+ 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

Detection of one attomole of [Arg8]-vasopressin by novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay)



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



One attomole of [Arg8]-vasopressin (AVP) was detected by a novel noncompetitive enzyme immunoassay (hetero-two-site complex transfer enzyme immunoassay). AVP was indirectly biotinylated using N-hydroxysuccinimidobiotin and trapped onto an anti-AVP IgG-coated polystyrene ball. After washing, biotinylated AVP was eluted from the polystyrene ball with HCl and was reacted with 2,4-dinitrophenyl-fluorescein disulfide-bovine serum albumin-rabbit anti-AVP IgG conjugate. The complex formed was trapped on [anti-2,4-dinitrophenyl group] IgG-coated polystyrene balls and, after washing, reacted with avidin-beta-D-galactosidase conjugate. The polystyrene balls were washed, and the complex of the three components was eluted with 2,4-dinitrophenyl-L-lysine and transferred to anti-fluorescein IgG-coated polystyrene balls. After washing, the complex was released from the polystyrene balls by reduction with 2-mercaptoethylamine and transferred to [anti-rabbit IgG] IgG-coated polystyrene balls. beta-D-Galactosidase activity bound to the last polystyrene balls was assayed by fluorometry. The detection limit of AVP was 1.1 fg (1 amol)/tube. Interference by proteins in biological fluids was eliminated by separation of peptides from proteins using a molecular sieve. The principle of the present method may be applicable to the measurement of haptens, including peptides, that can be derivatized so as to be bound simultaneously by both anti-hapten antibody and avidin molecules.

Please choose payment method:






(PDF emailed within 1 workday: $29.90)

Accession: 039776826

Download citation: RISBibTeXText

PMID: 1778973


Related references

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 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

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

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

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

Measurement of human immunodeficiency virus type 1 p24 in serum by an ultrasensitive enzyme immunoassay, the two-site immune complex transfer enzyme immunoassay. Journal of Clinical Microbiology 33(2): 298-303, 1995

Detection of antibody IgG to HIV-1 in urine by ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant p24 as antigen for diagnosis of HIV-1 infection. Journal of Clinical Laboratory Analysis 8(2): 86-95, 1994

Diagnosis of HIV-1 infection by detection of antibody IgG to HIV-1 in urine with ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens. Journal of Clinical Laboratory Analysis 8(4): 237-246, 1994

Detection of antibody IgG to HIV-1 in urine by sensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant proteins as antigens for diagnosis of HIV-1 infection. Journal of Clinical Laboratory Analysis 7(6): 353-364, 1993

Detection of Anti-Thyroglobulin Immunoglobulin G in Urine by Sensitive Enzyme Immunoassay (Immune Complex Transfer Enzyme Immunoassay) as a Diagnostic Aid for Autoimmune Thyroid Diseases. Analytical Letters 26(1): 17-31, 1993

Detection of anti-human immunodeficiency virus type 1 (HIV-1) immunoglobulin G in urine by an ultrasensitive enzyme immunoassay (immune complex transfer enzyme immunoassay) with recombinant reverse transcriptase as an antigen. Journal of Clinical Microbiology 32(3): 819-822, 1994

Sensitive detection of anti-human T-cell leukemia virus type I IgG in human serum by a novel enzyme immunoassay (immune complex transfer enzyme immunoassay) using recombinant gag-env hybrid protein as antigen. Journal of Virological Methods 31(1): 77-92, 1991

More reliable diagnosis of infection with human immunodeficiency virus type 1 (HIV-1) by detection of antibody IgGs to pol and gag proteins of HIV-1 and p24 antigen of HIV-1 in urine, saliva, and/or serum with highly sensitive and specific enzyme immunoassay (immune complex transfer enzyme immunoassay): A review. Journal of Clinical Laboratory Analysis 11(5): 267-286, 1997

Detection of circulating immune complexes by enzyme immunoassay. A basic study of C1q enzyme immunoassay and anti-C3 enzyme immunoassay using microplates. Arerugi 33(2): 117-126, 1984