Exploring carbohydrate binding module fusions and Fab fragments in a cellulose-based lateral flow immunoassay for detection of cystatin C.

Autor: Natarajan S; Healthcare Technology Innovation Centre, Indian Institute of Technology, Madras, Chennai, 600113, India., Joseph J; Healthcare Technology Innovation Centre, Indian Institute of Technology, Madras, Chennai, 600113, India.; Department of Electrical Engineering, Indian Institute of Technology, ESB 317, Measurements and Instrumentation Laboratory, Chennai, 600036, India., França Prazeres DM; Department of Bioengineering, iBB- Institute for Bioengineering and Biosciences, Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal. miguelprazeres@tecnico.ulisboa.pt.; Associate Laboratory i4HB-Institute for Health and Bioeconomy at Instituto Superior Técnico, Universidade de Lisboa, Av. Rovisco Pais, 1049-001, Lisbon, Portugal. miguelprazeres@tecnico.ulisboa.pt.
Jazyk: angličtina
Zdroj: Scientific reports [Sci Rep] 2022 Mar 31; Vol. 12 (1), pp. 5478. Date of Electronic Publication: 2022 Mar 31.
DOI: 10.1038/s41598-022-09454-9
Abstrakt: This paper presents a lateral flow assay (LFA) for the quantitative, fluorescence-based detection of the kidney biomarker cystatin C that features conjugates of capture antibodies and fusions of carbohydrate binding modules (CBM) with ZZ domains anchored on cellulose deposited over nitrocellulose (NC). The ZZ-CBM3 fusion provides a biomolecular interface between the cellulose layer and the Fc portion of the capture antibodies. By resorting to detection Fab fragments that lack the Fc portion we overcome the observed interference of full-length detection antibodies with the ZZ-CBM3 fusion at the test lines. Using the new LFA architecture, a linear concentration-response relationship was observed in the 0-10 ng/mL cystatin C concentration range, which is compatible with the clinically normal (5-120 ng/mL) and abnormal (> 250 ng/mL) levels of cystatin C, as long as proper dilutions are made. An inter assay CoV of 0.72% was obtained. Finally, mock urine samples characteristic of normal (100 ng/mL) and kidney tubular disease (4000 ng/mL) patients were successfully analyzed. Overall, we demonstrate an innovative LFA architecture that combines NC strips with layered cellulose, ZZ-CBM3 fusions and fluorescently labeled Fab fragments.
(© 2022. The Author(s).)
Databáze: MEDLINE
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