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Updated: Apr 25, 2026
Generation of Discriminative Human Monoclonal Antibodies from Rare Antigen-specific B Cells Circulating in Blood
Published on: February 6, 2018
A monoclonal antibody against PMEL
Fangyuan Shi1, Zhenjie Xu, Hongdong Chen
11 Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University , Xi'an, China .
Abstract:
PMEL, also known as Pmel17 or gp100, is a melanocyte-specific glycoprotein that is essential for the formation of stage II melanosomes. As it has a highly restricted expression pattern in normal tissues and a transient presence on the cell surface, PMEL is believed to be a potential target for antibody drug conjugate therapy in some pigmentary diseases. The production of a high specificity and high affinity monoclonal antibody against human PMEL was helpful for the antibody drug conjugate therapy study. In the present study, monoclonal antibodies (MAbs) against PMEL were obtained by immunizing BALB/c mice with the recombinant PMEL-GST fusion protein. Three mAbs (A3F, G11B, and J7E) with a titer of 1:6000, 1:10,000, and 1:3000, respectively, were obtained. Immunoglobulin subclass assay revealed that A3F was IgG2b, G11B was IgG1, and J7E was IgG2a. Specificity analysis by Western blotting demonstrated that A3F and J7E cross-reacted with GPNMB or LAMP; however, G11B reacted with PMEL only. Immunohistochemistry experiments showed that G11B could bind human PMEL antigen in normal skin. Flow cytometry assay demonstrated that G11B could bind to the surface of PMEL positive melanoma cells but not PMEL negative cells. Taken together, these results show that this G11B provides a useful tool for the antibody drug conjugate therapy study in some pigmentary diseases.
Insights
Researchers developed a specific monoclonal antibody, G11B, targeting PMEL (preliminary melanoma antigen). This antibody is crucial for advancing antibody drug conjugate therapies for pigmentary diseases.
Area of Science:
- Biochemistry
- Immunology
- Dermatology
Background:
- PMEL (preliminary melanoma antigen), also known as Pmel17 or gp100, is a melanocyte-specific glycoprotein vital for melanosome formation.
- Its restricted expression and transient cell surface presence make PMEL a potential target for antibody drug conjugate (ADC) therapy in pigmentary diseases.
Purpose of the Study:
- To generate high-specificity and high-affinity monoclonal antibodies (mAbs) against human PMEL for ADC therapy research.
- To characterize and validate the specificity and binding capabilities of newly developed anti-PMEL mAbs.
Main Methods:
- Immunization of BALB/c mice with recombinant PMEL-GST fusion protein to produce mAbs.
- Characterization of mAbs using immunoglobulin subclass assay, Western blotting, immunohistochemistry, and flow cytometry.
Main Results:
- Three anti-PMEL mAbs (A3F, G11B, J7E) were successfully generated with high titers.
- Specificity analysis revealed that G11B reacted exclusively with PMEL, while A3F and J7E showed cross-reactivity.
- G11B demonstrated specific binding to human PMEL in normal skin and on PMEL-positive melanoma cells via immunohistochemistry and flow cytometry.
Conclusions:
- The monoclonal antibody G11B exhibits high specificity for the PMEL antigen.
- G11B serves as a valuable tool for further research and development of antibody drug conjugate therapies targeting pigmentary diseases.
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