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Updated: May 10, 2026
Analyzing Tumor and Tissue Distribution of Target Antigen Specific Therapeutic Antibody
Published on: May 16, 2020
Renewed interest in basic and applied research involving monoclonal antibodies against an oncofetal Tn-antigen
1Department of Applied Biochemistry, Tokai University School of Engineering, Hiratsuka, Kanagawa, Japan. yokoyamaguchi@tokai-u.jp
Abstract:
Tn-antigen (GalNAcα-Ser/Thr) is one of the most common aberrations associated with cancer progression and metastasis, and thus is an excellent target for development of cancer diagnostics and therapeutics. MLS128 monoclonal antibody (mAb), derived from a mouse immunized with human colon carcinoma cells, was reported to bind to two or three consecutive Tn-antigens (Tn2 or Tn3) with one-order higher affinity for Tn3 than for Tn2. Our recent studies demonstrated that MLS128 significantly inhibits breast and colon cancer cell growth. Molecular cloning of the variable regions of heavy (VH) and light (VL) chains revealed that the VH sequence of MLS128 shared 97% nucleotide sequence identity with the VH of 83D4 mAb, derived from breast cancer-immunized mice, which has a similar affinity for Tn2/Tn3. MLS128 single-chain antibodies (scFv) and scFv-Fc were constructed to confirm the affinity for synthetic Tn2/Tn3 peptides. Thermodynamic studies on MLS128 binding to Tn2/Tn3 revealed its unique nature of temperature-dependent binding.
Insights
The Tn-antigen, a cancer biomarker, is targeted by the MLS128 antibody. This antibody shows significant inhibition of breast and colon cancer cell growth and unique temperature-dependent binding properties.
Area of Science:
- Immunology
- Oncology
- Biochemistry
Background:
- The Tn-antigen (GalNAcα-Ser/Thr) is a common cancer-associated aberration linked to tumor progression and metastasis.
- The Tn-antigen serves as a promising target for developing novel cancer diagnostics and therapeutics.
Purpose of the Study:
- To characterize the binding properties and therapeutic potential of the MLS128 monoclonal antibody (mAb) targeting Tn-antigens.
- To investigate the molecular characteristics and affinity of MLS128 for Tn-antigens.
Main Methods:
- Immunization of mice with human colon carcinoma cells to generate MLS128 mAb.
- Molecular cloning of variable regions (VH and VL) of MLS128.
- Construction of MLS128 single-chain antibodies (scFv) and scFv-Fc fragments.
- Affinity confirmation using synthetic Tn2/Tn3 peptides and thermodynamic studies.
Main Results:
- MLS128 mAb binds to Tn2 and Tn3 antigens with higher affinity for Tn3.
- MLS128 demonstrates significant inhibition of breast and colon cancer cell growth in vitro.
- VH sequence of MLS128 shares high identity with 83D4 mAb, another Tn-binding antibody.
- Thermodynamic studies revealed unique temperature-dependent binding of MLS128 to Tn2/Tn3.
Conclusions:
- MLS128 is a potent antibody targeting Tn-antigens, showing significant anti-cancer activity.
- The antibody's unique binding characteristics warrant further investigation for therapeutic and diagnostic applications in cancer.
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