Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuE...Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuEPO was covalently coupled with bovine serum albumin （BSA） and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked irmnunosorbent assay （ELISA）, SDS-PAGE and Western blot. Results The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1x10s L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work. Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.展开更多
Opioid addiction is one of the top challenges for society, particularly in China. To fight it, the key is to reveal its underlying mechanisms. Among the strategies to overcome the mental damage caused by opioids, inve...Opioid addiction is one of the top challenges for society, particularly in China. To fight it, the key is to reveal its underlying mechanisms. Among the strategies to overcome the mental damage caused by opioids, investigating native anti-opioid peptides derived from mammalian （including human） brains is an important option because of safety concerns. In 1983, diazepam-binding inhibitor （DBI）, a 10-kDa peptide, was first derived from rat brains （Guidotti et al., 1983）. After repeated treatment with morphine, the DBI level is enhanced in rodent brains （Katsura et al., 1998; Shibasaki et al., 2006）.展开更多
基金This work was supported by the National Natural Science Foundation of China (No.20675006).
文摘Objective To produce specific monoclonal antibody （mAb） against recombinant human erythropoietin （rHuEPO） for development of highly efficient methods for erythropoietin detection in biological fluids. Methods rHuEPO was covalently coupled with bovine serum albumin （BSA） and the conjugate was used to immunize mice to produce specific mAb against rHuEPO based on hybridoma technology. The obtained F3-mAb was characterized by enzyme-linked irmnunosorbent assay （ELISA）, SDS-PAGE and Western blot. Results The isotype of F3-mAb was found to be IgM with an affinity constant of 2.1x10s L/mol. The competitive ELISA using the obtained IgM showed a broader linear range and lower detection limit compared with previous work. Conclusions The modification of rHuEPO was proved to be successful in generating required specific mAb with high avidity to rHuEPO.
基金ACKNOWLEDGEMENTS We thank Dr. lain C Bruce for helpful comments on the manuscript. 77 and ZL were partially supported by grants from the National Basic Research Program （973 Program） （No. 2012CB518006）, and the National Natural Science Foundation of China （Grant Nos. 31228010, 31171026, 31100597, 31327901, 31221002, 31330024, 31400708, 31670843, 31521062, and SQ2011SF11B01041）. Yu-Zhen Chen, Xiao-Cun Li, Zhen-Quan Guo, Li Zhou, Zhuan Zhou, Song-vve no conflict of interest. This article does not contain any studies with human subjects performed by the any of the authors. All institutional and national guidelines for the care and use of laboratory animals were followed.
文摘Opioid addiction is one of the top challenges for society, particularly in China. To fight it, the key is to reveal its underlying mechanisms. Among the strategies to overcome the mental damage caused by opioids, investigating native anti-opioid peptides derived from mammalian （including human） brains is an important option because of safety concerns. In 1983, diazepam-binding inhibitor （DBI）, a 10-kDa peptide, was first derived from rat brains （Guidotti et al., 1983）. After repeated treatment with morphine, the DBI level is enhanced in rodent brains （Katsura et al., 1998; Shibasaki et al., 2006）.
