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Characterization of Two Novel mAbs Recognizing Different Epitopes on CD43
Soseul Kim1, Jeong Won Hong2, Woon-Dong Cho3
1Department of Pathology, College of Medicine, Chungbuk National University, Cheongju 361-763, Korea. ; Research Institute, DiNonA Inc, Iksan 570-912, Korea.
Insights
Researchers developed two new monoclonal antibodies (mAbs), 2C8 and 8E10, for quantitative detection of acute leukemia biomarkers. These mAbs, along with YG5, enable precise measurement of CD43, aiding leukemia diagnosis.
Area of Science:
- Immunology
- Biochemistry
- Hematology
Background:
- CD43 is a potential biomarker for acute leukemia diagnosis.
- Existing qualitative assays for leukemia-specific CD43 lack quantitative capabilities.
- Novel monoclonal antibodies (mAbs) are needed for precise CD43 detection.
Purpose of the Study:
- To develop novel mAbs for quantitative detection of leukemia-specific CD43.
- To characterize mAbs 2C8 and 8E10 for their binding specificities and epitopes on CD43.
- To establish a sandwich ELISA assay for detecting leukemia-specific CD43 using the developed mAbs.
Main Methods:
- Development of two novel mAbs, 2C8 and 8E10, targeting CD43.
- Sandwich ELISA and western blotting were used to characterize the mAbs and confirm antigen specificity.
- Enzyme assays and analysis of normal tissues were performed to investigate epitope characteristics and distribution.
Main Results:
- Two novel mAbs, 2C8 and 8E10, recognizing distinct epitopes on CD43 were successfully developed.
- The mAbs were confirmed to recognize CD43 and demonstrated differential epitope distribution on leukemic cell lines.
- Enzyme assays indicated that the recognized epitopes are sialic acid-dependent, with distinct patterns observed in normal tissues.
Conclusions:
- Monoclonal antibodies 2C8 and 8E10, along with YG5, can be utilized in a sandwich ELISA for quantitative detection of leukemia-specific CD43.
- These developed mAbs offer a promising tool for improving the diagnosis and monitoring of acute leukemia.
- The characterization of distinct and sialic acid-dependent epitopes provides valuable insights into CD43 biology in leukemia.
Abstract:
JL1, a specific epitope on CD43, is a potential biomarker for the diagnosis of acute leukemia. Although qualitative assays for detecting leukemia-specific CD43 exist, there is a need to develop quantitative assays for the same. Here, we developed two novel monoclonal antibodies (mAbs), 2C8 and 8E10, recognizing different epitopes on CD43. These clones are capable of pairing with YG5, another mAb against JL1 epitope, because they were selectively obtained using sandwich ELISA. Antigens recognized by 2C8 and 8E10 were confirmed as CD43 by western blotting using the CD43-hFC recombinant protein. When expression on various leukemic cell lines was investigated, 2C8 and 8E10 displayed a disparity in the distribution of the epitope. Enzyme assays revealed that these mAbs recognized a sialic acid-dependent epitope on CD43. Using normal thymus and lymph node paraffin-embedded tissues, we confirmed a difference in the epitopes recognized by the two mAbs that was predicted based on the maturity of the cells in the tissue. In summary, we developed and characterized two mAbs, 2C8 and 8E10, which can be used with YG5 in a sandwich ELISA for detecting leukemia-specific CD43.
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