Generation and characterization of a monoclonal antibody to the cytoplasmic tail of MUC16

TY - JOUR

T1 - Generation and characterization of a monoclonal antibody to the cytoplasmic tail of MUC16

AU - Gipson, Ilene K

AU - Mandel, Ulla

AU - Menon, Balaraj

AU - Michaud, Sandra

AU - Tisdale, Ann

AU - Campos, Diana

AU - Clausen, Henrik

N1 - © The Author 2017. Published by Oxford University Press. All rights reserved. For permissions, please e-mail: [email protected].

PY - 2017/10/1

Y1 - 2017/10/1

N2 - MUC16 is a large transmembrane mucin expressed on the apical surfaces of the epithelium covering the ocular surface, respiratory system and female reproductive tract. The transmembrane mucin is overexpressed by ovarian carcinomas, it is one of the most frequently used diagnostic markers for the disease and it is considered a promising target for immunotherapeutic intervention. Immunodetection of the mucin has to date been through antibodies that recognize its exceptionally large ectodomain. Similar to other membrane anchored mucins, MUC16 has a short cytoplasmic tail (CT), but studies of the biological relevance of the C-terminal domain of MUC16 has been limited by lack of availability of monoclonal antibodies that recognize the native CT. Here, we report the development of a novel monoclonal antibody to the CT region of the molecule that recognizes native MUC16 and its enzymatically released CT region. The antibody is useful for immunoprecipitation of the released CT domain as demonstrated with the OVCAR3 ovarian cancer cell line and can be used for detailed cytolocalization in cells as well as in frozen sections of ocular surface and uterine epithelium.

AB - MUC16 is a large transmembrane mucin expressed on the apical surfaces of the epithelium covering the ocular surface, respiratory system and female reproductive tract. The transmembrane mucin is overexpressed by ovarian carcinomas, it is one of the most frequently used diagnostic markers for the disease and it is considered a promising target for immunotherapeutic intervention. Immunodetection of the mucin has to date been through antibodies that recognize its exceptionally large ectodomain. Similar to other membrane anchored mucins, MUC16 has a short cytoplasmic tail (CT), but studies of the biological relevance of the C-terminal domain of MUC16 has been limited by lack of availability of monoclonal antibodies that recognize the native CT. Here, we report the development of a novel monoclonal antibody to the CT region of the molecule that recognizes native MUC16 and its enzymatically released CT region. The antibody is useful for immunoprecipitation of the released CT domain as demonstrated with the OVCAR3 ovarian cancer cell line and can be used for detailed cytolocalization in cells as well as in frozen sections of ocular surface and uterine epithelium.

KW - Journal Article

U2 - 10.1093/glycob/cwx054

DO - 10.1093/glycob/cwx054

M3 - Journal article

C2 - 28673046

SN - 0959-6658

VL - 27

SP - 920

EP - 926

JO - Glycobiology

JF - Glycobiology

IS - 10

ER -