[Preparation and characterization of a McAb against tissue factor pathway inhibitor].
OBJECTIVE
To prepare monoclonal antibody (McAb) against tissue factor pathway inhibitor (TFPI).


METHODS
Twice intrasplenic immunization of BALB/c mice with TFPI-binded nitrocellulose paper followed by an intraperitoneal boost with TFPI raised two hybridoma cell lines, 4F8 and 4F4, by hybridoma technique, 4F8 was studied in detail. The mode value of its chromosome is 93. The Ig class of McAb 4F8 was IgG1. The McAb was purified from ascites by a simple procedure of caprylic acid pre cipitation. The purified McAb was conjugated with horseradish peroxidase (HRP) by a modified periodate oxidation procedure.


RESULTS
Immunoblotting indicated that the McAb recognized a single band of TFPI with a molecular weight of 34800. The McAb was also proved to shorten markedly the dilute thromboplastin coagulation time of factor IX-deficient and normal plasma. The results of Sandwich ELISA using the HRP labeled McAb showed that the mean TFPI in normal human plasma is 103.2 +/- 11.5 microg/L, being similar to that measured with TFPI ELISA kit from America Diagnostica Inc. (98.4 +/- 10.3 microg/L) (r=0.92).


CONCLUSIONS
McAb 4F8 may be a useful reagent for TFPI research.
Yang
Y
Y
Department of Physiology, Hunan Medical University, Changsha 410078.
He
X
X
Li
J
J
He
S
S
D000818:Animals; D000911:Antibodies, Monoclonal / Q000032:analysis / Q000276:immunology* / Q000378:metabolism; D000918:Antibody Specificity; D015153:Blotting, Western; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D006825:Hybridomas / Q000166:cytology; D008074:Lipoproteins / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C
D000911:Antibodies, Monoclonal; D008074:Lipoproteins; C051928:lipoprotein-associated coagulation inhibitor