Rapid detection of pathogenic bacteria and screening of phage-derived peptides using microcantilevers.

We report the use of an array of microcantilevers to measure the specific binding of Salmonella to peptides derived from phage display libraries. Selectivity of these phage-derived peptides for Salmonella spp. and other pathogens ( Listeria monocytogenes and Escherichia coli ) are compared with a commercially available anti- Salmonella antibody and the antimicrobial peptide alamethicin. A Langmuir isotherm model was applied to determine the binding affinity constants of the peptides to the pathogens. One particular peptide, MSal 020417, demonstrated a higher binding affinity to Salmonella spp. than the commercially available antibody and is able to distinguish among eight Salmonella serovars on a microcantilever. A multiplexed screening system to quickly determine the binding affinities of various peptides to a particular pathogen highly improves the efficiency of the peptide screening process. Combined with phage-derived peptides, this microcantilever-based technique provides a novel biosensor to rapidly and accurately detect pathogens and holds potential to be further developed as a screening method to identify pathogen-specific recognition elements.

[1]  Christopher T. Elliott,et al.  Phage Display-Derived Binders Able to Distinguish Listeria monocytogenes from Other Listeria Species , 2013, PloS one.

[2]  C. Elliott,et al.  Production and evaluation of the utility of novel phage display‐derived peptide ligands to Salmonella spp. for magnetic separation , 2013, Journal of applied microbiology.

[3]  U. Tamer,et al.  Comparison of sensing strategies in SPR biosensor for rapid and sensitive enumeration of bacteria. , 2012, Biosensors & bioelectronics.

[4]  Michael C. McAlpine,et al.  Electrical detection of pathogenic bacteria via immobilized antimicrobial peptides , 2010, Proceedings of the National Academy of Sciences.

[5]  S. Biswal,et al.  Using microcantilevers to study the interactions of lipid bilayers with solid surfaces. , 2010, Analytical chemistry.

[6]  A. Ramanavičius,et al.  Comparative study of random and oriented antibody immobilization techniques on the binding capacity of immunosensor. , 2010, Analytical chemistry.

[7]  Khalil Arshak,et al.  An overview of foodborne pathogen detection: in the perspective of biosensors. , 2010, Biotechnology advances.

[8]  M. Sisido,et al.  A novel method for screening peptides that bind to proteins by using multiple fluorescent amino acids as fluorescent tags. , 2010, Chemical communications.

[9]  Etienne Weiss,et al.  Therapeutic antibodies: successes, limitations and hopes for the future , 2009, British journal of pharmacology.

[10]  Marek Piliarik,et al.  High-throughput SPR sensor for food safety. , 2009, Biosensors & bioelectronics.

[11]  Bong Hyun Chung,et al.  Recent advances in immobilization methods of antibodies on solid supports. , 2008, The Analyst.

[12]  V. Wu,et al.  Using oligonucleotide-functionalized Au nanoparticles to rapidly detect foodborne pathogens on a piezoelectric biosensor. , 2008, Journal of microbiological methods.

[13]  L. A. Pinnaduwage,et al.  An energy conservation approach to adsorbate-induced surface stress and the extraction of binding energy using nanomechanics , 2008, Nanotechnology.

[14]  T. Thundat,et al.  Detection of Cd(II) using antibody-modified microcantilever sensors. , 2007, Ultramicroscopy.

[15]  C. Vágvölgyi,et al.  The History of Alamethicin: A Review of the Most Extensively Studied Peptaibol , 2007, Chemistry & biodiversity.

[16]  Olivier Lazcka,et al.  Pathogen detection: a perspective of traditional methods and biosensors. , 2007, Biosensors & bioelectronics.

[17]  A. Majumdar,et al.  Nanomechanical detection of DNA melting on microcantilever surfaces. , 2006, Analytical chemistry.

[18]  Kiyoshi Toko,et al.  Preparation and characterization of a polyclonal antibody from rabbit for detection of trinitrotoluene by a surface plasmon resonance biosensor. , 2005, Talanta.

[19]  Heidi S Erickson,et al.  Layered peptide arrays: high-throughput antibody screening of clinical samples. , 2005, The Journal of molecular diagnostics : JMD.

[20]  H. Hoogenboom,et al.  Selecting and screening recombinant antibody libraries , 2005, Nature Biotechnology.

[21]  A. Bradbury,et al.  Antibodies from phage antibody libraries. , 2004, Journal of immunological methods.

[22]  R. Siegel,et al.  Yeast display of antibody fragments: a discovery and characterization platform. , 2004, Journal of immunological methods.

[23]  Tom Humphrey,et al.  Salmonella, stress responses and food safety , 2004, Nature Reviews Microbiology.

[24]  Pina M Fratamico,et al.  Comparison of culture, polymerase chain reaction (PCR), TaqMan Salmonella, and Transia Card Salmonella assays for detection of Salmonella spp. in naturally-contaminated ground chicken, ground turkey, and ground beef. , 2003, Molecular and cellular probes.

[25]  H. Lang,et al.  Multiple label-free biodetection and quantitative DNA-binding assays on a nanomechanical cantilever array , 2002, Proceedings of the National Academy of Sciences of the United States of America.

[26]  M. Zasloff.  Antimicrobial peptides of multicellular organisms , 2002, Nature.

[27]  K. Demnerova,et al.  Detection of Salmonella in food samples by the combination of immunomagnetic separation and PCR assay. , 2000, International microbiology : the official journal of the Spanish Society for Microbiology.

[28]  A P Turner,et al.  Biosensors--Sense and Sensitivity , 2000, Science.

[29]  Ihab Abdel-Hamid,et al.  Highly sensitive flow-injection immunoassay system for rapid detection of bacteria , 1999 .

[30]  Norio Miura,et al.  Highly selective and sensitive SPR immunosensor for detection of methamphetamine , 1999 .

[31]  I. Park,et al.  Thiolated Salmonella antibody immobilization onto the gold surface of piezoelectric quartz crystal. , 1998, Biosensors & bioelectronics.

[32]  Norio Miura,et al.  A surface plasmon resonance-based immunosensor for highly sensitive detection of morphine , 1998 .

[33]  N. Yamazoe,et al.  Evaluation of binding of human serum albumin (HSA) to monoclonal and polyclonal antibody by means of piezoelectric immunosensing technique , 1997 .

[34]  G. Lubrano,et al.  A quartz crystal microbalance displacement assay for Listeria monocytogenes , 1996 .

[35]  J H Luong,et al.  Development of a flow injection analysis (FIA) immunosensor for the detection of Escherichia coli. , 1995, International journal of food microbiology.

[36]  H R Hoogenboom,et al.  By-passing immunization. Human antibodies from V-gene libraries displayed on phage. , 1991, Journal of molecular biology.

[37]  R. Kuriyama,et al.  Differential pathways of recruitment for centrosomal antigens to the mitotic poles during bipolar spindle formation. , 1991, Journal of cell science.

[38]  R. Barrett,et al.  Peptides on phage: a vast library of peptides for identifying ligands. , 1990, Proceedings of the National Academy of Sciences of the United States of America.

[39]  G. Stoney.  The Tension of Metallic Films Deposited by Electrolysis , 1909 .