Transgenic Expression of a Variant of Human Tissue-Type Plasminogen Activator in Goat Milk: Purification and Characterization of the Recombinant Enzyme

1991 ◽  
Vol 9 (9) ◽  
pp. 839-843 ◽  
Author(s):  
Julie Denman ◽  
Michael Hayes ◽  
Christine O'Day ◽  
Timothy Edmunds ◽  
Catherine Bartlett ◽  
...  
Keyword(s):  
Plasminogen Activator ◽  
Human Tissue ◽  
Goat Milk ◽  
Recombinant Enzyme ◽  
Tissue Type ◽  
Purification And Characterization ◽  
Transgenic Expression ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator

Purification and Characterization of Tissue-Type Plasminogen Activator in Maxillary Mucosa with Chronic Inflammation

1985 ◽  
Vol 54 (02) ◽  
pp. 485-489 ◽  
Author(s):  
Yukiyoshi Hamaguchi ◽  
Masuichi Ohi ◽  
Yasuo Sakakura ◽  
Yasuro Miyoshi
Keyword(s):  
Plasminogen Activator ◽  
Chronic Inflammation ◽  
Gel Filtration ◽  
Potassium Acetate ◽  
Tissue Type ◽  
Purification And Characterization ◽  
Tissue Type Plasminogen Activator ◽  
Urokinase Inhibitor ◽  
Type Plasminogen Activator

SummaryTissue-type plasminogen activator (TPA) was purified from maxillary mucosa with chronic inflammation and compared with urokinase. Purification procedure consisted of the extraction from delipidated mucosa with 0.3M potassium acetate buffer (pH 4.2), 66% saturation of ammonium sulfate, zinc chelate-Sepharose, concanavalin A-Sepharose and Sephadex G-100 gel filtration chromatographies.The molecular weight of the TPA was approximately 58,000 ± 3,000. Its activity was enhanced in the presence of fibrin and was quenched by placental urokinase inhibitor, but not quenched by anti-urokinase antibody. The TPA made no precipitin line against anti-urokinase antibody, while urokinase did.All these findings indicate that the TPA in maxillary mucosa with chronic inflammation is immunologically dissimilar to urokinase and in its affinity for fibrin.

Characterization of Two Monoclonal Antibodies Against Human Tissue-Type Plasminogen Activator

1985 ◽  
Vol 53 (02) ◽  
pp. 170-175 ◽  
Author(s):  
Raymond R Schleef ◽  
Manjula Sinha ◽  
David J Loskutoff
Keyword(s):  
Monoclonal Antibodies ◽  
Plasminogen Activator ◽  
Human Tissue ◽  
Association Constant ◽  
Tissue Type ◽  
Tissue Type Plasminogen Activator ◽  
High Affinity ◽  
Standard Procedures ◽  
Type Plasminogen Activator

SummaryA series of hybridoma clones, each producing monoclonal antibodies to human tissue-type plasminogen activator (t-PA), were prepared from mice by standard procedures. Two of these clones were selected for further study. One HI72A1, produced antibodies that bound to t-PA and strongly inhibited its activity, whereas another, LI72D1, produced antibodies that bound to t-PA but did not affect its activity. The specificity of these antibodies was assessed in immunoabsorption experiments. Both immunoprecipitated 125I-labeled t-PA, and both were specific since only t-PA was recognized in conditioned media collected from Bowes melanoma cells cultured in the presence of 3H-leucine. Neither antibody recognized urokinase. t-PA was desorbed from antibody HI72A1-Sepharose columns with 0.5 M NaCl, consistent with its relatively low association constant (Ka = 9.37 × 107 M-1). In contrast, a strong chaotropic agent (i.e., 2 M KI) was required to elute t-PA from antibody LI72D1 columns (Ka = 2.08 × 109 M-1). This latter high affinity antibody was employed to develop an immunoradiometric assay for t-PA having a sensitivity of 0.5 ng/ml.

Characterization of a Panel of Monoclonal Antibodies Against Human Tissue-Type Plasminogen Activator

Hybridoma ◽  
1988 ◽  
Vol 7 (2) ◽  
pp. 177-184 ◽  
Author(s):  
THOMAS M. REILLY ◽  
SANDRA K. FLINT ◽  
BERNIE G. McHUGH ◽  
KATHLEEN M. WILSBACH-VOLCHECK ◽  
PIETER B.M.W.M. TIMMERMANS
Keyword(s):  
Monoclonal Antibodies ◽  
Plasminogen Activator ◽  
Human Tissue ◽  
Tissue Type ◽  
Tissue Type Plasminogen Activator ◽  
Type Plasminogen Activator
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