Establishment and preliminary application of indirect sandwich ELISA for MUC1 protein detection in peripheral blood
MA Jichun
LIU Zhonghui
FANG Fang
DOU Rui
ZHAO Xiaoxia
ZHANG Qingyong
CHEN Wenbo
TAI Guixiang
Abstract：To establish and apply an indirect sandwich ELISA kit for MUC1 protein detection in peripheral blood.The MUC1-GST and MUC1-MBP fusion protein were expressed and purified, and then anti-MUCl serum was obtained by immunization of rabbits and rats.In order to further purify rabbit and rat anti-human MUC1 polyclonal antibody, the serum was purified using saturated ammonium sulfate, protein A/G, and anti-GST and anti-MBP antibody absorption.The standard MUC1 peptide was prepared with MUC1-GST dissolved by thrombin.In the indirect sandwich ELISA kit, rabbit anti-human MUC1 antibodies and rat anti-MUC1 antibodies were used as coating an-tibodies and detecting antibodies, respectively.The sensitivity of the indirect sandwich ELISA kit was up to 0.2 ng/ml.It was detected that the levels of MUC1 protein in the peripheral blood serum of 32 cases of breast cancer patients and 20 healthy subjects, which means the positive detection rate of breast cancer patients was 53.1% (17 cases / 32 cases), while 0 in healthy control group.All the results support that the established anti-MUC1 polyclonal antibody indirect sandwich ELISA kit is sensitive and expected to be used in clinical diagnosis.
Keywords：MUC1Anti-MUCl polyclonal antibodyIndirect sandwich ELISAAdenocarcinoma diagnosis
Publication Date：2010-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：5( 49-52,55 )