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Quantitative determination of islet cell surface antibodies using 125I-protein A

Huen, A. H.J. ; Haneda, M. ; Freedman, Z. ; Lernmark, A. LU orcid and Rubenstein, A. H. (1983) In Diabetes 32(5 I). p.460-465
Abstract

A quantitative method to measure islet cell surface antibodies in human patients has been developed using 125I-protein A. Isolated, dispersed, viable rat islet cells prepared by collagenase digestion were fixed in 4% paraformaldehyde to allow storage for up to 7 wk at 4°C. Human sera, heat inactivated and adsorbed with rat liver and kidney powder (100 mg/ml), were incubated with the fixed cells (50 x 103) for 60 min at 37°C. Thereafter the cells were washed and exposed to 5 x 105 cpm 125I-protein A, which binds to IgG attached to the cell surface. Assay precision (14%) and reproducibility (16%) were established by repeated analysis of pooled sera from healthy individuals and IDDM patients... (More)

A quantitative method to measure islet cell surface antibodies in human patients has been developed using 125I-protein A. Isolated, dispersed, viable rat islet cells prepared by collagenase digestion were fixed in 4% paraformaldehyde to allow storage for up to 7 wk at 4°C. Human sera, heat inactivated and adsorbed with rat liver and kidney powder (100 mg/ml), were incubated with the fixed cells (50 x 103) for 60 min at 37°C. Thereafter the cells were washed and exposed to 5 x 105 cpm 125I-protein A, which binds to IgG attached to the cell surface. Assay precision (14%) and reproducibility (16%) were established by repeated analysis of pooled sera from healthy individuals and IDDM patients using pooled batches of islet cells. Using this method, islet cell surface antibodies were detected in 35% of insulin-dependent diabetic patients.

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author
; ; ; and
publishing date
type
Contribution to journal
publication status
published
in
Diabetes
volume
32
issue
5 I
pages
6 pages
publisher
American Diabetes Association Inc.
external identifiers
  • scopus:0020584785
  • pmid:6341129
ISSN
0012-1797
language
English
LU publication?
no
id
92c99692-3e1f-461e-90a4-d516e22c77e8
date added to LUP
2019-09-16 12:50:33
date last changed
2024-03-13 08:03:50
@article{92c99692-3e1f-461e-90a4-d516e22c77e8,
  abstract     = {{<p>A quantitative method to measure islet cell surface antibodies in human patients has been developed using <sup>125</sup>I-protein A. Isolated, dispersed, viable rat islet cells prepared by collagenase digestion were fixed in 4% paraformaldehyde to allow storage for up to 7 wk at 4°C. Human sera, heat inactivated and adsorbed with rat liver and kidney powder (100 mg/ml), were incubated with the fixed cells (50 x 10<sup>3</sup>) for 60 min at 37°C. Thereafter the cells were washed and exposed to 5 x 10<sup>5</sup> cpm <sup>125</sup>I-protein A, which binds to IgG attached to the cell surface. Assay precision (14%) and reproducibility (16%) were established by repeated analysis of pooled sera from healthy individuals and IDDM patients using pooled batches of islet cells. Using this method, islet cell surface antibodies were detected in 35% of insulin-dependent diabetic patients.</p>}},
  author       = {{Huen, A. H.J. and Haneda, M. and Freedman, Z. and Lernmark, A. and Rubenstein, A. H.}},
  issn         = {{0012-1797}},
  language     = {{eng}},
  month        = {{06}},
  number       = {{5 I}},
  pages        = {{460--465}},
  publisher    = {{American Diabetes Association Inc.}},
  series       = {{Diabetes}},
  title        = {{Quantitative determination of islet cell surface antibodies using <sup>125</sup>I-protein A}},
  volume       = {{32}},
  year         = {{1983}},
}