Anti-CD133 monoclonal antibodies and related compositions and methods
Inventors
Pfister, Thomas D. • Kinders, Robert J. • Navas, Tony
Assignees
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Publication Number
US-10711068-B2
Publication Date
2020-07-14
Expiration Date
2036-03-28
Abstract
Anti-CD133 monoclonal antibodies having advantageous properties; products, compositions and kits comprising the monoclonal antibodies; methods (processes) of making the monoclonal antibodies and related compositions, as well as methods of using the monoclonal antibodies in analytical, diagnostic and therapeutic applications.
Core Innovation
The invention describes novel anti-CD133 monoclonal antibodies characterized by high specificity and the ability to bind both glycosylated and unglycosylated forms of the CD133 protein. These antibodies are designed to recognize a range of CD133 variants and isoforms, regardless of posttranslational modifications such as glycosylation, and are suitable for a variety of analytical, diagnostic, and therapeutic applications.
The problem addressed by the invention arises from deficiencies in existing anti-CD133 monoclonal antibodies, which often recognize only certain glycosylation forms or epitopes of CD133 and are useful only within limited assay ranges. Early monoclonal antibodies may fail to detect CD133 during cell differentiation due to glycosylation changes or may only recognize unglycosylated epitopes, limiting their utility. There is a need for antibodies with reliable recognition of CD133 variants to enable broad application.
The invention provides methods of producing anti-CD133 monoclonal antibodies by immunizing with selected CD133 extracellular domain polypeptides lacking glycosylation sites, generating hybridomas, screening for specificity to domain A or domain B of CD133, and producing recombinant antibodies. The resultant antibodies bind specifically to defined epitopes on CD133, detect multiple glycosylated forms, and can be labeled or modified for diverse uses. Additionally, the invention covers compositions, kits, and processes for using these antibodies in detection and therapeutic contexts.
Claims Coverage
The patent claims cover several inventive features including a specific anti-CD133 monoclonal antibody with defined complementarity determining regions (CDRs), methods of detection, and methods of isolating CD133-positive cells. These features relate to the antibody structure, binding specificity, production methods, and applications.
Anti-CD133 monoclonal antibody with defined CDR sequences
An anti-CD133 monoclonal antibody having an antibody binding site comprising specific CDR-H1, CDR-H2, CDR-H3, CDR-L1, CDR-L2, and CDR-L3 residues as per SEQ ID NO:9 for the heavy chain and SEQ ID NO:11 for the light chain, allowing specific binding to CD133 domain A.
Specific binding to CD133 domain A but not domain B
The monoclonal antibody is capable of specifically binding domain A but not domain B of a CD133 polypeptide, independent of glycosylation state.
Recombinant production of the monoclonal antibody
The monoclonal antibody can be recombinantly produced, including variants with moieties not naturally occurring in antibodies such as labels or tags.
Method of detecting CD133 polypeptide in a sample
A method comprising contacting a sample with the claimed monoclonal antibody under binding conditions, detecting specific binding indicative of CD133 presence, and optionally determining amount of CD133 present.
Method of isolating CD133-positive cells
A method comprising contacting a sample containing cells with the monoclonal antibody to form complexes with CD133-positive cells and isolating these complexes from the sample, applicable to blood or marrow samples.
The claims define novel monoclonal antibodies with precise binding site sequences targeting CD133 domain A, their recombinant production, and their use in methods for detecting CD133 and isolating CD133-positive cells, emphasizing improved specificity and applicability in various biological samples.
Stated Advantages
High specificity of monoclonal antibodies enabling binding to multiple CD133 variants including glycosylated and unglycosylated forms.
Utility in a wide range of analytical and diagnostic assays such as immunofluorescence, Western blotting, ELISA, and flow cytometry.
Usefulness in therapeutic applications including cancer detection, monitoring, and treatment.
Improved performance compared to commercially available anti-CD133 antibodies, with enhanced sensitivity and specificity demonstrated in assays.
Documented Applications
Analytical and diagnostic detection of CD133 in various samples including cells, tissues, blood, and tumor biopsies.
Assessment, detection, and monitoring of cancer and cancer stem cells in subjects, including hematological and solid tumors.
Isolation and removal of CD133-positive cells from blood or marrow samples, potentially for preparative or therapeutic purposes.
Monitoring efficacy and optimizing cancer therapies in patients by measuring CD133 expression levels before, during, and after treatment.
Use as therapeutic agents, including neutralizing antibodies for treating various cancers such as breast cancer, colorectal cancer, glioblastoma, melanoma, lung cancer, ovarian cancer, acute leukemia, and others.
Use in cancer diagnostics through immunofluorescence and Western blotting of tumor samples and xenograft tumors.
Research applications including studies on CD133 expression related to cancer stem cell biology and tumor progression.
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