stress as compared to controls. For this study, the threshold criteria of acceptability required an osmolality variance of less than ± 2.5 mOsm / kg relative to control (unstressed). Notably, the industry standard is ± 4 mOsm / kg. Accordingly, the criteria selected for this analysis was more rigid. The results are summarized in Table 12.Table 12. Osmolality measurementsComposition Stress Osmolality (mOsm / kg) Fl control 74.3F2 control 54.7Fl freeze-thaw 73.3F2 freeze-thaw 55.0Fl gentle agitation 73.0F2 gentle agitation 54.7Fl vigorous agitation 72.7F2 vigorous agitation 54.7

[0164] As shown in Table 12, the osmolality of each of the IVSS samples remained within 2.5% of the relevant control sample regardless of the stress applied, which demonstrated the stability of these compositions. Similarly, the pH of each of these compositions exhibited remarkable stability after the application of stress, as each IVSS sample remained within 0.2 pH units of its target pH as shown in Table 13.Table 13. pH measurementsComposition Stress pHAtty. Docket No. 0282-0010W01Fl control 5.99F2 control 6.54Fl freeze-thaw 6.00F2 freeze-thaw 6.56Fl gentle agitation 5.99F2 gentle agitation 6.52Fl vigorous agitation 6.00F2 vigorous agitation 6.54Light Obscuration Analysis

[0165] Light obscuration measurements were performed on the IVSS following agitation / temperature stress and light stress. Under United States Pharmacopeia (USP) <788> requirements, injectable compositions must contain less than about 6,000 particles / mL of particles that are 10 microns or greater in size and less than about 600 particles / mL of particles that are 25 microns or greater in size. As shown in Tables 14a and 14b, all IVSS particle counts were within the industry standard and complied with USP requirements for particles having a particle size of 10 microns, or greater, and particles having a particle size of 25 microns, or greater. Samples where freeze-thaw cycles were applied had generally higher particle counts for particles less than 10 microns. However, the higher number of small particles did not translate to higher particle counts for the larger particles.

[0166] In addition, a FlowCam system was used to measure microfluidic imaging concurrently with light obscuration. The results are summarized in Tables 15a-16b. All IVSS tested were within the particle count criteria acceptable according to USP <788> standards, thus demonstrating the chemical stability of the IVSS of the disclosure in response to stress.Table 14a. Light Obscuration measurementsLight Obscuration Sub-visible particles / mL Composition / Stress >2μm >5μm >10μm >25μmFl control 1290 350 57 0Fl freeze-thaw 4903 570 67 3Fl gentle agitation 1277 107 30 0Fl vigorous agitation 917 310 67 0Atty. Docket No. 0282-0010W01F2 control 633 110 27 3 F2 freeze-thaw 2623 273 47 3 F2 gentle agitation 623 183 33 0 F2 vigorous agitation 1040 363 37 7Table 14b. Light Obscuration measurementsLight Obscuration Sub-visible particles / mL Composition / Stress >2μm >5μm >10μm >25μm Fl control 433 87 10 0 F2 control 600 130 13 3 Fl UV 493 140 30 0 F2 UV 247 53 7 0 Fl UV dark control 587 197 37 3 F2 UV dark control 293 93 23 3 Fl visible light 1157 303 30 3 F2 visible light 283 47 7 3 Fl visible light dark553 157 23 3 controlF2 visible light dark630 143 17 0 controlTable 15a. FlowCam microfluidic imagingFlowCam Microfluidic Imaging Sub-visible Particles ZmL Composition / Stress >2μm >5μm >10μm >25μm Fl control 389.86 343.66 103.96 17.33 Fl freeze-thaw 2550.46 2498.47 1195.8 228.18 Fl gentle agitation 418.82 404.38 138.64 37.55 Fl vigorous agitation 150.17 150.17 72.20 25.99 F2 control 115.54 112.65 40.44 11.55 F2 freeze-thaw 1772.81 1746.82 612.11 60.63 F2 gentle agitation 98.19 98.19 28.88 5.78 F2 vigorous agitation 92.43 89.54 34.66 17.33Table 15b. FlowCam microfluidic imagingAtty. Docket No. 0282-0010W01FlowCam Microfluidic Imaging Sub-visible Particles / ml Composition / Stress >2μm >5μm >10μm >25μm Fl control 271.41 256.97 69.30 11.55 F2 control 233.83 190.53 51.96 8.66 Fl UV 592.12 543.02 210.85 43.33 F2 UV 106.81 89.49 20.21 5.77 Fl UV dark control 349.56 326.45 109.78 26.00 F2 UV dark control 251.29 225.30 54.88 8.67 Fl visible light 513.94 441.76 69.30 25.99 F2 visible light 231.12 210.89 57.78 20.22 Fl visible light dark326.39 306.17 101.09 23.11 controlF2 visible light dark317.72 291.73 77.99 23.11 controlTable 16a. FlowCam light obscurationFlowCam Light Obscuration Sub-visible Particles / mL Composition / Stress >2μm >5μm >10μm >25μm Fl control 1436.44 644.06 211.30 44.70 Fl freeze-thaw 2455.86 802.37 144.22 6.09 Fl gentle agitation 542.59 233.70 40.64 2.03 Fl vigorous agitation 914.28 406.35 103.62 8.13 F2 control 621.85 254.02 48.77 2.03 F2 freeze-thaw 4962.50 2033.35 452.98 8.13 F2 gentle agitation 743.78 422.69 152.41 34.55 F2 vigorous agitation 886.03 487.72 148.35 6.10Table 16b. FlowCam light obscurationFlowCam Light Obscuration Sub-visible Particles / mL Composition / Stress >2μm >5μm >10μm >25μm Fl control 674.40 349.39 101.57 8.13 F2 control 720.96 320.88 58.90 18.28 Fl UV 1107.54 564.95 162.57 42.68 F2 UV 710.81 300.57 48.74 2.03Atty. Docket No. 0282-0010W01Fl UV dark control 886.03 438.95 99.58 22.35F2 UV dark control 516.17 254.02 54.87 4.06Fl visible light 1793.65 727.21 115.78 8.13F2 visible light 638.10 296.70 56.90 6.10Fl visible light dark524.30 237.76 71.13 6.10 controlF2 visible light dark723.30 294.60 52.83 6.10 controlExample 2. Long Term Stability of IVSS

[0167] To determine the chemical stability of IVSS during long term storage, each of the non-limiting, exemplary IVSS described above in Table 7 were stored at 5 °C, 25 °C, or 30 °C for up to 12 months. The test samples were taken at various timepoints (time 0, 1 month, 3 months, 6 months, 9 months, and 12 months) and tested for variations in pH, osmolality, and particle formation as compared to control samples. After visual inspection, all samples examined at each of the storage conditions were described as colorless, clean, and free from particles (data not shown). Further, each of the IVSS exhibited consistent pH (within ± 0.2 pH units of target pH) when stored at 5 °C for up to 12 months. However, IVSS Fl maintained its pH within target parameters even at temperatures as high as 25 °C. In addition, osmolality appeared consistent across all timepoints for each of the Fl, F2, and F3 sample compositions and at all storage temperatures. The results are summarized in Tables 17 and 18.Table 17. Long Term Stability of pH.Composition pH(storage 0 months 1 month 3 months 6 months 9 months 12 months temp)Fl (+5 °C)F2 (+5 °C) P655 -^-q-^^ 165?" >Fl (+25 °C) [N / A f'ftOl [ 5'99 f?87 ]To2 foi F2 (+25 °C) '" N / A T'55 ^55 >.................. p— ----- Fl (+30 °C) rN / A~""" """ — - Too""" """ """^ ^ / / T F2 (+30 °C) p^--------------TTT'''''''''''^^^^^^^Atty. Docket No. 0282-0010W01Table 18. Long Term Stability of OsmolalityComposition Osmolality (mOsm / kg)(storage 0 months 1 month 3 months 6 months 9 months 12 months temp)Fl (+5 °C) 73.33 73.30 75.00 74.00 74.00 74.00 F2 (+5 °C) 54.00 54.67 56.00 55.00 56.00 55.00 Fl (+25 °C) N / A 73.33 74.00 74.00 75.00 73.00 F2 (+25 °C) N / A 54.33 60.00 55.00 56.00 57.00 Fl (+30 °C) N / A 73.33 76.00 N / A N / A N / A F2 (+30 °C) N / A 54.33 56.00 N / A N / A N / A

[0168] Light obscuration measurements were performed on the sample IVSS following storage at 5 °C, 25 °C, and 30 °C for up to 12 months to measure particle formation. Each of the IVSS tested exhibited low particle counts at all storage temperatures that were well within USP requirements (less than 6,000 particles / mL of > 10 micron particles; less than 600 particles / mL of > 25 micron particles). The results are shown in Table 19. These results were confirmed by FlowCam analysis (data not shown).Table 19. Long Term Stability (Light Obscuration)Composition Particles / mL (+5°C)(particle 0 months 1 month 3 months 6 months 9 months 12 months size)Fl (> 10 pm) 0.00 13.33 30.00 50.00 10.00 37.00 F2 (> 10 pm) 16.67 10.00 30.00 13.00 20.00 10.00 Fl (> 25 pm) 0.00 6.67 0.00 23.00 7.00 0.00 F2 (> 10 pm) 0.00 3.33 0.00 7.00 3.00 0.00Particles / mL (+25°C)Fl (> 10 pm) 0.00 23.33 30.00 27.00 40.00 23.00 F2 (> 10 pm) 16.67 26.67 3.33 30.00 30.00 30.00 Fl (> 25 pm) 0.00 0.00 0.00 10.00 7.00 0.00 F2 (> 10 pm) 0.00 0.00 0.00 17.00 3.00 0.00Particles / mL (+30°C)Atty. Docket No. 0282-0010W01Fl (> 10 pm) 0.00 16.67 20.00 N / A N / A N / A F2 (> 10 pm) 16.67 46.67 53.33 N / A N / A N / A Fl (> 25 pm) 0.00 6.67 3.33 N / A N / A N / A F2 (> 10 pm) 0.00 6.67 0.00 N / A N / A N / A

[0169] In addition, the relative percentage of polyesters and monoesters of the IVSS Fl and F2 compositions during storage for up to 6 months at 5 °C and 25 °C was measured by reverse phase high protein liquid chromatography (RP-HPLC) as described in Kranz et al., Journal of Pharmaceutical Sciences 108:2022-2032 (2019).

[0170] A liquid chromatography-mass spectrometry (LC-MS)-based method was used to separate monoesters and polyesters to confirm the classification of the peaks. LC-MS analysis with longer cycle times (60 min) and by using the additional mass information was used for a more detailed and unambiguous differentiation between polyesters and monoesters. Best possible but arbitrary” peak integration limits, supported by LC-MS analysis, were set.

[0171] The results show that the relative area of both polyesters and monoesters did not vary significantly during this testing period demonstrating good chemical stability of both compositions. The results are shown in Table 20.Table 20. Long Term Stability (Polyesters and Monoesters)Composition Relative Area (%) Polyesters(temperature) 0 months 1 month | 3 months | 6 monthsFl (+5 °C) 71.47 71.50 71.79 71.11F2 (+5 °C) 71.76 71.47 71.60 71.21Fl (+25 °C) N / A 71.70 71.60 71.22F2 (+25 °C) N / A 71.44 71.67 70.85Relative Area (%) MonoestersFl (+5 °C) 28.53 28.50 28.21 28.89F2 (+5 °C) 28.24 28.53 28.40 28.78Fl (+25 °C) N / A 28.30 28.40 28.79F2 (+25 °C) N / A 28.56 28.33 29.15Atty. Docket No. 0282-0010W01

[0172] RP-HPLC-charged aerosol detector (CAD) analysis summarized above revealed that the IVSS Fl and F2 compositions exhibited chemical stability for up to 3 months regardless of the storage temperature. At 12 months, IVSS F2 shows some signs of oxidation at 25 °C. See FIG. 2. Surprisingly, however, the IVSS Fl composition showed good chemical stability of polysorbate 80 even up to 12 months. See FIG. 3 (complete overlay). Therefore, these results show that while each of the IVSS described above displayed chemical stability during long term storage, the IVSS Fl composition exhibited superior chemical stability even as compared to the IVSS F2 composition.Example 3. In-Use Stability Study

[0173] The IVSS Fl composition (see Table 7) was further tested for in-use stability. In particular, the IVSS Fl composition was used as medical bag protectant for soluble TCR proteins. The in-use stability of the IVSS Fl composition following the application of stress was measured as the percent recovery of the TCR protein as compared to the control samples. Two control samples were used. One of the controls samples was a medical IV bag using human serum albumin (HSA) as the protectant. The second control was the unstressed IVSS Fl composition. The test samples were compared to both controls.

[0174] Meso scale developability (MSD) assays were performed using an acceptance criteria of ±20%, which suggested that a sample with 100% recovery was taken to have a value of between about 80% and about 120%. Briefly, the testing consisted of two parts. In the first part, the percent recovery of a 0.2 pg dose of the TCR protein brenetafusp was measured after exposure of the IVSS to light stress and freeze-thaw stress (5 cycles). In the second part, the percent recovery of a 20 pg dose of the TCR protein tebentafusp was measured after exposure of the IVSS to excess Tween or terminal sterilization. Terminal sterilization consisted of autoclaving the sealed test vial containing the IVSS composition. The process began with vacuum and steam injection for 15 minutes followed by sterilization for 17 minutes at about 123 °C. The results are summarized in Table 21.Table 21. Percent Recovery of TCR proteinsSample Actual Recovery (%) for 0.2 mg dose of brenetafusp Composition Operator 1 Operator 1 Operator 2 Operator 2 (repeat) (repeat) HSA control 117.0 N / A 117.0 N / AAtty. Docket No. 0282-0010W01 120.1 N / A 116.0 N / A 115.9 / 112.3 98.9 111.0 / 101.0 99.0Fl unstressed*110.2 / 106.1 105.8 109.0 / 86.0 98.0117.3 N / A 118.0 N / AFl visible light115.6 N / A 109.0 N / A113.8 N / A 107.0 N / AFl (dark control)116.3 N / A 110.0 N / A107.4 N / A 98.0 N / AFl UV light87.9 N / A 93.0 N / A97.9 N / A 86.0 N / AFl (dark control)111.9 N / A 91.0 N / A103.3 N / A 101.0 N / AFl freeze-thaw102.8 N / A 95.0 N / A Sample Actual Recovery (%) for 20 mg dose of tebentafusp Composition Operator 1 (0 h) Operator 1 (6 d) Operator 2 (0 h) Operator 2 (6 d)94.4 93.7 97.6 N / AHSA control95.7 95.3 97.4 N / A91.2 80.5 93.0 N / AFl unstressed91.5 83.6 97.9 N / A92.1 N / A 93.2 N / AFl excess Tween100.7 N / A 103.5 N / AFl terminal 97.2 N / A 96.4 N / A sterilization 95.7 N / A 99.3 N / A+ The sample measurements in columns 2 and 4 were repeated by the operator the following day.

[0175] As shown above in Table 21 (top), the percent recovery for all brenetafusp samples was comparable when grouped by operator, which demonstrated that the unstressed, visible light stressed, and dark control sample IVSS Fl compositions were at least as effective at protecting the TCR protein from absorption to the container surfaces as compared to HSA. Moreover, when the unstressed sample measurements were repeated on the following day, the measurements were similar to the values obtained on the previous day, further demonstrating that the IVSS Fl composition is stable when used as a container protectant. Likewise, the data revealed that the freeze-thaw stress did not reduce the protective ability of the IVSS Fl composition. The UV light stress lowered the percentAtty. Docket No. 0282-0010W01recovery of the brenetafusp sample by only about 5% compared to controls, which indicated that UV light has little to no effect on the ability of the IVSS Fl composition to protect the TCR against surface absorption.

[0176] As shown in Table 21 (bottom), the percent recovery for all tebentafusp samples tested at time Oh. These results shown that the IVSS Fl composition is stable against various stresses and protects against surface absorption of soluble TCR proteins.

Claims

Atty. Docket No. 0282-0010W01CLAIMSWhat is claimed is:

1. A method of administering a low dose soluble protein to a subject in need thereof, the method comprising:a. adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a container to form a pretreated container, wherein the IVSS comprises:i. about 20 mM to about 30 mM sodium citrate;ii. about 0.5% (w / v) polysorbate 80; andiii. water,wherein the IVSS has a pH of about 5.8 to about 6.2,b. adding an amount from a protein stock solution to the pretreated container of step (a) to form a low dose protein solution; andc. administering the low dose protein solution of step (b) to the subject.

2. A method of administering an ultra-low dose soluble protein to a subject in need thereof, the method comprising:a. adding an intravenous solution stabilizer (“IVSS”) to an isotonic aqueous solution in a first container to form a first pretreated container, wherein the IVSS comprises: i. about 20 mM to about 30 mM sodium citrate;ii. about 0.5% (w / v) polysorbate 80; andiii. water,wherein the IVSS has a pH of about 5.8 to about 6.2;b. adding an amount of a protein stock solution to the first pretreated container of step (a) to form a low dose protein solution;c. adding an IVSS to an isotonic aqueous solution in a second container to form a second pretreated container;d. adding an amount of the low dose protein solution of step (b) to the second pretreated container of step (c) to form an ultra-low dose protein solution; and e. administering the ultra-low dose protein solution of step (d) to the subject.

3. The method of claim 1 or 2, wherein the protein stock solution comprises about 50 pg / mL to about 500 pg / mL of the protein.Atty. Docket No. 0282-0010W014. The method of any one of claims 1 to 3, wherein the low dose protein solution comprises 20 pg to about 800 pg of the protein.

5. The method of any one of claims 2 to 4, wherein the ultra-low dose protein solution comprises about 0.2 pg to about 20 pg of the protein.

6. The method of any one of claims 1 to 5, wherein the amount of the protein stock solution added to the pretreated container is at least 100 pL.

7. The method of any one of claims 2 to 6, wherein the amount of the low dose protein solution added to the second pretreated container is at least 100 pL.

8. The method of any one of claims 1 to 7, wherein the IVSS comprises 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate, and 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate.

9. The method of any one of claims 1 to 8, wherein the low dose or the ultra-low dose soluble protein is a soluble TCR protein.

10. The method of claim 9, wherein the soluble TCR protein is a heterodimeric TCR protein.

11. The method of claim 9 or 10, wherein the soluble TCR protein is a bispecific fusion protein.

12. The method of any one of claims 9 to 11, wherein the soluble TCR protein is a bispecific T-cell engager, such as a gplOO peptide-HLA-directed CD3 T-cell engager.

13. The method of claim 12, wherein the soluble TCR protein comprises an alpha chain complementarity determining region (CDR) sequence according to SEQ ID NOs: 6-8 and a beta chain CDR sequence according to SEQ ID NOs: 9-11.

14. The method of claim 12 or 13, wherein the soluble TCR protein is tebentafusp.

15. The method of claim 14, wherein the tebentafusp is added to the pretreated container from a stock solution at a concentration of about 50 pg / mL to about 500 pg / mL.

16. The method of claim 14 or 15, wherein the tebentafusp is administered to the subject at a dose amount of about 20 pg to about 800 pg.Atty. Docket No. 0282-0010W0117. The method of claim 14 or 15, wherein the tebentafusp is administered to the subject at a dose amount of about 0.2 pg to about 20 pg.

18. The method of any one of claims 9 to 11, wherein the soluble TCR protein targets preferentially expressed antigen of melanoma (PRAME).

19. The method of claim 18, wherein the soluble TCR protein comprises an alpha chain CDR sequence according to SEQ ID NOs: 31-33, SEQ ID NOs: 59-61, or SEQ ID NOs: 65-67; and a beta chain CDR sequence according to SEQ ID NOs: 39-41; SEQ ID NOs: 62-64; or SEQ ID NOs: 68-70.

20. The method of claim 18 or 19, wherein the soluble TCR protein is brenetafusp, IMC-P115C, or IMC-T119C.

21. The method of claim 20, wherein the soluble TCR protein is brenetafusp.

22. The method of claim 20 or 21, wherein the brenetafusp is added to the pretreated container from a stock solution at a concentration of about 50 pg / mL to about 500 pg / mL.

23. The method of claim 21 or 22, wherein the brenetafusp is administered to the subject at a dose amount of about 20 pg to about 800 pg.

24. The method of claim 21 or 22, wherein the brenetafusp is administered to the subject at a dose amount of about 0.2 pg to about 20 pg.

25. The method of any one of claims 9 to 11, wherein the soluble TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator.

26. The method of claim 25, wherein the soluble TCR protein comprises an alpha chain CDR sequence according to SEQ ID NOs: 71-73, and a beta chain CDR sequence according to SEQ ID NOs: 74-76.

27. The method of claim 25 or 26, wherein the PIWIL1 modulator is IMC-R117C.

28. The method of claim 27, wherein IMC-R117C is added to the pretreated container from a stock solution at a concentration of about 50 pg / mL to about 500 pg / mL.

29. The method of claim 27 or 28, wherein the IMC-R117C is administered to the subject at a dose amount of about 20 pg to about 800 pg.Atty. Docket No. 0282-0010W0130. The method of claim 27 or 28, wherein the IMC-R117C is administered to the subject at a dose amount of about 0.2 µg to about 20 µg.

31. The method of any one of claims 1 to 30, wherein the isotonic aqueous solution comprises one or more of normal saline, Ringer’s lactate solution, dextrose, or a combination thereof.

32. The method of any one of claims 1 to 31, wherein the IVSS comprises about 25 mM sodium citrate.

33. The method of any one of claims 1 to 32, wherein the protein stock solution is prepared from a concentrated protein solution.

34. The method of any one of claims 1 to 33, wherein the protein stock solution is prepared from a lyophilized TCR protein.

35. The method of claim 34, wherein the lyophilized TCR protein is reconstituted prior to adding to the pretreated container.

36. A pretreated container suitable for administering a low dose or an ultra-low dose protein, wherein the pretreated container comprises a composition comprising:a. an isotonic aqueous base;b. about 0.2 mM to about 0.3 mM sodium citrate; andc. about 0.005% (w / v) polysorbate 80;wherein the composition has a pH of about 5.8 to about 6.2.

37. The pretreated container of claim 36, wherein the composition further comprises the low dose or the ultra-low dose protein, wherein the low dose or the ultra-low dose protein is a soluble TCR protein.

38. The pretreated container of claim 37, wherein the soluble TCR protein is tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, or IMC-R117C.

39. The pretreated container of claim 37 or 38, wherein the composition comprises about 20 pg to about 800 pg of tebentafusp.

40. The pretreated container of claim 37 or 38, wherein the composition comprises about 0.2 pg to about 20 pg of tebentafusp.Atty. Docket No. 0282-0010W0141. The pretreated container of claim 37 or 38, wherein the composition comprises about 20 pg to about 800 pg of brenetafusp.

42. The pretreated container of claim 37 or 38, wherein the composition comprises about 0.2 pg to about 20 pg of brenetafusp.

43. The pretreated container of claim 37 or 38, wherein the composition comprises about 20 pg to about 800 pg of IMC-R117C.

44. The pretreated container of claim 37 or 38, wherein the composition comprises about 0.2 pg to about 20 pg of IMC-R117C.

45. An intravenous solution stabilizer (“IVSS”) for pretreating a container, the IVSS comprising:a. 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate;b. 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate; andc. 5 mg / mL PS80wherein the IVSS has a pH of about 5.8 to about 6.2.

46. A package comprising an intravenous solution stabilizer (“IVSS”), wherein the package comprises a container comprising the IVSS comprising:a. 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate;b. 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate; andc. 5 mg / mL PS80,wherein the IVSS has a pH of about 5.8 to about 6.2, andwherein the package does not transmit visible light or UV light.

47. The package of claim 46, wherein the package comprises a layer that blocks greater than 90% of UV light.

48. The package of claim 46 or claim 47, further comprising instructions for preparing a low dose or ultra-low dose protein solution, wherein the instructions comprise: (i) adding the IVSS to the container, wherein the container comprises an isotonic aqueous solution to form a pretreated container; (ii) adding a protein solution to the pretreated container to form a low dose protein solution; and, optionally, (iii) adding the low dose protein solution to a second pretreated container to form an ultra-low dose protein solution.Atty. Docket No. 0282-0010W0149. The package of any one of claims 46 to 48, wherein the IVSS further comprises the low dose or ultra-low dose protein, wherein the low dose or ultra-low dose protein is a soluble TCR protein.

50. The package of claim 49, wherein the soluble TCR protein is tebentafusp, brenetafusp, IMC-P115C, IMC-T119C, or IMC-R117C.

51. A kit comprising:a. a lyophilized T-cell receptor (“TCR”) protein;b. an intravenous solution stabilizer (“IVSS”), wherein the IVSS comprises:i. about 20 mM to about 30 mM sodium citrate;ii. about 0.5% (w / v) polysorbate 80; andiii. water,wherein the IVSS has a pH of about 6.0; andc. a container.

52. The kit of claim 51, wherein the IVSS comprises about 0.7 mg / mL to 0.8 mg / mL citric acid monohydrate.

53. The kit of claim 51, wherein the IVSS comprises 6.2 mg / mL to 6.3 mg / mL trisodium citrate dihydrate.

54. The kit of any one of claims 51 to 53, wherein the container is pre-treated with the IVSS.

55. The kit of any one of claims 51 to 54, wherein the lyophilized TCR protein is a bispecific T-cell engager, such as a gplOO peptide-HLA-directed CD3 T-cell engager.

56. The kit of claim 55, wherein the bispecific T-cell engager is tebentafusp.

57. The kit of claim 56, wherein the tebentafusp is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pre-treated containers.

58. The kit of any one of claims 51 to 54, wherein the lyophilized TCR protein is a PRAME targeting TCR protein.

59. The kit of claim 58, wherein the PRAME targeting TCR protein is selected from the group consisting of brenetafusp, IMC-P115C, and IMC-T119C.Atty. Docket No. 0282-0010W0160. The kit of claim 59, wherein the PRAME targeting TCR protein is brenetafusp.

61. The kit of claim 60, wherein the brenetafusp is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pre-treated containers.

62. The kit of any one of claims 51 to 54, wherein the lyophilized TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator.

63. The kit of claim 62, wherein the PIWIL1 modulator is IMC-R117C.

64. The kit of claim 63, wherein the IMC-R117C is in an amount that provides a dose of about 0.2 pg to about 800 pg when reconstituted in one or more pre-treated containers.

65. The kit of any one of claims 51 to 64, further comprising a medical grade plastic bag, wherein the IVSS is disposed within the medical grade bag.

66. The kit of claim 65, wherein the medical grade bag comprises a layer that does not transmit visible light or UV light.

67. The kit of claim 65 or 66, wherein the kit comprises a first medical grade bag and a second medical grade bag, wherein the IVSS is disposed in the first medical grade bag and the lyophilized TCR protein is disposed in the second medical grade bag.

68. A method of treating a disease or condition in a subject in need thereof comprising administering to the subject a soluble protein administered at low dose or ultra-low dose, i.e., a low dose protein or an ultra-low dose protein,wherein the low dose protein or the ultra-low dose protein is prepared with the use of an intravenous solution stabilizer (“IVSS”),wherein the IVSS comprises:i. about 20 mM to about 30 mM sodium citrate;ii. about 0.5% (w / v) polysorbate 80; andiii. water,wherein the IVSS has a pH of about 5.8 to about 6.2.

69. The method of claim 68, wherein the low dose protein or the ultra-low dose protein is a soluble TCR protein.Atty. Docket No. 0282-0010W0170. The method of claim 69, wherein the soluble TCR protein is a heterodimeric TCR protein.

71. The method of claim 69 or 70, wherein the soluble TCR protein is a bispecific fusion protein.

72. The method of any one of claims 69 to 71, wherein the soluble TCR protein is a bispecific CD3 T-cell engager.

73. The method of any one of claims 69 to 72, wherein the soluble TCR protein targets gplOO.

74. The method of claim 73, wherein the soluble TCR protein is tebentafusp.

75. The method of any one of claims 69 to 72, wherein the soluble TCR protein is a piwi like RNA-mediated gene silencing 1 (“PIWIL1”) modulator.

76. The method of claim 75, wherein the soluble TCR protein is IMC-R117C.

77. The method of any one of claims 69 to 72, wherein the soluble TCR protein targets preferentially expressed antigen of melanoma (PRAME).

78. The method of claim 77, wherein the soluble TCR protein is selected from the group consisting of brenetafusp, IMC-P115C, and IMC-T119C.

79. The method of any one of claims 68 to 78, wherein the disease is a PRAME-positive cancer.

80. A composition comprising:(a) an intravenous solution stabilizer (“IVSS”), wherein the IVSS comprises:i. about 20 mM to about 30 mM sodium citrate;ii. about 0.5% (w / v) polysorbate 80; andiii. water,wherein the IVSS has a pH of about 6.0; and(b) a low dose or an ultra-low dose soluble protein.

Images