Purification of aflatoxin B1 antibody for the development of aflatoxin biosensor

Aflatoxin B1 (AFB1) is produced from agricultural products especially peanuts overgrown with aspergillus flavus during the post-harvest process. Aflatoxin is classified as a highly toxic and carcinogenic substance to humans by the International Agency for Research on Cancer (IARC), WHO. This researc...

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Bibliographic Details
Main Authors: Prihantoro, E. A. B., Saepudin, E., Ivandini, T. A.
Format: Conference Proceeding
Language:English
Subjects:
Affinity
Biosensors
Carcinogens
Chromatography
Dimethylformamide
Ethanol
Identification methods
Peanuts
Protein A
Proteins
Purification
Serum albumin
Tetrahydrofuran
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Summary:Aflatoxin B1 (AFB1) is produced from agricultural products especially peanuts overgrown with aspergillus flavus during the post-harvest process. Aflatoxin is classified as a highly toxic and carcinogenic substance to humans by the International Agency for Research on Cancer (IARC), WHO. This research was conducted to develop the AFB1 biosensor using antibody that specifically binds to aflatoxin B1. This antibody was produced by injecting an AFB1 hapten-protein (immunogen) to a rabbit. Antibody was obtained from rabbit’s blood serum and purified using Protein A affinity chromatography and precipitation at the isoelectric point. The result showed that purification using protein A contains antibody of 4.0 mg/mL, whereas purification using precipitation at isoelectric pH contains antibody of 0.3 mg/mL. The pure antibody was tested for its specificity against AFB1, tetrahydrofuran (THF), dimethyl formamide (DMF), bovine serum albumin (BSA), and ethanol. The result revealed that THF, BSA, and ethanol were bound to antibody, while DMF showed no interaction. It was concluded that the polyclonal antibody which have been successfully purified from rabbit’s blood serum using protein A affinity chromatography and precipitation methods showed an unspecific identification.
ISSN:0094-243X
1551-7616
DOI:10.1063/1.4991180