Tools for molecular imaging of endometriosis

Abstract

An immunoconjugate is provided that selectively binds to endometriosis tissue preferentially over non-endometriosis tissue. Methods for imaging endometriosis tissue in vivo are also provided.

Description

TOOLS FOR MOLECULAR IMAGING OF ENDOMETRIOSISCROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims priority to, and is a non-provisional of, of U.S. PatentApplication 63 / 691,902 (filed September 6, 2024), the entirety of which is incorporatedherein by reference. STATEMENT OF FEDERALLY SPONSORED RESEARCH OR DEVELOPMENT

[0002] This invention was made with Government support under grant numberUL1TR002384 awarded by the National Center for Advancing Translational SciencesThe government has certain rights in the invention. REFERENCE TO A SEQUENCE LISTING

[0003] This application contains a Sequence Listing in computer readable form. Thecomputer readable form is incorporated herein by reference. The computer readable fileis named Seqeuence.xml and was created on November 2, 2024 (92 kB size).BACKGROUND OF THE INVENTION

[0004] Endometriosis is a chronic female reproductive disorder commonly definedas the growth of endometrial tissue outside of the uterus. Endometriosis effects roughly 10-15% of women of reproductive age, nearly 6.5 million women in the United States. The cardinal symptoms of endometriosis include chronic pelvic pain, infertility, and pain during menstruation, intercourse, and defecation. The social and financial costs of the disease are staggering: the pain from endometriosis can affect all aspects of a patient’s daily life, and the disorder is responsible for ~$70 billion per year in excess health expenditures in the U.S. alone. Endometriosis can manifest as superficial peritoneal lesions, ovarian endometriomas, or deep infiltrating endometriosis (DIE). Despite its ubiquity, endometriosis remains under-funded and under-researched. As a workinggroup assembled by the Society for Women’s Health Research’s recently noted, “scientific progress has been slow, and diagnostic and treatment options remain limited.”

[0005] For decades, the clinical diagnosis of endometriosis has been predicated onlaparoscopic surgery. Laparoscopic surgery is invasive, painful, expensive, and prone tosampling and operator biases. Indeed, the invasive and painful nature of laparoscopy are instrumental in creating the extensive diagnostic delays common in endometriosis. Furthermore, while the sensitivity and positive predictive value of laparoscopy are relatively high, the procedure’s negative predictive value and specificity are low. Thoughnon-invasive imaging methods, like magnetic resonance imaging (MRI) and ultrasound,can detect endometriomas and DIE, they often miss smaller endometriosis lesions that are the majority of cases. Not surprisingly, therapeutic paradigms for endometriosis havealso long been inadequate. Indeed, surgical resection stands as the principal clinicalmethod for the treatment of endometriosis. Surgeons’ reliance upon visual inspection forthe identification of disease during these procedures can lead to significant operator bias,the inadvertent removal of healthy tissue, and the accidental failure to remove diseased tissue.

[0006] Taken together, the data make it clear that new tools for diagnosingendometriosis are an urgent unmet clinical need. To date, no such tool is readily available. The discussion above is merely provided for general background information and is not intended to be used as an aid in determining the scope of the claimed subject matter. SUMMARY

[0007] This disclosure provides immunoconjugates that selectively bind toendometriosis tissue preferentially over non-endometriosis tissue. Methods for imagingendometriosis tissue in vivo are also provided. An advantage that may be realized in thepractice of some disclosed embodiments is the ability to non-invasively detect and / or image endometriosis.

[0008] In a first embodiment, an antibody is provided. The antibody comprising: aheavy chain (VH) selected from SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO: 46, SEQID NO: 64 and SEQ ID NO: 82; and a light chain (VL) selected from SEQ ID NO: 10,SEQ ID NO: 37, SEQ ID NO: 55, SEQ ID NO: 73 and SEQ ID NO: 91.

[0009] In a second embodiment, a composition of matter is provided. Thecomposition of matter comprising an antibody comprising: a heavy chain (VH) that is at least 80% identical to SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO: 46, SEQ ID NO: 64 or SEQ ID NO: 82; a light chain (VL) that is at least 80% identical to SEQ ID NO: 19,SEQ ID NO: 37, SEQ ID NO: 55, SEQ ID NO: 73 and SEQ ID NO: 91; and a labelcovalently conjugated to the antibody.

[0010] In a third embodiment, a method for imaging a patient is provided. Themethod comprising: administering to a human patient the composition of matter as recited in any one of claims 10-25; and imaging the label within the human patient.

[0011] This brief description of the invention is intended only to provide a briefoverview of subject matter disclosed herein according to one or more illustrative embodiments and does not serve as a guide to interpreting the claims or to define or limit the scope of the invention, which is defined only by the appended claims. This brief description is provided to introduce an illustrative selection of concepts in a simplified form that are further described below in the detailed description. This brief description is not intended to identify key features or essential features of the claimed subject matter, nor is it intended to be used as an aid in determining the scope of the claimed subject matter. The claimed subject matter is not limited to implementations that solve any or all disadvantages noted in the background. BRIEF DESCRIPTION OF THE DRAWINGS

[0012] The patent or application file contains at least one drawing executed in color.Copies of this patent or patent application publication with color drawing(s) will be provided by the Office upon request and payment of the necessary fee.

[0013] So that the manner in which the features of the invention can be understood, adetailed description of the invention may be had by reference to certain embodiments,some of which are illustrated in the accompanying drawings. It is to be noted, however,that the drawings illustrate only certain embodiments of this invention and are thereforenot to be considered limiting of its scope, for the scope of the invention encompasses other equally effective embodiments. The drawings are not necessarily to scale,emphasis generally being placed upon illustrating the features of certain embodiments ofthe invention. In the drawings, like numerals are used to indicate like parts throughout the various views. Thus, for further understanding of the invention, reference can be made to the following detailed description, read in connection with the drawings in which:

[0014] FIG. 1A shows a Western blotting of MMP26 in 12Z and SHT290 cells withan actin loading control.

[0015] FIG. 1B depicts confocal microscopy images of immunocytochemical stainingfor MMP26 of endometrial cell lines.

[0016] FIG. 1C depicts confocal microscopy images of immunocytochemical stainingfor MMP26 of normal human ovarian and uterine cell lines.

[0017] FIG. 2A depicts confocal microscopy images of histologicalimmunofluorescent staining for MMP26 of subcutaneous xenograft tissue of mixed 12Z and SHT290 cells.

[0018] FIG. 2B depicts confocal microscopy images of histologicalimmunofluorescent staining for MMP26 of two representative patients’ biopsiedendometriosis tissue (n=6).

[0019] FIG. 2C depicts confocal microscopy images of histologicalimmunofluorescent staining for MMP26 of two representative patients’ biopsied normalperimetrium tissue (negative for endometriosis) (n=4).

[0020] FIG. 3A is a graph depicting ELISA results of five MMP26-targeting mAbsbinding to MMP26.

[0021] FIG. 3B illustrates a table showing yes / no binding results via ELISA assays ofthe MMP26 targeting mAbs to six alternative matrix metalloproteinase enzymes. Percent identity with MMP26 of each alternative MMP is shown.

[0022] FIG. 3C is an ELISA assay graph showing the disclosed antibodies do notbind to MMP1.

[0023] FIG. 3D is an ELISA assay graph showing the disclosed antibodies do notbind to MMP3.

[0024] FIG. 3E is an ELISA assay graph showing the disclosed antibodies do notbind to MMP7.

[0025] FIG. 3F is an ELISA assay graph showing the disclosed antibodies do notbind to MMP10.

[0026] FIG. 3G is an ELISA assay graph showing the disclosed antibodies do notbind to MMP12.

[0027] FIG. 3H is an ELISA assay graph showing the disclosed antibodies do notbind to MMP13.

[0028] FIG. 3I is an ELISA assay graph showing the disclosed antibodies do not bindto MMP20.

[0029] FIG. 3J is an ELISA assay graph showing the disclosed antibodies do not bindto MMPMT1-MMP.

[0030] FIG. 4A is a graph depicting the results of an ELISA binding assay comparingnative mAb to the DFO-bearing immunoconjugate of 2F11.

[0031] FIG. 4B is a graph depicting the results of an ELISA binding assay comparingnative mAb to the DFO-bearing immunoconjugate of 6A5.

[0032] IG. 4C is a graph depicting the results of an ELISA binding assay comparingnative mAb to the DFO-bearing immunoconjugate of 11D2.

[0033] IG. 4D is a graph depicting the results of an ELISA binding assay comparingnative mAb to the DFO-bearing immunoconjugate of 15B9.

[0034] FIG. 4E is a graph depicting the results of an ELISA binding assay comparingnative mAb to the DFO-bearing immunoconjugate of 12E2.

[0035] FIG. 5A results the results of radio-iTLC post-radiolabeling with 89Zr of 2F11.

[0036] FIG. 5B results the results of radio-iTLC post-radiolabeling with 89Zr of 6A5.

[0037] FIG. 5C results the results of radio-iTLC post-radiolabeling with 89Zr of11D2.

[0038] FIG. 5D results the results of radio-iTLC post-radiolabeling with 89Zr of15B9.

[0039] FIG. 5E results the results of radio-iTLC post-radiolabeling with 89Zr of 12E2.

[0040] FIG. 6A is a graph showing the result of a 5-day stability study of the[89Zr]Zr-DFO-2F11 radioimmunoconjugates in human serum.

[0041] FIG. 6B is a graph showing the result of a 5-day stability study of the[89Zr]Zr-DFO-6A5 radioimmunoconjugates in human serum.

[0042] FIG. 6C is a graph showing the result of a 5-day stability study of the[89Zr]Zr-DFO-11D2 radioimmunoconjugates in human serum.

[0043] FIG. 6D is a graph showing the result of a 5-day stability study of the[89Zr]Zr-DFO-15B9 radioimmunoconjugates in human serum.

[0044] FIG. 6E is a graph showing the result of a 5-day stability study of the [89Zr]Zr-DFO-12E2 radioimmunoconjugates in human serum.

[0045] FIG. 7 is a schematic depiction of an antibody.DETAILED DESCRIPTION OF THE INVENTION

[0046] In order for the present invention to be more readily understood, certain termsare first defined below. Additional definitions for the following terms and other terms are set forth throughout the specification.

[0047] Animal: As used herein, the term “animal” refers to any member of theanimal kingdom. In some embodiments, “animal” refers to humans, at any stage of development. In some embodiments, “animal” refers to non-human animals, at any stage of development. In certain embodiments, the non-human animal is a mammal (e.g., a rodent, a mouse, a rat, a rabbit, a monkey, a dog, a cat, a sheep, cattle, a primate, and / or a pig). In some embodiments, animals include, but are not limited to, mammals, birds,reptiles, amphibians, fish, insects, and / or worms. In some embodiments, an animal may be a transgenic animal, genetically-engineered animal, and / or a clone.

[0048] Approximately or about: As used herein, the term “approximately” or“about,” as applied to one or more values of interest, refers to a value that is similar to a stated reference value. In certain embodiments, the term “approximately” or “about” refers to a range of values that fall within 25%, 20%, 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, or less in either direction (greater than or less than) of the stated reference value unless otherwise stated or otherwise evident from the context (except where such number would exceed 100% of a possible value).

[0049] In Vitro: As used herein, the term “in vitro” refers to events that occur in anartificial environment, e.g., in a test tube or reaction vessel, in cell culture, etc., rather than within a multi-cellular organism.

[0050] In Vivo: As used herein, the term “in vivo” refers to events that occur within amulti-cellular organism, such as a human and a non-human animal. In the context of cell- based systems, the term may be used to refer to events that occur within a living cell (as opposed to, for example, in vitro systems).

[0051] Isolated: As used herein, the term “isolated” refers to a substance and / orentity that has been (1) separated from at least some of the components with which it was associated when initially produced (whether in nature and / or in an experimental setting), and / or (2) produced, prepared, and / or manufactured by the hand of man. Isolated substances and / or entities may be separated from about 10%, about 20%, about 30%, about 40%, about 50%, about 60%, about 70%, about 80%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% of the other components with which they were initially associated. In some embodiments, isolated agents are about 80%, about 85%, about 90%, about 91%, about 92%, about 93%, about 94%, about 95%, about 96%, about 97%, about 98%, about 99%, or more than about 99% pure. As used herein, a substance is “pure” if it is substantially free of other components. As used herein, calculation of percent purity ofisolated substances and / or entities should not include excipients (e.g., buffer, solvent, water, etc.).

[0052] Patient: As used herein, the term “patient” or “subject” refers to any organismto which a provided composition may be administered, e.g., for experimental, diagnostic, prophylactic, cosmetic, and / or therapeutic purposes. Typical patients include animals (e.g., mammals such as mice, rats, rabbits, non-human primates, and / or humans). In some embodiments, a patient is a human. A human includes pre and post natal forms.

[0053] Pharmaceutically acceptable: The term “pharmaceutically acceptable” asused herein, refers to substances that, within the scope of sound medical judgment, are suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit / risk ratio.

[0054] Systemic distribution or delivery: As used herein, the terms “systemicdistribution,” “systemic delivery,” or grammatical equivalent, refer to a delivery or distribution mechanism or approach that affect the entire body or an entire organism. Typically, systemic distribution or delivery is accomplished via body's circulation system, e.g., blood stream. Compared to the definition of “local distribution or delivery.”

[0055] Subject: As used herein, the term “subject” refers to a human or any non-human animal (e.g., mouse, rat, rabbit, dog, cat, cattle, swine, sheep, horse or primate). Ahuman includes pre- and post-natal forms. In many embodiments, a subject is a humanbeing. A subject can be a patient, which refers to a human presenting to a medicalprovider for diagnosis or treatment of a disease. The term “subject” is used herein interchangeably with “individual” or “patient.” A subject can be afflicted with or is susceptible to a disease or disorder but may or may not display symptoms of the disease or disorder.

[0056] Substantially: As used herein, the term “substantially” refers to the qualitativecondition of exhibiting total or near-total extent or degree of a characteristic or property of interest. One of ordinary skill in the biological arts will understand that biological and chemical phenomena rarely, if ever, go to completion and / or proceed to completeness orachieve or avoid an absolute result. The term “substantially” is therefore used herein to capture the potential lack of completeness inherent in many biological and chemical phenomena.

[0057] Therapeutically effective amount: As used herein, the term “therapeuticallyeffective amount” of a therapeutic agent means an amount that is sufficient, when administered to a subject suffering from or susceptible to a disease, disorder, and / or condition, to treat, diagnose, prevent, and / or delay the onset of the symptom(s) of the disease, disorder, and / or condition. It will be appreciated by those of ordinary skill in the art that a therapeutically effective amount is typically administered via a dosing regimen comprising at least one unit dose.

[0058] Treating: As used herein, the term “treat,” “treatment,” or “treating” refers toany method used to partially or completely alleviate, ameliorate, relieve, inhibit, prevent,delay onset of, reduce severity of and / or reduce incidence of one or more symptoms orfeatures of a particular disease, disorder, and / or condition. Treatment may be administered to a subject who does not exhibit signs of a disease and / or exhibits only early signs of the disease for the purpose of decreasing the risk of developing pathology associated with the disease.

[0059] This disclosure provides an endometriosis-targeted Positron EmissionTomography (PET) imaging composition that improves diagnoses. Such anendometriosis-targeted intraoperative imaging composition could help surgeons safelyremove unhealthy tissue while sparing healthy organs. The target for the composition ismatrix metalloproteinase 26 (MMP26), a recently discovered member of the matrix metalloproteinase family. MMP26 makes a particularly attractive target for imagingbecause transcription data reveals that its mRNA is present in high levels in theendometrium but not within any other healthy tissues. Furthermore, IHC and ELISAhave confirmed the expression of high levels of MMP26 by healthy endometrium as well as endometriosis tissue.

[0060] MMP26 extracellularly expressed by endometrial cells

[0061] The experiments that were preliminarily performed were dedicated to theinvestigation of MMP26 expression and secretion by human endometrial cells and endometriosis. To this end, we began by performing western blotting of two human endometrial cell line lysates—endometrial epithelial cells (12Z) and endometrial stromalcells (SHT290). Three western blots were performed of each lystate (loading 50 g, 25g and 10 g of each) within Actin used as an internal control. MMP26 expression was found in both cell extracts, with notably higher expression within the 12Z cells(FIG. 1A).

[0062] As the disclosed radiotracer is based upon an antibody, which does notpenetrate the cell membrane, immunocytochemical (ICC) staining of the endometrial cell lines was conducted in order to validate the extracellular expression and secretion of MMP26. ICC and confocal microscopy results displayed that MMP26 was extracellularly expressed in 12Z cells, SHT290 cells, and the two cells co-cultured together.

[0063] When the co- -estradiol — ahormone that is upregulated in patients with endometriosis — prior to ICC staining, thecells exhibited even higher expression of MMP26 (FIG. 1B). Two other cell lines,H6036 (human primary ovarian epithelial cells) and HUtMEC (human uterine microvascular endothelial cells), were also stained for MMP26 with the goal being toshow little-to-no expression of the target antigen to ensure its specific expression inendometriosis lesions. ICC and confocal imaging of these cells displayed almost noexpression of MMP26 (FIG. 1C).

[0064] MMP26 expression is selective

[0065] A series of histological staining for MMP26 was then performed to furtherensure specific expression of the target antigen in human endometriosis tissue. First, 12Z and SHT290 cells were subcutaneously xenografted in nude athymic mice and allowed to grow for approximately two months. These xenografts were then extracted, flash-frozen in OCT, sliced and mounted onto microscope slides. Immunofluorescent staining andconfocal images of the tissue slices confirmed MMP26 expression by the xenograftedtissue (FIG. 2A, two representative images shown).

[0066] The same immunofluorescent staining was performed on biopsied humanendometriosis tissue (n=6). Five of the six human endometriosis tissues expressedMMP26, visualized via confocal imaging of the stained slides (FIG. 2B, tworepresentative images shown). Normal human perimetrium tissue biopsies that were deemed negative for endometriosis were also obtained and subsequently stained forMMP26 (n=4), and none of the tissues displayed expression of the antigen (FIG. 2C, tworepresentative images shown).

[0067] MMP26 Antibodies

[0068] Once MMP26 expression by endometrial cells and human endometriosis wasconfirmed, a small library of five monoclonal antibodies (mAb) that target MMP26 wasformed from rabbits via immunization with recombinant human MMP26. The resultantmAb constructs were named the following: 2F11, 6A5, 11D2, 15B9, and 12E2. These antibodies underwent ELISA binding assays using recombinant human MMP26 where allfive antibodies were confirmed in their binding to the MMP26 (SEQ ID NO: 1)(FIG. 3A). Each antibody preferentially binds to MMP26 by at least a factor of 10compared to one or more of MMP7, MMP13, MMP1, MMP10, MMP20, MIT1-MMP,MMP3 and MMP12 at a concentration of 1 per mL according to an ELISA assay(OD450). In one embodiment, each antibody preferentially binds to MMP26 by at least afactor of 10 compared to at least three or more of the aforementioned proteins. In another embodiment, antibody preferentially binds to MMP26 by at least a factor of 10 compared to at least five or more of the aforementioned proteins.

[0069] A series of ELISAs was also performed on the library of antibodies to testtheir binding to other recombinant matrix metalloproteinases in order to ensure specific binding of the antibodies to their target. Each recombinant MMP antigen was diluted to 0.5 µg / mL in sterile PBS, and 100 µL / well was coated onto an ELISA plate overnight at4ºC. MMP26 was also plated and simultaneously assessed for each ELISA as a positivecontrol. The following day, all wells were washed three times each with PBS + 0.05%Tween-20, and blocking was performed with PBS + 10% FBS. Immunoconjugates werediluted in blocking buffer at a serial dilution (1-0.00001 µg / mL) and 100 µL wasincubated in each well for 1 h at room temperature (22 ). The immunoconjugates weredetected using 1:10000 dilution of horseradish peroxidase (HRP)-labeled anti-mousesecondary IgG. After the final wash, 3,3’,5,5’-tetramethylbenzidine (TMB) substrate wasused to develop the bound HRP secondary antibody and after 12 minutes, the reactionwas quenched using 2 N H2SO4. A SpectraMax i3 plate reader was used to determineoptical densities at 450 nm and the binding data was collected in triplicate, normalized tobackground, and plotted. None of the antibodies displayed any significant binding toMMP7 (SEQ ID NO: 2), MMP13 (SEQ ID NO: 3), MMP1 (SEQ ID NO: 4), MMP10(SEQ ID NO: 5), MMP20 (SEQ ID NO: 6), or MMP12 (SEQ ID NO: 7) (FIG. 3B).FIGS. 3C-3J are ELISA assay graphs showing the disclosed antibodies are highlyselective for MMP26 and do not bind to other MMPs.

[0070] MMP26 immunoconjugates

[0071] The library of antibodies then underwent a random lysine-directedbioconjugation to the chelator desferrioxamine (DFO) and the resultant immunoconjugates’ degree of labeling (DOL) was determined via MALDI-TOF mass spectrometry. MALDI-TOF results revealed that the DOLs of 2F11, 6A5, 11D2, 15B9, 12E2 were 1.8, 2.3, 2.6, 1.6, and 1.6, respectively.

[0072] In other embodiments, other conjugation strategies are used such as randomlysine bioconjugation, maleimide-directed thiol conjugation, PODS-directed thiolconjugation, strain-promoted azide-aklyne cycloaddition (SPAAC) conjugation betweena cyclooctyne and an azide, strain-promoted oxidation-controlled quinone (SPOCQ)cycloaddition conjugation between a trans-cyclooctene and a 1,2-quinone, enzyme-mediated glycan modification bioconjugation (EndoS-GalT, ß-1,4-galactosidase-GalT,mushroom tyrosinase), peptide tag-directed conjugation (sortase, lipoic acid ligase), Z-domain photoactivatable BPA-directed conjugation, perfluorophenyl ester-directedconjugation, Fc-binding peptide-directed conjugation.

[0073] In other embodiments, other chelating agents are used such as DOTA and itsderivatives (CB-DO2A, 3p-C-DEPA, TCMC, Oxo-DO3A), TETA and its derivatives(TE2A, CB-TE2A, CB-TE1A1P, CB-TE2P, MM-TE2A, DM-TE2A), Diamsar, NOTA and its derivatives (NETA, TACN-TM, C-NETA, C-NE2TA, 3p-C-NETA, 3p-C-DEPA, TACN-HSB), DTPA and its derivatives (1B4M-DTPA, CHX-A’’-DTPA, CHX-B’’- DTPA), TRAP (PRP9), NOPO, PCTA, CP526, AAZTA and its derivative (DATA), pa family chelators (H2dedpa, H4octapa, H2azapa, H5decapa), HBED and its derivative (SHBED), BPCA, H6phospa, HEHA and its derivative (PEPA), NEC-SP, L1, BAT-TM,EC, SBAD, BAPEN, TACHPYR, L2, DiP-LICAM, and L3.

[0074] Due to the stochastic nature of the chosen bioconjugation strategy, additionalELISAs using recombinant MMP26 were performed directly comparing the binding ofthe native mAbs to that of the DFO-bearing immunoconjugates (FIGs. 4A-4E). Thisensured that the binding of the immunoconjugates to their target antigen was not perturbed due to the addition of the chelators.

[0075] The MMP26-targeting immunoconjugates were then radiolabeled with thepositron-emitting radiometal zirconium-89 (89Zr) according to standard published protocols and the purity of the radioimmunoconjugates was determined via radio-iTLC(FIGs. 5A-5E). Briefly, each immunoconjugate (0.5 mg) was diluted in Chelex-treatedPBS to a final concentration of 1 mg / mL. [89Zr]Zr4+ in 1.0 M oxalic acid was diluted inHEPES buffer so that the solution pH was adjusted to 7.0-7.5. The [89Zr]Zr4+ was addedto the solution of mAb, mixed thoroughly, and incubated on a ThermoMixer for 15 min at 500 rpm and 37 ºC. The progress of the reaction was monitored via radio-iTLC with an eluent of 50 mM EDTA, pH 5.0, an AR-2000 Radio-TLC plate reader, and Winscan Radio-TLC software. Once the reaction reached completion, free [89Zr]Zr4+was removed via size exclusion chromatography. The radiochemical purity of the final radiolabeled construct was assayed using radio-iTLC with an eluent of 50 mM EDTA, pH 5.0. All five radioimmunoconjugates were synthesized at >95% radiochemical yield and >95% purity with a specific activity of 3 mCi / mg, post-purification.

[0076] The radioimmunoconjugates were then placed into human serum and over thecourse of five days, their stability was evaluated over a 5-day period via radio-iTLC and size exclusion chromatography-HPLC. Following radiolabeling with [89Zr]Zr4+, 500 µCi of each radioimmunoconjugate were incubated in human serum at 37 ºC for 120 h. The stability of the radioimmunoconjugates to demetallation was measured every day viaradio-iTLC and every other day via SEC-HPLC. [89Zr]Zr-DFO-6A5, -11D2, -15B9, and -12E2 all exhibited >85% stability over the five days. [89Zr]Zr-DFO-2F11 exhibited somedegree of instability after 72 hours (<50%) (FIGS. 6A-E).

[0077] In other embodiments, other labels are used such as a radiolabel (e.g. Zr-89,Cu-60, Cu-61, Cu-62, Cu-64, Ga-68, Mn-52, Y-86, Ga-66, Sc-44, I-120, I-122, I-124[18F]AlF, etc.) or a fluorophore (green fluorescent proteins (GFP), yellow fluorescent proteins (YFP), red fluorescent proteins (RFP), xanthenes (fluorescein, fluoresceinisothiocyanate, rhodamine), cyanines, coumarins, pyrenes (cascade blue, etc.)), and thelike, may be used. Other suitable labels would be apparent to those skilled in the art afterbenefitting from reading this specification and such labels are also contemplated for use with the disclosed immunoconjugates.

[0078] FIG. 7 shows a schematic depiction of an antibody. Antibodies 2F11, 6A5,11D2, 15B9, and 12E2 are characterized by the variable region of the heavy chain (VH)and light chain (VL). Each of these include four framework regions (FR1, FR2, FR3 and FR4) that separate three respective complementary determining regions (CDR1, CDR2, CDR3). A constant region (CH1 and CLin FIG.7) follows the fourth framework region FR4. A fragment crystallizable (Fc) region comprises CH2 and CH3. Table 1: SEQ ID NOs of VH regionsTable 2: SEQ ID NOs of VL regions Antibody VL FR1 CDR1 FR2 CDR2 FR3 CDR3 FR4 CL, %,at least 85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 10 and SEQ IDNO: 19, respectively. In one such embodiment, the CDR1, CDR2 and CDR3 regions areat least 90%, at least 95% or are 100% identical with SEQ ID NOs: 12, 14, 16, 21, 23 and25 for the VH and VL regions, respectively. In one such embodiment, the FR1, FR2, FR3and FR4 regions are at least 80%, at least 85%, at least 90%, at least 95% or are 100%identical with SEQ ID NOs: 11, 13, 15, 17, 20, 22, 24 or 26 for the VH and VL regions,respectively. The constant regions, CH1 and CL are at least 80%, at least 85%, at least90%, at least 95% or 100% identical to SEQ ID NO: 18 and SEQ ID NO: 27, respectivelyand are directly connected to the respective VH and VL. With regard to the Fc regions,the isotype of 6A5 is mouse IgG2a / kappa based on the constant region.

[0080] In one embodiment, the antibody is 11D2 and both VH and VL are at least80%, at least 85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 28 andSEQ ID NO: 37, respectively. In one such embodiment, the CDR1, CDR2 and CDR3regions are at least 90%, at least 95% or are 100% identical with SEQ ID NOs: 30, 32,34, 39, 41 and 43 for the VH and VL regions, respectively. In one such embodiment, theFR1, FR2, FR3 and FR4 regions are at least 80%, at least 85%, at least 90%, at least 95%or are 100% identical with SEQ ID NOs: 29, 31, 33, 35, 38, 40, 42 or 44 for the VH andVLregions, respectively. The constant regions, CH1 and CLare at least 80%, at least85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 36 and SEQ ID NO:45, respectively and are directly connected to the respective VH and VL. The isotype of11D2 is mouse IgG2b / kappa based on the constant region.

[0081] In one embodiment, the antibody is 15B9 and both VH and VL are at least80%, at least 85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 46 andSEQ ID NO: 55, respectively. In one such embodiment, the CDR1, CDR2 and CDR3regions are at least 90%, at least 95% or are 100% identical with SEQ ID NOs: 48, 50,52, 57, 59 and 61 for the VH and VL regions, respectively. In one such embodiment, theFR1, FR2, FR3 and FR4 regions are at least 80%, at least 85%, at least 90%, at least 95%or are 100% identical with SEQ ID NOs: 47, 49, 51, 53, 56, 58, 60 or 62 for the VH andVL regions, respectively. The constant regions, CH1 and CL are at least 80%, at least85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 54 and SEQ ID NO:63, respectively and are directly connected to the respective VHand VL. The isotype of15B9 is mouse IgG2b / kappa based on the constant region.

[0082] In one embodiment, the antibody is 12E2 and both VH and VL are at least80%, at least 85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 64 andSEQ ID NO: 73, respectively. In one such embodiment, the CDR1, CDR2 and CDR3regions are at least 90%, at least 95% or are 100% identical with SEQ ID NOs: 66, 68,70, 75, 77 and 79 for the VH and VL regions, respectively. In one such embodiment, theFR1, FR2, FR3 and FR4 regions are at least 80%, at least 85%, at least 90%, at least 95%or are 100% identical with SEQ ID NOs: 65, 67, 69, 71, 74, 76, 78 or 80 for the VH andVL regions, respectively. The constant regions, CH1 and CL are at least 80%, at least85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 72 and SEQ ID NO:81, respectively and are directly connected to the respective VHand VL. The isotype of12E2 is mouse IgG2b / kappa based on the constant region.

[0083] In one embodiment, the antibody is 2F11 and both VH and VL are at least80%, at least 85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 82 andSEQ ID NO: 91, respectively. In one such embodiment, the CDR1, CDR2 and CDR3regions are at least 90%, at least 95% or are 100% identical with SEQ ID NOs: 84, 86,88, 93, 95 and 97for the VH and VL regions, respectively. In one such embodiment, theFR1, FR2, FR3 and FR4 regions are at least 80%, at least 85%, at least 90%, at least 95%or are 100% identical with SEQ ID NOs: 83, 85, 87, 89, 92, 94, 96 or 97 for the VH andVLregions, respectively. The constant regions, CH1 and CLare at least 80%, at least85%, at least 90%, at least 95% or 100% identical to SEQ ID NO: 90 and SEQ ID NO:99, respectively and are directly connected to the respective VH and VL. The isotype of2F11 is Mouse IgG1 / kappa based on the constant region.

[0084] The disclosed immunoconjugates can be used in conjunction with a variety ofmethods. For example, a saline solution of the immunoconjugate may be administeredintravenously (IV) to a human patient prior to imaging of the label. A variety ofconventional imaging techniques may be used to provide images of the label. Examplesinclude Computerized Axial Tomography (CAT) scans. The imaging technique may display a digital image of the human patient and the location of the label therein.

[0085] This written description uses examples to disclose the invention, including thebest mode, and also to enable any person skilled in the art to practice the invention, including making and using any devices or systems and performing any incorporated methods. The patentable scope of the invention is defined by the claims, and may include other examples that occur to those skilled in the art. Such other examples are intended to be within the scope of the claims if they have structural elements that do not differ from the literal language of the claims, or if they include equivalent structural elements with insubstantial differences from the literal language of the claims.SEQ ID NO: 1 MMP-26MQLVILRVTIFLPWCFAVPVPPAADHKGWDFVEGYFHQFFLTKKESPLLTQETQT QLLQQFHRNGTDLLDMQMHALLHQPHCGVPDGSDTSISPGRCKWNKHTLTYRII NYPHDMKPSAVKDSIYNAVSIWSNVTPLIFQQVQNGDADIKVSFWQWAHEDGW PFDGPGGILGHAFLPNSGNPGVVHFDKNEHWSASDTGYNLFLVATHEIGHSLGL QHSGNQSSIMYPTYWYHDPRTFQLSADDIQRIQHLYGEKCSSDIPSEQ ID NO: 2 MMP7MRLTVLCAVCLLPGSLALPLPQEAGGMSELQWEQAQDYLKRFYLYDSETKNAN SLEAKLKEMQKFFGLPITGMLNSRVIEIMQKPRCGVPDVAEYSLFPNSPKWTSKVVTYRIVSYTRDLPHITVDRLVSKALNMWGKEIPLHFRKVVWGTADIMIGFARGA HGDSYPFDGPGNTLAHAFAPGTGLGGDAHFDEDERWTDGSSLGINFLYAATHEL GHSLGMGHSSDPNAVMYPTYGNGDPQNFKLSQDDIKGIQKLYGKRSNSRKKSEQ ID NO: 3 MMP13MHPGVLAAFLFLSWTHCRALPLPSGGDEDDLSEEDLQFAERYLRSYYHPTNLAG ILKENAASSMTERLREMQSFFGLEVTGKLDDNTLDVMKKPRCGVPDVGEYNVFP RTLKWSKMNLTYRIVNYTPDMTHSEVEKAFKKAFKVWSDVTPLNFTRLHDGIA DIMISFGIKEHGDFYPFDGPSGLLAHAFPPGPNYGGDAHFDDDETWTSSSKGYNL FLVAAHEFGHSLGLDHSKDPGALMFPIYTYTGKSHFMLPDDDVQGIQSLYGPGD EDPNPKHPKTPDKCDPSLSLDAITSLRGETMIFKDRFFWRLHPQQVDAELFLTKSF WPELPNRIDAAYEHPSHDLIFIFRGRKFWALNGYDILEGYPKKISELGLPKEVKKI SAAVHFEDTGKTLLFSGNQVWRYDDTNHIMDKDYPRLIEEDFPGIGDKVDAVYE KNGYIYFFNGPIQFEYSIWSNRIVRVMPANSILWCSEQ ID NO: 4 MMP1MHSFPPLLLLLFWGVVSHSFPATLETQEQDVDLVQKYLEKYYNLKNDGRQVEK RRNSGPVVEKLKQMQEFFGLKVTGKPDAETLKVMKQPRCGVPDVAQFVLTEGN PRWEQTHLTYRIENYTPDLPRADVDHAIEKAFQLWSNVTPLTFTKVSEGQADIMI SFVRGDHRDNSPFDGPGGNLAHAFQPGPGIGGDAHFDEDERWTNNFREYNLHR VAAHELGHSLGLSHSTDIGALMYPSYTFSGDVQLAQDDIDGIQAIYGRSQNPVQP IGPQTPKACDSKLTFDAITTIRGEVMFFKDRFYMRTNPFYPEVELNFISVFWPQLP NGLEAAYEFADRDEVRFFKGNKYWAVQGQNVLHGYPKDIYSSFGFPRTVKHID AALSEENTGKTYFFVANKYWRYDEYKRSMDPGYPKMIAHDFPGIGHKVDAVFM KDGFFYFFHGTRQYKFDPKTKRILTLQKANSWFNCRKNSEQ ID NO: 5 MMP10MMHLAFLVLLCLPVCSAYPLSGAAKEEDSNKDLAQQYLEKYYNLEKDVKQFRR KDSNLIVKKIQGMQKFLGLEVTGKLDTDTLEVMRKPRCGVPDVGHFSSFPGMPKWRKTHLTYRIVNYTPDLPRDAVDSAIEKALKVWEEVTPLTFSRLYEGEADIMISF AVKEHGDFYSFDGPGHSLAHAYPPGPGLYGDIHFDDDEKWTEDASGTNLFLVA AHELGHSLGLFHSANTEALMYPLYNSFTELAQFRLSQDDVNGIQSLYGPPPASTE EPLVPTKSVPSGSEMPAKCDPALSFDAISTLRGEYLFFKDRYFWRRSHWNPEPEF HLISAFWPSLPSYLDAAYEVNSRDTVFIFKGNEFWAIRGNEVQAGYPRGIHTLGF PPTIRKIDAAVSDKEKKKTYFFAADKYWRFDENSQSMEQGFPRLIADDFPGVEPK VDAVLQAFGFFYFFSGSSQFEFDPNARMVTHILKSNSWLHCSEQ ID NO: 6 MMP20MKVLPASGLAVFLIMALKFSTAAPSLVAASPRTWRNNYRLAQAYLDKYYTNKE GHQIGEMVARGSNSMIRKIKELQAFFGLQVTGKLDQTTMNVIKKPRCGVPDVAN YRLFPGEPKWKKNTLTYRISKYTPSMSSVEVDKAVEMALQAWSSAVPLSFVRIN SGEADIMISFENGDHGDSYPFDGPRGTLAHAFAPGEGLGGDTHFDNAEKWTMGT NGFNLFTVAAHEFGHALGLAHSTDPSALMYPTYKYKNPYGFHLPKDDVKGIQA LYGPRKVFLGKPTLPHAPHHKPSIPDLCDSSSSFDAVTMLGKELLLFKDRIFWRR QVHLRTGIRPSTITSSFPQLMSNVDAAYEVAERGTAYFFKGPHYWITRGFQMQGP PRTIYDFGFPRHVQQIDAAVYLREPQKTLFFVGDEYYSYDERKRKMEKDYPKNT EEEFSGVNGQIDAAVELNGYIYFFSGPKTYKYDTEKEDVVSVVKSSSWSEQ ID NO: 7 MMP12MKFLLILLLQATASGALPLNSSTSLEKNNVLFGERYLEKFYGLEINKLPVTKMKY SGNLMKEKIQEMQHFLGLKVTGQLDTSTLEMMHAPRCGVPDVHHFREMPGGPV WRKHYITYRINNYTPDMNREDVDYAIRKAFQVWSNVTPLKFSKINTGMADILVV FARGAHGDFHAFDGKGGILAHAFGPGSGIGGDAHFDEDEFWTTHSGGTNLFLTA VHEIGHSLGLGHSSDPKAVMFPTYKYVDINTFRLSADDIRGIQSLYGDPKENQRL PNPDNSEPALCDPNLSFDAVTTVGNKIFFFKDRFFWLKVSERPKTSVNLISSLWPT LPSGIEAAYEIEARNQVFLFKDDKYWLISNLRPEPNYPKSIHSFGFPNFVKKIDAA VFNPRFYRTYFFVDNQYWRYDERRQMMDPGYPKLITKNFQGIGPKIDAVFYSKN KYYYFFQGSNQFEYDFLLQRITKTLKSNSWFGCSEQ ID NO: 8 MMP3 MKSLPILLLLCVAVCSAYPLDGAARGEDTSMNLVQKYLENYYDLKKDVKQFVR RKDSGPVVKKIREMQKFLGLEVTGKLDSDTLEVMRKPRCGVPDVGHFRTFPGIP KWRKTHLTYRIVNYTPDLPKDAVDSAVEKALKVWEEVTPLTFSRLYEGEADIMI SFAVREHGDFYPFDGPGNVLAHAYAPGPGINGDAHFDDDEQWTKDTTGTNLFL VAAHEIGHSLGLFHSANTEALMYPLYHSLTDLTRFRLSQDDINGIQSLYGPPPDSP ETPLVPTEPVPPEPGTPANCDPALSFDAVSTLRGEILIFKDRHFWRKSLRKLEPELH LISSFWPSLPSGVDAAYEVTSKDLVFIFKGNQFWAIRGNEVRAGYPRGIHTLGFPP TVRKIDAAISDKEKNKTYFFVEDKYWRFDEKRNSMEPGFPKQIAEDFPGIDSKID AVFEEFGFFYFFTGSSQLEFDPNAKKVTHTLKSNSWLNC SEQ ID NO: 9 MT1-MMP NPMATRIMETALLPRTEINASEPREPRPRTEINHMSAPIENSMSPAPRPPRCLLLPL LTLGTALASLGSAQSSSFSPEAWLQQYGYLPPGDLRTHTQRSPQSLSAAIAAMQK FYGLQVTGKADADTMKAMRRPRCGVPDKFGAEIKANVRRKRYAIQGLKWQHN EITFCIQNYTPKVGEYATYEAIRKAFRVWESATPLRFREVPYAYIREGHEKQADI MIFFAEGFHGDSTPFDGEGGFLAHAYFPGPNIGGDTHFDSAEPWTVRNEDLNGN DIFLVAVHELGHALGLEHSSDPSAIMAPFYQWMDTENFVLPDDDRRGIQQLYGG ESGFPTKMPPQPRTTSRPSVPDKPKNPTYGPNICDGNFDTVAMLRGEMFVFKER WFWRVRNNQVMDGYPMPIGQFWRGLPASINTAYERKDGKFVFFKGDKHWVFD EASLEPGYPKHIKELGRGLPTDKIDAALFWMPNGKTYFFRGNKYYRFNEELRAV DSEYPKNIKVWEGIPESPRGSFMGSDEVFTYFYKGNKYWKFNNQKLKVEPGYPK SALRDWMGCPSGGRPDEGTEEETEVIIIEVDEEGGGAVSAAAVVLPVLLLLLVLA VGLAVFFFRRHGTPRRLLYCQRSLLDKV 6A5 VHSEQ ID NO: 10 FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4EVQLQQSGAELVKPGASVKLSCTASGFNIKDTYINWVKQRPEQGLEWIGRIDPAN GNTKYDPKFQGKATITADTSSNTAYLQLSSLTSEDTAVYYCAREVLGPAWFAYW GQGTLVTVSA6A5 VH – FR1 SEQ ID NO: 11EVQLQQSGAELVKPGASVKLSCTASGFNIK6A5 VH – CDR1 SEQ ID NO: 12DTYIN6A5 VH – FR2 SEQ ID NO: 13WVKQRPEQGLEWIG6A5 VH – CDR2 SEQ ID NO: 14RIDPANGNTKYDPKFQG6A5 VH – FR3 SEQ ID NO: 15KATITADTSSNTAYLQLSSLTSEDTAVYYCAR6A5 VH – CDR3 SEQ ID NO: 16EVLGPAWFAY6A5 VH – FR4 SEQ ID NO: 17WGQGTLVTVSA6A5 CH1 – constant region SEQ ID NO: 18AKTTAPSVYPLAPVCGDTTGSSVTLGCLVKGYFPEPVTLTWNSGSLSSGVHTFPA VLQSDLYTLSSSVTVTSSTWPSQSITCNVAHPASSTKVDKKIEPRGPTIKPCPPCKC PAPNLLGGPSVFIFPPKIKDVLMISLSPIVTCVVVDVSEDDPDVQISWFVNNVEVH TAQTQTHREDYNSTLRVVSALPIQHQDWMSGKEFKCKVNNKDLPAPIERTISKP KGSVRAPQVYVLPPPEEEMTKKQVTLTCMVTDFMPEDIYVEWTNNGKTELNYK NTEPVLDSDGSYFMYSKLRVEKKNWVERNSYSCSVVHEGLHNHHTTKSFSRTPG K6A5 VL SEQ ID NO: 19FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4DIVLTQSPASLAVSLGQRATISCRASKSVSTSGYSYMHWYQQKPGQPPKLLIYLA SNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPFTFGSGTKLEIK6A5 VL – FR1 SEQ ID NO: 20DIVLTQSPASLAVSLGQRATISC6A5 VL – CDR1 SEQ ID NO: 21RASKSVSTSGYSYMH6A5 VL – FR2 SEQ ID NO: 22WYQQKPGQP PKLLIY6A5 VL – CDR2 SEQ ID NO: 23LASNLES6A5 VL – FR3 SEQ ID NO: 24GVPARFSGSGSGTDFTLNIHPVEEEDAATYYC6A5 VL – CDR3 SEQ ID NO: 25QHSRELPFT6A5 VL – FR4 SEQ ID NO: 26TFGSGTKLEIK6A5 CL – Constant region SEQ ID NO: 27RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNS WTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC11D2 VH SEQ ID NO: 28FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSWVRQTPEKRLEWVATISSG GSYTYYPDSVKGRFTISRDNAKNTLYLQMSSLRSEDTAMYYCARRPDGYFDVW GAGTTVTVSS11D2 VH – FR1 SEQ ID NO: 29EVMLVESGGGLVKPGGSLKLSCAASGFTFS11D2 VH – CDR1 SEQ ID NO: 30SYAMS11D2 VH – FR2 SEQ ID NO: 31WVRQTPEKRLEWVA11D2 VH – CDR2 SEQ ID NO: 32TISSGGSYTYYPDSVKG11D2 VH – FR3 SEQ ID NO: 33RFTISRDNAKNTLYLQMSSLRSEDTAMYYCAR11D2 VH – CDR3 SEQ ID NO: 34RPDGYFDV11D2 VH – FR4 SEQ ID NO: 35WGAGTTVTVSS11D2 CH1 – Constant region SEQ ID NO: 36AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPA LLQSGLYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPC KECHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVQISWFV NNVEVHTAQTQTHREDYNSTIRVVSTLPIQHQDWMSGKEFKCKVNNKDLPSPIE RTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEE NYKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSCNVRHEGLKNYYLKKTISR SPGK11D2 VL SEQ ID NO: 37FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 QIVLTQSPAIMSASLGERVTMTCTASSSVSSSYLHWYQQKPGSSPKLWIYSTSNL ASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCHQYHRSPLTFGAGTKLELK11D2 VL – FR1 SEQ ID NO: 38QIVLTQSPAIMSASLGERVTMTC11D2 VL – CDR1 SEQ ID NO: 39TASSSVSSSYLH11D2 VL – FR2 SEQ ID NO: 40WYQQKPGSSPKLWIY11D2 VL – CDR2 SEQ ID NO: 41STSNLAS11D2 VL – FR3 SEQ ID NO: 42GVPARFSGSGSGTSYSLTISSMEAEDAATYYC11D2 VL – CDR3 SEQ ID NO: 43HQYHRSPLT11D2 CL – FR4 SEQ ID NO: 44FGAGTKLELK11D2 CL – Constant region SEQ ID NO: 45RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNS WTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC15B9 VH SEQ ID NO: 46FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 EVRLQQSGPELVKPGASVKISCKASGYSFTGYYMHWVKQSHVKSLEWIGRINPY NGATRYNQNFKDKASLTVDKSSSTAYMELHSLTSEDSAVYYCAREERGYDARF DYWGQGTTLTVSS15B9 VH – FR1 SEQ ID NO: 47EVRLQQSGPELVKPGASVKISCKASGYSFT15B9 VH – CDR1 SEQ ID NO: 48GYYMH15B9 VH – FR2 SEQ ID NO: 49WVKQSHVKSLEWIG15B9 VH – CDR2 SEQ ID NO: 50RINPYNGATRYNQNFKD15B9 VH – FR3 SEQ ID NO: 51KASLTVDKSSSTAYMELHSLTSEDSAVYYCAR15B9 VH – CDR3 SEQ ID NO: 52EERGYDARFDY15B9 VH – FR4 SEQ ID NO: 53WGQGTTLTVSS15B9 CH1 – Constant Region SEQ ID NO: 54AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPA LLQSGLYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPC KECHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVQISWFV NNVEVHTAQTQTHREDYNSTIRVVSTLPIQHQDWMSGKEFKCKVNNKDLPSPIE RTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEE NYKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSCNVRHEGLKNYYLKKTISR SPGK15B9 VL SEQ ID NO: 55FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 DIVLTQSPASLAVSLGQRATISYRASKSVSTSGYSYMHWNQQKPGQPPRLLIYLV SNLESGVPARFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPLTFGAGTKLEL KRADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLN SWTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC15B9 VL – FR1 SEQ ID NO: 56DIVLTQSPASLAVSLGQRATISY15B9 VL – CDR1 SEQ ID NO: 57RASKSVSTSGYSYMH15B9 VL – FR2 SEQ ID NO: 58WNQQKPGQPPRLLIY15B9 VL – CDR2 SEQ ID NO: 59LVSNLES15B9 VL – FR3 SEQ ID NO: 60GVPARFSGSGSGTDFTLNIHPVEEEDAATYYC15B9 VL – CDR3 SEQ ID NO: 61QHSRELPLT15B9 VL – FR4 SEQ ID NO: 62FGAGTKLELK15B9 CL – Constant region SEQ ID NO: 63RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNS WTDQDSKDSTYSMSS TLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC12E2 VH SEQ ID NO: 64FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4- EVMLVESGGGLVKPGGSLKLSCAASGFTFSSYAMSTISSGGSYTYYPDSVKGTIS SGGSYTYYPDSVKGRFTISRDNAKNTLYLQMSSLRSEDTAMYYCARRPDGYFDV WGAGTTVTVSS12E2 VH – FR1 SEQ ID NO: 65EVMLVESGGGLVKPGGSLKLSCAASGFTFS12E2 VH – CDR1 SEQ ID NO: 66SYAMS12E2 VH – FR2 SEQ ID NO: 67WVRQTPEKRLEWVA12E2 VH – CDR2 SEQ ID NO: 68TISSGGSYTYYPDSVKG12E2 VH – FR3 SEQ ID NO: 69RFTISRDNAKNTLYLQMSSLRSEDTAMYYCAR12E2 VH – CDR3 SEQ ID NO: 70RPDGYFDV12E2 VH – FR4 SEQ ID NO: 71WGAGTTVTVSS12E2 CH1 – Constant region SEQ ID NO: 72AKTTPPSVYPLAPGCGDTTGSSVTLGCLVKGYFPESVTVTWNSGSLSSSVHTFPA LLQSGLYTMSSSVTVPSSTWPSQTVTCSVAHPASSTTVDKKLEPSGPISTINPCPPC KECHKCPAPNLEGGPSVFIFPPNIKDVLMISLTPKVTCVVVDVSEDDPDVQISWFVNNVEVHTAQTQTHREDYNSTIRVVSTLPIQHQDWMSGKEFKCKVNNKDLPSPIE RTISKIKGLVRAPQVYILPPPAEQLSRKDVSLTCLVVGFNPGDISVEWTSNGHTEE NYKDTAPVLDSDGSYFIYSKLNMKTSKWEKTDSFSCNVRHEGLKNYYLKKTISR SPGK12E2 VL SEQ ID NO: 73FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4 QIVLTQSPAIMSASLGERVTMTCTASSSVSSSYLHWYQQKPGSSPKLWIYSTSNL ASGVPARFSGSGSGTSYSLTISSMEAEDAATYYCHQYHRSPLTFGAGTKLELK12E2 VL – FR1 SEQ ID NO: 74QIVLTQSPAIMSASLGERVTMTC12E2 VL – CDR1 SEQ ID NO: 75TASSSVSSSYLH12E2 VL – FR2 SEQ ID NO: 76WYQQKPGSSPKLWIY12E2 VL – CDR2 SEQ ID NO: 77STSNLAS12E2 VL – FR3 SEQ ID NO: 78GVPARFSGSGSGTSYSLTISSMEAEDAATYYC12E2 VL – CDR3 SEQ ID NO: 79HQYHRSPLT12E2 VL – FR4 SEQ ID NO: 80FGAGTKLELK12E2 CL – Constant region SEQ ID NO:81RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNS WTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC2F11 VH SEQ ID NO: 82FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4EVQLQQSGPELVKPGASVKISCKASGYSFTGYYMHWVKQSHVKSLEWIGRINPY NGATRYNQNFKDKASLTVDKSSSTAYMELHSLTSEDSAVYYCAREERGYDARF DYWGQGTTLTVSS2F11 VH – FR1 SEQ ID NO:83EVQLQQSGPELVKPGASVKISCKASGYSFT2F11 VH – CDR1 SEQ ID NO: 84GYYMH2F11 VH – FR2 SEQ ID NO: 85WVKQSHVKSLEWIG2F11 VH – CDR2 SEQ ID NO: 86RINPYNGATRYNQNFKD2F11 VH – FR3 SEQ ID NO: 87KASLTVDKSSSTAYMELHSLTSEDSAVYYCAR2F11 VH – CDR3 SEQ ID NO: 88EERGYDARFDY2F11 VH – FR4 SEQ ID NO: 89WGQGTTLTVSS2F11 CH1 – Constant region SEQ ID NO: 90AKTTPPSVYPLAPGSAAQTNSMVTLGCLVKGYFPEPVTVTWNSGSLSSGVHTFP AVLQSDLYTLSSSVTVPSSTWPSETVTCNVAHPASSTKVDKKIVPRDCGCKPCIC TVPEVSSVFIFPPKPKDVLTITLTPKVTCVVVDISKDDPEVQFSWFVDDVEVHTAQ TQPREEQFNSTFRSVSELPIMHQDWLNGKEFKCRVNSAAFPAPIEKTISKTKGRPK APQVYTIPPPKEQMAKDKVSLTCMITDFFPEDITVEWQWNGQPAENYKNTQPIM DTDGSYFVYSKLNVQKSNWEAGNTFTCSVLHEGLHNHHTEKSLSHSPGK2F11 VL SEQ ID NO: 91FR1-CDR1-FR2-CDR2-FR3-CDR3-FR4-Constant regionDIVLTQSPASLAVSLGQRATISCRASKSVSTSVYSYMHWYQQKPGQPPKLLIYLA SNLESGVPGRFSGSGSGTDFTLNIHPVEEEDAATYYCQHSRELPLTFGAGTKLEL K2F11 VL – FR1 SEQ ID NO: 92DIVLTQSPASLAVSLGQRATISC2F11 VL – CDR1 SEQ ID NO: 93RASKSVSTSVYSYMH2F11 VL – FR2 SEQ ID NO: 94WYQQKPGQPPKLLIY2F11 VL – CDR2 SEQ ID NO: 95LASNLES2F11 VL – FR3 SEQ ID NO: 96GVPGRFSGSGSGTDFTLNIHPVEEEDAATYYC2F11 VL – CDR3 SEQ ID NO: 97QHSRELPLT2F11 VL – FR4 SEQ ID NO: 98FGAGTKLELK2F11 CL – Constant region SEQ ID NO: 99RADAAPTVSIFPPSSEQLTSGGASVVCFLNNFYPKDINVKWKIDGSERQNGVLNS WTDQDSKDSTYSMSSTLTLTKDEYERHNSYTCEATHKTSTSPIVKSFNRNEC

Claims

What is claimed is:

1. An antibody comprising:a heavy chain (VH) selected from SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO:46, SEQ ID NO: 64 and SEQ ID NO: 82; anda light chain (VL) selected from SEQ ID NO: 10, SEQ ID NO: 37, SEQ ID NO: 55, SEQ ID NO: 73 and SEQ ID NO: 91.

2. The antibody as recited in claim 1, wherein the heavy chain (VH) is SEQ ID NO:10 and the light chain (VL) is SEQ ID NO: 19.

3. The antibody as recited in claim 1, wherein the heavy chain (VH) is SEQ ID NO:46 and the light chain (VL) is SEQ ID NO: 55.

4. The antibody as recited in claim 1, wherein the heavy chain (VH) is SEQ ID NO:28 and the light chain (VL) is SEQ ID NO: 37.

5. The antibody as recited in claim 1, wherein the heavy chain (VH) is SEQ ID NO:64 and the light chain (VL) is SEQ ID NO: 73.

6. The antibody as recited in claim 1, wherein the heavy chain (VH) is SEQ ID NO:82 and the light chain (VL) is SEQ ID NO: 91.

7. The antibody as recited in any one of claims 1-6, further comprising a labelcovalently conjugated to the antibody.

8. The antibody as recited in claim 7, wherein the label is selected from the groupconsisting of a radiolabel and a fluorophore.

9. The antibody as recited in claim 7, wherein the label is a chelator bound to aradioactive metal ion.

10. A composition of matter comprisingan antibody comprising: a heavy chain (VH) that is at least 80% identical to SEQ ID NO: 10, SEQ ID NO: 28, SEQ ID NO: 46, SEQ ID NO: 64 or SEQ ID NO: 82; a light chain (VL) that is at least 80% identical to SEQ ID NO: 19, SEQ ID NO: 37, SEQ ID NO: 55, SEQ ID NO: 73 and SEQ ID NO: 91; and alabel covalently conjugated to the antibody.

11. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 85% identical to SEQ ID NO: 10 and the light chain (VL) is at least 85%identical to SEQ ID NO: 19.

12. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 90% identical to SEQ ID NO: 10 and the light chain (VL) is at least 85% identical to SEQ ID NO: 19.

13. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 95% identical to SEQ ID NO: 10 and the light chain (VL) is at least 85% identical to SEQ ID NO: 19.

14. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 85% identical to SEQ ID NO: 46 and the light chain (VL) is at least 85%identical to SEQ ID NO: 55.

15. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 90% identical to SEQ ID NO: 46 and the light chain (VL) is at least 85%identical to SEQ ID NO: 55.

16. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 95% identical to SEQ ID NO: 46 and the light chain (VL) is at least 85%identical to SEQ ID NO: 55.

17. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 85% identical to SEQ ID NO: 28 and the light chain (VL) is at least 85%identical to SEQ ID NO: 37.

18. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 90% identical to SEQ ID NO: 28 and the light chain (VL) is at least 85%identical to SEQ ID NO: 37.

19. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 95% identical to SEQ ID NO: 28 and the light chain (VL) is at least 85%identical to SEQ ID NO: 37.

20. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 85% identical to SEQ ID NO: 64 and the light chain (VL) is at least 85%identical to SEQ ID NO: 73.

21. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 90% identical to SEQ ID NO: 64 and the light chain (VL) is at least 85%identical to SEQ ID NO: 73.

22. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 95% identical to SEQ ID NO: 64 and the light chain (VL) is at least 85%identical to SEQ ID NO: 73.

23. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 85% identical to SEQ ID NO: 82 and the light chain (VL) is at least 85% identical to SEQ ID NO: 91.

24. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 90% identical to SEQ ID NO: 82 and the light chain (VL) is at least 85% identical to SEQ ID NO: 91.

25. The composition of matter as recited in claim 10, wherein the heavy chain (VH) isat least 95% identical to SEQ ID NO: 82 and the light chain (VL) is at least 85% identical to SEQ ID NO: 91.

26. The composition of matter as recited in any one of claims 10-25, wherein thelabel is selected from the group consisting of a radiolabel and a fluorophore.

27. A method for imaging a patient, the method comprising:administering to a human patient the composition of matter as recited in anyone of claims 10-25; andimaging the label within the human patient.

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