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Enrichment and Purging of Human Embryonic Stem Cells by Detection of Cell Surface Antigens Using the Monoclonal Antibodies TG30 and GCTM-2
Published on: December 6, 2013
Detection and characterization of 2-E2-specific surface protein in human pluripotent stem cells
Hana Kim1, Hong Seo Choi, Cheorl-Ho Kim
1Insitute of Bioscience, Department of Bioscience and Biotechnology, Sejong University, 98 Gunja-Dong, Gwangjin-Gu, Seoul, Korea.
Hybridoma (2005)
|August 20, 2011
Summary
Researchers identified a novel cell surface protein on human embryonic stem cells (hESCs) using monoclonal antibody 2-E2. This protein is involved in the early differentiation processes of hESCs.
Area of Science:
- Stem cell biology
- Immunology
- Cell surface markers
Background:
- Human embryonic stem cells (hESCs) possess unique cell surface antigens.
- Identifying these antigens is crucial for understanding pluripotency and differentiation.
Purpose of the Study:
- To generate and characterize novel monoclonal antibodies (MAbs) targeting hESC surface antigens.
- To investigate the expression patterns and potential roles of a newly identified antigen (recognized by MAb 2-E2) during hESC differentiation.
Main Methods:
- Generation of MAbs against undifferentiated hESCs using a decoy immunization strategy.
- Immunological assays (e.g., flow cytometry) to determine MAb binding specificity.
- Western blot analysis to identify the molecular weight of the target antigen.
- Analysis of antigen expression during induced differentiation (retinoic acid treatment, embryoid body formation).
Main Results:
- MAb 2-E2 specifically recognized an antigen on human pluripotent stem cells, but not on mouse pluripotent stem cells or fibroblasts.
- The 2-E2 antigen was also detected on some human differentiated cells, including monocytes and fibroblasts.
- Antigen expression decreased upon retinoic acid-induced differentiation but showed a dynamic change during embryoid body formation.
- The 2-E2 antigen corresponds to approximately 68 and 27 kDa surface proteins.
Conclusions:
- MAb 2-E2 identifies a novel cell surface protein expressed on human pluripotent stem cells.
- This protein's expression dynamics suggest a role in the early stages of hESC differentiation.
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