+ Site Statistics
+ Search Articles
+ PDF Full Text Service
How our service works
Request PDF Full Text
+ Follow Us
Follow on Facebook
Follow on Twitter
Follow on LinkedIn
+ Subscribe to Site Feeds
Most Shared
PDF Full Text
+ Translate
+ Recently Requested

Novel and sensitive noncompetitive (two-site) enzyme immunoassay for haptens with amino groups



Novel and sensitive noncompetitive (two-site) enzyme immunoassay for haptens with amino groups



Journal of Clinical Laboratory Analysis 4(3): 208-212



A novel and sensitive noncompetitive (two-site) enzyme immunoassay for haptens with amino groups is described. L-Thyroxine (T4) was used as a model hapten. T4 was indirectly biotinylated with glutathione as spacer between T4 and biotin molecules and trapped onto anti-(T4-bovine serum albumin) IgG-coated polystyrene balls. After washing the polystyrene balls to eliminate unreacted biotin and other biotinylated substances, biotinylated T4 was eluted from the polystyrene balls with HCl and was reacted with anti-(T4-bovine serum albumin) Fab'-horseradish peroxidase conjugate. The complex formed was trapped onto streptavidin-coated polystyrene balls. Peroxidase activity bound to the polystyrene balls was assayed by fluorimetry. The detection limit of T4 was 78 fg (0.1 fmol)/tube, which was 50-fold lower than the limit by competitive enzyme immunoassay using the same antibody and T4-peroxidase conjugate. This noncompetitive enzyme immunoassay was applied to the measurement of total T4 in serum. Serum levels of total T4 in 10 healthy subjects aged 25-40 yr were 94 +/- 13 (SD) micrograms/L (range, 78-114). The principle of this noncompetitive enzyme immunoassay may allow it to measure other haptens with amino groups more sensitively than can competitive immunoassays.

Please choose payment method:






(PDF emailed within 0-6 h: $19.90)

Accession: 040823876

Download citation: RISBibTeXText

PMID: 2191099

DOI: 10.1002/jcla.1860040312


Related references

Novel and sensitive noncompetitive (two-site) immunoassay for haptens with emphasis on peptides. Clinical Biochemistry 23(5): 445-453, 1990

Novel and sensitive noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for alpha-human atrial natriuretic peptide in plasma. Journal of Clinical Laboratory Analysis 5(5): 324-330, 1991

Novel and sensitive noncompetitive enzyme immunoassay hetero two site enzyme immunoassay for arginine vasopressin in plasma. Analytical Letters 24(7): 1109-1124, 1991

Novel and Sensitive Noncompetitive Enzyme Immunoassay (Hetero-Two-Site Enzyme Immunoassay) for Arginine Vasopressin. Analytical Letters 24(7): 1109-1123, 1991

Detection of one attomole of [Arg8]-vasopressin by novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay). Journal of Biochemistry 110(4): 486-492, 1991

Novel and ultrasensitive noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay) for alpha-human atrial natriuretic peptide. Journal of Clinical Laboratory Analysis 6(4): 201-208, 1992

Novel and sensitive noncompetitive enzyme immunoassay for peptides. Biochemical & Biophysical Research Communications 160(1): 40-45, 1989

Noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for gamma 2-melanocyte-stimulating hormone (gamma 2-MSH) and measurement of immunoreactive gamma 2-MSH in plasma of healthy subjects. Journal of Clinical Laboratory Analysis 9(6): 397-406, 1995

Novel and sensitive noncompetitive enzyme immunoassay for kassinin in rat plasma. Experientia 45(5): 470-472, 1989

A noncompetitive enzyme immunoassay (hetero-two-site enzyme immunoassay) for salmon calcitonin: determination of the bioavailability of subcutaneous salmon calcitonin and its correlation with the hypocalcemic activity in rats. Journal of Clinical Laboratory Analysis 10(2): 91-97, 1996

Novel and sensitive noncompetitive enzyme immunoassay to measure peptides by biotinylation. Analytical Letters 22(2): 353-364, 1989

A homogeneous noncompetitive immunoassay for the detection of small haptens. Analytical Chemistry 74(11): 2500-2504, 2002

Noncompetitive immunoassay detection system for haptens on the basis of antimetatype antibodies. Clinical Chemistry 61(4): 627-635, 2015

A sensitive two-site sandwich enzyme immunoassay for human angiotensin converting enzyme utilizing monoclonal antibodies. Journal of Immunological Methods 132(2): 263-273, 1990

A noncompetitive enzyme immunoassay for rat prolactin. Journal of Immunoassay 16(1): 81-96, 1995