title: Phage-displayed peptide that mimics aflatoxins and its application in immunoassay.
abstract: To search for an alternative to using protein conjugated aflatoxin as a coating antigen in aflatoxin detection by an ELISA method, a random-8-peptide library was constructed and used as a source of peptides that mimic aflatoxins (termed as mimotopes). Five mimotope peptides were obtained by panning-elution from the library and were successfully used in an indirect competitive ELISA for analyzing total aflatoxin concentration. The assay exhibited an IC50 value of 14 μg/kg in samples (with 1 in 7 dilution of sample extract) for aflatoxins. The linear range is 4-24 μg/kg. Further validation indicated relatively good recovery (60-120%) in peanut, rice and corn. Natural contaminated samples (peanut and feedstuff) were analyzed for aflatoxin concentration by both conventional ELISA and phage ELISA. The results showed good correlation. It can be concluded that the mimotope preparation is an effective substitute for the aflatoxin based coating antigen in ELISA and can be used in real sample analysis.
authors: Oil Crops Research Institute of the Chinese Academy of Agricultural Sciences , Wuhan 430062, P. R. China.
authors: Yanru
authors: Y
authors: Wang
authors: Hong
authors: H
authors: Wang
authors: Peiwu
authors: P
authors: Li
authors: Qi
authors: Q
authors: Zhang
authors: Hee Joo
authors: HJ
authors: Kim
authors: Shirley J
authors: SJ
authors: Gee
authors: Bruce D
authors: BD
authors: Hammock
mesh_terms: D000348:Aflatoxins / Q000032:analysis*; D010367:Arachis / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005506:Food Contamination / Q000032:analysis*; D012275:Oryza / Q000737:chemistry*; D019151:Peptide Library; D010455:Peptides / Q000032:analysis* / Q000235:genetics / Q000378:metabolism; D003313:Zea mays / Q000737:chemistry*
chemical_list: D000348:Aflatoxins; D019151:Peptide Library; D010455:Peptides
references: Nat Biotechnol. 1997 Apr;15(4):336-42
references: 9094134
references: J Agric Food Chem. 1999 Nov;47(11):4815-20
references: 10552895
references: Proc Natl Acad Sci U S A. 1997 Jan 21;94(2):412-7
references: 9012796
references: J Agric Food Chem. 2005 May 4;53(9):3364-8
references: 15853373
references: Appl Environ Microbiol. 1999 Aug;65(8):3279-86
references: 10427007
references: J Immunol Methods. 1996 Jun 10;192(1-2):73-85
references: 8699024
references: Gene. 1993 Jun 15;128(1):21-7
references: 7685299
references: Environ Sci Technol. 2008 Mar 15;42(6):2047-53
references: 18409635
references: Proc Natl Acad Sci U S A. 1990 Aug;87(16):6378-82
references: 2201029
references: Cell Adhes Commun. 1998 Jan;5(1):75-82
references: 9638343
references: J Appl Microbiol. 2001 Mar;90(3):330-6
references: 11298226
references: Toxicol Lett. 2002 Feb 28;127(1-3):19-28
references: 12052637
references: Anal Chim Acta. 2009 Mar 16;636(1):63-9
references: 19231357
references: Food Cosmet Toxicol. 1968 Dec;6(5):619-25
references: 5753534
references: Biotechnology (N Y). 1995 Feb;13(2):165-9
references: 9634758
references: Food Addit Contam. 2004 Feb;21(2):165-71
references: 14754639
references: Environ Sci Technol. 2005 Jun 1;39(11):4234-41
references: 15984805
references: Scand J Clin Lab Invest Suppl. 1991;205:105-12
references: 1947738
references: Nat Biotechnol. 1996 Jun;14(6):741-4
references: 9630982
references: J Assoc Off Anal Chem. 1987 Sep-Oct;70(5):854-7
references: 3119560
references: J Agric Food Chem. 2004 May 19;52(10):2746-55
references: 15137809
references: Science. 1990 Jul 27;249(4967):386-90
references: 1696028
references: Anal Chim Acta. 2011 Oct 3;703(1):64-9
references: 21843676