An enzyme immunoassay for mouse epidermal growth factor utilizing a liquid phase double-antibody system

An enzyme immunoassay for mouse epidermal growth factor (EGF) involving a liquid phase double-antibody system was developed. The EGF-beta-galactosidase conjugate prepared was stable for at least 8 months. By this method, EGF was detectable at a concentration as low as 20 pg per tube. The concentrations of EGF in various tissues of mice are also presented.     

An enzyme immunoassay for mouse epidermal growth factor utilizing a liquid phase double-antibody system

Summary An enzyme immunoassay for mouse epidermal growth factor (EGF) involving a liquid phase double-antibody system was developed. The EGF--galactosidase conjugate prepared was stable for at least 8 months. By this method, EGF was detectable at a concentration as low as 20 pg per tube. The concentrations of EGF in various tissues of mice are also presented.This work was supported in part by a Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

Synthesis of maleimide derivative of cortisol for enzyme coupling in cortisol enzyme immunoassay

Summary A new cortisol derivative, cortisol-21-m-maleimidobenzoate (CMB), was synthesized and conjugated with sulfhydryl groups of -galactosidase (BG). Both CMB-BG and CHS-BG conjugates have a high immunoreactivity to cortisol antibody, and although CHS-BG does not displace well with the added cortisol, CMB-BG does.Acknowledgments. This work was supported in part by Grant 884M from the Council for Tobacco Research. We wish to thank Miss Laura Morbach and Mrs Tatsue Monji for their assistance on this project.Address reprint requests to A. Castro.     

A novel method in enzyme immunoassay: Maleimide derivative of hapten for enzyme coupling

Summary Meta-maleimidobenzoyl derivative of L-thyroxine methyl ester (MBTM) was synthesized and coupled to -galactosidase at molar ratio of over 5 to 1. More than 97% of the enzyme was found to be labeled with MBTM. A thyroxine enzyme immunoassay was carried out with sensitivity in the 0–10 g/100 ml range.     

A novel method in enzyme immunoassay: maleimide derivative of hapten for enzyme coupling.

Meta-maleimidobenzoyl derivative of L-thyroxine methyl ester (MBTM) was synthesized and coupled to beta-galactosidase at molar ratio of over 5 to 1. More than 97% of the enzyme was found to be labeled with MBTM. A thyroxine enzyme immunoassay was carried out with sensitivity in 0-10 microgram/100 ml range.     

Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma

Summary A novel and sensitive noncompetitive enzyme immunoassay for kassinin is described. Kassinin was biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate. The biotinylated kassinin was trapped on antikassinin IgG-coated polystyrene balls and, after washing to eliminate other biotinylated substances, was eluted with HCl. The biotinylated kassinin eluted was reacted with anti-kassinin Fab'-peroxidase conjugate and trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of kassinin was 0.13 pg (0.1 fmol)/tube or 0.065 g/l of rat plasma, which was 750-fold or 15-fold lower than that for competitive radioimmunoassay.     

Quantification of digoxin by enzyme immunoassay: synthesis of a maleimide derivative of digoxigenin succinate for enzyme coupling

Summary We synthesized the m-maleimidobenzoyl derivative of digoxigenin-3-0-succinate (through a p-phenylenediamine bridge) as a hapten derivative directed towards coupling to sulfhydryl groups of -galactosidase. Prepared enzyme conjugate had about 97% of the enzyme labeled with the hapten derivative while retaining full enzyme activity. The enzyme immunoassay for digoxin we prepared showed a maximum sensitivity of 30 pg per assay (c.v.=3%) with minimal cross-reaction with digotoxin (3.8%). Our method for hapten conjugation to -galactosidase is highly efficient and is simple and easily replicated.     

Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma

A novel and sensitive noncompetitive enzyme immunoassay for kassinin is described. Kassinin was biotinylated using sulfosuccinimidyl-6-(biotinamido)hexanoate. The biotinylated kassinin was trapped on anti-kassinin IgG-coated polystyrene balls and, after washing to eliminate other biotinylated substances, was eluted with HCl. The biotinylated kassinin eluted was reacted with anti-kassinin Fab'-peroxidase conjugate and trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of kassinin was 0.13 pg (0.1 fmol)/tube or 0.065 microgram/l of rat plasma, which was 750-fold or 15-fold lower than that for competitive radioimmunoassay.     

Compared expression of murine plasmacytoma associated virus in tumor cells and in mouse embryo cells]

A DNA complementary to the viral genome of C-type particles produced by a Mouse myeloma derived cell line (MF2 cell line) was synthesized. This cDNA was used as a probe to study the viral genome expression among the total RNA and the poly (A)-rich RNA extracted from the MF2 and Balb/c embryonic cells. As evidenced by molecular hybridization experiments, the presence of at least one endogenous Balb/c virus in the MF2 virus stocks is suggested. In the productive cells, the viral RNA sequences are expressed in the poly (A)-rich RNA fraction.     

Suppression of carcinogen-induced rat mammary tumor formation by Actinomycin D

Zusammenfassung Sprague-Dawley-Ratten, die eine kombinierte Belastung mit 7,12-Dimethylbenz()anthrazen und Aktinomyzin D überleben, weisen eine drastisch erniedrigte tastbare Tumorinzidenz auf. Kontrolltiere erhielten nur das Karzinogen.