A highly stable, sensitive, regenerable and rapid immunoassay for detecting aflatoxin B1 in corn incorporating covalent AFB1 immobilization and a recombinant Fab antibody.
A highly robust immunoassay applicable for the detection of aflatoxin B1 (AFB1) using a Fab antibody fragment was developed. A key factor was the use of covalently immobilized AFB1 which allowed an almost three fold increase in sensitivity, reduced assay time and regeneration with retention of binding capacity. Various factors that might affect the sensitivity of the assay such as pH, organic solvents, storage stability and wash stringency were critically evaluated. It was also demonstrated that the assay was applicable for determination of AFB1 in corn samples at concentration within the European union regulatory limits.
Applied Biochemistry Group (ABG), School of Biotechnology, Dublin City University, Dublin 9, Ireland. Electronic address: soujanyaratna@gmail.com.
Soujanya Ratna
SR
Edupuganti
Om Prakash
OP
Edupuganti
Stephen
S
Hearty
Richard
R
O'Kennedy
D016604:Aflatoxin B1 / Q000032:analysis*; D000906:Antibodies / Q000737:chemistry*; D055910:Antibodies, Immobilized / Q000096:biosynthesis / Q000737:chemistry*; D000915:Antibody Affinity; D000918:Antibody Specificity; D005062:European Union; D006863:Hydrogen-Ion Concentration; D007118:Immunoassay; D007140:Immunoglobulin Fab Fragments / Q000096:biosynthesis / Q000737:chemistry*; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012997:Solvents; D003313:Zea mays / Q000737:chemistry*
D000906:Antibodies; D055910:Antibodies, Immobilized; D007140:Immunoglobulin Fab Fragments; D011994:Recombinant Proteins; D012997:Solvents; D016604:Aflatoxin B1
Amide bond; Covalent immobilization; Enzyme-linked immunosorbent assay (ELISA); Passive adsorption; Re-usability