Custom Anti-Tn Antibody Development for Tumor Biomarker Discovery

The reliable and early detection of cancer remains one of the most significant challenges in modern medicine. Researchers are continually seeking specific, sensitive, and reliable biomarkers—molecular signals that can indicate the presence of disease, predict its progression, and inform treatment decisions. In this pursuit, a class of molecules known as glycans (sugars) has emerged as a critical source of next-generation biomarkers. At Creative Biolabs, we offer researchers a comprehensive portfolio of custom anti-TACA antibody development services, including high-specificity anti-Tn (CD175) antibody products, enabling the creation of novel tools against this target. A high-quality anti-Tn antibody is an indispensable tool for tumor biomarker discovery, and our platform is dedicated to providing it. The reliable and early detection of cancer remains one of the greatest challenges in modern medicine. Researchers are continually seeking specific, sensitive, and reliable biomarkers that can indicate the presence of disease, predict its progression, and inform treatment decisions. In this pursuit, the Tn antigen has emerged as a critical target.

Background

The Tn Antigen (CD175): A Premier Pan-Carcinoma Marker

While the entire TF family is essential, the Tn antigen holds a unique position for biomarker discovery.

• Low Normal Expression: In healthy adult tissues, T-synthase is highly efficient. As soon as the Tn antigen is formed, it is immediately converted to the T antigen. Therefore, the Tn antigen is virtually absent from the surface of normal cells. Its expression is "cryptic," or hidden.
• High Cancer-Specific Expression: In many cancers (including breast, colon, pancreas, lung, and bladder carcinomas), the gene for T-synthase or its chaperone Cosmc is mutated or silenced. This is a catastrophic failure for the cell. Without a functional T-synthase, the assembly line stops at the very first step. The cell can no longer convert Tn to T.

The result is a massive accumulation and surface expression of the Tn antigen. This simple, single-sugar marker goes from being invisible to being one of the most dominant features of the cancer cell's glycocalyx. This expression pattern makes the Tn antigen (CD175) an ideal pan-carcinoma marker—a biomarker found across a wide variety of different cancer types. Its presence is a clear indication of malignancy, making it a powerful target for early diagnosis and prognosis.

The Challenge: Why High-Quality Anti-Tn Antibodies (CD175 Antibodies) Are Rare

If the Tn antigen is such a great target, why isn't it more widely used? The answer lies in the extreme technical difficulty of producing a good Tn antibody.

• Poor Immunogenicity: The Tn antigen is just a single sugar (GalNAc). This tiny molecule is a very poor immunogen, meaning it is difficult for the immune system (such as in a mouse or rabbit) to recognize it and produce an antibody against it.
• The Specificity Problem: The immune system must generate an antibody that only binds to Tn. It must not cross-react with:
  • T antigen (Tn with one extra sugar)
  • sTn antigen (Tn with a sialic acid cap, know more about Custom Anti-sTn Antibody)
  • Blood Group A antigen (which also contains a GalNAc residue)
• Cryptic Nature: Because Tn is technically a "self" molecule, the immune system may have tolerance mechanisms that prevent a strong response.

Our Custom Anti-Tn Antibody Development Workflow

Developing CD175 antibodies that are both highly sensitive and highly specific requires a specialized platform and deep expertise in glycobiology. This is precisely what our service provides. We offer a start-to-finish, fully customized service to develop the specific anti-Tn antibody your research requires. Our process is transparent, collaborative, and customizable.

Phase 1: Strategic Antigen Design and Synthesis

This is the most critical phase for targeting a glycan. A poor antigen will result in a useless antibody. We do not use "off-the-shelf" antigens; we design one specifically for your goal. We discuss these options with you to select the best strategy for your budget and application.

• Carrier Protein Conjugation: To overcome poor immunogenicity, the Tn antigen must be attached to a large, immunogenic carrier protein, such as Keyhole Limpet Hemocyanin (KLH) or Bovine Serum Albumin (BSA).
• Glycopeptide Synthesis: For the highest specificity, we strongly recommend using a synthetic glycopeptide. Instead of just the Tn sugar, we synthesize a short piece of a protein known to carry Tn in cancer with the Tn antigen attached at the correct serine or threonine. This presents the Tn antigen to the immune system in its most natural context, leading to superior antibodies.

Phase 2: Immunization and Antibody Generation

We employ optimized immunization protocols to break immune tolerance and generate a strong response against the glycan target. We offer multiple platforms for antibody generation:

• Hyperdoma™ Platform: The gold standard for generating robust, consistent CD175 antibodies. This involves immunizing mice and fusing their B-cells to create immortal hybridoma cell lines that continuously produce a single, specific antibody.
• Phage Display Library Screening: We can screen vast libraries of antibody fragments to screen for binders against the Tn antigen. This approach is often faster and enables the development of fully human antibodies.

Phase 3: Screening and Specificity Profiling

This phase separates a mediocre antibody from a research-grade tool. All clones are subjected to a multi-stage screening process to ensure absolute specificity. We use a "positive/negative" screening panel. Your Tn antigen antibody candidates must pass the following screening assays, and only the clones that pass this rigorous specificity check are advanced to the next stage.

Phase 4: Antibody Production, Purification, and Validation

Once you select your lead candidate, we scale up.

• Production: We produce the antibody in the quantity you need, from milligrams to grams.
• Purification: We use advanced chromatography to purify the antibody to >95% purity.
• Validation: We validate the final antibody in the application you intend to use. We can provide validation data for common applications, such as ELISA, WB, IHC, FC, and so on.

Applications for Your Custom Anti-Tn Antibody

A highly specific anti-Tn antibody (CD175 antibody) is a powerful tool for biomarker discovery and validation.

• Immunohistochemistry (IHC): The primary application. Use your antibody to stain tumor tissue sections to visualize Tn expression, assess its prevalence in different tumor grades, and correlate its presence with clinical outcomes.
• ELISA Development: Design sensitive sandwich or competitive ELISAs to detect soluble Tn-bearing proteins in patient serum or plasma, opening avenues for non-invasive liquid biopsy research.
• Flow Cytometry: Identify and isolate populations of circulating tumor cells based on their surface Tn expression.
• Fundamental Research: Probe the basic biology of Tn. Use your antibody to track its expression, study the effects of T-synthase knockdown, and map its location within the cell.
• More applications upon request

Ready-to-Use Anti-Tn Antibody Products

In addition to our bespoke custom development services, Creative Biolabs maintains a comprehensive catalog of ready-to-use anti-Tn antibody products. These reagents are ideal for researchers who require a validated, high-quality antibody for standard applications without the timeline of a full custom project. Our portfolio of CD175 antibodies has been developed and rigorously tested by our in-house glycobiology experts. Each product is validated for high specificity, ensuring minimal cross-reactivity with related T, sTn, or Blood Group A antigens, allowing you to be confident in your data. We invite you to explore our full selection of anti-Tn (CD175) antibodies, including but not limited to:

The Tn antigen represents a massive, untapped opportunity in cancer diagnostics and biomarker research. But accessing this target requires a tool that is precise, specific, and reliable. Standard, off-the-shelf antibodies often fail to meet the specificity needed for conclusive research. Do not let a poor-quality antibody compromise your data. Partner with the experts in glycobiology. We will collaborate with you to understand your research goals and deliver a custom antibody that is specifically designed for the Tn antigen. Contact us today to discuss your project with one of our glycan antibody specialists.

Published Data

A recent preclinical study validates the Tn antigen as a premier target for antibody-based therapeutics. This study demonstrated this by constructing an immunotoxin, SM3GRNLY, which links an anti-Tn scFv (from the SM3 mAb) to the lytic protein granulysin. This anti-Tn immunotoxin retained its high affinity for the MUC1-Tn antigen, with a K_D measured at 0.149 µM by Surface Plasmon Resonance (SPR). In cell-based assays using flow cytometry, the SM3GRNLY construct demonstrated highly specific binding. It successfully bound to the surface of known Tn-positive tumor cell lines (including CAPAN-2, PANC-1, H929, and Jurkat) while showing no binding to Tn-negative control cells (such as MDA-MB-231 and MIA-PACA-2). This targeted binding translated directly to enhanced function. The immunotoxin induced significantly more cell death in Tn-positive cells compared to granulysin alone. The most compelling results came from an in vivo xenograft model of human pancreatic cancer. Systemic treatment with SM3GRNLY reduced tumor volume by 42%, while granulysin alone had no therapeutic effect. Histological analysis confirmed the immunotoxin-induced apoptosis within the tumor tissue.

Fig.1 Specific binding of anti-Tn immunotoxins to tumor cell lines.¹

Reference:

1. Guerrero-Ochoa, Patricia, et al. "Preclinical studies of granulysin-based anti-MUC1-Tn immunotoxins as a new antitumoral treatment." Biomedicines 10.6 (2022): 1223. Distributed under Open Access license CC BY 4.0, without modification. https://doi.org/10.3390/biomedicines10061223

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