Detection of biological threats. A challenge for directed molecular evolution.
The probe technique originated from early attempts of Anton van Leeuwenhoek to contrast microorganisms under the microscope using plant juices, successful staining of tubercle bacilli with synthetic dyes by Paul Ehrlich and discovery of a stain for differentiation of gram-positive and gram-negative bacteria by Hans Christian Gram. The technique relies on the principle that pathogens have unique structural features, which can be recognized by specifically labeled organic molecules. A hundred years of extensive screening efforts led to discovery of a limited assortment of organic probes that are used for identification and differentiation of bacteria. A new challenge--continuous monitoring of biological threats--requires long lasting molecular probes capable of tight specific binding of pathogens in unfavorable conditions. To respond to the challenge, probe technology is being revolutionized by utilizing methods of combinatorial chemistry, phage display and directed molecular evolution. This review describes how molecular evolution methods are applied for development of peptide, antibody and phage probes, and summarizes the author's own data on development of landscape phage probes against Salmonella typhimurium. The performance of the probes in detection of Salmonella is illustrated by a precipitation test, enzyme-linked immunosorbent assay (ELISA), fluorescence-activated cell sorting (FACS) and fluorescent, optical and electron microscopy.
Petrenko
Valery A
VA
Department of Pathobiology, College of Veterinary Medicine, Auburn University, 253 Greene Hall, Auburn, AL 36849, USA. petreva@vetmed.auburn.edu
Sorokulova
Iryna B
IB
D000906:Antibodies; D023482:Bioterrorism / Q000517:prevention & control; D019020:Directed Molecular Evolution / Q000379:methods; D004783:Environmental Microbiology; D004784:Environmental Monitoring / Q000379:methods; D005516:Food Microbiology; D008828:Microbiological Techniques; D015336:Molecular Probe Techniques; D019151:Peptide Library; D012475:Salmonella / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification; D012478:Salmonella Food Poisoning / Q000517:prevention & control
D000906:Antibodies; D019151:Peptide Library