---
_id: '11462'
abstract:
- lang: eng
  text: Nanobodies (VHH) from camelid antibody libraries hold great promise as therapeutic
    agents and components of immunoassay systems. Synthetic antibody libraries that
    could be designed and generated once and for various applications could yield
    binders to virtually any targets, even for non-immunogenic or toxic ones, in a
    short term. One of the most difficult tasks is to obtain antibodies with a high
    affinity and specificity to polyglycosylated proteins. It requires antibody libraries
    with extremely high functional diversity and the use of sophisticated selection
    techniques. Here we report a development of a novel sandwich immunoassay involving
    a combination of the synthetic library-derived VHH-Fc fusion protein as a capture
    antibody and the immune single-chain fragment variable (scFv) as a tracer for
    the detection of pregnancy-associated glycoprotein (PAG) of cattle (Bos taurus).
    We succeeded in the generation of a number of specific scFv antibodies against
    PAG from the mouse immune library. Subsequent selection using the immobilized
    scFv-Fc capture antibody allowed to isolate 1.9 nM VHH binder from the diverse
    synthetic library without any overlapping with the capture antibody binding site.
    The prototype sandwich ELISA based on the synthetic VHH and the immune scFv was
    established. This is the first successful example of the combination of synthetic
    and immune antibody libraries in a single sandwich immunoassay. Thus, our approach
    could be used for the express isolation of antibody pairs and the development
    of sandwich immunoassays for challenging antigens.
acknowledgement: This study was financially supported by the State Committee on Science
  and Technology. We would like to thank Elena Tumar and Elena Kisileva at the Institute
  of Bioorganic Chemistry of NASB for their kind assistance with mouse immunizations.
article_processing_charge: No
article_type: original
author:
- first_name: Dmitri
  full_name: Dormeshkin, Dmitri
  last_name: Dormeshkin
- first_name: Michail
  full_name: Shapira, Michail
  last_name: Shapira
- first_name: Alena
  full_name: Karputs, Alena
  last_name: Karputs
- first_name: Anton
  full_name: Kavaleuski, Anton
  id: 62304f89-eb97-11eb-a6c2-8903dd183976
  last_name: Kavaleuski
  orcid: 0000-0003-2091-526X
- first_name: Ivan
  full_name: Kuzminski, Ivan
  last_name: Kuzminski
- first_name: Elena
  full_name: Stepanova, Elena
  last_name: Stepanova
- first_name: Andrei
  full_name: Gilep, Andrei
  last_name: Gilep
citation:
  ama: Dormeshkin D, Shapira M, Karputs A, et al. Combining of synthetic VHH and immune
    scFv libraries for pregnancy-associated glycoproteins ELISA development. <i>Applied
    Microbiology and Biotechnology</i>. 2022;106:5093-5103. doi:<a href="https://doi.org/10.1007/s00253-022-12022-w">10.1007/s00253-022-12022-w</a>
  apa: Dormeshkin, D., Shapira, M., Karputs, A., Kavaleuski, A., Kuzminski, I., Stepanova,
    E., &#38; Gilep, A. (2022). Combining of synthetic VHH and immune scFv libraries
    for pregnancy-associated glycoproteins ELISA development. <i>Applied Microbiology
    and Biotechnology</i>. Springer Nature. <a href="https://doi.org/10.1007/s00253-022-12022-w">https://doi.org/10.1007/s00253-022-12022-w</a>
  chicago: Dormeshkin, Dmitri, Michail Shapira, Alena Karputs, Anton Kavaleuski, Ivan
    Kuzminski, Elena Stepanova, and Andrei Gilep. “Combining of Synthetic VHH and
    Immune ScFv Libraries for Pregnancy-Associated Glycoproteins ELISA Development.”
    <i>Applied Microbiology and Biotechnology</i>. Springer Nature, 2022. <a href="https://doi.org/10.1007/s00253-022-12022-w">https://doi.org/10.1007/s00253-022-12022-w</a>.
  ieee: D. Dormeshkin <i>et al.</i>, “Combining of synthetic VHH and immune scFv libraries
    for pregnancy-associated glycoproteins ELISA development,” <i>Applied Microbiology
    and Biotechnology</i>, vol. 106. Springer Nature, pp. 5093–5103, 2022.
  ista: Dormeshkin D, Shapira M, Karputs A, Kavaleuski A, Kuzminski I, Stepanova E,
    Gilep A. 2022. Combining of synthetic VHH and immune scFv libraries for pregnancy-associated
    glycoproteins ELISA development. Applied Microbiology and Biotechnology. 106,
    5093–5103.
  mla: Dormeshkin, Dmitri, et al. “Combining of Synthetic VHH and Immune ScFv Libraries
    for Pregnancy-Associated Glycoproteins ELISA Development.” <i>Applied Microbiology
    and Biotechnology</i>, vol. 106, Springer Nature, 2022, pp. 5093–103, doi:<a href="https://doi.org/10.1007/s00253-022-12022-w">10.1007/s00253-022-12022-w</a>.
  short: D. Dormeshkin, M. Shapira, A. Karputs, A. Kavaleuski, I. Kuzminski, E. Stepanova,
    A. Gilep, Applied Microbiology and Biotechnology 106 (2022) 5093–5103.
date_created: 2022-06-26T22:01:34Z
date_published: 2022-08-01T00:00:00Z
date_updated: 2023-10-10T07:15:02Z
day: '01'
department:
- _id: GradSch
- _id: LeSa
doi: 10.1007/s00253-022-12022-w
external_id:
  isi:
  - '000813677500001'
  pmid:
  - '35723693'
intvolume: '       106'
isi: 1
language:
- iso: eng
month: '08'
oa_version: None
page: 5093-5103
pmid: 1
publication: Applied Microbiology and Biotechnology
publication_identifier:
  eissn:
  - 1432-0614
  issn:
  - 0175-7598
publication_status: published
publisher: Springer Nature
quality_controlled: '1'
scopus_import: '1'
status: public
title: Combining of synthetic VHH and immune scFv libraries for pregnancy-associated
  glycoproteins ELISA development
type: journal_article
user_id: 2DF688A6-F248-11E8-B48F-1D18A9856A87
volume: 106
year: '2022'
...