Protein-chip for autoantibodies profile detection

We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solu...

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Bibliographic Details
Published in:Shengwu gongcheng xuebao Vol. 24; no. 8; p. 1496
Main Authors: Wang, Guoqiang, Gao, Xueqin, Han, Jinxiang, Pan, Jihong, Huang, Hainan
Format: Journal Article
Language:Chinese
Published: China 01.08.2008
Subjects:
Autoantibodies - blood
Autoantibodies - immunology
Autoimmune Diseases - blood
Autoimmune Diseases - immunology
Humans
Protein Array Analysis - instrumentation
Protein Array Analysis - methods
Sensitivity and Specificity
ISSN:1000-3061
Online Access:Get more information
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Summary:We selected 12 antigens corresponding to commonly used autoantibodies in clinical practice to prepare antigen microarray. We chose NBT/BCIP color reaction as the end detection strategy to develop a new autoantibody protein chip detection system. Using this system, we optimized the best spotting solution, spotting concentration of the 12 antigens and the dilution of serum. We prepared a protein chip that could detect 12 autoantibodies simultaneously using the optimized antigen concentration. We established a new method to determine the cutoff of each autoantibodies by evaluation of 678 positive and 120 negative serum of clinical sample. We also evaluated the sensitivity and specificity of our new detection system. The optimal spotting solution was 0.1% TBST, the dilution of serum was 1:4 and the best spotting concentration of the 12 antigens were ANA 520 microg/mL, Ro-60/SSa 465 microg/mL, La/SSb 530 microg/mL, Jo-1 530 microg/mL, Scl-70 525 microg/mL, Sm 520 microg/mL, Ro-52/SSa 615 microg/mL, RF 340 microg/m
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ISSN:1000-3061