Abstract
The current work describes the elaboration of a simple, sensitive and reliable β-casein modified glass beads, for the detection and quantification of its specific antibody anti β-casein. This is an elementary receptor without electronic part, developed by grafting glass bead surface with the antigenic β-casein via 3-aminopropyltriethoxy silane and then glutaraldehyde as cross-linker. The whole is realized by a classical process, called in two steps and in mild conditions where chemical protocol is optimized for β-casein use. The detection and quantification of the specific reaction antibody–receptor is carried out by the technique of the second antibody labeled with horse radish peroxidase (HRP). Our receptor can detect the β-casein antibody present in the serum at dilutions up to a factor 10 7 in strong ionic strength medium. The same antibody of the same serum and in the same conditions can be detected by ELISA test at dilutions up to a factor 10 5. The whole test, after our receptor realization, takes about 5 h.
Highlights
The current work describes the elaboration of a simple, sensitive and reliable -casein modified glass beads, for the detection and quantification of its specific antibody anti -casein
From the results we obtained during the quantification of the reaction between antibody anti -casein and antigen fixed on the glass beads, that the grafting is functional
The receptor can detect very low antibody concentrations compared to ELISA test
Summary
The current work describes the elaboration of a simple, sensitive and reliable -casein modified glass beads, for the detection and quantification of its specific antibody anti -casein. It was an interesting challenge to develop a simple and fast quantitative receptor, using antigenic -casein from cow’s milk, for specific antibodies’ titration and give rise to screening tests via enzyme immunoassay (EIA) measurements In this purpose, we grafted on a glass substrate (5 mm diameter glass bead) [11,12] the -casein by a cross-linking reaction in two steps [13]. We optimized and tested this functional grafting on the glass beads in 96-well EIA plate thanks to the specific reaction of this receptor formed with its corresponding antibody anti -casein The quantification of this specific reaction is carried out by the technique of the second labeled antibody. We compared the sensitivity of our elementary receptor with that of ELISA test versus the same antibody
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