title: A monoclonal antibody against PMEL.
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.
authors.affiliation: 1 Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Science, Northwest University , Xi'an, China .
authors.forename: Fangyuan
authors.initials: F
authors.lastname: Shi
authors.forename: Zhenjie
authors.initials: Z
authors.lastname: Xu
authors.forename: Hongdong
authors.initials: H
authors.lastname: Chen
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Jihong
authors.initials: J
authors.lastname: Cui
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Xin
authors.initials: X
authors.lastname: Xie
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D019556:COS Cells; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D003429:Cross Reactions / Q000276:immunology; D006801:Humans; D007136:Immunoglobulins / Q000276:immunology; D019169:Jurkat Cells; D008544:Melanocytes / Q000276:immunology; D008545:Melanoma / Q000276:immunology; D020460:Melanosomes / Q000276:immunology; D008562:Membrane Glycoproteins / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C; D058951:gp100 Melanoma Antigen / Q000276:immunology*
chemical_list: D000911:Antibodies, Monoclonal; D007136:Immunoglobulins; D008562:Membrane Glycoproteins; D058951:gp100 Melanoma Antigen