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

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.

Download Full-text

A Highly Sensitive Enzyme Immunoassay of Anti-Insulin Antibodies in Guinea Pig Serum

The Journal of Biochemistry ◽

10.1093/oxfordjournals.jbchem.a121830 ◽

1986 ◽

Vol 100 (5) ◽

pp. 1247-1251 ◽

Cited By ~ 31

Author(s):

Takeyuki KOHNO ◽

Eiji ISHIKAWA

Keyword(s):

Guinea Pig ◽

Enzyme Immunoassay ◽

Insulin Antibodies ◽

Highly Sensitive ◽

Pig Serum

Download Full-text

Development of a highly sensitive and selective microplate chemiluminescence enzyme immunoassay for the determination of free thyroxine in human serum

International Journal of Biological Sciences ◽

10.7150/ijbs.3.274 ◽

2007 ◽

pp. 274-280 ◽

Cited By ~ 14

Author(s):

Xu Wang ◽

Hui Chen ◽

Jin-Ming Lin ◽

Xitang Ying

Keyword(s):

Human Serum ◽

Enzyme Immunoassay ◽

Free Thyroxine ◽

Highly Sensitive ◽

Chemiluminescence Enzyme Immunoassay

Download Full-text

Highly sensitive amperometric enzyme immunoassay for α-fetoprotein in human serum

Journal of Immunological Methods ◽

10.1016/0022-1759(96)00044-0 ◽

1996 ◽

Vol 193 (1) ◽

pp. 51-62 ◽

Cited By ~ 10

Author(s):

Leopoldo Della Ciana ◽

Giuliana Bernacca ◽

Concetta De Nitti ◽

Anilla Massaglia

Keyword(s):

Human Serum ◽

Enzyme Immunoassay ◽

Highly Sensitive

Download Full-text

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

Analytical Letters ◽

10.1080/00032718308069543 ◽

1983 ◽

Vol 16 (19) ◽

pp. 1509-1523 ◽

Cited By ~ 64

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

Download Full-text

Solid-phase enzyme immunoassay for serum ferritin.

Clinical Chemistry ◽

10.1093/clinchem/25.8.1426 ◽

1979 ◽

Vol 25 (8) ◽

pp. 1426-1431 ◽

Cited By ~ 22

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.

Download Full-text

A Highly Sensitive Enzyme Immunoassay of Anti-Insulin Antibodies in Guinea Pig Serum

Analytical Letters ◽

10.1080/00032718608066284 ◽

1986 ◽

Vol 19 (9-10) ◽

pp. 1083-1095 ◽

Cited By ~ 4

Author(s):

Takeyuki Kohno ◽

Ke-He Ruan ◽

Eiji Ishikawa

Keyword(s):

Guinea Pig ◽

Enzyme Immunoassay ◽

Insulin Antibodies ◽

Highly Sensitive ◽

Pig Serum

Download Full-text

A highly sensitive sandwich enzyme immunoassay of urinary growth hormone in children with short stature, Turner's syndrome, and simple obesity

Acta Endocrinologica ◽

10.1530/acta.0.1210513 ◽

1989 ◽

Vol 121 (4) ◽

pp. 513-519 ◽

Cited By ~ 6

Author(s):

Hiroshi Tomita ◽

Masamichi Ogawa ◽

Takashi Kamijo ◽

Osamu Mori ◽

Eiji Ishikawa ◽

...

Keyword(s):

Short Stature ◽

Enzyme Immunoassay ◽

Gh Deficiency ◽

Urine Samples ◽

Turner's Syndrome ◽

Turner’S Syndrome ◽

Highly Sensitive ◽

Significant Difference ◽

Simple Obesity ◽

Sandwich Enzyme Immunoassay

Abstract. GH values were determined by a highly sensitive sandwich enzyme immunoassay in the 1st morning and/or 24-h accumulated urine samples in 94 children (short stature 70, including 14 with complete GH deficiency, 9 with partial GH deficiency, and 47 with GH-normal short stature; Turner's syndrome, 10, and simple obesity, 14). GH values were also determined in the 2nd to 4th urine samples taken on the same day together with the 1st morning urine in 5 of them. GH values in the 1st morning urine correlated significantly with those of the 24-h urine and with serum peak and mean GH values during nocturnal sleep as a physiological GH secretion test. The 2nd to 4th urines had lower GH concentrations than the 1st morning urine. The GH value of the 1st morning urine in complete GH deficiency was significantly lower than those in GH-normal short stature, partial GH deficiency and Turner's syndrome. However, no significant difference was detected in urinary GH values between complete GH deficiency and simple obesity. We conclude that 1st morning urinary GH estimation may be useful for differentiation of complete GH deficiency from other causes of short stature, but may be difficult for the distinction between complete GH deficiency and obesity with normal GH secretory ability.

Download Full-text