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Functional Mutations in and Characterization of VHH Against Helicobacter pylori Urease

Applied Biochemistry and Biotechnology volume 172, pages 3079–3091 (2014)Cite this article

Abstract

Manipulation of clinically significant antibodies can effectively improve the processes of diagnosis and treatment. Affinity maturation process has a significant role in improvement of antibodies efficiency. Error-prone PCR technique is one of the proposed methods for improvement of the affinity of antibodies. In the present research, a method was applied to camel heavy-chain antibody (VHH, nanobody) raised against UreC subunit of urease enzyme from Helicobacter pylori. This VHH was used as a starting molecule to construct a highly diversified phage displayed VHH library. The constructed library of nanobody mutants was subjected to several rounds of panning against UreC antigen. High-affinity mutant was selected. Our VHH (HMR23) showed 1.5-fold higher binding activity than the parental VHH. In addition, the mutant VHH presented a better performance in inhibition of urease activity at low concentrations retaining its specificity and thermal stability.

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Author information

Affiliations

  1. Department of Biology, Faculty of Basic Science, Shahed University, Opposite Imam Khomeini’s Shrine, Tehran-Qom Express way, Tehran, 3319118651, Iran

    Reyhaneh Hoseinpoor, Seyed Latif Mousavi Gargari, Iraj Rasooli & Bahareh Shahi

  2. Department of Clinical Biochemistry, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran

    Masoumeh Rajabibazl

Authors
  1. Reyhaneh Hoseinpoor
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  2. Seyed Latif Mousavi Gargari
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  3. Iraj Rasooli
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  4. Masoumeh Rajabibazl
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  5. Bahareh Shahi
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Correspondence to Seyed Latif Mousavi Gargari.

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Hoseinpoor, R., Mousavi Gargari, S.L., Rasooli, I. et al. Functional Mutations in and Characterization of VHH Against Helicobacter pylori Urease. Appl Biochem Biotechnol 172, 3079–3091 (2014). https://doi.org/10.1007/s12010-014-0750-4

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  • DOI: https://doi.org/10.1007/s12010-014-0750-4

Keywords

  • Affinity maturation
  • Error-prone PCR
  • Helicobacter pylori
  • Random library
  • Urease
  • VHH