Abstract
Autoantibody detection is a useful and obligatory tool for clinicians and researchers in diagnosing autoimmune diseases. Line blotting is a simple, sensitive, and flexible technique for fast semiquantitative detection of multiple antibodies. Line blotting enables the detection of antibodies on membrane strips coated with thin parallel lines of several purified, biochemically characterized antigens, which are fixed onto a synthetic support. Each strip can contain more than ten antigens, thus allowing simultaneous screening for multiple antibodies. Here, we describe the principle of line blotting and how the buffer composition can interfere with the results of autoantibody detection.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Baslund B, Wieslander J (1994) A rapid and semi-quantitative method for the detection of autoantibodies by multiple spot immunoassay. J Immunol Methods 169(2):183–193
Noya O, Losada S, Toledo M, Gauna A, Lorenzo MA, Bermúdez H, Alarcón de Noya B (2015) Improvements and variants of the multiple antigen blot assay-MABA: an immunoenzymatic technique for simultaneous antigen and antibody screening. Methods Mol Biol. 1312:301–319
Stott DI (2000) Immunoblotting, dot-blotting, and ELISPOT assays: methods and applications. J Immunoassay 21(2–3):273–296
Mishra M, Tiwari S, Gomes AV (2017) Protein purification and analysis: next generation Western blotting techniques. Expert Rev Proteomics 14(11):1037–1053
Kurien BT, Scofield RH (2015) Western blotting: an introduction. Methods Mol Biol. 1312:17–30
Stott DI (1989) Immunoblotting and dot blotting. J Immunol Methods 119(2):153–187
Noya O, Losada S, Toledo M, Alarcón de Noya B (2009) The multiple antigen blot assay: a simple, versatile and multipurpose immunoenzymatic technique. Methods Mol Biol. 536:237–251
Dorri Y (2015) Immunoblotting of antigens: whole, strip, and new-line nitrocellulose membrane immunoblotting using the chemiluminescence technique. Methods Mol Biol. 1314:79–94
Kurien BT (2009) Strip immunoblotting of multiple antigenic peptides to nitrocellulose membrane. Methods Mol Biol 536:191–199
Gallo-Oller G, Ordoñez R, Dotor J (2018) A new background subtraction method for Western blot densitometry band quantification through image analysis software. J Immunol Methods 467:1–5
Gassmann M, Grenacher B, Rohde B, Vogel J (2009) Quantify Western blots: pitfalls of densitometry. Electrophoresis 30:1845–1855
Murphy K, Janeway CA, Travers P, Walport M, Mowart A, Weaver CT (2012) Janeway’s immuno biology, 8th edn. Garland Science, New York
Borg EJ, Risselada AP, Kelder JC (2011) Relation of systemic autoantibodies to the number of extraglandular manifestations in primary Sjögren’s syndrome: a retrospective analysis of 65 patients in the Netherlands. Semin Arthritis Rheum 40(6):547–551
Ciechomska M, Laar JV, O’Reilly S (2015) Current frontiers in systemic sclerosis pathogenesis. Exp Dermatol 24(6):401–406
Saito H, Takahashi A, Abe K, Okai K, Katsushima F, Monoe K, Kanno Y, Ohira H (2012) Autoantibodies by line immunoassay in patients with primary biliary cirrhosis. Fukushima J Med Sci 58(2):107–116
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2019 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Sternbæk, L., Friis, T., Houen, G. (2019). Determination of Autoantibodies by Line Immunoblotting. In: Houen, G. (eds) Autoantibodies. Methods in Molecular Biology, vol 1901. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8949-2_22
Download citation
DOI: https://doi.org/10.1007/978-1-4939-8949-2_22
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-8948-5
Online ISBN: 978-1-4939-8949-2
eBook Packages: Springer Protocols