Solid-phase enzyme immunoassay for serum ferritin.

1979 ◽  
Vol 25 (8) ◽  
pp. 1426-1431 ◽  
Author(s):  
S Anaokar ◽  
P J Garry ◽  
J C Standefer
Keyword(s):  
Horseradish Peroxidase ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Serum Ferritin ◽  
Solid Phase ◽  
Highly Sensitive

Abstract We describe a solid-phase enzyme immunoassay for ferritin in human serum, with use of a horseradish peroxidase-labeled antibody and a highly sensitive chromogen, 2,2'-azino-di(3-ethyl-benzthiazoline-6-sulfonate). The assay requires only 10 microL of serum per assay, relatively less time and labor than other assys for ferritin, and as little as 10 pg of ferritin can be measured. We believe this assay offers a reliable alternative to radioimmunoassay for serum ferritin.

A solid-phase enzyme immunoassay for serum ferritin.

1977 ◽  
Vol 23 (11) ◽  
pp. 2142-2144 ◽  
Author(s):  
L Thériault ◽  
M Pagé
Keyword(s):  
Alkaline Phosphatase ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Serum Ferritin ◽  
Solid Phase ◽  
Men And Women ◽  
Detectable Concentration ◽  
Human Ferritin ◽  
Antibody Conjugate ◽  
Normal Men

Abstract We describe an enzyme immunoassay for human serum ferritin in which antibody adsorbed on polystyrene tubes is used. Adsorbed gamma-globulins against human ferritin were first allowed to react with ferritin and a second antiferritin antibody, labeled with alkaline phosphatase, was added. The amount of bound enzyme/antibody conjugate was proportional to the ferritin titer in the assay. This method offers stable reagents that can be kept for many months at 4 degrees C. The average values for ferritin in normal men and women were, respectively, 58 and 43 microgram/liter. The lowest detectable concentration was 5 microgram/liter.

A Highly Sensitive Sandwich Enzyme Immunoassay for Insulin in Human Serum Developed Using Capybara Anti-Insulin Fab' -Horseradish Peroxidase Conjugate

1983 ◽  
Vol 16 (19) ◽  
pp. 1509-1523 ◽  
Author(s):  
Masayoshi Imagawa ◽  
Seiichi Hashida ◽  
Eiji Ishikawa ◽  
Hiroyuki Mori ◽  
Chiharu Nakai ◽  
...  
Keyword(s):  
Horseradish Peroxidase ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Highly Sensitive ◽  
Sandwich Enzyme Immunoassay ◽  
Peroxidase Conjugate

A highly sensitive enzyme immunoassay of anti-insulin antibodies in human serum

1987 ◽  
Vol 1 (2) ◽  
pp. 170-174 ◽  
Author(s):  
Takeyuki Kohno ◽  
Eiji Ishikawa ◽  
Satoru Sugiyama ◽  
Syuji Nakamura ◽  
Yoshimasa Kanemaru
Keyword(s):  
Human Serum ◽  
Enzyme Immunoassay ◽  
Insulin Antibodies ◽  
Highly Sensitive

Solid-phase enzymoimmunoassay for osteocalcin in human serum or plasma, with use of a monoclonal antibody.

1989 ◽  
Vol 35 (10) ◽  
pp. 2087-2092 ◽  
Author(s):  
M J Power ◽  
P F Fottrell
Keyword(s):  
Monoclonal Antibody ◽  
Detection Limit ◽  
Horseradish Peroxidase ◽  
Human Serum ◽  
Solid Phase ◽  
Sample Volume ◽  
Microtiter Plates ◽  
Analytical Recovery ◽  
Peroxidase Conjugate

Abstract In this solid-phase enzymoimmunoassay on microtiter plates for osteocalcin in serum or plasma, we use an osteocalcin-horseradish-peroxidase conjugate and a monoclonal antibody raised against bovine osteocalcin. We thoroughly standardized the assay for measurement of osteocalcin in both serum and plasma, demonstrating independence of sample volume, and determining the analytical recovery and within-and between-assay CVs. The detection limit was between 0.6 and 1.1 micrograms/L and the ED50 was 16 micrograms/L for a 5-microL sample volume. The intra-assay CV over the range 3 to 74 micrograms/L was less than or equal to 15%. The interassay CV over the range 3.6 to 46 micrograms/L was less than or equal to 16%. Results by this assay and by an in-house radioimmunoassay in which the same monoclonal antibody was used correlated well (r2 = 0.948). Osteocalcin concentrations in serum and plasma as measured with the present assay agreed well with published values.

Specific enzyme immunoassay for human chorionic gonadotrophin

1981 ◽  
Vol 97 (4) ◽  
pp. 562-568 ◽  
Author(s):  
Tatsuhiro Sekiya ◽  
Yoshihito Furuhashi ◽  
Setsuko Goto ◽  
Shigeaki Kaseki ◽  
Yutaka Tomoda ◽  
...  
Keyword(s):  
Enzyme Immunoassay ◽  
Solid Phase ◽  
Chorionic Gonadotrophin ◽  
Human Chorionic Gonadotrophin ◽  
Serum Samples ◽  
Coefficients Of Variation ◽  
Highly Sensitive ◽  
Assay Tube ◽  
Sandwich Type ◽  
Serial Samples

Abstract. A sandwich-type enzyme immunoassay system specific for human chorionic gonadotrophin (hCG) was prepared with the antibody Fab'-β-D-galactosidase complex and the antibody F(ab')2-immobilized silicone rubber solid phase by using a purified antibody to β subunit of hCG (hCGβ). The assay system cross-reacted less than 4% with human luteinizing hormone (hLH) and human follicular stimulating hormone (hFSH), and proved to be highly sensitive with hCG measurable at levels as low as 0.3 mIU per assay tube. Using 50 μl of serum sample, 6–600 mIU/ml of hCG in serum could be determined specifically with the same degree of precision as in radioimmunoassay but without sample interference with the assay. The coefficients of variation within-run and between-run were 8.6–8.9%, and 4.9– 10.7%, respectively. Values obtained with the enzyme immunoassay correlated well with those of radioimmunoassay ([unk] = 0.98, slope = 0.94, y-intercept = 10.2 mIU/ml for 75 serum samples). Results of the immunoassay of hCG levels in serial samples of serum from healthy women and patients with choriocarcinoma show that this method is useful in the clinical diagnosis of trophoblastic disease.

scholarly journals Solid-phase enzyme immunoassay for immunoglobulin M antibodies to cytomegalovirus

1977 ◽  
Vol 5 (6) ◽  
pp. 629-634
Author(s):  
H Schmitz ◽  
H W Doerr ◽  
D Kampa ◽  
A Vogt
Keyword(s):  
Alkaline Phosphatase ◽  
Horseradish Peroxidase ◽  
Enzyme Immunoassay ◽  
Indirect Immunofluorescence ◽  
Solid Phase ◽  
Immunoglobulin M ◽  
Nuclear Antigen ◽  
Immunofluorescence Method ◽  
Infected Cells

A sensitive, solid-phase enzyme immunoassay for the detection of immunoglobulin M antibodies to cytomegalovirus is described. The results of the enzyme immunoassay correlated well with those obtained by an indirect immunofluorescence method. Horseradish peroxidase proved to be a more sensitive label than alkaline phosphatase. Nonspecific reactions, occurring with commercially available cytomegalovirus antigens, could be avoided by using a nuclear antigen prepared from sonically disrupted nuclei of cytomegalovirus-infected cells.

Rapid, competitive enzymoimmunoassay for albumin in urine.

1986 ◽  
Vol 32 (4) ◽  
pp. 669-671 ◽  
Author(s):  
J Chesham ◽  
S W Anderton ◽  
C F Kingdon
Keyword(s):  
Diabetic Nephropathy ◽  
Human Serum Albumin ◽  
Horseradish Peroxidase ◽  
Serum Albumin ◽  
Human Serum ◽  
Human Urine ◽  
Solid Phase ◽  
Low Concentrations ◽  
Initiation Of Therapy ◽  
Assay Protocol

Abstract In this solid-phase competitive enzymoimmunoassay for albumin in human urine, antiserum to human serum albumin labeled with horseradish peroxidase (EC 1.11.1.7) is incubated with solid-phase-bound human serum albumin in the presence of sample or standard. Results obtained correlate well (r = 0.96) with those of an established fluoroimmunoassay. The present assay covers the range 0.9 to 200 mg/L and can be performed within 1 h. These characteristics, together with the simplicity of the assay protocol, make it very useful for monitoring low concentrations of albumin in urine. Detection of such minimal albuminuria allows initiation of therapy that may prevent development of clinical proteinuria and associated diabetic nephropathy.

Highly sensitive enzyme immunoassay of proinsulin immunoreactivity with use of two monoclonal antibodies

1993 ◽  
Vol 39 (10) ◽  
pp. 2146-2150 ◽  
Author(s):  
L L Kjems ◽  
M E Røder ◽  
B Dinesen ◽  
S G Hartling ◽  
P N Jørgensen ◽  
...  
Keyword(s):  
Monoclonal Antibodies ◽  
Detection Limit ◽  
Human Serum ◽  
Enzyme Immunoassay ◽  
Healthy Subjects ◽  
Sandwich Elisa ◽  
C Peptide ◽  
Peptide Antibody ◽  
Analytical Recovery ◽  
Highly Sensitive

Abstract A highly sensitive two-site sandwich ELISA measuring total proinsulin immunoreactive material in serum or plasma was developed. The assay was based on two monoclonal antibodies, an anti-C-peptide antibody bound to a microtest plate and a biotin-labeled anti-insulin antibody. The detection limit (3 SD above zero value) in buffer was 0.05 pmol/L, corresponding to 0.25 pmol/L in human serum (diluted 1:5). The linear calibrator range was 0.05-20 pmol/L. Interassay CVs were 4.7% at a median (range) of 2.3 pmol/L (1.4-2.8 pmol/L, n = 8), 6.7% at 5.1 pmol/L (3.3-8.0 pmol/L, n = 8), and 8.7% at 10.0 pmol/L (8-12 pmol/L, n = 10). Mean analytical recovery of added human proinsulin (hPI) (2, 5, and 10 pmol/L) to serum was 84% (range 68-128%, n = 9). Human insulin and human C-peptide did not cross-react at 5000 and 10,000 pmol/L, respectively. The four major proinsulin conversion intermediates reacted 65-99%: split(32-33)hPI 74%, des-(31,32)hPI 65%, split(65-66)hPI 78%, and des(64,65)hPI 99%. All serum values from 38 fasting healthy subjects were above the detection limit: median (range) 4.0 (2.1-12.6) pmol/L.

scholarly journals Sensitive Sandwich Enzyme Immunoassay for Serum Ferritin on Microtitre Plates

1981 ◽  
Vol 18 (1) ◽  
pp. 48-53 ◽  
Author(s):  
Sukanya Linpisarn ◽  
Larry J Kricka ◽  
John H Kennedy ◽  
Thomas P Whitehead
Keyword(s):  
Enzyme Immunoassay ◽  
Serum Ferritin ◽  
Clinical Laboratory ◽  
Solid Phase ◽  
Good Precision ◽  
Ferritin Concentration ◽  
Assay Procedure ◽  
Serum Ferritin Concentration ◽  
Sandwich Enzyme Immunoassay

A solid-phase sandwich enzyme immunoassay for serum ferritin is described. The assay procedure was simple, showed good precision, and was suitable for the routine determination of serum ferritin concentration in a clinical laboratory. The assay compared well with a conventional radioimmunoassay for ferritin. The assay was extremely sensitive and had a detection limit of 2·2 pg.

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