McCormack, Christine C. and Hobson, Audrey H. and Doyle, Sean and Jackson, John and Kilty, Cormac and Whitehead, Alexander S. (1996) Generation of soluble recombinant human acute phase serum amyloid A2 protein and its use in the development of an A-SAA specific ELISA. Journal of Immunological Methods, 198 (1). pp. 101-110. ISSN 0022-1759

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Abstract

Human acute phase serum amyloid A (the A-SAA2 isoform) was expressed at high levels using the pGEX bacterial expression system. A-SAA2 protein was expressed in E. coli NM544 as part of a fusion protein facilitating rapid purification. A-SAA2 was cleaved from the fusion moiety in the presence of a non-ionic detergent (Triton X-100) to release a soluble A-SAA2. Further purification using ion exchange chromatography yielded a pure A-SAA2 (3 mg per litre of culture). Antibodies generated against recombinant A-SAA2 were specific for the acute phase SAAs, A-SAA1 and A-SAA2 and showed no cross-reactivity with the constitutively expressed SAA (C-SAA). These antibodies were used to develop a rapid enzyme-linked immunosorbent assay (ELISA) specific for the measurement of A-SAA in serum.

Item Type:	Article
Keywords:	Recombinant; Serum amyloid A protein; Acute phase protein; Amyloidosis; ELISA;
Academic Unit:	Faculty of Science and Engineering > Biology
Item ID:	7405
Identification Number:	https://doi.org/10.1016/0022-1759(96)00149-4
Depositing User:	Dr. Sean Doyle
Date Deposited:	29 Aug 2016 15:29
Journal or Publication Title:	Journal of Immunological Methods
Publisher:	Elsevier
Refereed:	Yes
URI:	http://www.sciencedirect.com
Use Licence:	This item is available under a Creative Commons Attribution Non Commercial Share Alike Licence (CC BY-NC-SA). Details of this licence are available here

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