Development of a competitive liposome-based lateral flow assay for the rapid detection of the allergenic peanut protein Ara h1.
A competitive lateral flow assay for detecting the major peanut allergen, Ara h1, has been developed. The detector reagents are Ara h1-tagged liposomes, and the capture reagents are anti-Ara h1 polyclonal antibodies. Two types of rabbit polyclonal antibodies were raised either against the entire Ara h1 molecules (anti-Ara h1 Ab) or against an immunodominant epitope on Ara h1 (anti-peptide Ab). All of them reacted specifically with Ara h1 in Western Blot against crude peanut proteins. Moreover, the anti-Ara h1 Ab was chosen for this assay development because of its highest immunoactivity to Ara h1-tagged liposomes in the lateral flow assay. The calculated limit of detection (LOD) of this assay is 0.45 microg mL(-1) of Ara h1 with a dynamic range between 0.1 and 10 microg mL(-1) of Ara h1 in buffer. Additionally, the visually determined detection range is from 1 to 10 microg mL(-1) of Ara h1 in buffer. Results using this assay can be obtained within 30 min without the need of sophisticated equipment or techniques; therefore, this lateral flow assay has the potential to be a cost-effective, fast, simple, and sensitive method for on-site screening of peanut allergens.
Wen
Hsiao-Wei
HW
Department of Food Science and Technology, Cornell University, 630 West North Street, Geneva, NY 14456-0462, USA.
Borejsza-Wysocki
Wlodzimierz
W
DeCory
Thomas R
TR
Durst
Richard A
RA
D000485:Allergens / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D000595:Amino Acid Sequence; D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D000917:Antibody Formation; D052179:Antigens, Plant; D010367:Arachis / Q000276:immunology*; D015153:Blotting, Western; D003001:Cloning, Molecular; D004591:Electrophoresis, Polyacrylamide Gel; D004797:Enzyme-Linked Immunosorbent Assay; D006023:Glycoproteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D007202:Indicators and Reagents; D008081:Liposomes; D008565:Membrane Proteins; D008969:Molecular Sequence Data; D010940:Plant Proteins / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D011817:Rabbits; D012680:Sensitivity and Specificity
D000485:Allergens; D000906:Antibodies; D052179:Antigens, Plant; C500889:Ara h 1 protein, Arachis hypogaea; D006023:Glycoproteins; D007202:Indicators and Reagents; D008081:Liposomes; D008565:Membrane Proteins; D010940:Plant Proteins