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Six hybridoma cell lines (AP1-AP6) secreting monoclonal antibodies (McAb) against PAI-1 were obtained by fusing the murine myeloma cell line SP2/0 with the spleen cells from Balb/c mouse immunized with recombinant PAI-1 expressed in E. coli. These antibodies were purified by SPA affinity chromatography. All McAbs recognized rPAI-1 and PAI-1 from the human hepatoma cell line HepG2. The titers of ascites were more than 10(6). The antibody-antigen affinity constants (Kaff) for anti-PAI-1 McAb measured by EIA were between 3.45 x 10(7)-1.05 x 10(10) M. AP2 and AP3 McAbs were effective in quenching the activity of PAI-1. Partial quenching of PAI-1 activity was achieved with AP4, AP5 and AP6 McAbs respectively. AP1 McAb had no effect upon PAI-1 activity. Three of the six McAbs (AP1, AP4 and AP5) bound to the PAI-1/t-PA complex, while the others did not. The PAI-1 was purified 51 folds to homogeneity from serum free medium of HepG2 with the recovery rate of 92% by one-step procedure using Sepharose 4B conjugated with anti-PAI-1 McAb (AP1, AP3 and AP4). A sandwich ELISA for the measurement of PAI-1 antigen in human plasma was developed, based on anti-PAI-1 McAb against non-overlapping epitopes. The mean value of plasma PAI-1 for the healthy donors was 24.7 +/- 7.75 ng/ml measured by ELISA.
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
October 2004,
Hybridoma and hybridomics,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
January 2005,
Zhonghua bing li xue za zhi = Chinese journal of pathology,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
February 2002,
Zhonghua bing li xue za zhi = Chinese journal of pathology,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
August 2001,
Hybridoma,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
July 2008,
Sichuan da xue xue bao. Yi xue ban = Journal of Sichuan University. Medical science edition,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
January 1991,
Toxicon : official journal of the International Society on Toxinology,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
June 2009,
Hybridoma (2005),
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
June 2008,
Beijing da xue xue bao. Yi xue ban = Journal of Peking University. Health sciences,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
August 2013,
Monoclonal antibodies in immunodiagnosis and immunotherapy,
H Ding, and
X Wu, and
X Zhou, and
Y Zhu, and
H Song
September 2014,
Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology,
