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Application of recombinant antibody fragments for troponin I measurements

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Abstract

The cardiac isoform of troponin I is a reliable biomarker of damaged cardiomyocytes that accompanies such severe cardiovascular diseases as myocardial infarction. Monoclonal antibody 19C7 recognizes troponin I in the blood-stream with high affinity and specificity. Recombinant antibodies can be used to improve detection systems based on monoclonal antibodies produced with hybridoma technology. In the present study, we compare the properties of monoclonal anti-body 19C7 and its recombinant fragments. It is shown that the recombinant antibody fragments demonstrate similar affinity values as monoclonal antibodies and can be applied for troponin I detection.

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Abbreviations

BITC:

biotin isothiocyanate

eFab:

Fab fragments of MAb 19C7 produced by enzymatic cleavage of fullsized MAb by papain

FIA:

fluorescent immunoassay

hcTnI:

human cardiac troponin I isoform

IC and ICT:

double and triple troponin complexes formed by TnI and TnC and TnI TnC and TnT respectively

MAb:

monoclonal antibody

MAb 19C7-BITC:

MAb 19C7 biotinylated with BITC

MPB:

maleimide-PEG11-biotin

rAb:

recombinant antibody

rFab:

recombinant Fab fragments of MAb 19C7

scFv:

single-chained fragments of MAb 19C7 composed of variable domains of light and heavy chains of MAb that are linked with Gly-Ser linker

scFv-BITC:

scFv-fragments of MAb 19C7 biotinylated with BITC

scFv-MPB:

scFv-fragments of MAb 19C7 biotinylated with MPB

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Authors and Affiliations

  1. Faculty of Biology, Lomonosov Moscow State University, 119991, Moscow, Russia

    E. P. Altshuler, I. A. Katrukha, A. V. Bereznikova & D. V. Serebryanaya

  2. Bialexa Ltd., Simferopolsky Ave. 8, 117149, Moscow, Russia

    A. V. Vylegzhanina

Authors
  1. E. P. Altshuler
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  2. A. V. Vylegzhanina
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  3. I. A. Katrukha
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  4. A. V. Bereznikova
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  5. D. V. Serebryanaya
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Corresponding author

Correspondence to E. P. Altshuler.

Additional information

Published in Russian in Biokhimiya, 2012, Vol. 77, No. 12, pp. 1653–1660.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM12-242, September 30, 2012.

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Altshuler, E.P., Vylegzhanina, A.V., Katrukha, I.A. et al. Application of recombinant antibody fragments for troponin I measurements. Biochemistry Moscow 77, 1362–1367 (2012). https://doi.org/10.1134/S0006297912120036

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  • DOI: https://doi.org/10.1134/S0006297912120036

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