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Covalent Binding of Antibodies to Cellulose Paper Discs and Their Applications in Naked-eye Colorimetric Immunoassays
- Publication Year :
- 2016
- Publisher :
- MyJove Corporation, 2016.
-
Abstract
- This report presents two methods for the covalent immobilization of capture antibodies on cellulose filter paper grade No. 1 (medium-flow filter paper) discs and grade No. 113 (fast-flow filter paper) discs. These cellulose paper discs were grafted with amine functional groups through a silane coupling technique before the antibodies were immobilized on them. Periodate oxidation and glutaraldehyde cross-linking methods were used to graft capture antibodies on the cellulose paper discs. In order to ensure the maximum binding capacity of the capture antibodies to their targets after immobilization, the effects of various concentrations of sodium periodate, glutaraldehyde, and capture antibodies on the surface of the paper discs were investigated. The antibodies that were coated on the amine-functionalized cellulose paper discs through a glutaraldehyde cross-linking agent showed enhanced binding activity to the target when compared to the periodate oxidation method. IgG (in mouse reference serum) was used as a reference target in this study to test the application of covalently immobilized antibodies through glutaraldehyde. A new paper-based, enzyme-linked immunosorbent assay (ELISA) was successfully developed and validated for the detection of IgG. This method does not require equipment, and it can detect 100 ng/ml of IgG. The fast-flow filter paper was more sensitive than the medium-flow filter paper. The incubation period of this assay was short and required small sample volumes. This naked-eye, colorimetric immunoassay can be extended to detect other targets that are identified with conventional ELISA.
- Subjects :
- Paper
General Chemical Engineering
Enzyme-Linked Immunosorbent Assay
02 engineering and technology
01 natural sciences
Biochemistry
General Biochemistry, Genetics and Molecular Biology
Antibodies
chemistry.chemical_compound
Mice
medicine
Animals
Cellulose
Chromatography
General Immunology and Microbiology
Filter paper
medicine.diagnostic_test
Chemistry
Sodium periodate
General Neuroscience
010401 analytical chemistry
Periodate
021001 nanoscience & nanotechnology
0104 chemical sciences
Covalent bond
Immunoassay
Colorimetry
Glutaraldehyde
Naked eye
0210 nano-technology
Subjects
Details
- Language :
- English
- Database :
- OpenAIRE
- Accession number :
- edsair.doi.dedup.....35e13da118c21e811e440b4f6960bc1b