Chemistry: molecular biology and microbiology – Enzyme – proenzyme; compositions thereof; process for... – Hydrolase
Patent
1995-05-23
1997-12-30
Weber, Jon P.
Chemistry: molecular biology and microbiology
Enzyme , proenzyme; compositions thereof; process for...
Hydrolase
435183, C12N 948, C12N 900
Patent
active
057029377
ABSTRACT:
A method for potentiating the activity of tissue plasminogen activator proteins by adding certain amounts of beta-lactoglobulin A is claimed. In particular, the invention includes an enhanced solid-phase immunoassay for measuring tissue plasminogen activator (nt-PA) activity in a mammalian serum sample with sufficient sensitivity to detect neutralizing activity in samples containing .mu.g/ml levels of total nt-PA antibody. Neutralizing activity can be easily detected in serum dilutions containing as little as 0.038 to 0.48 .mu.g of total nt-PA antibody in a 20 .mu.l sample. The assay has a sensitivity of 1 pM of nt-PA (67 pg/ml) in 5 hr.
REFERENCES:
Burck et al. (1990) Journal Biol. Chem., 265(9), "Characterization of a Modified human Tissue Plasminogen Activator Comprising a Kringle-2 and a Protease Domain", pp. 5170-5177.
Buessecker, et al., 1993, Enzyme-linked immunosorbent assays for plasminogen activators; Journal of Immunological Methods 162: 193-200.
Verheijen, et. al., 1982, Activation of Plasminogen by Tissue Activator is Increased Specifically In The Presence Of Certain Soluble Fibrin(ogen) Fragments; Thrombosis Research 27: 377-385.
Ranby, et. al., 1982, A Sensitive Assay For Tissue Plasminogen Activator; Thrombosis Research 27: 743-749.
Gyzander, et al., A Sensitive Assay For Tissue Plasminogen Activator Activity In Plasma, Using Adsorption On Lysine-Sepharose, Thrombosis Research 35: 547-558, 1984.
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Takada, et al., The Physiology of the Fibrinolytic System; Japanese Journal of Physiology 43: 1-19.
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Zwickl, Immunogenicity of Biosynthetic Human LysPro Insulin Compared to Native-sequence Human and Purified Porcine Insulins in Rhesus Monkeys Immunized Over a 6-Week Period; Arzneum-Forsch/Drug Res. 45, 524-530 (1995).
Johnson, et al, Improved Technique Utilizing Nonfat Dry Milk for Analysis of Proteins and Nucleic Acids Transferred to Nitrocellulose, Gene Anal Techn 1:3-8.
Boone David E.
Eli Lilly and Company
Leeds James P.
Maciak Ronald S.
Weber Jon P.
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