Isolation of recombinant antibodies against EspA and intimin of Escherichia coli O157:H7

Journal of Clinical Microbiology
Sarah A KühneKevin C Gough

Abstract

Intimin, Tir, and EspA proteins are expressed by attaching-effacing Escherichia coli, which include enteropathogenic and enterohemorrhagic E. coli pathotypes. EspA proteins are part of the type three secretion system needle complex that delivers Tir to the host epithelial cell, while surface arrayed intimin docks the bacterium to the translocated Tir. This intimate attachment leads to attaching and effacing lesions. Recombinant forms of these effector proteins from enterohemorrhagic E. coli O157:H7 were produced by using E. coli expression vectors. Binding of intimin and Tir fragments in enzyme-linked immunosorbent assay (ELISAs) demonstrated the interaction of intimin fragments containing the C-terminal 282 or 188 amino acids to a Tir fragment containing amino acid residues 258 to 361. Recombinant intimin and EspA proteins were used to elicit immune responses in rabbits and immune phage-display antibody libraries were produced. Screening of these immune libraries by conventional phage-antibody panning and colony filter screening produced a panel of antibodies with specificity for EspA or intimin. Antibodies recognizing different C-terminal epitopes on intimin bound specifically to the gamma intimin of O157:H7 and not to other ...Continue Reading

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Citations

Dec 10, 2009·Theoretical Biology & Medical Modelling·Jafar AmaniAli H Salmanian
Feb 4, 2011·BMC Research Notes·Márcio A MenezesRoxane Mf Piazza
Feb 24, 2016·Protein Engineering, Design & Selection : PEDS·Teruyo Ojima-KatoHideo Nakano
Mar 23, 2011·Plant Science : an International Journal of Experimental Plant Biology·Jafar AmaniAli Hatef Salmanian
Jun 24, 2017·Journal of Biotechnology·Mieko Kato, Yoshiro Hanyu
Feb 14, 2008·International Journal of Cancer. Journal International Du Cancer·Alessandra VillaDario Neri
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Feb 6, 2018·Clinical and Experimental Vaccine Research·Zahra Sadat HosseiniFatemeh Shafighian
Oct 28, 2006·The Journal of Biological Chemistry·Rémy RobertGisèle Clofent-Sanchez

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