A novel LIPS assay for insulin autoantibodies

Daniela Liberati; Rebecca C. Wyatt; Cristina Brigatti; Ilaria Marzinotto; Maurizio Ferrari; Elena Bazzigaluppi; Emanuele Bosi; Ben T. Gillard; Kathleen M. Gillespie; Frans Gorus; Ilse Weets; Eric Balti; Lorenzo Piemonti; Peter Achenbach; Alistair J.K. Williams; Vito Lampasona

doi:10.1007/s00592-017-1082-y

A novel LIPS assay for insulin autoantibodies

Daniela Liberati, Rebecca C. Wyatt, Cristina Brigatti, Ilaria Marzinotto, Maurizio Ferrari, Elena Bazzigaluppi, Emanuele Bosi, Ben T. Gillard, Kathleen M. Gillespie, Frans Gorus, Ilse Weets, Eric Balti, Lorenzo Piemonti, Peter Achenbach, Alistair J.K. Williams, Vito Lampasona^*

^*Corresponding author for this work

Research output: Contribution to journal › Article (Academic Journal) › peer-review

40 Citations (Scopus)

658 Downloads (Pure)

Abstract

Aims: Insulin autoantibodies (IAA) are often the first marker of autoimmunity detected in children in the preclinical phase of type 1 diabetes (T1D). Currently, the vast majority of laboratories adopt the radiobinding micro-assay (RBA) for measuring IAA. Our aim was to replace RBA with a novel non-radioactive IAA Luciferase Immuno Precipitation System (LIPS) assay with improved performance. Methods: We developed (pro)insulin antigens with alternative placements of a NanoLuc™ luciferase reporter (NLuc). Performance in LIPS was evaluated by testing sera from new onset T1D (n = 80), blood donors (n = 123), schoolchildren (n = 186), first-degree relatives (FDRs) from the Bart’s Oxford family study (n = 53) and from the Belgian Diabetes Registry (n = 136), coded sera from the Islet Autoantibody Standardization Program (IASP) (T1D n = 50, blood donors n = 90). Results: IAA LIPS based on B chain-NLuc proinsulin or B chain-NLuc insulin, in which NLuc was fused at the C-terminus of the insulin B chain, required only 2 μL of serum and a short incubation time, showed high concordance with RBA (Spearman r = 0.866 and 0.833, respectively), high assay performance (B chain-NLuc proinsulin ROC-AUC = 0.894 and B chain-NLuc insulin ROC-AUC = 0.916), and an adjusted sensitivity at 95% specificity ranking on par with the best assays submitted to the two most recent IASP workshops. In FDRs, the IAA LIPS showed improved discrimination of progressors to T1D compared to RBA. Conclusions: We established a novel high-performance non-radioactive IAA LIPS that might replace the current gold standard RBA and find wide application in the study of the IAA response in T1D.

Original language	English
Pages (from-to)	263-270
Number of pages	8
Journal	Acta Diabetologica
Volume	55
Early online date	5 Jan 2018
DOIs	https://doi.org/10.1007/s00592-017-1082-y
Publication status	Published - 1 Mar 2018

Keywords

Autoantibodies
Immunoassay
Insulin
LIPS
Prediction
T1D

UN SDGs

This output contributes to the following UN Sustainable Development Goals (SDGs)

Access to Document

10.1007/s00592-017-1082-y

Full-text PDF (accepted author manuscript)
This is the author accepted manuscript (AAM). The final published version (version of record) is available online via Springer at https://link.springer.com/article/10.1007%2Fs00592-017-1082-y. Please refer to any applicable terms of use of the publisher.
Accepted author manuscript, 1.6 MB
Supplementary information PDFAccepted author manuscript, 3.29 MB

Handle.net

Persistent link

Cite this

@article{5bcc138a8b5c491896121c004b428080,

title = "A novel LIPS assay for insulin autoantibodies",

abstract = "Aims: Insulin autoantibodies (IAA) are often the first marker of autoimmunity detected in children in the preclinical phase of type 1 diabetes (T1D). Currently, the vast majority of laboratories adopt the radiobinding micro-assay (RBA) for measuring IAA. Our aim was to replace RBA with a novel non-radioactive IAA Luciferase Immuno Precipitation System (LIPS) assay with improved performance. Methods: We developed (pro)insulin antigens with alternative placements of a NanoLuc{\texttrademark} luciferase reporter (NLuc). Performance in LIPS was evaluated by testing sera from new onset T1D (n = 80), blood donors (n = 123), schoolchildren (n = 186), first-degree relatives (FDRs) from the Bart{\textquoteright}s Oxford family study (n = 53) and from the Belgian Diabetes Registry (n = 136), coded sera from the Islet Autoantibody Standardization Program (IASP) (T1D n = 50, blood donors n = 90). Results: IAA LIPS based on B chain-NLuc proinsulin or B chain-NLuc insulin, in which NLuc was fused at the C-terminus of the insulin B chain, required only 2 μL of serum and a short incubation time, showed high concordance with RBA (Spearman r = 0.866 and 0.833, respectively), high assay performance (B chain-NLuc proinsulin ROC-AUC = 0.894 and B chain-NLuc insulin ROC-AUC = 0.916), and an adjusted sensitivity at 95% specificity ranking on par with the best assays submitted to the two most recent IASP workshops. In FDRs, the IAA LIPS showed improved discrimination of progressors to T1D compared to RBA. Conclusions: We established a novel high-performance non-radioactive IAA LIPS that might replace the current gold standard RBA and find wide application in the study of the IAA response in T1D.",

keywords = "Autoantibodies, Immunoassay, Insulin, LIPS, Prediction, T1D",

author = "Daniela Liberati and Wyatt, {Rebecca C.} and Cristina Brigatti and Ilaria Marzinotto and Maurizio Ferrari and Elena Bazzigaluppi and Emanuele Bosi and Gillard, {Ben T.} and Gillespie, {Kathleen M.} and Frans Gorus and Ilse Weets and Eric Balti and Lorenzo Piemonti and Peter Achenbach and Williams, {Alistair J.K.} and Vito Lampasona",

year = "2018",

month = mar,

day = "1",

doi = "10.1007/s00592-017-1082-y",

language = "English",

volume = "55",

pages = "263--270",

journal = "Acta Diabetologica",

issn = "0940-5429",

publisher = "Springer Verlag",

}

TY - JOUR

T1 - A novel LIPS assay for insulin autoantibodies

AU - Liberati, Daniela

AU - Wyatt, Rebecca C.

AU - Brigatti, Cristina

AU - Marzinotto, Ilaria

AU - Ferrari, Maurizio

AU - Bazzigaluppi, Elena

AU - Bosi, Emanuele

AU - Gillard, Ben T.

AU - Gillespie, Kathleen M.

AU - Gorus, Frans

AU - Weets, Ilse

AU - Balti, Eric

AU - Piemonti, Lorenzo

AU - Achenbach, Peter

AU - Williams, Alistair J.K.

AU - Lampasona, Vito

PY - 2018/3/1

Y1 - 2018/3/1

N2 - Aims: Insulin autoantibodies (IAA) are often the first marker of autoimmunity detected in children in the preclinical phase of type 1 diabetes (T1D). Currently, the vast majority of laboratories adopt the radiobinding micro-assay (RBA) for measuring IAA. Our aim was to replace RBA with a novel non-radioactive IAA Luciferase Immuno Precipitation System (LIPS) assay with improved performance. Methods: We developed (pro)insulin antigens with alternative placements of a NanoLuc™ luciferase reporter (NLuc). Performance in LIPS was evaluated by testing sera from new onset T1D (n = 80), blood donors (n = 123), schoolchildren (n = 186), first-degree relatives (FDRs) from the Bart’s Oxford family study (n = 53) and from the Belgian Diabetes Registry (n = 136), coded sera from the Islet Autoantibody Standardization Program (IASP) (T1D n = 50, blood donors n = 90). Results: IAA LIPS based on B chain-NLuc proinsulin or B chain-NLuc insulin, in which NLuc was fused at the C-terminus of the insulin B chain, required only 2 μL of serum and a short incubation time, showed high concordance with RBA (Spearman r = 0.866 and 0.833, respectively), high assay performance (B chain-NLuc proinsulin ROC-AUC = 0.894 and B chain-NLuc insulin ROC-AUC = 0.916), and an adjusted sensitivity at 95% specificity ranking on par with the best assays submitted to the two most recent IASP workshops. In FDRs, the IAA LIPS showed improved discrimination of progressors to T1D compared to RBA. Conclusions: We established a novel high-performance non-radioactive IAA LIPS that might replace the current gold standard RBA and find wide application in the study of the IAA response in T1D.

AB - Aims: Insulin autoantibodies (IAA) are often the first marker of autoimmunity detected in children in the preclinical phase of type 1 diabetes (T1D). Currently, the vast majority of laboratories adopt the radiobinding micro-assay (RBA) for measuring IAA. Our aim was to replace RBA with a novel non-radioactive IAA Luciferase Immuno Precipitation System (LIPS) assay with improved performance. Methods: We developed (pro)insulin antigens with alternative placements of a NanoLuc™ luciferase reporter (NLuc). Performance in LIPS was evaluated by testing sera from new onset T1D (n = 80), blood donors (n = 123), schoolchildren (n = 186), first-degree relatives (FDRs) from the Bart’s Oxford family study (n = 53) and from the Belgian Diabetes Registry (n = 136), coded sera from the Islet Autoantibody Standardization Program (IASP) (T1D n = 50, blood donors n = 90). Results: IAA LIPS based on B chain-NLuc proinsulin or B chain-NLuc insulin, in which NLuc was fused at the C-terminus of the insulin B chain, required only 2 μL of serum and a short incubation time, showed high concordance with RBA (Spearman r = 0.866 and 0.833, respectively), high assay performance (B chain-NLuc proinsulin ROC-AUC = 0.894 and B chain-NLuc insulin ROC-AUC = 0.916), and an adjusted sensitivity at 95% specificity ranking on par with the best assays submitted to the two most recent IASP workshops. In FDRs, the IAA LIPS showed improved discrimination of progressors to T1D compared to RBA. Conclusions: We established a novel high-performance non-radioactive IAA LIPS that might replace the current gold standard RBA and find wide application in the study of the IAA response in T1D.

KW - Autoantibodies

KW - Immunoassay

KW - Insulin

KW - LIPS

KW - Prediction

KW - T1D

UR - http://www.scopus.com/inward/record.url?scp=85040029477&partnerID=8YFLogxK

U2 - 10.1007/s00592-017-1082-y

DO - 10.1007/s00592-017-1082-y

M3 - Article (Academic Journal)

C2 - 29305766

AN - SCOPUS:85040029477

SN - 0940-5429

VL - 55

SP - 263

EP - 270

JO - Acta Diabetologica

JF - Acta Diabetologica

ER -

A novel LIPS assay for insulin autoantibodies

Abstract

Keywords

UN SDGs

Access to Document

Handle.net

Other files and links

Fingerprint

Cite this