A sensitive paper-based lateral flow immunoassay platform using engineered cellulose-binding protein linker fused with antibody-binding domains

Jong Min Yang, Kyeong Rok Kim, Sangmin Jeon, Hyung Joon Cha, Chang Sup Kim

Research output: Contribution to journalArticlepeer-review

26 Scopus citations

Abstract

An advanced paper-based LFIA using engineered cellulose-binding protein linker fused with antibody-binding domains was developed to resolve problems associated with conventional LFIA strip, including the limitation of quantitative analysis and low sensitivity. CBP31-BC linker was prepared by genetically fusing the cellulose-binding modules of family 3 (CBM3) and family 1 (CBM1) with antibody-binding B and C domains of Protein A. Cellulose-binding analysis showed that the addition of two CBMs led to high cellulose-binding capacity in CBP31-BC. Transmission electron microscopy and quartz crystal microbalance analyses demonstrated that the cellulose-binding ability of CBP31-BC enabled the BC domain to be efficiently exposed, allowing about 6-fold higher antibody-binding efficiency (∼32.5 %) than that (∼5.5 %) of sole BC. To evaluate the feasibility of the CBP31-BC linker-employed LFIA platform, an immunoassay of prostate-specific antigen (PSA) was performed as a model reaction. The CBP31-BC-employed paper-based LFIA detected PSA at levels as low as 0.25 ng/mL in 20 min, which is about 10-fold more sensitive to conventional LFIAs that use simple physical adsorption. CBP31-BC-employed LFIA also exhibited a lower detection range of 0.25–2.5 ng/mL when compared to conventional LFIAs. These results might be due to efficient oriented immobilization of the capture antibody by CBP31-BC linker on cellulose membrane. Thus, our developed CBP31-BC linker-employed paper-based LFIA platform holds great promise for the detection of target markers due to significantly enhanced sensitivity.

Original languageEnglish
Article number129099
JournalSensors and Actuators B: Chemical
Volume329
DOIs
StatePublished - 15 Feb 2021

Keywords

  • Antibody-binding domain
  • Cellulose membrane
  • Cellulose-binding module
  • Lateral flow immunoassay
  • Oriented immobilization
  • Sensitivity

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