Simultaneous measurement of Bacillus thuringiensis Cry1Ab and Cry3B proteins in corn extracts.
Sammons-DL; Biagini-RE; Smith-JP; MacKenzie-BA; Striley-CAF; Robertson-SA; Snawder-JE; Ferguson-BS; Larkin-KA
Rational Environmental Management of Agrochemicals: Risk Assessment, Monitoring, and Remedial Action. Kennedy, IR; Solomon KR; Gee SJ; Crossan AN; Wang S; Sanchez-Bayo-F, eds., Washington, DC: American Chemical Society, 2007 Aug; :274-285
Transgenic com hybrids have been developed which contain synthetic genes for the production of Bacillus thuringiensis insecticidal proteins. Two of these proteins are Cry1Ab and Cry3B. Enzyme-linked immunosorbent assays (ELISA) and immunochromatographic lateral flow assays, which are sensitive, specific and easy to perform, have been developed for Cry1Ab and Cry3B proteins in corn extracts. In the present work, we describe, a sandwich fluorescent covalent microsphere immunoassay (FCMIA) to simultaneously measure Cry1Ab and Cry3B proteins in corn extracts. Mixtures of genetically modified organism (GMO) and nonGMO corn were prepared and extracted. The extracts were diluted and Cry1Ab and Cry3B were measured by FCMIA. Measurement of pure Cry1Ab protein by FCMIA gave a minimum detectable concentration of 0.1 ng/mL. There were no significant differences between the assays when performed singly vs. multiplexed (P>0.588). When the observed dilutions from the four parameter logistic fits were compared to the known dilutions, highly linear relationships were observed (r = 0.984 and 0.983, P<0.001) over a dilution range of 1 to 100. These data give proof of principle that FCMIA can be used to simultaneously measure multiple GMO proteins in com and may be a valuable adjunct to existing assays when the number of GMO pesticidal proteins to be measured becomes large.
Genetic-engineering; Insecticides; Pesticides; Analytical-chemistry; Analytical-processes; Bacteria
National Institute for Occupational Safety and Health, Division of Applied Research and Technology, Biomonitoring and Health Assessment Branch, 4676 Columbia Parkway, Cincinnati, OH 45226
Book or book chapter
Kennedy-IR; Solomon-KR; Gee-SJ; Crossan-AN; Wang-S; Sanchez-Bayo-F
Agriculture, Forestry and Fishing; Manufacturing; Construction
Rational Environmental Management of Agrochemicals: Risk Assessment, Monitoring, and Remedial Action