Antigenic epitopes on the outer membrane protein A of Escherichia coli identified with single-chain variable fragment (scFv) antibodies

2019 ◽  
Vol 103 (13) ◽  
pp. 5285-5299 ◽  
Author(s):  
Pharaoh Fellow Mwale ◽  
Chi-Hsin Lee ◽  
Sy-Jye Leu ◽  
Yu-Ching Lee ◽  
Hsueh-Hsia Wu ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein A ◽  
Single Chain Variable Fragment ◽  
Single Chain ◽  
Outer Membrane Protein A ◽  
Antigenic Epitopes

Overexpression of Outer Membrane Protein A Is a Risk Factor for Mortality in Escherichia coli Bloodstream Infections

2019 ◽  
Author(s):  
Ángel Rodríguez-Villodres ◽  
Rocío Álvarez-Marín ◽  
María Antonia Pérez-Moreno ◽  
Andrea Miró-Canturri ◽  
Marco Durán Lobato ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Risk Factor ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein A ◽  
Bloodstream Infections ◽  
Outer Membrane Protein A

scholarly journals Recombinant outer membrane protein A fragments protect against Escherichia coli meningitis

2016 ◽  
Vol 49 (3) ◽  
pp. 329-334 ◽  
Author(s):  
Wen-Shyang Hsieh ◽  
Yi-Yuan Yang ◽  
Hsin-Yi Yang ◽  
Yu-Shan Huang ◽  
Hsueh-Hsia Wu
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein A ◽  
Outer Membrane Protein A

scholarly journals Allelic Variation in Outer Membrane Protein A and Its Influence on Attachment of Escherichia coli to Corn Stover

2017 ◽  
Vol 8 ◽  
Author(s):  
Chunyu Liao ◽  
Xiao Liang ◽  
Fan Yang ◽  
Michelle L. Soupir ◽  
Adina C. Howe ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Corn Stover ◽  
Outer Membrane Protein ◽  
Allelic Variation ◽  
Protein A ◽  
Outer Membrane Protein A

scholarly journals Cloning and Expression of the Escherichia coli K1 Outer Membrane Protein A Receptor, a gp96 Homologue

2003 ◽  
Vol 71 (4) ◽  
pp. 1680-1688 ◽  
Author(s):  
Nemani V. Prasadarao ◽  
Pramod K. Srivastava ◽  
Rajyalakshmi S. Rudrabhatla ◽  
Kwang Sik Kim ◽  
Sheng-he Huang ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein A ◽  
Tumor Rejection ◽  
Cloning And Expression ◽  
Cdna Expression Library ◽  
E Coli ◽  
Outer Membrane Protein A

ABSTRACT Escherichia coli is one of the most common gram-negative bacteria that cause meningitis in neonates. Our previous studies have shown that outer membrane protein A (OmpA) of E. coli interacts with a 95-kDa human brain microvascular endothelial cell (HBMEC) glycoprotein, Ecgp, for invasion. Here, we report the identification of a gene that encodes Ecgp by screening of an HBMEC cDNA expression library as well as by 5′ rapid amplification of cDNA ends. The sequence of the Ecgp gene shows that it is highly similar to gp96, a tumor rejection antigen-1, and contains an endoplasmic reticulum retention signal, KDEL. Overexpression of either Ecgp or gp96 in both HBMECs and CHO cells increases E. coli binding and invasion. We further show that Ecgp gene-transfected HBMECs express Ecgp on the cell surface despite the presence of the KDEL motif. Northern blot analysis of total RNA from various eukaryotic cells indicates that Ecgp is significantly expressed in HBMECs. Recombinant His-tagged Ecgp blocked E. coli invasion efficiently by binding directly to the bacteria. These results suggest that OmpA of E. coli K1 interacts with a gp96-like molecule on HBMECs for invasion.

scholarly journals Immunoprotective evaluation of Escherichia coli outer membrane protein A against the main pathogens of animal mastitis

2020 ◽  
Vol 19 (1) ◽  
pp. 155-162
Author(s):  
Chen Chen ◽  
Nana Wu ◽  
Na Rong ◽  
Chao Kang ◽  
Chunlin Chen ◽  
...  
Keyword(s):  
Escherichia Coli ◽  
Membrane Protein ◽  
Outer Membrane ◽  
Outer Membrane Protein ◽  
Protein A ◽  
Enzyme Linked Immunosorbent Assay ◽  
Protective Function ◽  
E Coli ◽  
Ompa Protein ◽  
Outer Membrane Protein A

Purpose: To evaluate prokaryotic expression of the Escherichia coli (E. coli) outer membrane protein A (OmpA) and its immunoprotective function against the main pathogens of animal mastitis.Methods: A molecular cloning method was used to develop a prokaryotic strain expressing OmpA protein, which was purified by Ni-affinity  chromatography. Polyclonal antiserum was generated in mice immunized with OmpA protein. Enzyme-linked immunosorbent assay (ELISA) and western blotting were used to determine the titer and verify anti-OmpA serum specificity, respectively. Interaction between OmpA antiserum and main pathogens of animal mastitis was verified by ELISA and a pull-down method. The immune protective function of OmpA protein was evaluated in mice challenged with pathogens of animal mastitis. Optimal fermentation conditions to produce OmpA protein were determined by the L9(34) orthogonal test.Results: A prokaryotic strain expressing OmpA protein was developed, and purified OmpA was used to develop a mouse polyclonal antibody. The anti-OmpA serum exhibited high specificity and a titer of 1:1600. Anti-OmpA serum directly interacted with E. coli and Staphylococcus aureus (S. aureus). OmpA demonstrated a significant immune protective function of 58.33 % against E. coli and 46.15 % against S. aureus. The optimal conditions for expressing fermentation OmpA were a strain absorbance of 0.5 at a wavelength of 600 nm, IPTG final concentration of 0.3 mmol/L, induction time of 12 h, and induction temperature of 28 °C.Conclusion: OmpA possesses selective immunogenicity and a significant immune protective effect against the main pathogens of animal mastitis. The results suggest that OmpA may potentially be used as a vaccine for animal mastitis. Keywords: E. coli, OmpA protein, Immunoprotection, Animal mastitis, Protein fermentation

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