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Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays

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Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays. / Williams, AJK; Curnock, R; Reed, CR; Easton, P; Rokni, S; Bingley, PJ.

In: Journal of Immunological Methods, Vol. 31;362(1-2), 10.2010, p. 199 - 203.

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Williams, AJK ; Curnock, R ; Reed, CR ; Easton, P ; Rokni, S ; Bingley, PJ. / Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays. In: Journal of Immunological Methods. 2010 ; Vol. 31;362(1-2). pp. 199 - 203.

Bibtex

@article{6e7ec1ce125f4ed281fd950bdfdf4b6b,
title = "Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays",
abstract = "Insulin autoantibodies (IAA) are usually the first risk-markers detected during the type 1 diabetes prodrome, but precise measurement is difficult as insulin binding is often low. Non-specific binding (NSB) of (125)I-labelled insulin necessitates competitive displacement with unlabelled insulin to demonstrate specificity. NSB varies with different batches of label, suggesting that it is caused by impurities in the label. Addition of bovine serum albumin (BSA) can reduce NSB, so we investigated whether BSA antibodies cause lack of specificity in IAA assays. Samples from patients with newly-diagnosed type 1 diabetes, healthy schoolchildren previously found to have raised (125)I-insulin binding (≥ 0.4 units) and IAA-negative schoolchildren were re-assayed for IAA by radiobinding microassay using commercial (125)I-insulin with and without 1g/dl BSA added to the buffer. Of 100 patients, 68 were IAA-positive on re-assay with BSA compared to 72 without BSA (p=0.125). Of 154 schoolchildren who previously had raised (125)I-insulin binding, only 45 had (125)I-insulin binding ≥ 0.4 units on re-assay with BSA compared to 90 without BSA (p",
author = "AJK Williams and R Curnock and CR Reed and P Easton and S Rokni and PJ Bingley",
year = "2010",
month = "10",
doi = "10.1016/j.jim.2010.09.009",
language = "English",
volume = "31;362(1-2)",
pages = "199 -- 203",
journal = "Journal of Immunological Methods",
issn = "0022-1759",
publisher = "Elsevier B.V.",

}

RIS - suitable for import to EndNote

TY - JOUR

T1 - Anti-BSA antibodies are a major cause of non-specific binding in insulin autoantibody radiobinding assays

AU - Williams, AJK

AU - Curnock, R

AU - Reed, CR

AU - Easton, P

AU - Rokni, S

AU - Bingley, PJ

PY - 2010/10

Y1 - 2010/10

N2 - Insulin autoantibodies (IAA) are usually the first risk-markers detected during the type 1 diabetes prodrome, but precise measurement is difficult as insulin binding is often low. Non-specific binding (NSB) of (125)I-labelled insulin necessitates competitive displacement with unlabelled insulin to demonstrate specificity. NSB varies with different batches of label, suggesting that it is caused by impurities in the label. Addition of bovine serum albumin (BSA) can reduce NSB, so we investigated whether BSA antibodies cause lack of specificity in IAA assays. Samples from patients with newly-diagnosed type 1 diabetes, healthy schoolchildren previously found to have raised (125)I-insulin binding (≥ 0.4 units) and IAA-negative schoolchildren were re-assayed for IAA by radiobinding microassay using commercial (125)I-insulin with and without 1g/dl BSA added to the buffer. Of 100 patients, 68 were IAA-positive on re-assay with BSA compared to 72 without BSA (p=0.125). Of 154 schoolchildren who previously had raised (125)I-insulin binding, only 45 had (125)I-insulin binding ≥ 0.4 units on re-assay with BSA compared to 90 without BSA (p

AB - Insulin autoantibodies (IAA) are usually the first risk-markers detected during the type 1 diabetes prodrome, but precise measurement is difficult as insulin binding is often low. Non-specific binding (NSB) of (125)I-labelled insulin necessitates competitive displacement with unlabelled insulin to demonstrate specificity. NSB varies with different batches of label, suggesting that it is caused by impurities in the label. Addition of bovine serum albumin (BSA) can reduce NSB, so we investigated whether BSA antibodies cause lack of specificity in IAA assays. Samples from patients with newly-diagnosed type 1 diabetes, healthy schoolchildren previously found to have raised (125)I-insulin binding (≥ 0.4 units) and IAA-negative schoolchildren were re-assayed for IAA by radiobinding microassay using commercial (125)I-insulin with and without 1g/dl BSA added to the buffer. Of 100 patients, 68 were IAA-positive on re-assay with BSA compared to 72 without BSA (p=0.125). Of 154 schoolchildren who previously had raised (125)I-insulin binding, only 45 had (125)I-insulin binding ≥ 0.4 units on re-assay with BSA compared to 90 without BSA (p

U2 - 10.1016/j.jim.2010.09.009

DO - 10.1016/j.jim.2010.09.009

M3 - Article

VL - 31;362(1-2)

SP - 199

EP - 203

JO - Journal of Immunological Methods

JF - Journal of Immunological Methods

SN - 0022-1759

ER -