Phage display-derived antibody fragments against conserved regions of VacA toxin of Helicobacter pylori

Phage display-derived antibody fragments against conserved regions of VacA toxin of Helicobacter... Infection with Helicobacter pylori may result in the emergence of gastric adenocarcinoma. Among various toxins assisting pathogenesis of H. pylori, the vacuolating cytotoxin A (VacA) is one of the most potent toxins known as the major cause of the peptic ulcer and gastric adenocarcinoma. To isolate single-chain variable fragments (scFvs) against two conserved regions of VacA, we capitalized on the phage display technology and a solution-phase biopanning (SPB). Characterization of scFvs was carried out by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and surface plasmon resonance (SPR). Bioinformatics analyses were also performed in order to characterize the structural and functional properties of the isolated scFvs and the interaction(s) between the isolated antibodies (Ab)-antigen (Ag). After four rounds of biopanning, the positive colonies detected by scFv ELISA were harvested to extract the plasmids and perform sequencing. Of several colonies, three colonies showed high affinity to the VacA1 and two colonies for the VacA2. Further complementary examinations (e.g., sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, SPR, and flow cytometry) displayed the high affinity and specificity of the isolated scFvs to the VacA. Docking results revealed the interaction of the complementarity-determining regions (CDRs) with the VacA peptide. In conclusion, for the first time, we report on the isolation of several scFvs against conserved residues of VacA toxin with high affinity and specificity, which may be used as novel diagnostic/therapeutic tool in the H. pylori infection. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png Applied Microbiology and Biotechnology Springer Journals

Phage display-derived antibody fragments against conserved regions of VacA toxin of Helicobacter pylori

15 pages
Read Article

Loading next page...
 
/lp/springer_journal/phage-display-derived-antibody-fragments-against-conserved-regions-of-DCMdPIBhx0
Publisher
Springer Journals
Copyright
Copyright © 2018 by Springer-Verlag GmbH Germany, part of Springer Nature
Subject
Life Sciences; Microbiology; Microbial Genetics and Genomics; Biotechnology
ISSN
0175-7598
eISSN
1432-0614
D.O.I.
10.1007/s00253-018-9068-4
Publisher site
See Article on Publisher Site

Abstract

Infection with Helicobacter pylori may result in the emergence of gastric adenocarcinoma. Among various toxins assisting pathogenesis of H. pylori, the vacuolating cytotoxin A (VacA) is one of the most potent toxins known as the major cause of the peptic ulcer and gastric adenocarcinoma. To isolate single-chain variable fragments (scFvs) against two conserved regions of VacA, we capitalized on the phage display technology and a solution-phase biopanning (SPB). Characterization of scFvs was carried out by enzyme-linked immunosorbent assay (ELISA), immunoblotting, and surface plasmon resonance (SPR). Bioinformatics analyses were also performed in order to characterize the structural and functional properties of the isolated scFvs and the interaction(s) between the isolated antibodies (Ab)-antigen (Ag). After four rounds of biopanning, the positive colonies detected by scFv ELISA were harvested to extract the plasmids and perform sequencing. Of several colonies, three colonies showed high affinity to the VacA1 and two colonies for the VacA2. Further complementary examinations (e.g., sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), western blot, SPR, and flow cytometry) displayed the high affinity and specificity of the isolated scFvs to the VacA. Docking results revealed the interaction of the complementarity-determining regions (CDRs) with the VacA peptide. In conclusion, for the first time, we report on the isolation of several scFvs against conserved residues of VacA toxin with high affinity and specificity, which may be used as novel diagnostic/therapeutic tool in the H. pylori infection.

Journal

Applied Microbiology and Biotechnology – Springer Journals

Published: Jun 3, 2018

References

  • Helicobacter pylori and oral pathology: relationship with the gastric infection
    Adler, I; Muino, A; Aguas, S; Harada, L; Diaz, M; Lence, A; Labbrozzi, M; Muino, JM; Elsner, B; Avagnina, A; Denninghoff, V
  • Phage display as a promising approach for vaccine development
    Aghebati-Maleki, L; Bakhshinejad, B; Baradaran, B; Motallebnezhad, M; Aghebati-Maleki, A; Nickho, H; Yousefi, M; Majidi, J
  • scFv antibody: principles and clinical application
    Ahmad, ZA; Yeap, SK; Ali, AM; Ho, WY; Alitheen, NB; Hamid, M
  • High-throughput screening of single-chain antibodies using multiplexed flow cytometry
    Ayriss, J; Woods, T; Bradbury, A; Pavlik, P
  • A fast mutagenesis procedure to recover soluble and functional scFvs containing amber stop codons from synthetic and semisynthetic antibody libraries
    Barderas, R; Shochat, S; Martinez-Torrecuadrada, J; Altschuh, D; Meloen, R; Ignacio Casal, J
  • Human polymeric IgA is superior to IgG and single-chain Fv of the same monoclonal specificity to inhibit urease activity associated with Helicobacter pylori
    Berdoz, J; Corthesy, B
  • Helicobacter pylori infection and stem cells at the origin of gastric cancer
    Bessede, E; Dubus, P; Megraud, F; Varon, C
  • A threading-based method (FINDSITE) for ligand-binding site prediction and functional annotation
    Brylinski, M; Skolnick, J
  • Predicting protein ligand binding sites by combining evolutionary sequence conservation and 3D structure
    Capra, JA; Laskowski, RA; Thornton, JM; Singh, M; Funkhouser, TA
  • Corpus-predominant gastritis as a risk factor for false-negative 13C-urea breath test results
    Capurso, G; Carnuccio, A; Lahner, E; Panzuto, F; Baccini, F; Delle Fave, G; Annibale, B
  • Upper gastrointestinal histopathological findings in children and adolescents with nonulcer dyspepsia with Helicobacter pylori infection
    Carvalho, MA; Machado, NC; Ortolan, EV; Rodrigues, MA
  • American College of Gastroenterology guideline on the management of Helicobacter pylori infection
    Chey, WD; Wong, BC
  • Verification of protein structures: patterns of nonbonded atomic interactions
    Colovos, C; Yeates, TO
  • Helicobacter pylori research: from bench to bedside
    Cover, TL; Holland, RL; Blanke, SR
  • CASTp: computed atlas of surface topography of proteins with structural and topographical mapping of functionally annotated residues
    Dundas, J; Ouyang, Z; Tseng, J; Binkowski, A; Turpaz, Y; Liang, J
  • VERIFY3D: assessment of protein models with three-dimensional profiles
    Eisenberg, D; Luthy, R; Bowie, JU
  • Pleiotropic cytotoxicity of VacA toxin in host cells and its impact on immunotherapy
    Fahimi, F; Tohidkia, MR; Fouladi, M; Aghabeygi, R; Samadi, N; Omidi, Y
  • Histological variety of gastric carcinoma and Helicobacter pylori cagA and vacA polymorphism
    Figura, N; Valassina, M; Moretti, E; Vindigni, C; Collodel, G; Iacoponi, F; Giordano, N; Roviello, F; Marrelli, D
  • Dissecting antibodies with regards to linear and conformational epitopes
    Forsstrom, B; Axnas, BB; Rockberg, J; Danielsson, H; Bohlin, A; Uhlen, M
  • Crystal structure of the Helicobacter pylori vacuolating toxin p55 domain
    Gangwer, KA; Mushrush, DJ; Stauff, DL; Spiller, B; McClain, MS; Cover, TL; Lacy, DB
  • A review of Helicobacter pylori diagnosis, treatment, and methods to detect eradication
    Garza-Gonzalez, E; Perez-Perez, GI; Maldonado-Garza, HJ; Bosques-Padilla, FJ
  • Design and screening of M13 phage display cDNA libraries
    Georgieva, Y; Konthur, Z
  • Helicobacter pylori infection: new pathogenetic and clinical aspects
    Hagymasi, K; Tulassay, Z
  • The use of phage display to generate conformation-sensor recombinant antibodies
    Haque, A; Tonks, NK
  • Mitogenic autoantibodies in Helicobacter pylori-associated stomach cancerogenesis
    Hensel, F; Knorr, C; Hermann, R; Krenn, V; Muller-Hermelink, HK; Vollmers, HP
  • Double-liposome-based dual-drug delivery system as vectors for effective management of peptic ulcer
    Jain, AK; Agarwal, A; Agrawal, H; Agrawal, GP
  • Development and characterization of nanolipobeads-based dual drug delivery system for H. pylori targeting
    Jain, AK; Jain, SK
  • Functional improvement of antibody fragments using a novel phage coat protein III fusion system
    Jensen, KB; Larsen, M; Pedersen, JS; Christensen, PA; Alvarez-Vallina, L; Goletz, S; Clark, BF; Kristensen, P
  • Princeton_TIGRESS: protein geometry refinement using simulations and support vector machines
    Khoury, GA; Tamamis, P; Pinnaduwage, N; Smadbeck, J; Kieslich, CA; Floudas, CA
  • Continuous cultures of fused cells secreting antibody of predefined specificity
    Kohler, G; Milstein, C
  • Increasing the precision of comparative models with YASARA NOVA—a self-parameterizing force field
    Krieger, E; Koraimann, G; Vriend, G
  • YASARA view—molecular graphics for all devices—from smartphones to workstations
    Krieger, E; Vriend, G
  • Helicobacter pylori-related diffuse large B-cell lymphoma of the stomach: a distinct entity with lower aggressiveness and higher chemosensitivity
    Kuo, SH; Yeh, KH; Chen, LT; Lin, CW; Hsu, PN; Hsu, C; Wu, MS; Tzeng, YS; Tsai, HJ; Wang, HP; Cheng, AL
  • Irregular meal timing is associated with Helicobacter pylori infection and gastritis
    Lim, SL; Canavarro, C; Zaw, MH; Zhu, F; Loke, WC; Chan, YH; Yeoh, KG
  • The history of monoclonal antibody development—progress, remaining challenges and future innovations
    Liu, JK
  • The role of Helicobacter pylori outer membrane proteins in adherence and pathogenesis
    Oleastro, M; Menard, A
  • Vacuolating cytotoxin A (VacA), a key toxin for Helicobacter pylori pathogenesis
    Palframan, SL; Kwok, T; Gabriel, K
  • In search of the Helicobacter pylori VacA mechanism of action
    Papini, E; Zoratti, M; Cover, TL
  • Protein Frustratometer 2: a tool to localize energetic frustration in protein molecules, now with electrostatics
    Parra, RG; Schafer, NP; Radusky, LG; Tsai, MY; Guzovsky, AB; Wolynes, PG; Ferreiro, DU
  • Diagnosis of Helicobacter pylori: what should be the gold standard?
    Patel, SK; Pratap, CB; Jain, AK; Gulati, AK; Nath, G
  • Diagnosis of Helicobacter pylori infection by invasive and noninvasive tests
    Pourakbari, B; Ghazi, M; Mahmoudi, S; Mamishi, S; Azhdarkosh, H; Najafi, M; Kazemi, B; Salavati, A; Mirsalehian, A
  • Helicobacter pylori and gastric cancer
    Pritchard, DM; Crabtree, JE
  • Marketed therapeutic antibodies compendium
    Reichert, JM
  • Interaction of Helicobacter pylori with host cells: function of secreted and translocated molecules
    Rieder, G; Fischer, W; Haas, R
  • I-TASSER: a unified platform for automated protein structure and function prediction
    Roy, A; Kucukural, A; Zhang, Y
  • COFACTOR: an accurate comparative algorithm for structure-based protein function annotation
    Roy, A; Yang, J; Zhang, Y
  • Life in the human stomach: persistence strategies of the bacterial pathogen Helicobacter pylori
    Salama, NR; Hartung, ML; Muller, A
  • Fully blind peptide-protein docking with pepATTRACT
    Schindler, CE; Vries, SJ; Zacharias, M
  • Monoclonal antibodies as diagnostics; an appraisal
    Siddiqui, MZ
  • Engineering M13 for phage display
    Sidhu, SS
  • Phage display on the base of filamentous bacteriophages: application for recombinant antibodies selection
    Tikunova, NV; Morozova, VV
  • Molecular considerations for development of phage antibody libraries
    Tohidkia, MR; Barar, J; Asadi, F; Omidi, Y
  • Functional characterization of two scFv-Fc antibodies from an HIV controller selected on soluble HIV-1 Env complexes: a neutralizing V3- and a trimer-specific gp41 antibody
    Trott, M; Weibeta, S; Antoni, S; Koch, J; Briesen, H; Hust, M; Dietrich, U
  • Antibody engineering: comparison of bacterial, yeast, insect and mammalian expression systems
    Verma, R; Boleti, E; George, AJ
  • Enhancement of scFv fragment reactivity with target antigens in binding assays following mixing with anti-tag monoclonal antibodies
    Wang, X; Campoli, M; Ko, E; Luo, W; Ferrone, S
  • ProSA-web: interactive web service for the recognition of errors in three-dimensional structures of proteins
    Wiederstein, M; Sippl, MJ
  • LOMETS: a local meta-threading-server for protein structure prediction
    Wu, S; Zhang, Y
  • A comparative assessment and analysis of 20 representative sequence alignment methods for protein structure prediction
    Yan, R; Xu, D; Yang, J; Walker, S; Zhang, Y
  • Protein-ligand binding site recognition using complementary binding-specific substructure comparison and sequence profile alignment
    Yang, J; Roy, A; Zhang, Y
  • The I-TASSER Suite: protein structure and function prediction
    Yang, J; Yan, R; Roy, A; Xu, D; Poisson, J; Zhang, Y
  • I-TASSER server: new development for protein structure and function predictions
    Yang, J; Zhang, Y
  • I-TASSER server for protein 3D structure prediction
    Zhang, Y
  • SPICKER: a clustering approach to identify near-native protein folds
    Zhang, Y; Skolnick, J
  • Phage antibody display libraries: a powerful antibody discovery platform for immunotherapy
    Zhao, A; Tohidkia, MR; Siegel, DL; Coukos, G; Omidi, Y

You’re reading a free preview. Subscribe to read the entire article.

Try 2 weeks free now

DeepDyve is your
personal research library

It’s your single place to instantly
discover and read the research
that matters to you.

Enjoy affordable access to
over 18 million articles from more than
15,000 peer-reviewed journals.

All for just $49/month

Explore the DeepDyve Library

or browse the journals available

Search

Query the DeepDyve database, plus search all of PubMed and Google Scholar seamlessly

Organize

Save any article or search result from DeepDyve, PubMed, and Google Scholar... all in one place.

Access

Get unlimited, online access to over 18 million full-text articles from more than 15,000 scientific journals.

Your journals are on DeepDyve

Read from thousands of the leading scholarly journals from SpringerNature, Elsevier, Wiley-Blackwell, Oxford University Press and more.

All the latest content is available, no embargo periods.

See the journals in your area

DeepDyve

Freelancer

DeepDyve

Pro

Price

FREE

$49/month
$360/year

Save searches from
Google Scholar,
PubMed

Create folders to
organize your research

Export folders, citations

Read DeepDyve articles

Abstract access only

Unlimited access to over
18 million full-text articles

Print

20 pages / month

PDF Discount

20% off

Sign up
for free
Start 14 day
Free Trial